Post on 08-Dec-2021
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PHYSICAL CHARACTERISTIC AND GERMINATION OF HIBUSCUS SABDARIFFA LFOLLOWING
HYDRATION TREATMENTS
Syazwanie Binti Dzulhaimi
SB 413 86
Bachelor of Science with HonoursS981 (Plant Resource Science and Management)2011
2012
J I Pusat Khidmat MakJuUUlt Akademik UNlVERSm MALAYSIA SARAWAK
PHYSICAL CHARACTERISTIC AND GERMINATION OF HIBISCUS
SABDARIFFA L FOLLOWING HYDRATION TREATMENTS
P~i~liiimi 1000235560
SYAZW ANIE BINTI DZULHAIMI
This project work is submitted in partial fulfilment ofthe requirement for the Degree of
Bachelor of Science with Honours in Plant Resource Science and Management
Faculty of Resource Science and Technology
UNIVERSITI MALAYSIA SARA W AK
2012
ttl
APPROVAL SHEET
Name of candidate Syazwanie Binti Dzulhaimi
Matric no 25214
Title of thesis Physical characteristic and germination of Hibiscus sabdarifJa L
following hydration treatments
Approved by
(Assoc Prof Dr Petrus Bulan)
Supervisor
Plant Resource Science and Management
Faculty of Resource Science and Technology
Date
(Dr Siti Rubiah Zainudin)
Coordinator
Plant Resource Science and Management
Faculty of Resource Science and Technology
Date
ii
t I
DECLARATION
I hereby declare that no portion of this project work has been submitted in support of an
application for another degree of qualification of this or any other university or institution
of higher learning
(Syazwanie Binti Dzulhaimi)
Plant Resource Science and Management
Department of Environment Science and Ecology
Faculty of Resource Science and Technology
Universiti Malaysia Sarawak
iii
middot
ACKNOWLEDGEMENT
First and foremost I would like to give my highest praise to God for giving me the
strength patience intelligence and integrity to complete this research study
Alhamdulillah
My gratitude and sincere appreciation to Assoc Prof Or Petrus Bulan as my supervisor
for his continuous help support guidance advice and constant assisstance
I would also like to give my deepest thanks to my beloved parents Mr Dzulhaimi Khalid
and Mdm Khariyah Ahmad for their support understanding and encouragement
throughout my three years journey as an undergraduate student in Universiti Malaysia
Sarawak Not forgetting my other families whose been always supporting me along the
way
Thanks to the people that have contributed tremendously in my final year project Mdm
Fatimah Daud the laboratory assisstant of Cryopreservation Laboratory and all other
Faculty of Resource Science and Technology staff
Last but not least my appreciation goes to all my dearest friends especially to
Norakliliriana Abdul Rahman Siti Nadiah Salleh and to all my Plant Science coursemates
for their assistance and support throughout this study
iv
Pusat Khidmat Maklumat Akademik I
UNlVERSm MALAYSIA SARAWAK
TABLE OF CONTENTS
TITLE AND FRONT COVER I
APPROVAL SHEET II
DECLARATION III
ACKNOWLEDGEMENT IV
TABLE OF CONTENTS V
LIST OF FIGURES AND PLATES VII
ABSTRACT IX
10 INTRODUCTION 1 11 Background 1 12 Commercial Production 2 13 Medicinal Uses 3 14 Problem Statement 3 15 Objectives 4
20 LITERATURE REVIEW 5 21 Characteristic of Seed 5 22 Seed Germination 6 23 Seed Moisture Content 7 24 Seed Vigour 7 25 Seed Hydration 8 26 Seed Storage 9 27 Seed Specific Density 10 28 Seed Deterioration 10
30 MATERIAL AND METHOD 12 31 Material 12 32 Method 12
321 Preliminary Evaluation 12 322 Moisture Content Test 13 323 Germination Test 13 324 Seed Characteristic 14 325 Hydration of Seeds 14 326 Data Analysis 14
40 RESULT AND DISCUSSION 15 41 Preliminary Test 15 42 Seed Calibration 15 43 Seed Hydration 16
43 1 Seed Moisture Content 16 v
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432 Seed Germination 22 433 Gennination Analysis 28
50 CONCLUSIONS AND RECOMMENDATION 29
60 REFERENCES 30
70 APPENDICES 31
vi
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LIST OF FIGURES AND PLATES
Figure Page
Moisture content of Hsabdariffa seeds for different period of 15
hydration at 28degC
2 Moisture content of Hsabdariffa seeds for different period of 16
hydration at 30degC
3 Moisture content of Hsabdariffa seeds for different period of 17
hydration at 35degC
4 Moisture content of Hsabdariffa seeds for different period of 18
hydration at 40degC
5 Moisture content of Hsabdariffa seeds for different period of 19
hydration at 28 30 35 and 40degC
6 Gennination of Hsabdariffa seeds for different period of 20
hydration at 28degC
7 Gennination of Hsabdariffa seeds for different period of 21
hydration at 30degC
8 Gennination ofHsabdariffa seeds for different period of 22
hydration at 35degC
Gennination of Hsabdariffa seeds for different period of
hydration at 40degC
9 23
vii
10 24
Germination of HsabdarifJa seeds for different period of
hydration at 28 30 35 and 40degC
Plate
Fresh fruit of H sabdarifJa 11
2 Fresh fruit of H sabdarifJa with seeds 11
3 Fresh seeds of H sabdarifJa 16
4 Dried seeds of H sabdarifJa 16
viii
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I
I
Physical characteristic and germination of Hibiscus Sabdariffa L following hydration treatments
Syazwanie Binti Dzulbaimi
Plant Resource and Management Programme Faculty of Science and Technology
Universiti Malaysia Sarawak
ABSTRACT
A study was conducted to determine the physical characteristics and to assess and evaluate the effectiveness of hydration techniques in improving the performance of HibisclIs sabdariffa L seeds The average length width and thickness obtained for H sabdariffa seeds were 539 mm 455 mm and 234 mm respectively The average weight for a single seed was 002 g and it was estimated that one kg weight contained approximately 50000 seeds This indicated that the seeds used in this experiment were small In hydration treatment seeds of H sabdariffa were soaked in water at temperature 28 30 35 and 40degC for 0246 8 10 and 12 hours and were evaluated for moisture content and germination Results indicated that hydration treatment was not suitable for H sabdariffa seeds as it did not improved the germination The highest germination percentage of H sabdarifJa seeds following hydration treatment was 29 which was obtained for 2 hours hydration at 30degC
Key words Hibiscus sabdarifJa calibration hydration temperature
ABSTRAK
Satu kajian telah dijalankan IInt1lk menentukan ciri-ciri jizikal dall mengkaji serta menentukan keberkesanan teknik rawatall hidrasi dalam meningkatkan kadar pertllmbllhan biji benih Hibiscus sabdariffa Dalam penentlkuran biji benih pIrata panjang lebar dan ketebalan biji benih H sabdariffa yang diperoleh adalah 539 mm 455 mm dan 234 mm Purata berat sebiji benih adalah 002 g dan dianggarkan bahawa satll kilogram mengandungi lebih kurang 50000 biji benih Ini memmjukkan yang biji benih yang digllnakan unIlk kajian ini agak keci Dalam rawatan hidrasi biji benih H sabdariffa direndam di dalam air pada suhu 28degC 30degC 35degC dan 40degC Bagi setiap suhu yang digunakan biji benih direndam selama 0 2 4 6 8 10 dan 12 jam dan selepas itu kandungan kelembapan dan percambahan biji benih dinilai Keputlsan yang diperoleh menllnjukkan rawatan hidrasi adalah tidak sesuai bagi biji benih H sabdariffa memalldangkan peratls percambahan tidak meningkat berbanding sebelum rawatan Peratlls percambahan tertinggi bagi biji benih H sabdariffa selepas rawatan hidrasi adalah 29 iaitll bagi hidrasi selama 2 jam pada slhu 30degC
Kata kllnciHibisclS sabdarifJa kalibrasi hidrasi suhl
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10 INTRODUCTION
11 Background
Hibiscus sabdarifJa Linn or also known as Roselle or red sorrel belongs to the family
Malvaceae It is also known as asam paya asam susur and asam kumbang locally It is a
new commercial crop in Malaysia where it was brought from India (Amin et al 2008)
Since Roselle plantation only had started in early 1990s it is considered as a new crop in
Malaysia Two introduced varieties in Malaysia are Arab and Terengganu variety
However in 2009 Universiti Kebangsaan Malaysia had launched three new varieties
named UKMR-l UKMR-2 and UKMR-3 to help promote the Roselle industry Recently
Department of Agriculture had collaborated with F AMA entrepreneurs and growers in
order to strengthen the H sabdarifJa industry in Malaysia by expanding and marketing this
product locally and in foreign market
According to Qi et al (2005) H sabdarifJa is an annual mostly brannched and erect
shrub Its reddish stem can achieve 35m tall The leaves are dark green to red alternate
glabrous long - petiolate palmately divided into 3 - 7 lobes with serrate margins Having
both male and female organs the flower are red to yellow in color with a dark centre
containing short peduncles
H sabdarifJa can survive in a wann and humid tropical and subtropical climate It can be
grown in a greenhouse with litle shade but the best condition for growing it is under the
full sunlight (Qi et aI 2005) Roselle plants are suitable for tropical climates with wellshy
distributed rain - fall of 1500 - 2000 mm per year from sea level to about 600 m in
altitude (Amin et aI 2008) It can adapt to variety of soils but prefered a friable sandy
loam with humus
1
Current production of H sabdarifJa in Malaysia is about 240 tonnes annually Reported in
unpublished result from Agricultural Department of Terengganu in the process of
removing the calyces out of 3 tonnes of raw materials about 15 tonnes of the velvety
capsules containing the seeds are being disposed as a by - product and unexploited
(Halimatul Amin Mohd Esa Nawalyah amp Siti 2007) Amin et at (2008) mentioned that
the plant takes about three to four months to reach the commercial stage of maturity before
the flowers are harvested
12 Commercial Production
Many parts of H sabdariffa including seeds leaves fruits and roots are very useful
especially the fleshy red calyces They are used fresh for making wine juice jam jelly
syrup gelatin pudding cakes ice cream and flavors and also dried and brewed into tea
spice and used for butter pies sauces tarts and other desserts (Qi et al 2005) In a study
about commercial practice of Roselle beverage production Bolade et at (2009) reported
that the dried calyceswater ratios involved in the commercial practice ranged between
157 and 171 (wv) while the sweetness level of the beverage ranged between 112 and
133 degBrix The red calyces which contains high amount of vitamin C was used to make
those healthy beverages In Malaysia we also consume the leaves as vegetables There are
previous study shows that H sabdarifJa seeds can be used as a protein source Instead of
using protein from conventional sources protein isolates or concentrates from Roselle
seeds might be useful as low cost source of protein substitute in dietary supplement or food
ingredient in food industry thus alleviating the problem of protein scarcity (Halimatul et
al 2007)
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13 Medicinal Uses
H sabdariffa is said to be useful in traditional medicine as a digestive agent purgative and
diuretic among others Having two diuretic ingredients ascorbic acid and glycolic acid it
has the ability to increase urination and also valued for its mild laxative effect Containing
citric acid it also can be used as cooling herb The tonic tea produce from the leaves and
flowers is good for digestive and kidney function The heated leaves are applied to cracks
in the feet and on boils and ulcers to speed maturation (Qi et aI 2005) Other roselle s part
have also been reported to be folk remedy for cancer obesity diabetes and hypertension
(Bolad et aI 2009) Amin et al (2008) also added that the flower and fleshy fruits are
used in pharmaceutical industry to relieve symptoms of bronchitis and coughs Roselle s
lotion products are useful for treating sores and wounds
Another nutritional value of Roselle discovered by UKM was the high content of
hydroxycitric acid (HCA) which is widely used as the main ingredients in many
commercial weight loss and slimming products Recent studies by local and international
scientist have proven the effectiveness of HCA in reducing body weight This
characteristic makes Roselle a very valuable crop
14 Problem Statement
The demand for H sabdariffa production nowadays had increase due to its commercial and
medicinal values To fulfill these demands more good quality seeds are needed
Furthermore not many research had been done on improving the H sabdarifJa seeds as a
planting material eventhough the best propagation method for H sabdariffa is by seeds
Other than that disease has been reported as a limiting factor to the production of Roselle
worldwide (Sie R S et aI 2011)
3
1S Objectives
1 To detennine the physical characteristics of H sabdariffa seeds for 10 100 and
1000 seeds weight and for length width and thickness of the seed
2 To evaluate the germination of H sabdariffa seeds following hydration at different
temperatures
3 To determine the effectiveness of hydration technique in improving the gennination
of H sabdariffa seeds
4
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Pusat Khidmat Maklumat Akadtmik VNIVERSm MALAYSIA SARAWAK
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20 LITERATURE REVIEW
21 Characteristic of Seed
Seeds quality can be affected by changes in environmental temperature and relative
humidity The characteristic of seed must be determine because it is important to ensure
high quality yield in field planting Seed germination can be affected by number of factors
such as level of oxygen temperature and moisture content Growth performance of seed
also can be related to seed size Therefore grading is important to obtain seed that can
grows better Some useful criteria for visual separation of seed includes the seed size
shape colour weight thickness density and surface texture
According to Jain and Bal (1997) roselle seeds are having average principal dimensions of
298-336 186-224 and 170-201 mm Omobuwajo et al (2000) found that the average of
roselle seeds length width and thickness were 558 521 and 281 mm respectively The
seed was reported to be important for its oil in some parts of Africa its native origin From
previous research by scientist roselle seed was among the highest protein-containing seeds
when compared with other seeds like passion fruit (Passiflora edulis) Amaranthus seeds
and Pisum sativum seeds Latest findings by Hainida et al (2008) found that the seed from
Malaysia are composed of 99 moisture 335 protein 221 lipids 130 available
carbohydrate 183 total dietary fibers and 75 ash
H sabdarifJa can be grown easily in most of the tropical country Like other hibiscus
species it is quite hard for roselle to be affected by pest and disease and maintaining this
plant in the field is not so difficult The seeds were sown in a suitable container and the
seedlings will be ready for planting at the age of 10-15 days However it is also important
to ensure that the seeds to be sown were in a good condition and quality to avoid losses
5
22 Seed Germination
Germination is the emergence and development from the seed embryo of those essential
structures which are indicative of the ability to produce a normal plant under favorable
conditions (AOSA 1991) According to Smith (2011) the sum of germination events
beginning with hydration and culminating with root emergence Several stages involved in
germination are imbibition of water activation of enzyme systems metabolism of storage
products and their transport and finally the emergence of the radicle and growth of the
seedling Seed germination is described as the emergence and development from the seed
embryo of those essential structures which for the kind of seed in question are indicative
of the ability to produce a normal plant under favorable conditions (AOSA 1978) In a
simpler contcxt germination is the emergence of the radicle through the seed coat
Depending on the species seeds can germinate as quick as a few days or for as long as
several years Due to the high respiration rate and some exudation and leakage through the
seed coat the germinating seedling will undergoes a net loss in dry weight before the
germination For a particular kind of species the germination condition is mostly affected
by temperature type of substrate on which to germinate the seed light condition and
moisture level
The germination test is commonly used to determine seed viability It is an analytical
procedure to evaluate seed germination under standardized favorable conditions that are
seldom if ever encountered in the field The germination test is merely an estimate and
has certain limitations as a universal estimate of seed quality However if these limitations
are recognized the germination test is a useful viability index The need for germination
test is to estimate how the seed will perform in the field
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23 Seed Moisture Content
Amount of water in the seed that usually expressed in a percentage fonn is the seed
moisture content Agrawal (1980) found that the amount of moisture in the seed is
probably the most important factor influencing seed viability during storage By
determining the seeds moisture content the possible storage life for the seeds can be
predicted According to McDonald (2005) selecting the appropriate salt to achieve a seed
moisture content of 5-6 is the recommended method for achieving optimum seed
storage and minimizing seed deterioration It is necessary to dry seeds to the safe moisture
contents since the life of a seed and its span largely revolves around its moisture content
and the suitable moisture content for starchy seeds are less than 14 while for oily seeds
are less than 11 (Agrawal 1980)
24 Seed Vigour
According to Geneve (2005) seed vigour is defined by the Asociation of official Seed
analysts as those seed properties which detennine the potential for rapid unifonn
emergence and development of nonnal seedlings under the wide range of field conditions
Seed vigour is closely related with seed storage environment and duration During seed
storage the vigour of a seed lot is reduced prior to seed viability as indicated by standard
germination (Hampton amp TeKrony 1995) Major factors influencing the seed vigour in
stored seed lots are the environmental conditions during seed storage and the length of time
in storage Tetrazolium test is one of the method for detennining the seeds vigour To
minimize the decline in vigour and germination seeds should be undamaged
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25 Seed Hydration
A process of supplying water to seed to maintain the fluid or moisture content in the seed
is called hydration According to Copeland and McDonald (1995) seed hydration is a
process whereby seeds are hydrated using various protocols and then redried to permit
routine handling This process results in increased germination rate more uniform
emergence germination under a broader range of environments and improved sedling
vigour and growth It is one of the priming treatments of seed which usually applied to
strengthen or increase vitality of seed Priming process can increase a rate of germination
as the treatment improves the seed vigor Hydration is one component of priming which
can increase germination with more range of environment condition The objective of seed
hydration technology is to increase the percentage and rate of germination expand the
range of temperatures over which the seed will germinate and increase the uniformity of
stand establishment To accomplish these objectives seeds must be hydrated in some way
at a moisture level sufficient to initiate the early events of germination but not sufficient to
permit radicle protrusion (Akers and Holley 1986) However the performance of the seed
might also being influence by other factors like the species type of seed and the seeds
quality
Water can be used for the hydration treatment because it had proved to enhance
germination emergence growth and yield of seed However other mediwns such as the
solution of potassium hydrophosphate (KH2P04) monobasic polyethylene glycol (PEG)
potassium chloride (KCl) potassiwn nitrate (KN03) potassium phoshate (K3P04)
magnesium sulphate (MgS04) sodiwn chloride (NaCl) glycerol and mannitol also can be
used to immerse the seed in hydration process The benefit of such salts is to supply the
seed with nitrogen and other nutrients essential for protein synthesis during germination
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26 Seed Storage
A basic requirement in seed production is a good seed storage The purpose of seed storage
is to preserve planting stocks from one season to the next (Larry amp McDonald 1995)
Generally we stored seeds for the commercial purpose for carry - over seeds or for genn
plasm seeds Maintaining the seed quality for the longest duration possible is the main
objective in seed storage Loss of gennination during storage cannot be stopped but it
could be reduce by giving good storage conditions According to Agrawal (1980) the
seeds are considered to be in storage from the moment they reach physiological maturity
until they genninate or until they are thrown away because they are dead or otherwise
worthless
The seed storage must be properly handled because it can affect the seed quality There are
some factors that can affect the seed longevity in storage such as the variety of the seed
initial seed quality moisture content relative humidity and storage temperature It is
necessary for the seed storage condition to be dry and cool effective in pest control and
have a proper sanitation Other than than storing only high quality of seed also can help in
better production From all of the factors temperature is one of the most important element
which influence seed viability and vigour during storage Agrawal (1980) mentioned that
the lower the temperature the longer the seeds maintain gennination capacity so
decreasing temperature and seed moisture is an effective means of maintaining seed quality
in storage Every seed lot should be gennination - tested when it is received for storage to
record the seed quality Subsequent periodic checks also should be made to detect
deterioration and if it occurs storage condition should be checked to discover the reasons
and to take corrective measures
9
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27 Seed Specific Density
Density of seed refers to it mass per unit volume The seed size and density will affect the
yield One of the method for grouping seeds into some specific density is the liquid density
separation system It is one of the easier and cheaper method compared to using a machine
When seed is loaded into a liquid some will float and some will sink That mean the
floating seeds density is lower than the liquids density Vice versa the sinking seeds
density is higher than the liquids density Wang and Alyarez (2008) explained that this
method of separating seeds works by adjusting the specific gravity of the liquid in the
reservoir so that a first portion of the seeds float near the surface of the liquid and a second
portion of the seeds sink near the bottom of the reservoir and providing an outlet flow of
the liquid from a location proximal to the top of the reservoir above the location of the
second portion of seeds By this way seeds of lower density can be separated from higher
density Kwong Sellman Jalink amp Schoor (2005) illustrated that separation by density is
commonly incorporated as part of the routine cleaning and grading process in flower seed
production procedures Other than grouping seeds according to their specific density this
density separation also can be used to remove broken partially filled empty or immature
seeds
28 Seed Deterioration
McDonald (2005) defined flower seed deterioration as deteriorative changes occuring with
time that increase the seeds vulnerability to external challenges and decrease the ability of
the seed to survive Important factors contribute to seed deterioration are genetics seed
structure seed chemistry physicalphysiological quality seed treatments relative humidity
10
and temperature Relative humidity is important because it will influence the moisture
content of seeds in storage while temperature determines the amount of moisture in the air
and influences the rate of deteriorative reactions in seeds According to Agrawal (1980)
the rate of deterioration increases as the seed moisture content increases This could be due
to the mold growth in and on the seed
11
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30 MATERIAL AND METHOD
31 Material
Fresh matured fruits of Hibiscus sabdarifJa L (Plate I) were obtained from local fanns in
Kota Samarahan Seeds were extracted from these fruits cleaned and dried under shade
Dried seeds were dusted with Captan 80 and then placed in air-tight bottle to keep it in a
safe place before use in the subsequent experiment
Plate 1 Fresh fruit ofH sabdarifJa Plate 2 Fresh fruit of H sabdarifJa with seeds
32 Method
321 Preliminary Evaluation
Moisture content and germination was conducted as an initial evaluation to assess the
quality of seeds used in this research The procedures were those prescribed by Association
ofOfficial Seeds Analysts (AOSA 1985)
12
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322 Moisture Content Test
Four replicates of 10 seeds each were placed into saucers (Plate 5) and calibrated to get the
wet weight The seeds then were placed in an oven at 60degC for 48 hours After that the
seeds were weighed again to get the dry weight The percentage of moisture content of the
seeds was calculated based on the formula below (AOSA 1985)
Moisture content () = b - c x 100
b-a
where a = weight of empty saucer
b = weight of (a) + weight of seeds before drying
c = weight of (a) + weight of seeds after drying in the oven
323 Germination Test
Germination test was conducted base on the AOSA (1985) where 25 seeds with four
replications were used For every replicates seeds were put into a petry dish on 3 layers of
moist Whatman filter paper (Plate 6) The seeds were left to germinate by incubating it in a
Plant Growth Chamber (28OC) and was checked daily until 10 days Seed was considered
as germinated if the radicles were about 5mm length emerged from the seed coat The
germination percentage was calculated base on AOSA 1985 formula below
13
-----------------------------------1
Gennination () a x 100
b
where a = total number of germinated seeds
b = total number of seeds used
324 Seed Characteristic
The size of the H sabdariffa seeds was calibrated for its length width and thickness Ten
replicates of seeds were used for calibrating the physical characteristics Then the seed
weights of 10 100 and 1000 seeds were calibrated Four replicates of 10 100 and 1000
seed-weight classes were used
325 Hydration of Seeds
Seed of H sabdariffa was placed in distilled water for 0 2 4 6 8 10 and 12 hours at
temperatures of 28 30 35 and 40degC Then seeds were analyzed for the moisture content
and gennination
326 Data Analysis
The experimental units were arranged randomly based on Completely Randomized Design
(CRD) and data was analyzed by using Analysis of Variance (ANOV A) If there any
significant differences the mean were discriminated using Least Significant Different
(LSD)
14
J I Pusat Khidmat MakJuUUlt Akademik UNlVERSm MALAYSIA SARAWAK
PHYSICAL CHARACTERISTIC AND GERMINATION OF HIBISCUS
SABDARIFFA L FOLLOWING HYDRATION TREATMENTS
P~i~liiimi 1000235560
SYAZW ANIE BINTI DZULHAIMI
This project work is submitted in partial fulfilment ofthe requirement for the Degree of
Bachelor of Science with Honours in Plant Resource Science and Management
Faculty of Resource Science and Technology
UNIVERSITI MALAYSIA SARA W AK
2012
ttl
APPROVAL SHEET
Name of candidate Syazwanie Binti Dzulhaimi
Matric no 25214
Title of thesis Physical characteristic and germination of Hibiscus sabdarifJa L
following hydration treatments
Approved by
(Assoc Prof Dr Petrus Bulan)
Supervisor
Plant Resource Science and Management
Faculty of Resource Science and Technology
Date
(Dr Siti Rubiah Zainudin)
Coordinator
Plant Resource Science and Management
Faculty of Resource Science and Technology
Date
ii
t I
DECLARATION
I hereby declare that no portion of this project work has been submitted in support of an
application for another degree of qualification of this or any other university or institution
of higher learning
(Syazwanie Binti Dzulhaimi)
Plant Resource Science and Management
Department of Environment Science and Ecology
Faculty of Resource Science and Technology
Universiti Malaysia Sarawak
iii
middot
ACKNOWLEDGEMENT
First and foremost I would like to give my highest praise to God for giving me the
strength patience intelligence and integrity to complete this research study
Alhamdulillah
My gratitude and sincere appreciation to Assoc Prof Or Petrus Bulan as my supervisor
for his continuous help support guidance advice and constant assisstance
I would also like to give my deepest thanks to my beloved parents Mr Dzulhaimi Khalid
and Mdm Khariyah Ahmad for their support understanding and encouragement
throughout my three years journey as an undergraduate student in Universiti Malaysia
Sarawak Not forgetting my other families whose been always supporting me along the
way
Thanks to the people that have contributed tremendously in my final year project Mdm
Fatimah Daud the laboratory assisstant of Cryopreservation Laboratory and all other
Faculty of Resource Science and Technology staff
Last but not least my appreciation goes to all my dearest friends especially to
Norakliliriana Abdul Rahman Siti Nadiah Salleh and to all my Plant Science coursemates
for their assistance and support throughout this study
iv
Pusat Khidmat Maklumat Akademik I
UNlVERSm MALAYSIA SARAWAK
TABLE OF CONTENTS
TITLE AND FRONT COVER I
APPROVAL SHEET II
DECLARATION III
ACKNOWLEDGEMENT IV
TABLE OF CONTENTS V
LIST OF FIGURES AND PLATES VII
ABSTRACT IX
10 INTRODUCTION 1 11 Background 1 12 Commercial Production 2 13 Medicinal Uses 3 14 Problem Statement 3 15 Objectives 4
20 LITERATURE REVIEW 5 21 Characteristic of Seed 5 22 Seed Germination 6 23 Seed Moisture Content 7 24 Seed Vigour 7 25 Seed Hydration 8 26 Seed Storage 9 27 Seed Specific Density 10 28 Seed Deterioration 10
30 MATERIAL AND METHOD 12 31 Material 12 32 Method 12
321 Preliminary Evaluation 12 322 Moisture Content Test 13 323 Germination Test 13 324 Seed Characteristic 14 325 Hydration of Seeds 14 326 Data Analysis 14
40 RESULT AND DISCUSSION 15 41 Preliminary Test 15 42 Seed Calibration 15 43 Seed Hydration 16
43 1 Seed Moisture Content 16 v
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t I shy
432 Seed Germination 22 433 Gennination Analysis 28
50 CONCLUSIONS AND RECOMMENDATION 29
60 REFERENCES 30
70 APPENDICES 31
vi
r---~~------------------------------------------------------------------~
LIST OF FIGURES AND PLATES
Figure Page
Moisture content of Hsabdariffa seeds for different period of 15
hydration at 28degC
2 Moisture content of Hsabdariffa seeds for different period of 16
hydration at 30degC
3 Moisture content of Hsabdariffa seeds for different period of 17
hydration at 35degC
4 Moisture content of Hsabdariffa seeds for different period of 18
hydration at 40degC
5 Moisture content of Hsabdariffa seeds for different period of 19
hydration at 28 30 35 and 40degC
6 Gennination of Hsabdariffa seeds for different period of 20
hydration at 28degC
7 Gennination of Hsabdariffa seeds for different period of 21
hydration at 30degC
8 Gennination ofHsabdariffa seeds for different period of 22
hydration at 35degC
Gennination of Hsabdariffa seeds for different period of
hydration at 40degC
9 23
vii
10 24
Germination of HsabdarifJa seeds for different period of
hydration at 28 30 35 and 40degC
Plate
Fresh fruit of H sabdarifJa 11
2 Fresh fruit of H sabdarifJa with seeds 11
3 Fresh seeds of H sabdarifJa 16
4 Dried seeds of H sabdarifJa 16
viii
--_shy
I
I
Physical characteristic and germination of Hibiscus Sabdariffa L following hydration treatments
Syazwanie Binti Dzulbaimi
Plant Resource and Management Programme Faculty of Science and Technology
Universiti Malaysia Sarawak
ABSTRACT
A study was conducted to determine the physical characteristics and to assess and evaluate the effectiveness of hydration techniques in improving the performance of HibisclIs sabdariffa L seeds The average length width and thickness obtained for H sabdariffa seeds were 539 mm 455 mm and 234 mm respectively The average weight for a single seed was 002 g and it was estimated that one kg weight contained approximately 50000 seeds This indicated that the seeds used in this experiment were small In hydration treatment seeds of H sabdariffa were soaked in water at temperature 28 30 35 and 40degC for 0246 8 10 and 12 hours and were evaluated for moisture content and germination Results indicated that hydration treatment was not suitable for H sabdariffa seeds as it did not improved the germination The highest germination percentage of H sabdarifJa seeds following hydration treatment was 29 which was obtained for 2 hours hydration at 30degC
Key words Hibiscus sabdarifJa calibration hydration temperature
ABSTRAK
Satu kajian telah dijalankan IInt1lk menentukan ciri-ciri jizikal dall mengkaji serta menentukan keberkesanan teknik rawatall hidrasi dalam meningkatkan kadar pertllmbllhan biji benih Hibiscus sabdariffa Dalam penentlkuran biji benih pIrata panjang lebar dan ketebalan biji benih H sabdariffa yang diperoleh adalah 539 mm 455 mm dan 234 mm Purata berat sebiji benih adalah 002 g dan dianggarkan bahawa satll kilogram mengandungi lebih kurang 50000 biji benih Ini memmjukkan yang biji benih yang digllnakan unIlk kajian ini agak keci Dalam rawatan hidrasi biji benih H sabdariffa direndam di dalam air pada suhu 28degC 30degC 35degC dan 40degC Bagi setiap suhu yang digunakan biji benih direndam selama 0 2 4 6 8 10 dan 12 jam dan selepas itu kandungan kelembapan dan percambahan biji benih dinilai Keputlsan yang diperoleh menllnjukkan rawatan hidrasi adalah tidak sesuai bagi biji benih H sabdariffa memalldangkan peratls percambahan tidak meningkat berbanding sebelum rawatan Peratlls percambahan tertinggi bagi biji benih H sabdariffa selepas rawatan hidrasi adalah 29 iaitll bagi hidrasi selama 2 jam pada slhu 30degC
Kata kllnciHibisclS sabdarifJa kalibrasi hidrasi suhl
ix
I bull
10 INTRODUCTION
11 Background
Hibiscus sabdarifJa Linn or also known as Roselle or red sorrel belongs to the family
Malvaceae It is also known as asam paya asam susur and asam kumbang locally It is a
new commercial crop in Malaysia where it was brought from India (Amin et al 2008)
Since Roselle plantation only had started in early 1990s it is considered as a new crop in
Malaysia Two introduced varieties in Malaysia are Arab and Terengganu variety
However in 2009 Universiti Kebangsaan Malaysia had launched three new varieties
named UKMR-l UKMR-2 and UKMR-3 to help promote the Roselle industry Recently
Department of Agriculture had collaborated with F AMA entrepreneurs and growers in
order to strengthen the H sabdarifJa industry in Malaysia by expanding and marketing this
product locally and in foreign market
According to Qi et al (2005) H sabdarifJa is an annual mostly brannched and erect
shrub Its reddish stem can achieve 35m tall The leaves are dark green to red alternate
glabrous long - petiolate palmately divided into 3 - 7 lobes with serrate margins Having
both male and female organs the flower are red to yellow in color with a dark centre
containing short peduncles
H sabdarifJa can survive in a wann and humid tropical and subtropical climate It can be
grown in a greenhouse with litle shade but the best condition for growing it is under the
full sunlight (Qi et aI 2005) Roselle plants are suitable for tropical climates with wellshy
distributed rain - fall of 1500 - 2000 mm per year from sea level to about 600 m in
altitude (Amin et aI 2008) It can adapt to variety of soils but prefered a friable sandy
loam with humus
1
Current production of H sabdarifJa in Malaysia is about 240 tonnes annually Reported in
unpublished result from Agricultural Department of Terengganu in the process of
removing the calyces out of 3 tonnes of raw materials about 15 tonnes of the velvety
capsules containing the seeds are being disposed as a by - product and unexploited
(Halimatul Amin Mohd Esa Nawalyah amp Siti 2007) Amin et at (2008) mentioned that
the plant takes about three to four months to reach the commercial stage of maturity before
the flowers are harvested
12 Commercial Production
Many parts of H sabdariffa including seeds leaves fruits and roots are very useful
especially the fleshy red calyces They are used fresh for making wine juice jam jelly
syrup gelatin pudding cakes ice cream and flavors and also dried and brewed into tea
spice and used for butter pies sauces tarts and other desserts (Qi et al 2005) In a study
about commercial practice of Roselle beverage production Bolade et at (2009) reported
that the dried calyceswater ratios involved in the commercial practice ranged between
157 and 171 (wv) while the sweetness level of the beverage ranged between 112 and
133 degBrix The red calyces which contains high amount of vitamin C was used to make
those healthy beverages In Malaysia we also consume the leaves as vegetables There are
previous study shows that H sabdarifJa seeds can be used as a protein source Instead of
using protein from conventional sources protein isolates or concentrates from Roselle
seeds might be useful as low cost source of protein substitute in dietary supplement or food
ingredient in food industry thus alleviating the problem of protein scarcity (Halimatul et
al 2007)
2
middot
13 Medicinal Uses
H sabdariffa is said to be useful in traditional medicine as a digestive agent purgative and
diuretic among others Having two diuretic ingredients ascorbic acid and glycolic acid it
has the ability to increase urination and also valued for its mild laxative effect Containing
citric acid it also can be used as cooling herb The tonic tea produce from the leaves and
flowers is good for digestive and kidney function The heated leaves are applied to cracks
in the feet and on boils and ulcers to speed maturation (Qi et aI 2005) Other roselle s part
have also been reported to be folk remedy for cancer obesity diabetes and hypertension
(Bolad et aI 2009) Amin et al (2008) also added that the flower and fleshy fruits are
used in pharmaceutical industry to relieve symptoms of bronchitis and coughs Roselle s
lotion products are useful for treating sores and wounds
Another nutritional value of Roselle discovered by UKM was the high content of
hydroxycitric acid (HCA) which is widely used as the main ingredients in many
commercial weight loss and slimming products Recent studies by local and international
scientist have proven the effectiveness of HCA in reducing body weight This
characteristic makes Roselle a very valuable crop
14 Problem Statement
The demand for H sabdariffa production nowadays had increase due to its commercial and
medicinal values To fulfill these demands more good quality seeds are needed
Furthermore not many research had been done on improving the H sabdarifJa seeds as a
planting material eventhough the best propagation method for H sabdariffa is by seeds
Other than that disease has been reported as a limiting factor to the production of Roselle
worldwide (Sie R S et aI 2011)
3
1S Objectives
1 To detennine the physical characteristics of H sabdariffa seeds for 10 100 and
1000 seeds weight and for length width and thickness of the seed
2 To evaluate the germination of H sabdariffa seeds following hydration at different
temperatures
3 To determine the effectiveness of hydration technique in improving the gennination
of H sabdariffa seeds
4
-
Pusat Khidmat Maklumat Akadtmik VNIVERSm MALAYSIA SARAWAK
I J bull bull
20 LITERATURE REVIEW
21 Characteristic of Seed
Seeds quality can be affected by changes in environmental temperature and relative
humidity The characteristic of seed must be determine because it is important to ensure
high quality yield in field planting Seed germination can be affected by number of factors
such as level of oxygen temperature and moisture content Growth performance of seed
also can be related to seed size Therefore grading is important to obtain seed that can
grows better Some useful criteria for visual separation of seed includes the seed size
shape colour weight thickness density and surface texture
According to Jain and Bal (1997) roselle seeds are having average principal dimensions of
298-336 186-224 and 170-201 mm Omobuwajo et al (2000) found that the average of
roselle seeds length width and thickness were 558 521 and 281 mm respectively The
seed was reported to be important for its oil in some parts of Africa its native origin From
previous research by scientist roselle seed was among the highest protein-containing seeds
when compared with other seeds like passion fruit (Passiflora edulis) Amaranthus seeds
and Pisum sativum seeds Latest findings by Hainida et al (2008) found that the seed from
Malaysia are composed of 99 moisture 335 protein 221 lipids 130 available
carbohydrate 183 total dietary fibers and 75 ash
H sabdarifJa can be grown easily in most of the tropical country Like other hibiscus
species it is quite hard for roselle to be affected by pest and disease and maintaining this
plant in the field is not so difficult The seeds were sown in a suitable container and the
seedlings will be ready for planting at the age of 10-15 days However it is also important
to ensure that the seeds to be sown were in a good condition and quality to avoid losses
5
22 Seed Germination
Germination is the emergence and development from the seed embryo of those essential
structures which are indicative of the ability to produce a normal plant under favorable
conditions (AOSA 1991) According to Smith (2011) the sum of germination events
beginning with hydration and culminating with root emergence Several stages involved in
germination are imbibition of water activation of enzyme systems metabolism of storage
products and their transport and finally the emergence of the radicle and growth of the
seedling Seed germination is described as the emergence and development from the seed
embryo of those essential structures which for the kind of seed in question are indicative
of the ability to produce a normal plant under favorable conditions (AOSA 1978) In a
simpler contcxt germination is the emergence of the radicle through the seed coat
Depending on the species seeds can germinate as quick as a few days or for as long as
several years Due to the high respiration rate and some exudation and leakage through the
seed coat the germinating seedling will undergoes a net loss in dry weight before the
germination For a particular kind of species the germination condition is mostly affected
by temperature type of substrate on which to germinate the seed light condition and
moisture level
The germination test is commonly used to determine seed viability It is an analytical
procedure to evaluate seed germination under standardized favorable conditions that are
seldom if ever encountered in the field The germination test is merely an estimate and
has certain limitations as a universal estimate of seed quality However if these limitations
are recognized the germination test is a useful viability index The need for germination
test is to estimate how the seed will perform in the field
6
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23 Seed Moisture Content
Amount of water in the seed that usually expressed in a percentage fonn is the seed
moisture content Agrawal (1980) found that the amount of moisture in the seed is
probably the most important factor influencing seed viability during storage By
determining the seeds moisture content the possible storage life for the seeds can be
predicted According to McDonald (2005) selecting the appropriate salt to achieve a seed
moisture content of 5-6 is the recommended method for achieving optimum seed
storage and minimizing seed deterioration It is necessary to dry seeds to the safe moisture
contents since the life of a seed and its span largely revolves around its moisture content
and the suitable moisture content for starchy seeds are less than 14 while for oily seeds
are less than 11 (Agrawal 1980)
24 Seed Vigour
According to Geneve (2005) seed vigour is defined by the Asociation of official Seed
analysts as those seed properties which detennine the potential for rapid unifonn
emergence and development of nonnal seedlings under the wide range of field conditions
Seed vigour is closely related with seed storage environment and duration During seed
storage the vigour of a seed lot is reduced prior to seed viability as indicated by standard
germination (Hampton amp TeKrony 1995) Major factors influencing the seed vigour in
stored seed lots are the environmental conditions during seed storage and the length of time
in storage Tetrazolium test is one of the method for detennining the seeds vigour To
minimize the decline in vigour and germination seeds should be undamaged
7
bull t I I
25 Seed Hydration
A process of supplying water to seed to maintain the fluid or moisture content in the seed
is called hydration According to Copeland and McDonald (1995) seed hydration is a
process whereby seeds are hydrated using various protocols and then redried to permit
routine handling This process results in increased germination rate more uniform
emergence germination under a broader range of environments and improved sedling
vigour and growth It is one of the priming treatments of seed which usually applied to
strengthen or increase vitality of seed Priming process can increase a rate of germination
as the treatment improves the seed vigor Hydration is one component of priming which
can increase germination with more range of environment condition The objective of seed
hydration technology is to increase the percentage and rate of germination expand the
range of temperatures over which the seed will germinate and increase the uniformity of
stand establishment To accomplish these objectives seeds must be hydrated in some way
at a moisture level sufficient to initiate the early events of germination but not sufficient to
permit radicle protrusion (Akers and Holley 1986) However the performance of the seed
might also being influence by other factors like the species type of seed and the seeds
quality
Water can be used for the hydration treatment because it had proved to enhance
germination emergence growth and yield of seed However other mediwns such as the
solution of potassium hydrophosphate (KH2P04) monobasic polyethylene glycol (PEG)
potassium chloride (KCl) potassiwn nitrate (KN03) potassium phoshate (K3P04)
magnesium sulphate (MgS04) sodiwn chloride (NaCl) glycerol and mannitol also can be
used to immerse the seed in hydration process The benefit of such salts is to supply the
seed with nitrogen and other nutrients essential for protein synthesis during germination
8
t I amp
26 Seed Storage
A basic requirement in seed production is a good seed storage The purpose of seed storage
is to preserve planting stocks from one season to the next (Larry amp McDonald 1995)
Generally we stored seeds for the commercial purpose for carry - over seeds or for genn
plasm seeds Maintaining the seed quality for the longest duration possible is the main
objective in seed storage Loss of gennination during storage cannot be stopped but it
could be reduce by giving good storage conditions According to Agrawal (1980) the
seeds are considered to be in storage from the moment they reach physiological maturity
until they genninate or until they are thrown away because they are dead or otherwise
worthless
The seed storage must be properly handled because it can affect the seed quality There are
some factors that can affect the seed longevity in storage such as the variety of the seed
initial seed quality moisture content relative humidity and storage temperature It is
necessary for the seed storage condition to be dry and cool effective in pest control and
have a proper sanitation Other than than storing only high quality of seed also can help in
better production From all of the factors temperature is one of the most important element
which influence seed viability and vigour during storage Agrawal (1980) mentioned that
the lower the temperature the longer the seeds maintain gennination capacity so
decreasing temperature and seed moisture is an effective means of maintaining seed quality
in storage Every seed lot should be gennination - tested when it is received for storage to
record the seed quality Subsequent periodic checks also should be made to detect
deterioration and if it occurs storage condition should be checked to discover the reasons
and to take corrective measures
9
middot
27 Seed Specific Density
Density of seed refers to it mass per unit volume The seed size and density will affect the
yield One of the method for grouping seeds into some specific density is the liquid density
separation system It is one of the easier and cheaper method compared to using a machine
When seed is loaded into a liquid some will float and some will sink That mean the
floating seeds density is lower than the liquids density Vice versa the sinking seeds
density is higher than the liquids density Wang and Alyarez (2008) explained that this
method of separating seeds works by adjusting the specific gravity of the liquid in the
reservoir so that a first portion of the seeds float near the surface of the liquid and a second
portion of the seeds sink near the bottom of the reservoir and providing an outlet flow of
the liquid from a location proximal to the top of the reservoir above the location of the
second portion of seeds By this way seeds of lower density can be separated from higher
density Kwong Sellman Jalink amp Schoor (2005) illustrated that separation by density is
commonly incorporated as part of the routine cleaning and grading process in flower seed
production procedures Other than grouping seeds according to their specific density this
density separation also can be used to remove broken partially filled empty or immature
seeds
28 Seed Deterioration
McDonald (2005) defined flower seed deterioration as deteriorative changes occuring with
time that increase the seeds vulnerability to external challenges and decrease the ability of
the seed to survive Important factors contribute to seed deterioration are genetics seed
structure seed chemistry physicalphysiological quality seed treatments relative humidity
10
and temperature Relative humidity is important because it will influence the moisture
content of seeds in storage while temperature determines the amount of moisture in the air
and influences the rate of deteriorative reactions in seeds According to Agrawal (1980)
the rate of deterioration increases as the seed moisture content increases This could be due
to the mold growth in and on the seed
11
~
30 MATERIAL AND METHOD
31 Material
Fresh matured fruits of Hibiscus sabdarifJa L (Plate I) were obtained from local fanns in
Kota Samarahan Seeds were extracted from these fruits cleaned and dried under shade
Dried seeds were dusted with Captan 80 and then placed in air-tight bottle to keep it in a
safe place before use in the subsequent experiment
Plate 1 Fresh fruit ofH sabdarifJa Plate 2 Fresh fruit of H sabdarifJa with seeds
32 Method
321 Preliminary Evaluation
Moisture content and germination was conducted as an initial evaluation to assess the
quality of seeds used in this research The procedures were those prescribed by Association
ofOfficial Seeds Analysts (AOSA 1985)
12
middot
322 Moisture Content Test
Four replicates of 10 seeds each were placed into saucers (Plate 5) and calibrated to get the
wet weight The seeds then were placed in an oven at 60degC for 48 hours After that the
seeds were weighed again to get the dry weight The percentage of moisture content of the
seeds was calculated based on the formula below (AOSA 1985)
Moisture content () = b - c x 100
b-a
where a = weight of empty saucer
b = weight of (a) + weight of seeds before drying
c = weight of (a) + weight of seeds after drying in the oven
323 Germination Test
Germination test was conducted base on the AOSA (1985) where 25 seeds with four
replications were used For every replicates seeds were put into a petry dish on 3 layers of
moist Whatman filter paper (Plate 6) The seeds were left to germinate by incubating it in a
Plant Growth Chamber (28OC) and was checked daily until 10 days Seed was considered
as germinated if the radicles were about 5mm length emerged from the seed coat The
germination percentage was calculated base on AOSA 1985 formula below
13
-----------------------------------1
Gennination () a x 100
b
where a = total number of germinated seeds
b = total number of seeds used
324 Seed Characteristic
The size of the H sabdariffa seeds was calibrated for its length width and thickness Ten
replicates of seeds were used for calibrating the physical characteristics Then the seed
weights of 10 100 and 1000 seeds were calibrated Four replicates of 10 100 and 1000
seed-weight classes were used
325 Hydration of Seeds
Seed of H sabdariffa was placed in distilled water for 0 2 4 6 8 10 and 12 hours at
temperatures of 28 30 35 and 40degC Then seeds were analyzed for the moisture content
and gennination
326 Data Analysis
The experimental units were arranged randomly based on Completely Randomized Design
(CRD) and data was analyzed by using Analysis of Variance (ANOV A) If there any
significant differences the mean were discriminated using Least Significant Different
(LSD)
14
ttl
APPROVAL SHEET
Name of candidate Syazwanie Binti Dzulhaimi
Matric no 25214
Title of thesis Physical characteristic and germination of Hibiscus sabdarifJa L
following hydration treatments
Approved by
(Assoc Prof Dr Petrus Bulan)
Supervisor
Plant Resource Science and Management
Faculty of Resource Science and Technology
Date
(Dr Siti Rubiah Zainudin)
Coordinator
Plant Resource Science and Management
Faculty of Resource Science and Technology
Date
ii
t I
DECLARATION
I hereby declare that no portion of this project work has been submitted in support of an
application for another degree of qualification of this or any other university or institution
of higher learning
(Syazwanie Binti Dzulhaimi)
Plant Resource Science and Management
Department of Environment Science and Ecology
Faculty of Resource Science and Technology
Universiti Malaysia Sarawak
iii
middot
ACKNOWLEDGEMENT
First and foremost I would like to give my highest praise to God for giving me the
strength patience intelligence and integrity to complete this research study
Alhamdulillah
My gratitude and sincere appreciation to Assoc Prof Or Petrus Bulan as my supervisor
for his continuous help support guidance advice and constant assisstance
I would also like to give my deepest thanks to my beloved parents Mr Dzulhaimi Khalid
and Mdm Khariyah Ahmad for their support understanding and encouragement
throughout my three years journey as an undergraduate student in Universiti Malaysia
Sarawak Not forgetting my other families whose been always supporting me along the
way
Thanks to the people that have contributed tremendously in my final year project Mdm
Fatimah Daud the laboratory assisstant of Cryopreservation Laboratory and all other
Faculty of Resource Science and Technology staff
Last but not least my appreciation goes to all my dearest friends especially to
Norakliliriana Abdul Rahman Siti Nadiah Salleh and to all my Plant Science coursemates
for their assistance and support throughout this study
iv
Pusat Khidmat Maklumat Akademik I
UNlVERSm MALAYSIA SARAWAK
TABLE OF CONTENTS
TITLE AND FRONT COVER I
APPROVAL SHEET II
DECLARATION III
ACKNOWLEDGEMENT IV
TABLE OF CONTENTS V
LIST OF FIGURES AND PLATES VII
ABSTRACT IX
10 INTRODUCTION 1 11 Background 1 12 Commercial Production 2 13 Medicinal Uses 3 14 Problem Statement 3 15 Objectives 4
20 LITERATURE REVIEW 5 21 Characteristic of Seed 5 22 Seed Germination 6 23 Seed Moisture Content 7 24 Seed Vigour 7 25 Seed Hydration 8 26 Seed Storage 9 27 Seed Specific Density 10 28 Seed Deterioration 10
30 MATERIAL AND METHOD 12 31 Material 12 32 Method 12
321 Preliminary Evaluation 12 322 Moisture Content Test 13 323 Germination Test 13 324 Seed Characteristic 14 325 Hydration of Seeds 14 326 Data Analysis 14
40 RESULT AND DISCUSSION 15 41 Preliminary Test 15 42 Seed Calibration 15 43 Seed Hydration 16
43 1 Seed Moisture Content 16 v
I
L
t I shy
432 Seed Germination 22 433 Gennination Analysis 28
50 CONCLUSIONS AND RECOMMENDATION 29
60 REFERENCES 30
70 APPENDICES 31
vi
r---~~------------------------------------------------------------------~
LIST OF FIGURES AND PLATES
Figure Page
Moisture content of Hsabdariffa seeds for different period of 15
hydration at 28degC
2 Moisture content of Hsabdariffa seeds for different period of 16
hydration at 30degC
3 Moisture content of Hsabdariffa seeds for different period of 17
hydration at 35degC
4 Moisture content of Hsabdariffa seeds for different period of 18
hydration at 40degC
5 Moisture content of Hsabdariffa seeds for different period of 19
hydration at 28 30 35 and 40degC
6 Gennination of Hsabdariffa seeds for different period of 20
hydration at 28degC
7 Gennination of Hsabdariffa seeds for different period of 21
hydration at 30degC
8 Gennination ofHsabdariffa seeds for different period of 22
hydration at 35degC
Gennination of Hsabdariffa seeds for different period of
hydration at 40degC
9 23
vii
10 24
Germination of HsabdarifJa seeds for different period of
hydration at 28 30 35 and 40degC
Plate
Fresh fruit of H sabdarifJa 11
2 Fresh fruit of H sabdarifJa with seeds 11
3 Fresh seeds of H sabdarifJa 16
4 Dried seeds of H sabdarifJa 16
viii
--_shy
I
I
Physical characteristic and germination of Hibiscus Sabdariffa L following hydration treatments
Syazwanie Binti Dzulbaimi
Plant Resource and Management Programme Faculty of Science and Technology
Universiti Malaysia Sarawak
ABSTRACT
A study was conducted to determine the physical characteristics and to assess and evaluate the effectiveness of hydration techniques in improving the performance of HibisclIs sabdariffa L seeds The average length width and thickness obtained for H sabdariffa seeds were 539 mm 455 mm and 234 mm respectively The average weight for a single seed was 002 g and it was estimated that one kg weight contained approximately 50000 seeds This indicated that the seeds used in this experiment were small In hydration treatment seeds of H sabdariffa were soaked in water at temperature 28 30 35 and 40degC for 0246 8 10 and 12 hours and were evaluated for moisture content and germination Results indicated that hydration treatment was not suitable for H sabdariffa seeds as it did not improved the germination The highest germination percentage of H sabdarifJa seeds following hydration treatment was 29 which was obtained for 2 hours hydration at 30degC
Key words Hibiscus sabdarifJa calibration hydration temperature
ABSTRAK
Satu kajian telah dijalankan IInt1lk menentukan ciri-ciri jizikal dall mengkaji serta menentukan keberkesanan teknik rawatall hidrasi dalam meningkatkan kadar pertllmbllhan biji benih Hibiscus sabdariffa Dalam penentlkuran biji benih pIrata panjang lebar dan ketebalan biji benih H sabdariffa yang diperoleh adalah 539 mm 455 mm dan 234 mm Purata berat sebiji benih adalah 002 g dan dianggarkan bahawa satll kilogram mengandungi lebih kurang 50000 biji benih Ini memmjukkan yang biji benih yang digllnakan unIlk kajian ini agak keci Dalam rawatan hidrasi biji benih H sabdariffa direndam di dalam air pada suhu 28degC 30degC 35degC dan 40degC Bagi setiap suhu yang digunakan biji benih direndam selama 0 2 4 6 8 10 dan 12 jam dan selepas itu kandungan kelembapan dan percambahan biji benih dinilai Keputlsan yang diperoleh menllnjukkan rawatan hidrasi adalah tidak sesuai bagi biji benih H sabdariffa memalldangkan peratls percambahan tidak meningkat berbanding sebelum rawatan Peratlls percambahan tertinggi bagi biji benih H sabdariffa selepas rawatan hidrasi adalah 29 iaitll bagi hidrasi selama 2 jam pada slhu 30degC
Kata kllnciHibisclS sabdarifJa kalibrasi hidrasi suhl
ix
I bull
10 INTRODUCTION
11 Background
Hibiscus sabdarifJa Linn or also known as Roselle or red sorrel belongs to the family
Malvaceae It is also known as asam paya asam susur and asam kumbang locally It is a
new commercial crop in Malaysia where it was brought from India (Amin et al 2008)
Since Roselle plantation only had started in early 1990s it is considered as a new crop in
Malaysia Two introduced varieties in Malaysia are Arab and Terengganu variety
However in 2009 Universiti Kebangsaan Malaysia had launched three new varieties
named UKMR-l UKMR-2 and UKMR-3 to help promote the Roselle industry Recently
Department of Agriculture had collaborated with F AMA entrepreneurs and growers in
order to strengthen the H sabdarifJa industry in Malaysia by expanding and marketing this
product locally and in foreign market
According to Qi et al (2005) H sabdarifJa is an annual mostly brannched and erect
shrub Its reddish stem can achieve 35m tall The leaves are dark green to red alternate
glabrous long - petiolate palmately divided into 3 - 7 lobes with serrate margins Having
both male and female organs the flower are red to yellow in color with a dark centre
containing short peduncles
H sabdarifJa can survive in a wann and humid tropical and subtropical climate It can be
grown in a greenhouse with litle shade but the best condition for growing it is under the
full sunlight (Qi et aI 2005) Roselle plants are suitable for tropical climates with wellshy
distributed rain - fall of 1500 - 2000 mm per year from sea level to about 600 m in
altitude (Amin et aI 2008) It can adapt to variety of soils but prefered a friable sandy
loam with humus
1
Current production of H sabdarifJa in Malaysia is about 240 tonnes annually Reported in
unpublished result from Agricultural Department of Terengganu in the process of
removing the calyces out of 3 tonnes of raw materials about 15 tonnes of the velvety
capsules containing the seeds are being disposed as a by - product and unexploited
(Halimatul Amin Mohd Esa Nawalyah amp Siti 2007) Amin et at (2008) mentioned that
the plant takes about three to four months to reach the commercial stage of maturity before
the flowers are harvested
12 Commercial Production
Many parts of H sabdariffa including seeds leaves fruits and roots are very useful
especially the fleshy red calyces They are used fresh for making wine juice jam jelly
syrup gelatin pudding cakes ice cream and flavors and also dried and brewed into tea
spice and used for butter pies sauces tarts and other desserts (Qi et al 2005) In a study
about commercial practice of Roselle beverage production Bolade et at (2009) reported
that the dried calyceswater ratios involved in the commercial practice ranged between
157 and 171 (wv) while the sweetness level of the beverage ranged between 112 and
133 degBrix The red calyces which contains high amount of vitamin C was used to make
those healthy beverages In Malaysia we also consume the leaves as vegetables There are
previous study shows that H sabdarifJa seeds can be used as a protein source Instead of
using protein from conventional sources protein isolates or concentrates from Roselle
seeds might be useful as low cost source of protein substitute in dietary supplement or food
ingredient in food industry thus alleviating the problem of protein scarcity (Halimatul et
al 2007)
2
middot
13 Medicinal Uses
H sabdariffa is said to be useful in traditional medicine as a digestive agent purgative and
diuretic among others Having two diuretic ingredients ascorbic acid and glycolic acid it
has the ability to increase urination and also valued for its mild laxative effect Containing
citric acid it also can be used as cooling herb The tonic tea produce from the leaves and
flowers is good for digestive and kidney function The heated leaves are applied to cracks
in the feet and on boils and ulcers to speed maturation (Qi et aI 2005) Other roselle s part
have also been reported to be folk remedy for cancer obesity diabetes and hypertension
(Bolad et aI 2009) Amin et al (2008) also added that the flower and fleshy fruits are
used in pharmaceutical industry to relieve symptoms of bronchitis and coughs Roselle s
lotion products are useful for treating sores and wounds
Another nutritional value of Roselle discovered by UKM was the high content of
hydroxycitric acid (HCA) which is widely used as the main ingredients in many
commercial weight loss and slimming products Recent studies by local and international
scientist have proven the effectiveness of HCA in reducing body weight This
characteristic makes Roselle a very valuable crop
14 Problem Statement
The demand for H sabdariffa production nowadays had increase due to its commercial and
medicinal values To fulfill these demands more good quality seeds are needed
Furthermore not many research had been done on improving the H sabdarifJa seeds as a
planting material eventhough the best propagation method for H sabdariffa is by seeds
Other than that disease has been reported as a limiting factor to the production of Roselle
worldwide (Sie R S et aI 2011)
3
1S Objectives
1 To detennine the physical characteristics of H sabdariffa seeds for 10 100 and
1000 seeds weight and for length width and thickness of the seed
2 To evaluate the germination of H sabdariffa seeds following hydration at different
temperatures
3 To determine the effectiveness of hydration technique in improving the gennination
of H sabdariffa seeds
4
-
Pusat Khidmat Maklumat Akadtmik VNIVERSm MALAYSIA SARAWAK
I J bull bull
20 LITERATURE REVIEW
21 Characteristic of Seed
Seeds quality can be affected by changes in environmental temperature and relative
humidity The characteristic of seed must be determine because it is important to ensure
high quality yield in field planting Seed germination can be affected by number of factors
such as level of oxygen temperature and moisture content Growth performance of seed
also can be related to seed size Therefore grading is important to obtain seed that can
grows better Some useful criteria for visual separation of seed includes the seed size
shape colour weight thickness density and surface texture
According to Jain and Bal (1997) roselle seeds are having average principal dimensions of
298-336 186-224 and 170-201 mm Omobuwajo et al (2000) found that the average of
roselle seeds length width and thickness were 558 521 and 281 mm respectively The
seed was reported to be important for its oil in some parts of Africa its native origin From
previous research by scientist roselle seed was among the highest protein-containing seeds
when compared with other seeds like passion fruit (Passiflora edulis) Amaranthus seeds
and Pisum sativum seeds Latest findings by Hainida et al (2008) found that the seed from
Malaysia are composed of 99 moisture 335 protein 221 lipids 130 available
carbohydrate 183 total dietary fibers and 75 ash
H sabdarifJa can be grown easily in most of the tropical country Like other hibiscus
species it is quite hard for roselle to be affected by pest and disease and maintaining this
plant in the field is not so difficult The seeds were sown in a suitable container and the
seedlings will be ready for planting at the age of 10-15 days However it is also important
to ensure that the seeds to be sown were in a good condition and quality to avoid losses
5
22 Seed Germination
Germination is the emergence and development from the seed embryo of those essential
structures which are indicative of the ability to produce a normal plant under favorable
conditions (AOSA 1991) According to Smith (2011) the sum of germination events
beginning with hydration and culminating with root emergence Several stages involved in
germination are imbibition of water activation of enzyme systems metabolism of storage
products and their transport and finally the emergence of the radicle and growth of the
seedling Seed germination is described as the emergence and development from the seed
embryo of those essential structures which for the kind of seed in question are indicative
of the ability to produce a normal plant under favorable conditions (AOSA 1978) In a
simpler contcxt germination is the emergence of the radicle through the seed coat
Depending on the species seeds can germinate as quick as a few days or for as long as
several years Due to the high respiration rate and some exudation and leakage through the
seed coat the germinating seedling will undergoes a net loss in dry weight before the
germination For a particular kind of species the germination condition is mostly affected
by temperature type of substrate on which to germinate the seed light condition and
moisture level
The germination test is commonly used to determine seed viability It is an analytical
procedure to evaluate seed germination under standardized favorable conditions that are
seldom if ever encountered in the field The germination test is merely an estimate and
has certain limitations as a universal estimate of seed quality However if these limitations
are recognized the germination test is a useful viability index The need for germination
test is to estimate how the seed will perform in the field
6
I bull bull
23 Seed Moisture Content
Amount of water in the seed that usually expressed in a percentage fonn is the seed
moisture content Agrawal (1980) found that the amount of moisture in the seed is
probably the most important factor influencing seed viability during storage By
determining the seeds moisture content the possible storage life for the seeds can be
predicted According to McDonald (2005) selecting the appropriate salt to achieve a seed
moisture content of 5-6 is the recommended method for achieving optimum seed
storage and minimizing seed deterioration It is necessary to dry seeds to the safe moisture
contents since the life of a seed and its span largely revolves around its moisture content
and the suitable moisture content for starchy seeds are less than 14 while for oily seeds
are less than 11 (Agrawal 1980)
24 Seed Vigour
According to Geneve (2005) seed vigour is defined by the Asociation of official Seed
analysts as those seed properties which detennine the potential for rapid unifonn
emergence and development of nonnal seedlings under the wide range of field conditions
Seed vigour is closely related with seed storage environment and duration During seed
storage the vigour of a seed lot is reduced prior to seed viability as indicated by standard
germination (Hampton amp TeKrony 1995) Major factors influencing the seed vigour in
stored seed lots are the environmental conditions during seed storage and the length of time
in storage Tetrazolium test is one of the method for detennining the seeds vigour To
minimize the decline in vigour and germination seeds should be undamaged
7
bull t I I
25 Seed Hydration
A process of supplying water to seed to maintain the fluid or moisture content in the seed
is called hydration According to Copeland and McDonald (1995) seed hydration is a
process whereby seeds are hydrated using various protocols and then redried to permit
routine handling This process results in increased germination rate more uniform
emergence germination under a broader range of environments and improved sedling
vigour and growth It is one of the priming treatments of seed which usually applied to
strengthen or increase vitality of seed Priming process can increase a rate of germination
as the treatment improves the seed vigor Hydration is one component of priming which
can increase germination with more range of environment condition The objective of seed
hydration technology is to increase the percentage and rate of germination expand the
range of temperatures over which the seed will germinate and increase the uniformity of
stand establishment To accomplish these objectives seeds must be hydrated in some way
at a moisture level sufficient to initiate the early events of germination but not sufficient to
permit radicle protrusion (Akers and Holley 1986) However the performance of the seed
might also being influence by other factors like the species type of seed and the seeds
quality
Water can be used for the hydration treatment because it had proved to enhance
germination emergence growth and yield of seed However other mediwns such as the
solution of potassium hydrophosphate (KH2P04) monobasic polyethylene glycol (PEG)
potassium chloride (KCl) potassiwn nitrate (KN03) potassium phoshate (K3P04)
magnesium sulphate (MgS04) sodiwn chloride (NaCl) glycerol and mannitol also can be
used to immerse the seed in hydration process The benefit of such salts is to supply the
seed with nitrogen and other nutrients essential for protein synthesis during germination
8
t I amp
26 Seed Storage
A basic requirement in seed production is a good seed storage The purpose of seed storage
is to preserve planting stocks from one season to the next (Larry amp McDonald 1995)
Generally we stored seeds for the commercial purpose for carry - over seeds or for genn
plasm seeds Maintaining the seed quality for the longest duration possible is the main
objective in seed storage Loss of gennination during storage cannot be stopped but it
could be reduce by giving good storage conditions According to Agrawal (1980) the
seeds are considered to be in storage from the moment they reach physiological maturity
until they genninate or until they are thrown away because they are dead or otherwise
worthless
The seed storage must be properly handled because it can affect the seed quality There are
some factors that can affect the seed longevity in storage such as the variety of the seed
initial seed quality moisture content relative humidity and storage temperature It is
necessary for the seed storage condition to be dry and cool effective in pest control and
have a proper sanitation Other than than storing only high quality of seed also can help in
better production From all of the factors temperature is one of the most important element
which influence seed viability and vigour during storage Agrawal (1980) mentioned that
the lower the temperature the longer the seeds maintain gennination capacity so
decreasing temperature and seed moisture is an effective means of maintaining seed quality
in storage Every seed lot should be gennination - tested when it is received for storage to
record the seed quality Subsequent periodic checks also should be made to detect
deterioration and if it occurs storage condition should be checked to discover the reasons
and to take corrective measures
9
middot
27 Seed Specific Density
Density of seed refers to it mass per unit volume The seed size and density will affect the
yield One of the method for grouping seeds into some specific density is the liquid density
separation system It is one of the easier and cheaper method compared to using a machine
When seed is loaded into a liquid some will float and some will sink That mean the
floating seeds density is lower than the liquids density Vice versa the sinking seeds
density is higher than the liquids density Wang and Alyarez (2008) explained that this
method of separating seeds works by adjusting the specific gravity of the liquid in the
reservoir so that a first portion of the seeds float near the surface of the liquid and a second
portion of the seeds sink near the bottom of the reservoir and providing an outlet flow of
the liquid from a location proximal to the top of the reservoir above the location of the
second portion of seeds By this way seeds of lower density can be separated from higher
density Kwong Sellman Jalink amp Schoor (2005) illustrated that separation by density is
commonly incorporated as part of the routine cleaning and grading process in flower seed
production procedures Other than grouping seeds according to their specific density this
density separation also can be used to remove broken partially filled empty or immature
seeds
28 Seed Deterioration
McDonald (2005) defined flower seed deterioration as deteriorative changes occuring with
time that increase the seeds vulnerability to external challenges and decrease the ability of
the seed to survive Important factors contribute to seed deterioration are genetics seed
structure seed chemistry physicalphysiological quality seed treatments relative humidity
10
and temperature Relative humidity is important because it will influence the moisture
content of seeds in storage while temperature determines the amount of moisture in the air
and influences the rate of deteriorative reactions in seeds According to Agrawal (1980)
the rate of deterioration increases as the seed moisture content increases This could be due
to the mold growth in and on the seed
11
~
30 MATERIAL AND METHOD
31 Material
Fresh matured fruits of Hibiscus sabdarifJa L (Plate I) were obtained from local fanns in
Kota Samarahan Seeds were extracted from these fruits cleaned and dried under shade
Dried seeds were dusted with Captan 80 and then placed in air-tight bottle to keep it in a
safe place before use in the subsequent experiment
Plate 1 Fresh fruit ofH sabdarifJa Plate 2 Fresh fruit of H sabdarifJa with seeds
32 Method
321 Preliminary Evaluation
Moisture content and germination was conducted as an initial evaluation to assess the
quality of seeds used in this research The procedures were those prescribed by Association
ofOfficial Seeds Analysts (AOSA 1985)
12
middot
322 Moisture Content Test
Four replicates of 10 seeds each were placed into saucers (Plate 5) and calibrated to get the
wet weight The seeds then were placed in an oven at 60degC for 48 hours After that the
seeds were weighed again to get the dry weight The percentage of moisture content of the
seeds was calculated based on the formula below (AOSA 1985)
Moisture content () = b - c x 100
b-a
where a = weight of empty saucer
b = weight of (a) + weight of seeds before drying
c = weight of (a) + weight of seeds after drying in the oven
323 Germination Test
Germination test was conducted base on the AOSA (1985) where 25 seeds with four
replications were used For every replicates seeds were put into a petry dish on 3 layers of
moist Whatman filter paper (Plate 6) The seeds were left to germinate by incubating it in a
Plant Growth Chamber (28OC) and was checked daily until 10 days Seed was considered
as germinated if the radicles were about 5mm length emerged from the seed coat The
germination percentage was calculated base on AOSA 1985 formula below
13
-----------------------------------1
Gennination () a x 100
b
where a = total number of germinated seeds
b = total number of seeds used
324 Seed Characteristic
The size of the H sabdariffa seeds was calibrated for its length width and thickness Ten
replicates of seeds were used for calibrating the physical characteristics Then the seed
weights of 10 100 and 1000 seeds were calibrated Four replicates of 10 100 and 1000
seed-weight classes were used
325 Hydration of Seeds
Seed of H sabdariffa was placed in distilled water for 0 2 4 6 8 10 and 12 hours at
temperatures of 28 30 35 and 40degC Then seeds were analyzed for the moisture content
and gennination
326 Data Analysis
The experimental units were arranged randomly based on Completely Randomized Design
(CRD) and data was analyzed by using Analysis of Variance (ANOV A) If there any
significant differences the mean were discriminated using Least Significant Different
(LSD)
14
t I
DECLARATION
I hereby declare that no portion of this project work has been submitted in support of an
application for another degree of qualification of this or any other university or institution
of higher learning
(Syazwanie Binti Dzulhaimi)
Plant Resource Science and Management
Department of Environment Science and Ecology
Faculty of Resource Science and Technology
Universiti Malaysia Sarawak
iii
middot
ACKNOWLEDGEMENT
First and foremost I would like to give my highest praise to God for giving me the
strength patience intelligence and integrity to complete this research study
Alhamdulillah
My gratitude and sincere appreciation to Assoc Prof Or Petrus Bulan as my supervisor
for his continuous help support guidance advice and constant assisstance
I would also like to give my deepest thanks to my beloved parents Mr Dzulhaimi Khalid
and Mdm Khariyah Ahmad for their support understanding and encouragement
throughout my three years journey as an undergraduate student in Universiti Malaysia
Sarawak Not forgetting my other families whose been always supporting me along the
way
Thanks to the people that have contributed tremendously in my final year project Mdm
Fatimah Daud the laboratory assisstant of Cryopreservation Laboratory and all other
Faculty of Resource Science and Technology staff
Last but not least my appreciation goes to all my dearest friends especially to
Norakliliriana Abdul Rahman Siti Nadiah Salleh and to all my Plant Science coursemates
for their assistance and support throughout this study
iv
Pusat Khidmat Maklumat Akademik I
UNlVERSm MALAYSIA SARAWAK
TABLE OF CONTENTS
TITLE AND FRONT COVER I
APPROVAL SHEET II
DECLARATION III
ACKNOWLEDGEMENT IV
TABLE OF CONTENTS V
LIST OF FIGURES AND PLATES VII
ABSTRACT IX
10 INTRODUCTION 1 11 Background 1 12 Commercial Production 2 13 Medicinal Uses 3 14 Problem Statement 3 15 Objectives 4
20 LITERATURE REVIEW 5 21 Characteristic of Seed 5 22 Seed Germination 6 23 Seed Moisture Content 7 24 Seed Vigour 7 25 Seed Hydration 8 26 Seed Storage 9 27 Seed Specific Density 10 28 Seed Deterioration 10
30 MATERIAL AND METHOD 12 31 Material 12 32 Method 12
321 Preliminary Evaluation 12 322 Moisture Content Test 13 323 Germination Test 13 324 Seed Characteristic 14 325 Hydration of Seeds 14 326 Data Analysis 14
40 RESULT AND DISCUSSION 15 41 Preliminary Test 15 42 Seed Calibration 15 43 Seed Hydration 16
43 1 Seed Moisture Content 16 v
I
L
t I shy
432 Seed Germination 22 433 Gennination Analysis 28
50 CONCLUSIONS AND RECOMMENDATION 29
60 REFERENCES 30
70 APPENDICES 31
vi
r---~~------------------------------------------------------------------~
LIST OF FIGURES AND PLATES
Figure Page
Moisture content of Hsabdariffa seeds for different period of 15
hydration at 28degC
2 Moisture content of Hsabdariffa seeds for different period of 16
hydration at 30degC
3 Moisture content of Hsabdariffa seeds for different period of 17
hydration at 35degC
4 Moisture content of Hsabdariffa seeds for different period of 18
hydration at 40degC
5 Moisture content of Hsabdariffa seeds for different period of 19
hydration at 28 30 35 and 40degC
6 Gennination of Hsabdariffa seeds for different period of 20
hydration at 28degC
7 Gennination of Hsabdariffa seeds for different period of 21
hydration at 30degC
8 Gennination ofHsabdariffa seeds for different period of 22
hydration at 35degC
Gennination of Hsabdariffa seeds for different period of
hydration at 40degC
9 23
vii
10 24
Germination of HsabdarifJa seeds for different period of
hydration at 28 30 35 and 40degC
Plate
Fresh fruit of H sabdarifJa 11
2 Fresh fruit of H sabdarifJa with seeds 11
3 Fresh seeds of H sabdarifJa 16
4 Dried seeds of H sabdarifJa 16
viii
--_shy
I
I
Physical characteristic and germination of Hibiscus Sabdariffa L following hydration treatments
Syazwanie Binti Dzulbaimi
Plant Resource and Management Programme Faculty of Science and Technology
Universiti Malaysia Sarawak
ABSTRACT
A study was conducted to determine the physical characteristics and to assess and evaluate the effectiveness of hydration techniques in improving the performance of HibisclIs sabdariffa L seeds The average length width and thickness obtained for H sabdariffa seeds were 539 mm 455 mm and 234 mm respectively The average weight for a single seed was 002 g and it was estimated that one kg weight contained approximately 50000 seeds This indicated that the seeds used in this experiment were small In hydration treatment seeds of H sabdariffa were soaked in water at temperature 28 30 35 and 40degC for 0246 8 10 and 12 hours and were evaluated for moisture content and germination Results indicated that hydration treatment was not suitable for H sabdariffa seeds as it did not improved the germination The highest germination percentage of H sabdarifJa seeds following hydration treatment was 29 which was obtained for 2 hours hydration at 30degC
Key words Hibiscus sabdarifJa calibration hydration temperature
ABSTRAK
Satu kajian telah dijalankan IInt1lk menentukan ciri-ciri jizikal dall mengkaji serta menentukan keberkesanan teknik rawatall hidrasi dalam meningkatkan kadar pertllmbllhan biji benih Hibiscus sabdariffa Dalam penentlkuran biji benih pIrata panjang lebar dan ketebalan biji benih H sabdariffa yang diperoleh adalah 539 mm 455 mm dan 234 mm Purata berat sebiji benih adalah 002 g dan dianggarkan bahawa satll kilogram mengandungi lebih kurang 50000 biji benih Ini memmjukkan yang biji benih yang digllnakan unIlk kajian ini agak keci Dalam rawatan hidrasi biji benih H sabdariffa direndam di dalam air pada suhu 28degC 30degC 35degC dan 40degC Bagi setiap suhu yang digunakan biji benih direndam selama 0 2 4 6 8 10 dan 12 jam dan selepas itu kandungan kelembapan dan percambahan biji benih dinilai Keputlsan yang diperoleh menllnjukkan rawatan hidrasi adalah tidak sesuai bagi biji benih H sabdariffa memalldangkan peratls percambahan tidak meningkat berbanding sebelum rawatan Peratlls percambahan tertinggi bagi biji benih H sabdariffa selepas rawatan hidrasi adalah 29 iaitll bagi hidrasi selama 2 jam pada slhu 30degC
Kata kllnciHibisclS sabdarifJa kalibrasi hidrasi suhl
ix
I bull
10 INTRODUCTION
11 Background
Hibiscus sabdarifJa Linn or also known as Roselle or red sorrel belongs to the family
Malvaceae It is also known as asam paya asam susur and asam kumbang locally It is a
new commercial crop in Malaysia where it was brought from India (Amin et al 2008)
Since Roselle plantation only had started in early 1990s it is considered as a new crop in
Malaysia Two introduced varieties in Malaysia are Arab and Terengganu variety
However in 2009 Universiti Kebangsaan Malaysia had launched three new varieties
named UKMR-l UKMR-2 and UKMR-3 to help promote the Roselle industry Recently
Department of Agriculture had collaborated with F AMA entrepreneurs and growers in
order to strengthen the H sabdarifJa industry in Malaysia by expanding and marketing this
product locally and in foreign market
According to Qi et al (2005) H sabdarifJa is an annual mostly brannched and erect
shrub Its reddish stem can achieve 35m tall The leaves are dark green to red alternate
glabrous long - petiolate palmately divided into 3 - 7 lobes with serrate margins Having
both male and female organs the flower are red to yellow in color with a dark centre
containing short peduncles
H sabdarifJa can survive in a wann and humid tropical and subtropical climate It can be
grown in a greenhouse with litle shade but the best condition for growing it is under the
full sunlight (Qi et aI 2005) Roselle plants are suitable for tropical climates with wellshy
distributed rain - fall of 1500 - 2000 mm per year from sea level to about 600 m in
altitude (Amin et aI 2008) It can adapt to variety of soils but prefered a friable sandy
loam with humus
1
Current production of H sabdarifJa in Malaysia is about 240 tonnes annually Reported in
unpublished result from Agricultural Department of Terengganu in the process of
removing the calyces out of 3 tonnes of raw materials about 15 tonnes of the velvety
capsules containing the seeds are being disposed as a by - product and unexploited
(Halimatul Amin Mohd Esa Nawalyah amp Siti 2007) Amin et at (2008) mentioned that
the plant takes about three to four months to reach the commercial stage of maturity before
the flowers are harvested
12 Commercial Production
Many parts of H sabdariffa including seeds leaves fruits and roots are very useful
especially the fleshy red calyces They are used fresh for making wine juice jam jelly
syrup gelatin pudding cakes ice cream and flavors and also dried and brewed into tea
spice and used for butter pies sauces tarts and other desserts (Qi et al 2005) In a study
about commercial practice of Roselle beverage production Bolade et at (2009) reported
that the dried calyceswater ratios involved in the commercial practice ranged between
157 and 171 (wv) while the sweetness level of the beverage ranged between 112 and
133 degBrix The red calyces which contains high amount of vitamin C was used to make
those healthy beverages In Malaysia we also consume the leaves as vegetables There are
previous study shows that H sabdarifJa seeds can be used as a protein source Instead of
using protein from conventional sources protein isolates or concentrates from Roselle
seeds might be useful as low cost source of protein substitute in dietary supplement or food
ingredient in food industry thus alleviating the problem of protein scarcity (Halimatul et
al 2007)
2
middot
13 Medicinal Uses
H sabdariffa is said to be useful in traditional medicine as a digestive agent purgative and
diuretic among others Having two diuretic ingredients ascorbic acid and glycolic acid it
has the ability to increase urination and also valued for its mild laxative effect Containing
citric acid it also can be used as cooling herb The tonic tea produce from the leaves and
flowers is good for digestive and kidney function The heated leaves are applied to cracks
in the feet and on boils and ulcers to speed maturation (Qi et aI 2005) Other roselle s part
have also been reported to be folk remedy for cancer obesity diabetes and hypertension
(Bolad et aI 2009) Amin et al (2008) also added that the flower and fleshy fruits are
used in pharmaceutical industry to relieve symptoms of bronchitis and coughs Roselle s
lotion products are useful for treating sores and wounds
Another nutritional value of Roselle discovered by UKM was the high content of
hydroxycitric acid (HCA) which is widely used as the main ingredients in many
commercial weight loss and slimming products Recent studies by local and international
scientist have proven the effectiveness of HCA in reducing body weight This
characteristic makes Roselle a very valuable crop
14 Problem Statement
The demand for H sabdariffa production nowadays had increase due to its commercial and
medicinal values To fulfill these demands more good quality seeds are needed
Furthermore not many research had been done on improving the H sabdarifJa seeds as a
planting material eventhough the best propagation method for H sabdariffa is by seeds
Other than that disease has been reported as a limiting factor to the production of Roselle
worldwide (Sie R S et aI 2011)
3
1S Objectives
1 To detennine the physical characteristics of H sabdariffa seeds for 10 100 and
1000 seeds weight and for length width and thickness of the seed
2 To evaluate the germination of H sabdariffa seeds following hydration at different
temperatures
3 To determine the effectiveness of hydration technique in improving the gennination
of H sabdariffa seeds
4
-
Pusat Khidmat Maklumat Akadtmik VNIVERSm MALAYSIA SARAWAK
I J bull bull
20 LITERATURE REVIEW
21 Characteristic of Seed
Seeds quality can be affected by changes in environmental temperature and relative
humidity The characteristic of seed must be determine because it is important to ensure
high quality yield in field planting Seed germination can be affected by number of factors
such as level of oxygen temperature and moisture content Growth performance of seed
also can be related to seed size Therefore grading is important to obtain seed that can
grows better Some useful criteria for visual separation of seed includes the seed size
shape colour weight thickness density and surface texture
According to Jain and Bal (1997) roselle seeds are having average principal dimensions of
298-336 186-224 and 170-201 mm Omobuwajo et al (2000) found that the average of
roselle seeds length width and thickness were 558 521 and 281 mm respectively The
seed was reported to be important for its oil in some parts of Africa its native origin From
previous research by scientist roselle seed was among the highest protein-containing seeds
when compared with other seeds like passion fruit (Passiflora edulis) Amaranthus seeds
and Pisum sativum seeds Latest findings by Hainida et al (2008) found that the seed from
Malaysia are composed of 99 moisture 335 protein 221 lipids 130 available
carbohydrate 183 total dietary fibers and 75 ash
H sabdarifJa can be grown easily in most of the tropical country Like other hibiscus
species it is quite hard for roselle to be affected by pest and disease and maintaining this
plant in the field is not so difficult The seeds were sown in a suitable container and the
seedlings will be ready for planting at the age of 10-15 days However it is also important
to ensure that the seeds to be sown were in a good condition and quality to avoid losses
5
22 Seed Germination
Germination is the emergence and development from the seed embryo of those essential
structures which are indicative of the ability to produce a normal plant under favorable
conditions (AOSA 1991) According to Smith (2011) the sum of germination events
beginning with hydration and culminating with root emergence Several stages involved in
germination are imbibition of water activation of enzyme systems metabolism of storage
products and their transport and finally the emergence of the radicle and growth of the
seedling Seed germination is described as the emergence and development from the seed
embryo of those essential structures which for the kind of seed in question are indicative
of the ability to produce a normal plant under favorable conditions (AOSA 1978) In a
simpler contcxt germination is the emergence of the radicle through the seed coat
Depending on the species seeds can germinate as quick as a few days or for as long as
several years Due to the high respiration rate and some exudation and leakage through the
seed coat the germinating seedling will undergoes a net loss in dry weight before the
germination For a particular kind of species the germination condition is mostly affected
by temperature type of substrate on which to germinate the seed light condition and
moisture level
The germination test is commonly used to determine seed viability It is an analytical
procedure to evaluate seed germination under standardized favorable conditions that are
seldom if ever encountered in the field The germination test is merely an estimate and
has certain limitations as a universal estimate of seed quality However if these limitations
are recognized the germination test is a useful viability index The need for germination
test is to estimate how the seed will perform in the field
6
I bull bull
23 Seed Moisture Content
Amount of water in the seed that usually expressed in a percentage fonn is the seed
moisture content Agrawal (1980) found that the amount of moisture in the seed is
probably the most important factor influencing seed viability during storage By
determining the seeds moisture content the possible storage life for the seeds can be
predicted According to McDonald (2005) selecting the appropriate salt to achieve a seed
moisture content of 5-6 is the recommended method for achieving optimum seed
storage and minimizing seed deterioration It is necessary to dry seeds to the safe moisture
contents since the life of a seed and its span largely revolves around its moisture content
and the suitable moisture content for starchy seeds are less than 14 while for oily seeds
are less than 11 (Agrawal 1980)
24 Seed Vigour
According to Geneve (2005) seed vigour is defined by the Asociation of official Seed
analysts as those seed properties which detennine the potential for rapid unifonn
emergence and development of nonnal seedlings under the wide range of field conditions
Seed vigour is closely related with seed storage environment and duration During seed
storage the vigour of a seed lot is reduced prior to seed viability as indicated by standard
germination (Hampton amp TeKrony 1995) Major factors influencing the seed vigour in
stored seed lots are the environmental conditions during seed storage and the length of time
in storage Tetrazolium test is one of the method for detennining the seeds vigour To
minimize the decline in vigour and germination seeds should be undamaged
7
bull t I I
25 Seed Hydration
A process of supplying water to seed to maintain the fluid or moisture content in the seed
is called hydration According to Copeland and McDonald (1995) seed hydration is a
process whereby seeds are hydrated using various protocols and then redried to permit
routine handling This process results in increased germination rate more uniform
emergence germination under a broader range of environments and improved sedling
vigour and growth It is one of the priming treatments of seed which usually applied to
strengthen or increase vitality of seed Priming process can increase a rate of germination
as the treatment improves the seed vigor Hydration is one component of priming which
can increase germination with more range of environment condition The objective of seed
hydration technology is to increase the percentage and rate of germination expand the
range of temperatures over which the seed will germinate and increase the uniformity of
stand establishment To accomplish these objectives seeds must be hydrated in some way
at a moisture level sufficient to initiate the early events of germination but not sufficient to
permit radicle protrusion (Akers and Holley 1986) However the performance of the seed
might also being influence by other factors like the species type of seed and the seeds
quality
Water can be used for the hydration treatment because it had proved to enhance
germination emergence growth and yield of seed However other mediwns such as the
solution of potassium hydrophosphate (KH2P04) monobasic polyethylene glycol (PEG)
potassium chloride (KCl) potassiwn nitrate (KN03) potassium phoshate (K3P04)
magnesium sulphate (MgS04) sodiwn chloride (NaCl) glycerol and mannitol also can be
used to immerse the seed in hydration process The benefit of such salts is to supply the
seed with nitrogen and other nutrients essential for protein synthesis during germination
8
t I amp
26 Seed Storage
A basic requirement in seed production is a good seed storage The purpose of seed storage
is to preserve planting stocks from one season to the next (Larry amp McDonald 1995)
Generally we stored seeds for the commercial purpose for carry - over seeds or for genn
plasm seeds Maintaining the seed quality for the longest duration possible is the main
objective in seed storage Loss of gennination during storage cannot be stopped but it
could be reduce by giving good storage conditions According to Agrawal (1980) the
seeds are considered to be in storage from the moment they reach physiological maturity
until they genninate or until they are thrown away because they are dead or otherwise
worthless
The seed storage must be properly handled because it can affect the seed quality There are
some factors that can affect the seed longevity in storage such as the variety of the seed
initial seed quality moisture content relative humidity and storage temperature It is
necessary for the seed storage condition to be dry and cool effective in pest control and
have a proper sanitation Other than than storing only high quality of seed also can help in
better production From all of the factors temperature is one of the most important element
which influence seed viability and vigour during storage Agrawal (1980) mentioned that
the lower the temperature the longer the seeds maintain gennination capacity so
decreasing temperature and seed moisture is an effective means of maintaining seed quality
in storage Every seed lot should be gennination - tested when it is received for storage to
record the seed quality Subsequent periodic checks also should be made to detect
deterioration and if it occurs storage condition should be checked to discover the reasons
and to take corrective measures
9
middot
27 Seed Specific Density
Density of seed refers to it mass per unit volume The seed size and density will affect the
yield One of the method for grouping seeds into some specific density is the liquid density
separation system It is one of the easier and cheaper method compared to using a machine
When seed is loaded into a liquid some will float and some will sink That mean the
floating seeds density is lower than the liquids density Vice versa the sinking seeds
density is higher than the liquids density Wang and Alyarez (2008) explained that this
method of separating seeds works by adjusting the specific gravity of the liquid in the
reservoir so that a first portion of the seeds float near the surface of the liquid and a second
portion of the seeds sink near the bottom of the reservoir and providing an outlet flow of
the liquid from a location proximal to the top of the reservoir above the location of the
second portion of seeds By this way seeds of lower density can be separated from higher
density Kwong Sellman Jalink amp Schoor (2005) illustrated that separation by density is
commonly incorporated as part of the routine cleaning and grading process in flower seed
production procedures Other than grouping seeds according to their specific density this
density separation also can be used to remove broken partially filled empty or immature
seeds
28 Seed Deterioration
McDonald (2005) defined flower seed deterioration as deteriorative changes occuring with
time that increase the seeds vulnerability to external challenges and decrease the ability of
the seed to survive Important factors contribute to seed deterioration are genetics seed
structure seed chemistry physicalphysiological quality seed treatments relative humidity
10
and temperature Relative humidity is important because it will influence the moisture
content of seeds in storage while temperature determines the amount of moisture in the air
and influences the rate of deteriorative reactions in seeds According to Agrawal (1980)
the rate of deterioration increases as the seed moisture content increases This could be due
to the mold growth in and on the seed
11
~
30 MATERIAL AND METHOD
31 Material
Fresh matured fruits of Hibiscus sabdarifJa L (Plate I) were obtained from local fanns in
Kota Samarahan Seeds were extracted from these fruits cleaned and dried under shade
Dried seeds were dusted with Captan 80 and then placed in air-tight bottle to keep it in a
safe place before use in the subsequent experiment
Plate 1 Fresh fruit ofH sabdarifJa Plate 2 Fresh fruit of H sabdarifJa with seeds
32 Method
321 Preliminary Evaluation
Moisture content and germination was conducted as an initial evaluation to assess the
quality of seeds used in this research The procedures were those prescribed by Association
ofOfficial Seeds Analysts (AOSA 1985)
12
middot
322 Moisture Content Test
Four replicates of 10 seeds each were placed into saucers (Plate 5) and calibrated to get the
wet weight The seeds then were placed in an oven at 60degC for 48 hours After that the
seeds were weighed again to get the dry weight The percentage of moisture content of the
seeds was calculated based on the formula below (AOSA 1985)
Moisture content () = b - c x 100
b-a
where a = weight of empty saucer
b = weight of (a) + weight of seeds before drying
c = weight of (a) + weight of seeds after drying in the oven
323 Germination Test
Germination test was conducted base on the AOSA (1985) where 25 seeds with four
replications were used For every replicates seeds were put into a petry dish on 3 layers of
moist Whatman filter paper (Plate 6) The seeds were left to germinate by incubating it in a
Plant Growth Chamber (28OC) and was checked daily until 10 days Seed was considered
as germinated if the radicles were about 5mm length emerged from the seed coat The
germination percentage was calculated base on AOSA 1985 formula below
13
-----------------------------------1
Gennination () a x 100
b
where a = total number of germinated seeds
b = total number of seeds used
324 Seed Characteristic
The size of the H sabdariffa seeds was calibrated for its length width and thickness Ten
replicates of seeds were used for calibrating the physical characteristics Then the seed
weights of 10 100 and 1000 seeds were calibrated Four replicates of 10 100 and 1000
seed-weight classes were used
325 Hydration of Seeds
Seed of H sabdariffa was placed in distilled water for 0 2 4 6 8 10 and 12 hours at
temperatures of 28 30 35 and 40degC Then seeds were analyzed for the moisture content
and gennination
326 Data Analysis
The experimental units were arranged randomly based on Completely Randomized Design
(CRD) and data was analyzed by using Analysis of Variance (ANOV A) If there any
significant differences the mean were discriminated using Least Significant Different
(LSD)
14
middot
ACKNOWLEDGEMENT
First and foremost I would like to give my highest praise to God for giving me the
strength patience intelligence and integrity to complete this research study
Alhamdulillah
My gratitude and sincere appreciation to Assoc Prof Or Petrus Bulan as my supervisor
for his continuous help support guidance advice and constant assisstance
I would also like to give my deepest thanks to my beloved parents Mr Dzulhaimi Khalid
and Mdm Khariyah Ahmad for their support understanding and encouragement
throughout my three years journey as an undergraduate student in Universiti Malaysia
Sarawak Not forgetting my other families whose been always supporting me along the
way
Thanks to the people that have contributed tremendously in my final year project Mdm
Fatimah Daud the laboratory assisstant of Cryopreservation Laboratory and all other
Faculty of Resource Science and Technology staff
Last but not least my appreciation goes to all my dearest friends especially to
Norakliliriana Abdul Rahman Siti Nadiah Salleh and to all my Plant Science coursemates
for their assistance and support throughout this study
iv
Pusat Khidmat Maklumat Akademik I
UNlVERSm MALAYSIA SARAWAK
TABLE OF CONTENTS
TITLE AND FRONT COVER I
APPROVAL SHEET II
DECLARATION III
ACKNOWLEDGEMENT IV
TABLE OF CONTENTS V
LIST OF FIGURES AND PLATES VII
ABSTRACT IX
10 INTRODUCTION 1 11 Background 1 12 Commercial Production 2 13 Medicinal Uses 3 14 Problem Statement 3 15 Objectives 4
20 LITERATURE REVIEW 5 21 Characteristic of Seed 5 22 Seed Germination 6 23 Seed Moisture Content 7 24 Seed Vigour 7 25 Seed Hydration 8 26 Seed Storage 9 27 Seed Specific Density 10 28 Seed Deterioration 10
30 MATERIAL AND METHOD 12 31 Material 12 32 Method 12
321 Preliminary Evaluation 12 322 Moisture Content Test 13 323 Germination Test 13 324 Seed Characteristic 14 325 Hydration of Seeds 14 326 Data Analysis 14
40 RESULT AND DISCUSSION 15 41 Preliminary Test 15 42 Seed Calibration 15 43 Seed Hydration 16
43 1 Seed Moisture Content 16 v
I
L
t I shy
432 Seed Germination 22 433 Gennination Analysis 28
50 CONCLUSIONS AND RECOMMENDATION 29
60 REFERENCES 30
70 APPENDICES 31
vi
r---~~------------------------------------------------------------------~
LIST OF FIGURES AND PLATES
Figure Page
Moisture content of Hsabdariffa seeds for different period of 15
hydration at 28degC
2 Moisture content of Hsabdariffa seeds for different period of 16
hydration at 30degC
3 Moisture content of Hsabdariffa seeds for different period of 17
hydration at 35degC
4 Moisture content of Hsabdariffa seeds for different period of 18
hydration at 40degC
5 Moisture content of Hsabdariffa seeds for different period of 19
hydration at 28 30 35 and 40degC
6 Gennination of Hsabdariffa seeds for different period of 20
hydration at 28degC
7 Gennination of Hsabdariffa seeds for different period of 21
hydration at 30degC
8 Gennination ofHsabdariffa seeds for different period of 22
hydration at 35degC
Gennination of Hsabdariffa seeds for different period of
hydration at 40degC
9 23
vii
10 24
Germination of HsabdarifJa seeds for different period of
hydration at 28 30 35 and 40degC
Plate
Fresh fruit of H sabdarifJa 11
2 Fresh fruit of H sabdarifJa with seeds 11
3 Fresh seeds of H sabdarifJa 16
4 Dried seeds of H sabdarifJa 16
viii
--_shy
I
I
Physical characteristic and germination of Hibiscus Sabdariffa L following hydration treatments
Syazwanie Binti Dzulbaimi
Plant Resource and Management Programme Faculty of Science and Technology
Universiti Malaysia Sarawak
ABSTRACT
A study was conducted to determine the physical characteristics and to assess and evaluate the effectiveness of hydration techniques in improving the performance of HibisclIs sabdariffa L seeds The average length width and thickness obtained for H sabdariffa seeds were 539 mm 455 mm and 234 mm respectively The average weight for a single seed was 002 g and it was estimated that one kg weight contained approximately 50000 seeds This indicated that the seeds used in this experiment were small In hydration treatment seeds of H sabdariffa were soaked in water at temperature 28 30 35 and 40degC for 0246 8 10 and 12 hours and were evaluated for moisture content and germination Results indicated that hydration treatment was not suitable for H sabdariffa seeds as it did not improved the germination The highest germination percentage of H sabdarifJa seeds following hydration treatment was 29 which was obtained for 2 hours hydration at 30degC
Key words Hibiscus sabdarifJa calibration hydration temperature
ABSTRAK
Satu kajian telah dijalankan IInt1lk menentukan ciri-ciri jizikal dall mengkaji serta menentukan keberkesanan teknik rawatall hidrasi dalam meningkatkan kadar pertllmbllhan biji benih Hibiscus sabdariffa Dalam penentlkuran biji benih pIrata panjang lebar dan ketebalan biji benih H sabdariffa yang diperoleh adalah 539 mm 455 mm dan 234 mm Purata berat sebiji benih adalah 002 g dan dianggarkan bahawa satll kilogram mengandungi lebih kurang 50000 biji benih Ini memmjukkan yang biji benih yang digllnakan unIlk kajian ini agak keci Dalam rawatan hidrasi biji benih H sabdariffa direndam di dalam air pada suhu 28degC 30degC 35degC dan 40degC Bagi setiap suhu yang digunakan biji benih direndam selama 0 2 4 6 8 10 dan 12 jam dan selepas itu kandungan kelembapan dan percambahan biji benih dinilai Keputlsan yang diperoleh menllnjukkan rawatan hidrasi adalah tidak sesuai bagi biji benih H sabdariffa memalldangkan peratls percambahan tidak meningkat berbanding sebelum rawatan Peratlls percambahan tertinggi bagi biji benih H sabdariffa selepas rawatan hidrasi adalah 29 iaitll bagi hidrasi selama 2 jam pada slhu 30degC
Kata kllnciHibisclS sabdarifJa kalibrasi hidrasi suhl
ix
I bull
10 INTRODUCTION
11 Background
Hibiscus sabdarifJa Linn or also known as Roselle or red sorrel belongs to the family
Malvaceae It is also known as asam paya asam susur and asam kumbang locally It is a
new commercial crop in Malaysia where it was brought from India (Amin et al 2008)
Since Roselle plantation only had started in early 1990s it is considered as a new crop in
Malaysia Two introduced varieties in Malaysia are Arab and Terengganu variety
However in 2009 Universiti Kebangsaan Malaysia had launched three new varieties
named UKMR-l UKMR-2 and UKMR-3 to help promote the Roselle industry Recently
Department of Agriculture had collaborated with F AMA entrepreneurs and growers in
order to strengthen the H sabdarifJa industry in Malaysia by expanding and marketing this
product locally and in foreign market
According to Qi et al (2005) H sabdarifJa is an annual mostly brannched and erect
shrub Its reddish stem can achieve 35m tall The leaves are dark green to red alternate
glabrous long - petiolate palmately divided into 3 - 7 lobes with serrate margins Having
both male and female organs the flower are red to yellow in color with a dark centre
containing short peduncles
H sabdarifJa can survive in a wann and humid tropical and subtropical climate It can be
grown in a greenhouse with litle shade but the best condition for growing it is under the
full sunlight (Qi et aI 2005) Roselle plants are suitable for tropical climates with wellshy
distributed rain - fall of 1500 - 2000 mm per year from sea level to about 600 m in
altitude (Amin et aI 2008) It can adapt to variety of soils but prefered a friable sandy
loam with humus
1
Current production of H sabdarifJa in Malaysia is about 240 tonnes annually Reported in
unpublished result from Agricultural Department of Terengganu in the process of
removing the calyces out of 3 tonnes of raw materials about 15 tonnes of the velvety
capsules containing the seeds are being disposed as a by - product and unexploited
(Halimatul Amin Mohd Esa Nawalyah amp Siti 2007) Amin et at (2008) mentioned that
the plant takes about three to four months to reach the commercial stage of maturity before
the flowers are harvested
12 Commercial Production
Many parts of H sabdariffa including seeds leaves fruits and roots are very useful
especially the fleshy red calyces They are used fresh for making wine juice jam jelly
syrup gelatin pudding cakes ice cream and flavors and also dried and brewed into tea
spice and used for butter pies sauces tarts and other desserts (Qi et al 2005) In a study
about commercial practice of Roselle beverage production Bolade et at (2009) reported
that the dried calyceswater ratios involved in the commercial practice ranged between
157 and 171 (wv) while the sweetness level of the beverage ranged between 112 and
133 degBrix The red calyces which contains high amount of vitamin C was used to make
those healthy beverages In Malaysia we also consume the leaves as vegetables There are
previous study shows that H sabdarifJa seeds can be used as a protein source Instead of
using protein from conventional sources protein isolates or concentrates from Roselle
seeds might be useful as low cost source of protein substitute in dietary supplement or food
ingredient in food industry thus alleviating the problem of protein scarcity (Halimatul et
al 2007)
2
middot
13 Medicinal Uses
H sabdariffa is said to be useful in traditional medicine as a digestive agent purgative and
diuretic among others Having two diuretic ingredients ascorbic acid and glycolic acid it
has the ability to increase urination and also valued for its mild laxative effect Containing
citric acid it also can be used as cooling herb The tonic tea produce from the leaves and
flowers is good for digestive and kidney function The heated leaves are applied to cracks
in the feet and on boils and ulcers to speed maturation (Qi et aI 2005) Other roselle s part
have also been reported to be folk remedy for cancer obesity diabetes and hypertension
(Bolad et aI 2009) Amin et al (2008) also added that the flower and fleshy fruits are
used in pharmaceutical industry to relieve symptoms of bronchitis and coughs Roselle s
lotion products are useful for treating sores and wounds
Another nutritional value of Roselle discovered by UKM was the high content of
hydroxycitric acid (HCA) which is widely used as the main ingredients in many
commercial weight loss and slimming products Recent studies by local and international
scientist have proven the effectiveness of HCA in reducing body weight This
characteristic makes Roselle a very valuable crop
14 Problem Statement
The demand for H sabdariffa production nowadays had increase due to its commercial and
medicinal values To fulfill these demands more good quality seeds are needed
Furthermore not many research had been done on improving the H sabdarifJa seeds as a
planting material eventhough the best propagation method for H sabdariffa is by seeds
Other than that disease has been reported as a limiting factor to the production of Roselle
worldwide (Sie R S et aI 2011)
3
1S Objectives
1 To detennine the physical characteristics of H sabdariffa seeds for 10 100 and
1000 seeds weight and for length width and thickness of the seed
2 To evaluate the germination of H sabdariffa seeds following hydration at different
temperatures
3 To determine the effectiveness of hydration technique in improving the gennination
of H sabdariffa seeds
4
-
Pusat Khidmat Maklumat Akadtmik VNIVERSm MALAYSIA SARAWAK
I J bull bull
20 LITERATURE REVIEW
21 Characteristic of Seed
Seeds quality can be affected by changes in environmental temperature and relative
humidity The characteristic of seed must be determine because it is important to ensure
high quality yield in field planting Seed germination can be affected by number of factors
such as level of oxygen temperature and moisture content Growth performance of seed
also can be related to seed size Therefore grading is important to obtain seed that can
grows better Some useful criteria for visual separation of seed includes the seed size
shape colour weight thickness density and surface texture
According to Jain and Bal (1997) roselle seeds are having average principal dimensions of
298-336 186-224 and 170-201 mm Omobuwajo et al (2000) found that the average of
roselle seeds length width and thickness were 558 521 and 281 mm respectively The
seed was reported to be important for its oil in some parts of Africa its native origin From
previous research by scientist roselle seed was among the highest protein-containing seeds
when compared with other seeds like passion fruit (Passiflora edulis) Amaranthus seeds
and Pisum sativum seeds Latest findings by Hainida et al (2008) found that the seed from
Malaysia are composed of 99 moisture 335 protein 221 lipids 130 available
carbohydrate 183 total dietary fibers and 75 ash
H sabdarifJa can be grown easily in most of the tropical country Like other hibiscus
species it is quite hard for roselle to be affected by pest and disease and maintaining this
plant in the field is not so difficult The seeds were sown in a suitable container and the
seedlings will be ready for planting at the age of 10-15 days However it is also important
to ensure that the seeds to be sown were in a good condition and quality to avoid losses
5
22 Seed Germination
Germination is the emergence and development from the seed embryo of those essential
structures which are indicative of the ability to produce a normal plant under favorable
conditions (AOSA 1991) According to Smith (2011) the sum of germination events
beginning with hydration and culminating with root emergence Several stages involved in
germination are imbibition of water activation of enzyme systems metabolism of storage
products and their transport and finally the emergence of the radicle and growth of the
seedling Seed germination is described as the emergence and development from the seed
embryo of those essential structures which for the kind of seed in question are indicative
of the ability to produce a normal plant under favorable conditions (AOSA 1978) In a
simpler contcxt germination is the emergence of the radicle through the seed coat
Depending on the species seeds can germinate as quick as a few days or for as long as
several years Due to the high respiration rate and some exudation and leakage through the
seed coat the germinating seedling will undergoes a net loss in dry weight before the
germination For a particular kind of species the germination condition is mostly affected
by temperature type of substrate on which to germinate the seed light condition and
moisture level
The germination test is commonly used to determine seed viability It is an analytical
procedure to evaluate seed germination under standardized favorable conditions that are
seldom if ever encountered in the field The germination test is merely an estimate and
has certain limitations as a universal estimate of seed quality However if these limitations
are recognized the germination test is a useful viability index The need for germination
test is to estimate how the seed will perform in the field
6
I bull bull
23 Seed Moisture Content
Amount of water in the seed that usually expressed in a percentage fonn is the seed
moisture content Agrawal (1980) found that the amount of moisture in the seed is
probably the most important factor influencing seed viability during storage By
determining the seeds moisture content the possible storage life for the seeds can be
predicted According to McDonald (2005) selecting the appropriate salt to achieve a seed
moisture content of 5-6 is the recommended method for achieving optimum seed
storage and minimizing seed deterioration It is necessary to dry seeds to the safe moisture
contents since the life of a seed and its span largely revolves around its moisture content
and the suitable moisture content for starchy seeds are less than 14 while for oily seeds
are less than 11 (Agrawal 1980)
24 Seed Vigour
According to Geneve (2005) seed vigour is defined by the Asociation of official Seed
analysts as those seed properties which detennine the potential for rapid unifonn
emergence and development of nonnal seedlings under the wide range of field conditions
Seed vigour is closely related with seed storage environment and duration During seed
storage the vigour of a seed lot is reduced prior to seed viability as indicated by standard
germination (Hampton amp TeKrony 1995) Major factors influencing the seed vigour in
stored seed lots are the environmental conditions during seed storage and the length of time
in storage Tetrazolium test is one of the method for detennining the seeds vigour To
minimize the decline in vigour and germination seeds should be undamaged
7
bull t I I
25 Seed Hydration
A process of supplying water to seed to maintain the fluid or moisture content in the seed
is called hydration According to Copeland and McDonald (1995) seed hydration is a
process whereby seeds are hydrated using various protocols and then redried to permit
routine handling This process results in increased germination rate more uniform
emergence germination under a broader range of environments and improved sedling
vigour and growth It is one of the priming treatments of seed which usually applied to
strengthen or increase vitality of seed Priming process can increase a rate of germination
as the treatment improves the seed vigor Hydration is one component of priming which
can increase germination with more range of environment condition The objective of seed
hydration technology is to increase the percentage and rate of germination expand the
range of temperatures over which the seed will germinate and increase the uniformity of
stand establishment To accomplish these objectives seeds must be hydrated in some way
at a moisture level sufficient to initiate the early events of germination but not sufficient to
permit radicle protrusion (Akers and Holley 1986) However the performance of the seed
might also being influence by other factors like the species type of seed and the seeds
quality
Water can be used for the hydration treatment because it had proved to enhance
germination emergence growth and yield of seed However other mediwns such as the
solution of potassium hydrophosphate (KH2P04) monobasic polyethylene glycol (PEG)
potassium chloride (KCl) potassiwn nitrate (KN03) potassium phoshate (K3P04)
magnesium sulphate (MgS04) sodiwn chloride (NaCl) glycerol and mannitol also can be
used to immerse the seed in hydration process The benefit of such salts is to supply the
seed with nitrogen and other nutrients essential for protein synthesis during germination
8
t I amp
26 Seed Storage
A basic requirement in seed production is a good seed storage The purpose of seed storage
is to preserve planting stocks from one season to the next (Larry amp McDonald 1995)
Generally we stored seeds for the commercial purpose for carry - over seeds or for genn
plasm seeds Maintaining the seed quality for the longest duration possible is the main
objective in seed storage Loss of gennination during storage cannot be stopped but it
could be reduce by giving good storage conditions According to Agrawal (1980) the
seeds are considered to be in storage from the moment they reach physiological maturity
until they genninate or until they are thrown away because they are dead or otherwise
worthless
The seed storage must be properly handled because it can affect the seed quality There are
some factors that can affect the seed longevity in storage such as the variety of the seed
initial seed quality moisture content relative humidity and storage temperature It is
necessary for the seed storage condition to be dry and cool effective in pest control and
have a proper sanitation Other than than storing only high quality of seed also can help in
better production From all of the factors temperature is one of the most important element
which influence seed viability and vigour during storage Agrawal (1980) mentioned that
the lower the temperature the longer the seeds maintain gennination capacity so
decreasing temperature and seed moisture is an effective means of maintaining seed quality
in storage Every seed lot should be gennination - tested when it is received for storage to
record the seed quality Subsequent periodic checks also should be made to detect
deterioration and if it occurs storage condition should be checked to discover the reasons
and to take corrective measures
9
middot
27 Seed Specific Density
Density of seed refers to it mass per unit volume The seed size and density will affect the
yield One of the method for grouping seeds into some specific density is the liquid density
separation system It is one of the easier and cheaper method compared to using a machine
When seed is loaded into a liquid some will float and some will sink That mean the
floating seeds density is lower than the liquids density Vice versa the sinking seeds
density is higher than the liquids density Wang and Alyarez (2008) explained that this
method of separating seeds works by adjusting the specific gravity of the liquid in the
reservoir so that a first portion of the seeds float near the surface of the liquid and a second
portion of the seeds sink near the bottom of the reservoir and providing an outlet flow of
the liquid from a location proximal to the top of the reservoir above the location of the
second portion of seeds By this way seeds of lower density can be separated from higher
density Kwong Sellman Jalink amp Schoor (2005) illustrated that separation by density is
commonly incorporated as part of the routine cleaning and grading process in flower seed
production procedures Other than grouping seeds according to their specific density this
density separation also can be used to remove broken partially filled empty or immature
seeds
28 Seed Deterioration
McDonald (2005) defined flower seed deterioration as deteriorative changes occuring with
time that increase the seeds vulnerability to external challenges and decrease the ability of
the seed to survive Important factors contribute to seed deterioration are genetics seed
structure seed chemistry physicalphysiological quality seed treatments relative humidity
10
and temperature Relative humidity is important because it will influence the moisture
content of seeds in storage while temperature determines the amount of moisture in the air
and influences the rate of deteriorative reactions in seeds According to Agrawal (1980)
the rate of deterioration increases as the seed moisture content increases This could be due
to the mold growth in and on the seed
11
~
30 MATERIAL AND METHOD
31 Material
Fresh matured fruits of Hibiscus sabdarifJa L (Plate I) were obtained from local fanns in
Kota Samarahan Seeds were extracted from these fruits cleaned and dried under shade
Dried seeds were dusted with Captan 80 and then placed in air-tight bottle to keep it in a
safe place before use in the subsequent experiment
Plate 1 Fresh fruit ofH sabdarifJa Plate 2 Fresh fruit of H sabdarifJa with seeds
32 Method
321 Preliminary Evaluation
Moisture content and germination was conducted as an initial evaluation to assess the
quality of seeds used in this research The procedures were those prescribed by Association
ofOfficial Seeds Analysts (AOSA 1985)
12
middot
322 Moisture Content Test
Four replicates of 10 seeds each were placed into saucers (Plate 5) and calibrated to get the
wet weight The seeds then were placed in an oven at 60degC for 48 hours After that the
seeds were weighed again to get the dry weight The percentage of moisture content of the
seeds was calculated based on the formula below (AOSA 1985)
Moisture content () = b - c x 100
b-a
where a = weight of empty saucer
b = weight of (a) + weight of seeds before drying
c = weight of (a) + weight of seeds after drying in the oven
323 Germination Test
Germination test was conducted base on the AOSA (1985) where 25 seeds with four
replications were used For every replicates seeds were put into a petry dish on 3 layers of
moist Whatman filter paper (Plate 6) The seeds were left to germinate by incubating it in a
Plant Growth Chamber (28OC) and was checked daily until 10 days Seed was considered
as germinated if the radicles were about 5mm length emerged from the seed coat The
germination percentage was calculated base on AOSA 1985 formula below
13
-----------------------------------1
Gennination () a x 100
b
where a = total number of germinated seeds
b = total number of seeds used
324 Seed Characteristic
The size of the H sabdariffa seeds was calibrated for its length width and thickness Ten
replicates of seeds were used for calibrating the physical characteristics Then the seed
weights of 10 100 and 1000 seeds were calibrated Four replicates of 10 100 and 1000
seed-weight classes were used
325 Hydration of Seeds
Seed of H sabdariffa was placed in distilled water for 0 2 4 6 8 10 and 12 hours at
temperatures of 28 30 35 and 40degC Then seeds were analyzed for the moisture content
and gennination
326 Data Analysis
The experimental units were arranged randomly based on Completely Randomized Design
(CRD) and data was analyzed by using Analysis of Variance (ANOV A) If there any
significant differences the mean were discriminated using Least Significant Different
(LSD)
14
Pusat Khidmat Maklumat Akademik I
UNlVERSm MALAYSIA SARAWAK
TABLE OF CONTENTS
TITLE AND FRONT COVER I
APPROVAL SHEET II
DECLARATION III
ACKNOWLEDGEMENT IV
TABLE OF CONTENTS V
LIST OF FIGURES AND PLATES VII
ABSTRACT IX
10 INTRODUCTION 1 11 Background 1 12 Commercial Production 2 13 Medicinal Uses 3 14 Problem Statement 3 15 Objectives 4
20 LITERATURE REVIEW 5 21 Characteristic of Seed 5 22 Seed Germination 6 23 Seed Moisture Content 7 24 Seed Vigour 7 25 Seed Hydration 8 26 Seed Storage 9 27 Seed Specific Density 10 28 Seed Deterioration 10
30 MATERIAL AND METHOD 12 31 Material 12 32 Method 12
321 Preliminary Evaluation 12 322 Moisture Content Test 13 323 Germination Test 13 324 Seed Characteristic 14 325 Hydration of Seeds 14 326 Data Analysis 14
40 RESULT AND DISCUSSION 15 41 Preliminary Test 15 42 Seed Calibration 15 43 Seed Hydration 16
43 1 Seed Moisture Content 16 v
I
L
t I shy
432 Seed Germination 22 433 Gennination Analysis 28
50 CONCLUSIONS AND RECOMMENDATION 29
60 REFERENCES 30
70 APPENDICES 31
vi
r---~~------------------------------------------------------------------~
LIST OF FIGURES AND PLATES
Figure Page
Moisture content of Hsabdariffa seeds for different period of 15
hydration at 28degC
2 Moisture content of Hsabdariffa seeds for different period of 16
hydration at 30degC
3 Moisture content of Hsabdariffa seeds for different period of 17
hydration at 35degC
4 Moisture content of Hsabdariffa seeds for different period of 18
hydration at 40degC
5 Moisture content of Hsabdariffa seeds for different period of 19
hydration at 28 30 35 and 40degC
6 Gennination of Hsabdariffa seeds for different period of 20
hydration at 28degC
7 Gennination of Hsabdariffa seeds for different period of 21
hydration at 30degC
8 Gennination ofHsabdariffa seeds for different period of 22
hydration at 35degC
Gennination of Hsabdariffa seeds for different period of
hydration at 40degC
9 23
vii
10 24
Germination of HsabdarifJa seeds for different period of
hydration at 28 30 35 and 40degC
Plate
Fresh fruit of H sabdarifJa 11
2 Fresh fruit of H sabdarifJa with seeds 11
3 Fresh seeds of H sabdarifJa 16
4 Dried seeds of H sabdarifJa 16
viii
--_shy
I
I
Physical characteristic and germination of Hibiscus Sabdariffa L following hydration treatments
Syazwanie Binti Dzulbaimi
Plant Resource and Management Programme Faculty of Science and Technology
Universiti Malaysia Sarawak
ABSTRACT
A study was conducted to determine the physical characteristics and to assess and evaluate the effectiveness of hydration techniques in improving the performance of HibisclIs sabdariffa L seeds The average length width and thickness obtained for H sabdariffa seeds were 539 mm 455 mm and 234 mm respectively The average weight for a single seed was 002 g and it was estimated that one kg weight contained approximately 50000 seeds This indicated that the seeds used in this experiment were small In hydration treatment seeds of H sabdariffa were soaked in water at temperature 28 30 35 and 40degC for 0246 8 10 and 12 hours and were evaluated for moisture content and germination Results indicated that hydration treatment was not suitable for H sabdariffa seeds as it did not improved the germination The highest germination percentage of H sabdarifJa seeds following hydration treatment was 29 which was obtained for 2 hours hydration at 30degC
Key words Hibiscus sabdarifJa calibration hydration temperature
ABSTRAK
Satu kajian telah dijalankan IInt1lk menentukan ciri-ciri jizikal dall mengkaji serta menentukan keberkesanan teknik rawatall hidrasi dalam meningkatkan kadar pertllmbllhan biji benih Hibiscus sabdariffa Dalam penentlkuran biji benih pIrata panjang lebar dan ketebalan biji benih H sabdariffa yang diperoleh adalah 539 mm 455 mm dan 234 mm Purata berat sebiji benih adalah 002 g dan dianggarkan bahawa satll kilogram mengandungi lebih kurang 50000 biji benih Ini memmjukkan yang biji benih yang digllnakan unIlk kajian ini agak keci Dalam rawatan hidrasi biji benih H sabdariffa direndam di dalam air pada suhu 28degC 30degC 35degC dan 40degC Bagi setiap suhu yang digunakan biji benih direndam selama 0 2 4 6 8 10 dan 12 jam dan selepas itu kandungan kelembapan dan percambahan biji benih dinilai Keputlsan yang diperoleh menllnjukkan rawatan hidrasi adalah tidak sesuai bagi biji benih H sabdariffa memalldangkan peratls percambahan tidak meningkat berbanding sebelum rawatan Peratlls percambahan tertinggi bagi biji benih H sabdariffa selepas rawatan hidrasi adalah 29 iaitll bagi hidrasi selama 2 jam pada slhu 30degC
Kata kllnciHibisclS sabdarifJa kalibrasi hidrasi suhl
ix
I bull
10 INTRODUCTION
11 Background
Hibiscus sabdarifJa Linn or also known as Roselle or red sorrel belongs to the family
Malvaceae It is also known as asam paya asam susur and asam kumbang locally It is a
new commercial crop in Malaysia where it was brought from India (Amin et al 2008)
Since Roselle plantation only had started in early 1990s it is considered as a new crop in
Malaysia Two introduced varieties in Malaysia are Arab and Terengganu variety
However in 2009 Universiti Kebangsaan Malaysia had launched three new varieties
named UKMR-l UKMR-2 and UKMR-3 to help promote the Roselle industry Recently
Department of Agriculture had collaborated with F AMA entrepreneurs and growers in
order to strengthen the H sabdarifJa industry in Malaysia by expanding and marketing this
product locally and in foreign market
According to Qi et al (2005) H sabdarifJa is an annual mostly brannched and erect
shrub Its reddish stem can achieve 35m tall The leaves are dark green to red alternate
glabrous long - petiolate palmately divided into 3 - 7 lobes with serrate margins Having
both male and female organs the flower are red to yellow in color with a dark centre
containing short peduncles
H sabdarifJa can survive in a wann and humid tropical and subtropical climate It can be
grown in a greenhouse with litle shade but the best condition for growing it is under the
full sunlight (Qi et aI 2005) Roselle plants are suitable for tropical climates with wellshy
distributed rain - fall of 1500 - 2000 mm per year from sea level to about 600 m in
altitude (Amin et aI 2008) It can adapt to variety of soils but prefered a friable sandy
loam with humus
1
Current production of H sabdarifJa in Malaysia is about 240 tonnes annually Reported in
unpublished result from Agricultural Department of Terengganu in the process of
removing the calyces out of 3 tonnes of raw materials about 15 tonnes of the velvety
capsules containing the seeds are being disposed as a by - product and unexploited
(Halimatul Amin Mohd Esa Nawalyah amp Siti 2007) Amin et at (2008) mentioned that
the plant takes about three to four months to reach the commercial stage of maturity before
the flowers are harvested
12 Commercial Production
Many parts of H sabdariffa including seeds leaves fruits and roots are very useful
especially the fleshy red calyces They are used fresh for making wine juice jam jelly
syrup gelatin pudding cakes ice cream and flavors and also dried and brewed into tea
spice and used for butter pies sauces tarts and other desserts (Qi et al 2005) In a study
about commercial practice of Roselle beverage production Bolade et at (2009) reported
that the dried calyceswater ratios involved in the commercial practice ranged between
157 and 171 (wv) while the sweetness level of the beverage ranged between 112 and
133 degBrix The red calyces which contains high amount of vitamin C was used to make
those healthy beverages In Malaysia we also consume the leaves as vegetables There are
previous study shows that H sabdarifJa seeds can be used as a protein source Instead of
using protein from conventional sources protein isolates or concentrates from Roselle
seeds might be useful as low cost source of protein substitute in dietary supplement or food
ingredient in food industry thus alleviating the problem of protein scarcity (Halimatul et
al 2007)
2
middot
13 Medicinal Uses
H sabdariffa is said to be useful in traditional medicine as a digestive agent purgative and
diuretic among others Having two diuretic ingredients ascorbic acid and glycolic acid it
has the ability to increase urination and also valued for its mild laxative effect Containing
citric acid it also can be used as cooling herb The tonic tea produce from the leaves and
flowers is good for digestive and kidney function The heated leaves are applied to cracks
in the feet and on boils and ulcers to speed maturation (Qi et aI 2005) Other roselle s part
have also been reported to be folk remedy for cancer obesity diabetes and hypertension
(Bolad et aI 2009) Amin et al (2008) also added that the flower and fleshy fruits are
used in pharmaceutical industry to relieve symptoms of bronchitis and coughs Roselle s
lotion products are useful for treating sores and wounds
Another nutritional value of Roselle discovered by UKM was the high content of
hydroxycitric acid (HCA) which is widely used as the main ingredients in many
commercial weight loss and slimming products Recent studies by local and international
scientist have proven the effectiveness of HCA in reducing body weight This
characteristic makes Roselle a very valuable crop
14 Problem Statement
The demand for H sabdariffa production nowadays had increase due to its commercial and
medicinal values To fulfill these demands more good quality seeds are needed
Furthermore not many research had been done on improving the H sabdarifJa seeds as a
planting material eventhough the best propagation method for H sabdariffa is by seeds
Other than that disease has been reported as a limiting factor to the production of Roselle
worldwide (Sie R S et aI 2011)
3
1S Objectives
1 To detennine the physical characteristics of H sabdariffa seeds for 10 100 and
1000 seeds weight and for length width and thickness of the seed
2 To evaluate the germination of H sabdariffa seeds following hydration at different
temperatures
3 To determine the effectiveness of hydration technique in improving the gennination
of H sabdariffa seeds
4
-
Pusat Khidmat Maklumat Akadtmik VNIVERSm MALAYSIA SARAWAK
I J bull bull
20 LITERATURE REVIEW
21 Characteristic of Seed
Seeds quality can be affected by changes in environmental temperature and relative
humidity The characteristic of seed must be determine because it is important to ensure
high quality yield in field planting Seed germination can be affected by number of factors
such as level of oxygen temperature and moisture content Growth performance of seed
also can be related to seed size Therefore grading is important to obtain seed that can
grows better Some useful criteria for visual separation of seed includes the seed size
shape colour weight thickness density and surface texture
According to Jain and Bal (1997) roselle seeds are having average principal dimensions of
298-336 186-224 and 170-201 mm Omobuwajo et al (2000) found that the average of
roselle seeds length width and thickness were 558 521 and 281 mm respectively The
seed was reported to be important for its oil in some parts of Africa its native origin From
previous research by scientist roselle seed was among the highest protein-containing seeds
when compared with other seeds like passion fruit (Passiflora edulis) Amaranthus seeds
and Pisum sativum seeds Latest findings by Hainida et al (2008) found that the seed from
Malaysia are composed of 99 moisture 335 protein 221 lipids 130 available
carbohydrate 183 total dietary fibers and 75 ash
H sabdarifJa can be grown easily in most of the tropical country Like other hibiscus
species it is quite hard for roselle to be affected by pest and disease and maintaining this
plant in the field is not so difficult The seeds were sown in a suitable container and the
seedlings will be ready for planting at the age of 10-15 days However it is also important
to ensure that the seeds to be sown were in a good condition and quality to avoid losses
5
22 Seed Germination
Germination is the emergence and development from the seed embryo of those essential
structures which are indicative of the ability to produce a normal plant under favorable
conditions (AOSA 1991) According to Smith (2011) the sum of germination events
beginning with hydration and culminating with root emergence Several stages involved in
germination are imbibition of water activation of enzyme systems metabolism of storage
products and their transport and finally the emergence of the radicle and growth of the
seedling Seed germination is described as the emergence and development from the seed
embryo of those essential structures which for the kind of seed in question are indicative
of the ability to produce a normal plant under favorable conditions (AOSA 1978) In a
simpler contcxt germination is the emergence of the radicle through the seed coat
Depending on the species seeds can germinate as quick as a few days or for as long as
several years Due to the high respiration rate and some exudation and leakage through the
seed coat the germinating seedling will undergoes a net loss in dry weight before the
germination For a particular kind of species the germination condition is mostly affected
by temperature type of substrate on which to germinate the seed light condition and
moisture level
The germination test is commonly used to determine seed viability It is an analytical
procedure to evaluate seed germination under standardized favorable conditions that are
seldom if ever encountered in the field The germination test is merely an estimate and
has certain limitations as a universal estimate of seed quality However if these limitations
are recognized the germination test is a useful viability index The need for germination
test is to estimate how the seed will perform in the field
6
I bull bull
23 Seed Moisture Content
Amount of water in the seed that usually expressed in a percentage fonn is the seed
moisture content Agrawal (1980) found that the amount of moisture in the seed is
probably the most important factor influencing seed viability during storage By
determining the seeds moisture content the possible storage life for the seeds can be
predicted According to McDonald (2005) selecting the appropriate salt to achieve a seed
moisture content of 5-6 is the recommended method for achieving optimum seed
storage and minimizing seed deterioration It is necessary to dry seeds to the safe moisture
contents since the life of a seed and its span largely revolves around its moisture content
and the suitable moisture content for starchy seeds are less than 14 while for oily seeds
are less than 11 (Agrawal 1980)
24 Seed Vigour
According to Geneve (2005) seed vigour is defined by the Asociation of official Seed
analysts as those seed properties which detennine the potential for rapid unifonn
emergence and development of nonnal seedlings under the wide range of field conditions
Seed vigour is closely related with seed storage environment and duration During seed
storage the vigour of a seed lot is reduced prior to seed viability as indicated by standard
germination (Hampton amp TeKrony 1995) Major factors influencing the seed vigour in
stored seed lots are the environmental conditions during seed storage and the length of time
in storage Tetrazolium test is one of the method for detennining the seeds vigour To
minimize the decline in vigour and germination seeds should be undamaged
7
bull t I I
25 Seed Hydration
A process of supplying water to seed to maintain the fluid or moisture content in the seed
is called hydration According to Copeland and McDonald (1995) seed hydration is a
process whereby seeds are hydrated using various protocols and then redried to permit
routine handling This process results in increased germination rate more uniform
emergence germination under a broader range of environments and improved sedling
vigour and growth It is one of the priming treatments of seed which usually applied to
strengthen or increase vitality of seed Priming process can increase a rate of germination
as the treatment improves the seed vigor Hydration is one component of priming which
can increase germination with more range of environment condition The objective of seed
hydration technology is to increase the percentage and rate of germination expand the
range of temperatures over which the seed will germinate and increase the uniformity of
stand establishment To accomplish these objectives seeds must be hydrated in some way
at a moisture level sufficient to initiate the early events of germination but not sufficient to
permit radicle protrusion (Akers and Holley 1986) However the performance of the seed
might also being influence by other factors like the species type of seed and the seeds
quality
Water can be used for the hydration treatment because it had proved to enhance
germination emergence growth and yield of seed However other mediwns such as the
solution of potassium hydrophosphate (KH2P04) monobasic polyethylene glycol (PEG)
potassium chloride (KCl) potassiwn nitrate (KN03) potassium phoshate (K3P04)
magnesium sulphate (MgS04) sodiwn chloride (NaCl) glycerol and mannitol also can be
used to immerse the seed in hydration process The benefit of such salts is to supply the
seed with nitrogen and other nutrients essential for protein synthesis during germination
8
t I amp
26 Seed Storage
A basic requirement in seed production is a good seed storage The purpose of seed storage
is to preserve planting stocks from one season to the next (Larry amp McDonald 1995)
Generally we stored seeds for the commercial purpose for carry - over seeds or for genn
plasm seeds Maintaining the seed quality for the longest duration possible is the main
objective in seed storage Loss of gennination during storage cannot be stopped but it
could be reduce by giving good storage conditions According to Agrawal (1980) the
seeds are considered to be in storage from the moment they reach physiological maturity
until they genninate or until they are thrown away because they are dead or otherwise
worthless
The seed storage must be properly handled because it can affect the seed quality There are
some factors that can affect the seed longevity in storage such as the variety of the seed
initial seed quality moisture content relative humidity and storage temperature It is
necessary for the seed storage condition to be dry and cool effective in pest control and
have a proper sanitation Other than than storing only high quality of seed also can help in
better production From all of the factors temperature is one of the most important element
which influence seed viability and vigour during storage Agrawal (1980) mentioned that
the lower the temperature the longer the seeds maintain gennination capacity so
decreasing temperature and seed moisture is an effective means of maintaining seed quality
in storage Every seed lot should be gennination - tested when it is received for storage to
record the seed quality Subsequent periodic checks also should be made to detect
deterioration and if it occurs storage condition should be checked to discover the reasons
and to take corrective measures
9
middot
27 Seed Specific Density
Density of seed refers to it mass per unit volume The seed size and density will affect the
yield One of the method for grouping seeds into some specific density is the liquid density
separation system It is one of the easier and cheaper method compared to using a machine
When seed is loaded into a liquid some will float and some will sink That mean the
floating seeds density is lower than the liquids density Vice versa the sinking seeds
density is higher than the liquids density Wang and Alyarez (2008) explained that this
method of separating seeds works by adjusting the specific gravity of the liquid in the
reservoir so that a first portion of the seeds float near the surface of the liquid and a second
portion of the seeds sink near the bottom of the reservoir and providing an outlet flow of
the liquid from a location proximal to the top of the reservoir above the location of the
second portion of seeds By this way seeds of lower density can be separated from higher
density Kwong Sellman Jalink amp Schoor (2005) illustrated that separation by density is
commonly incorporated as part of the routine cleaning and grading process in flower seed
production procedures Other than grouping seeds according to their specific density this
density separation also can be used to remove broken partially filled empty or immature
seeds
28 Seed Deterioration
McDonald (2005) defined flower seed deterioration as deteriorative changes occuring with
time that increase the seeds vulnerability to external challenges and decrease the ability of
the seed to survive Important factors contribute to seed deterioration are genetics seed
structure seed chemistry physicalphysiological quality seed treatments relative humidity
10
and temperature Relative humidity is important because it will influence the moisture
content of seeds in storage while temperature determines the amount of moisture in the air
and influences the rate of deteriorative reactions in seeds According to Agrawal (1980)
the rate of deterioration increases as the seed moisture content increases This could be due
to the mold growth in and on the seed
11
~
30 MATERIAL AND METHOD
31 Material
Fresh matured fruits of Hibiscus sabdarifJa L (Plate I) were obtained from local fanns in
Kota Samarahan Seeds were extracted from these fruits cleaned and dried under shade
Dried seeds were dusted with Captan 80 and then placed in air-tight bottle to keep it in a
safe place before use in the subsequent experiment
Plate 1 Fresh fruit ofH sabdarifJa Plate 2 Fresh fruit of H sabdarifJa with seeds
32 Method
321 Preliminary Evaluation
Moisture content and germination was conducted as an initial evaluation to assess the
quality of seeds used in this research The procedures were those prescribed by Association
ofOfficial Seeds Analysts (AOSA 1985)
12
middot
322 Moisture Content Test
Four replicates of 10 seeds each were placed into saucers (Plate 5) and calibrated to get the
wet weight The seeds then were placed in an oven at 60degC for 48 hours After that the
seeds were weighed again to get the dry weight The percentage of moisture content of the
seeds was calculated based on the formula below (AOSA 1985)
Moisture content () = b - c x 100
b-a
where a = weight of empty saucer
b = weight of (a) + weight of seeds before drying
c = weight of (a) + weight of seeds after drying in the oven
323 Germination Test
Germination test was conducted base on the AOSA (1985) where 25 seeds with four
replications were used For every replicates seeds were put into a petry dish on 3 layers of
moist Whatman filter paper (Plate 6) The seeds were left to germinate by incubating it in a
Plant Growth Chamber (28OC) and was checked daily until 10 days Seed was considered
as germinated if the radicles were about 5mm length emerged from the seed coat The
germination percentage was calculated base on AOSA 1985 formula below
13
-----------------------------------1
Gennination () a x 100
b
where a = total number of germinated seeds
b = total number of seeds used
324 Seed Characteristic
The size of the H sabdariffa seeds was calibrated for its length width and thickness Ten
replicates of seeds were used for calibrating the physical characteristics Then the seed
weights of 10 100 and 1000 seeds were calibrated Four replicates of 10 100 and 1000
seed-weight classes were used
325 Hydration of Seeds
Seed of H sabdariffa was placed in distilled water for 0 2 4 6 8 10 and 12 hours at
temperatures of 28 30 35 and 40degC Then seeds were analyzed for the moisture content
and gennination
326 Data Analysis
The experimental units were arranged randomly based on Completely Randomized Design
(CRD) and data was analyzed by using Analysis of Variance (ANOV A) If there any
significant differences the mean were discriminated using Least Significant Different
(LSD)
14
I
L
t I shy
432 Seed Germination 22 433 Gennination Analysis 28
50 CONCLUSIONS AND RECOMMENDATION 29
60 REFERENCES 30
70 APPENDICES 31
vi
r---~~------------------------------------------------------------------~
LIST OF FIGURES AND PLATES
Figure Page
Moisture content of Hsabdariffa seeds for different period of 15
hydration at 28degC
2 Moisture content of Hsabdariffa seeds for different period of 16
hydration at 30degC
3 Moisture content of Hsabdariffa seeds for different period of 17
hydration at 35degC
4 Moisture content of Hsabdariffa seeds for different period of 18
hydration at 40degC
5 Moisture content of Hsabdariffa seeds for different period of 19
hydration at 28 30 35 and 40degC
6 Gennination of Hsabdariffa seeds for different period of 20
hydration at 28degC
7 Gennination of Hsabdariffa seeds for different period of 21
hydration at 30degC
8 Gennination ofHsabdariffa seeds for different period of 22
hydration at 35degC
Gennination of Hsabdariffa seeds for different period of
hydration at 40degC
9 23
vii
10 24
Germination of HsabdarifJa seeds for different period of
hydration at 28 30 35 and 40degC
Plate
Fresh fruit of H sabdarifJa 11
2 Fresh fruit of H sabdarifJa with seeds 11
3 Fresh seeds of H sabdarifJa 16
4 Dried seeds of H sabdarifJa 16
viii
--_shy
I
I
Physical characteristic and germination of Hibiscus Sabdariffa L following hydration treatments
Syazwanie Binti Dzulbaimi
Plant Resource and Management Programme Faculty of Science and Technology
Universiti Malaysia Sarawak
ABSTRACT
A study was conducted to determine the physical characteristics and to assess and evaluate the effectiveness of hydration techniques in improving the performance of HibisclIs sabdariffa L seeds The average length width and thickness obtained for H sabdariffa seeds were 539 mm 455 mm and 234 mm respectively The average weight for a single seed was 002 g and it was estimated that one kg weight contained approximately 50000 seeds This indicated that the seeds used in this experiment were small In hydration treatment seeds of H sabdariffa were soaked in water at temperature 28 30 35 and 40degC for 0246 8 10 and 12 hours and were evaluated for moisture content and germination Results indicated that hydration treatment was not suitable for H sabdariffa seeds as it did not improved the germination The highest germination percentage of H sabdarifJa seeds following hydration treatment was 29 which was obtained for 2 hours hydration at 30degC
Key words Hibiscus sabdarifJa calibration hydration temperature
ABSTRAK
Satu kajian telah dijalankan IInt1lk menentukan ciri-ciri jizikal dall mengkaji serta menentukan keberkesanan teknik rawatall hidrasi dalam meningkatkan kadar pertllmbllhan biji benih Hibiscus sabdariffa Dalam penentlkuran biji benih pIrata panjang lebar dan ketebalan biji benih H sabdariffa yang diperoleh adalah 539 mm 455 mm dan 234 mm Purata berat sebiji benih adalah 002 g dan dianggarkan bahawa satll kilogram mengandungi lebih kurang 50000 biji benih Ini memmjukkan yang biji benih yang digllnakan unIlk kajian ini agak keci Dalam rawatan hidrasi biji benih H sabdariffa direndam di dalam air pada suhu 28degC 30degC 35degC dan 40degC Bagi setiap suhu yang digunakan biji benih direndam selama 0 2 4 6 8 10 dan 12 jam dan selepas itu kandungan kelembapan dan percambahan biji benih dinilai Keputlsan yang diperoleh menllnjukkan rawatan hidrasi adalah tidak sesuai bagi biji benih H sabdariffa memalldangkan peratls percambahan tidak meningkat berbanding sebelum rawatan Peratlls percambahan tertinggi bagi biji benih H sabdariffa selepas rawatan hidrasi adalah 29 iaitll bagi hidrasi selama 2 jam pada slhu 30degC
Kata kllnciHibisclS sabdarifJa kalibrasi hidrasi suhl
ix
I bull
10 INTRODUCTION
11 Background
Hibiscus sabdarifJa Linn or also known as Roselle or red sorrel belongs to the family
Malvaceae It is also known as asam paya asam susur and asam kumbang locally It is a
new commercial crop in Malaysia where it was brought from India (Amin et al 2008)
Since Roselle plantation only had started in early 1990s it is considered as a new crop in
Malaysia Two introduced varieties in Malaysia are Arab and Terengganu variety
However in 2009 Universiti Kebangsaan Malaysia had launched three new varieties
named UKMR-l UKMR-2 and UKMR-3 to help promote the Roselle industry Recently
Department of Agriculture had collaborated with F AMA entrepreneurs and growers in
order to strengthen the H sabdarifJa industry in Malaysia by expanding and marketing this
product locally and in foreign market
According to Qi et al (2005) H sabdarifJa is an annual mostly brannched and erect
shrub Its reddish stem can achieve 35m tall The leaves are dark green to red alternate
glabrous long - petiolate palmately divided into 3 - 7 lobes with serrate margins Having
both male and female organs the flower are red to yellow in color with a dark centre
containing short peduncles
H sabdarifJa can survive in a wann and humid tropical and subtropical climate It can be
grown in a greenhouse with litle shade but the best condition for growing it is under the
full sunlight (Qi et aI 2005) Roselle plants are suitable for tropical climates with wellshy
distributed rain - fall of 1500 - 2000 mm per year from sea level to about 600 m in
altitude (Amin et aI 2008) It can adapt to variety of soils but prefered a friable sandy
loam with humus
1
Current production of H sabdarifJa in Malaysia is about 240 tonnes annually Reported in
unpublished result from Agricultural Department of Terengganu in the process of
removing the calyces out of 3 tonnes of raw materials about 15 tonnes of the velvety
capsules containing the seeds are being disposed as a by - product and unexploited
(Halimatul Amin Mohd Esa Nawalyah amp Siti 2007) Amin et at (2008) mentioned that
the plant takes about three to four months to reach the commercial stage of maturity before
the flowers are harvested
12 Commercial Production
Many parts of H sabdariffa including seeds leaves fruits and roots are very useful
especially the fleshy red calyces They are used fresh for making wine juice jam jelly
syrup gelatin pudding cakes ice cream and flavors and also dried and brewed into tea
spice and used for butter pies sauces tarts and other desserts (Qi et al 2005) In a study
about commercial practice of Roselle beverage production Bolade et at (2009) reported
that the dried calyceswater ratios involved in the commercial practice ranged between
157 and 171 (wv) while the sweetness level of the beverage ranged between 112 and
133 degBrix The red calyces which contains high amount of vitamin C was used to make
those healthy beverages In Malaysia we also consume the leaves as vegetables There are
previous study shows that H sabdarifJa seeds can be used as a protein source Instead of
using protein from conventional sources protein isolates or concentrates from Roselle
seeds might be useful as low cost source of protein substitute in dietary supplement or food
ingredient in food industry thus alleviating the problem of protein scarcity (Halimatul et
al 2007)
2
middot
13 Medicinal Uses
H sabdariffa is said to be useful in traditional medicine as a digestive agent purgative and
diuretic among others Having two diuretic ingredients ascorbic acid and glycolic acid it
has the ability to increase urination and also valued for its mild laxative effect Containing
citric acid it also can be used as cooling herb The tonic tea produce from the leaves and
flowers is good for digestive and kidney function The heated leaves are applied to cracks
in the feet and on boils and ulcers to speed maturation (Qi et aI 2005) Other roselle s part
have also been reported to be folk remedy for cancer obesity diabetes and hypertension
(Bolad et aI 2009) Amin et al (2008) also added that the flower and fleshy fruits are
used in pharmaceutical industry to relieve symptoms of bronchitis and coughs Roselle s
lotion products are useful for treating sores and wounds
Another nutritional value of Roselle discovered by UKM was the high content of
hydroxycitric acid (HCA) which is widely used as the main ingredients in many
commercial weight loss and slimming products Recent studies by local and international
scientist have proven the effectiveness of HCA in reducing body weight This
characteristic makes Roselle a very valuable crop
14 Problem Statement
The demand for H sabdariffa production nowadays had increase due to its commercial and
medicinal values To fulfill these demands more good quality seeds are needed
Furthermore not many research had been done on improving the H sabdarifJa seeds as a
planting material eventhough the best propagation method for H sabdariffa is by seeds
Other than that disease has been reported as a limiting factor to the production of Roselle
worldwide (Sie R S et aI 2011)
3
1S Objectives
1 To detennine the physical characteristics of H sabdariffa seeds for 10 100 and
1000 seeds weight and for length width and thickness of the seed
2 To evaluate the germination of H sabdariffa seeds following hydration at different
temperatures
3 To determine the effectiveness of hydration technique in improving the gennination
of H sabdariffa seeds
4
-
Pusat Khidmat Maklumat Akadtmik VNIVERSm MALAYSIA SARAWAK
I J bull bull
20 LITERATURE REVIEW
21 Characteristic of Seed
Seeds quality can be affected by changes in environmental temperature and relative
humidity The characteristic of seed must be determine because it is important to ensure
high quality yield in field planting Seed germination can be affected by number of factors
such as level of oxygen temperature and moisture content Growth performance of seed
also can be related to seed size Therefore grading is important to obtain seed that can
grows better Some useful criteria for visual separation of seed includes the seed size
shape colour weight thickness density and surface texture
According to Jain and Bal (1997) roselle seeds are having average principal dimensions of
298-336 186-224 and 170-201 mm Omobuwajo et al (2000) found that the average of
roselle seeds length width and thickness were 558 521 and 281 mm respectively The
seed was reported to be important for its oil in some parts of Africa its native origin From
previous research by scientist roselle seed was among the highest protein-containing seeds
when compared with other seeds like passion fruit (Passiflora edulis) Amaranthus seeds
and Pisum sativum seeds Latest findings by Hainida et al (2008) found that the seed from
Malaysia are composed of 99 moisture 335 protein 221 lipids 130 available
carbohydrate 183 total dietary fibers and 75 ash
H sabdarifJa can be grown easily in most of the tropical country Like other hibiscus
species it is quite hard for roselle to be affected by pest and disease and maintaining this
plant in the field is not so difficult The seeds were sown in a suitable container and the
seedlings will be ready for planting at the age of 10-15 days However it is also important
to ensure that the seeds to be sown were in a good condition and quality to avoid losses
5
22 Seed Germination
Germination is the emergence and development from the seed embryo of those essential
structures which are indicative of the ability to produce a normal plant under favorable
conditions (AOSA 1991) According to Smith (2011) the sum of germination events
beginning with hydration and culminating with root emergence Several stages involved in
germination are imbibition of water activation of enzyme systems metabolism of storage
products and their transport and finally the emergence of the radicle and growth of the
seedling Seed germination is described as the emergence and development from the seed
embryo of those essential structures which for the kind of seed in question are indicative
of the ability to produce a normal plant under favorable conditions (AOSA 1978) In a
simpler contcxt germination is the emergence of the radicle through the seed coat
Depending on the species seeds can germinate as quick as a few days or for as long as
several years Due to the high respiration rate and some exudation and leakage through the
seed coat the germinating seedling will undergoes a net loss in dry weight before the
germination For a particular kind of species the germination condition is mostly affected
by temperature type of substrate on which to germinate the seed light condition and
moisture level
The germination test is commonly used to determine seed viability It is an analytical
procedure to evaluate seed germination under standardized favorable conditions that are
seldom if ever encountered in the field The germination test is merely an estimate and
has certain limitations as a universal estimate of seed quality However if these limitations
are recognized the germination test is a useful viability index The need for germination
test is to estimate how the seed will perform in the field
6
I bull bull
23 Seed Moisture Content
Amount of water in the seed that usually expressed in a percentage fonn is the seed
moisture content Agrawal (1980) found that the amount of moisture in the seed is
probably the most important factor influencing seed viability during storage By
determining the seeds moisture content the possible storage life for the seeds can be
predicted According to McDonald (2005) selecting the appropriate salt to achieve a seed
moisture content of 5-6 is the recommended method for achieving optimum seed
storage and minimizing seed deterioration It is necessary to dry seeds to the safe moisture
contents since the life of a seed and its span largely revolves around its moisture content
and the suitable moisture content for starchy seeds are less than 14 while for oily seeds
are less than 11 (Agrawal 1980)
24 Seed Vigour
According to Geneve (2005) seed vigour is defined by the Asociation of official Seed
analysts as those seed properties which detennine the potential for rapid unifonn
emergence and development of nonnal seedlings under the wide range of field conditions
Seed vigour is closely related with seed storage environment and duration During seed
storage the vigour of a seed lot is reduced prior to seed viability as indicated by standard
germination (Hampton amp TeKrony 1995) Major factors influencing the seed vigour in
stored seed lots are the environmental conditions during seed storage and the length of time
in storage Tetrazolium test is one of the method for detennining the seeds vigour To
minimize the decline in vigour and germination seeds should be undamaged
7
bull t I I
25 Seed Hydration
A process of supplying water to seed to maintain the fluid or moisture content in the seed
is called hydration According to Copeland and McDonald (1995) seed hydration is a
process whereby seeds are hydrated using various protocols and then redried to permit
routine handling This process results in increased germination rate more uniform
emergence germination under a broader range of environments and improved sedling
vigour and growth It is one of the priming treatments of seed which usually applied to
strengthen or increase vitality of seed Priming process can increase a rate of germination
as the treatment improves the seed vigor Hydration is one component of priming which
can increase germination with more range of environment condition The objective of seed
hydration technology is to increase the percentage and rate of germination expand the
range of temperatures over which the seed will germinate and increase the uniformity of
stand establishment To accomplish these objectives seeds must be hydrated in some way
at a moisture level sufficient to initiate the early events of germination but not sufficient to
permit radicle protrusion (Akers and Holley 1986) However the performance of the seed
might also being influence by other factors like the species type of seed and the seeds
quality
Water can be used for the hydration treatment because it had proved to enhance
germination emergence growth and yield of seed However other mediwns such as the
solution of potassium hydrophosphate (KH2P04) monobasic polyethylene glycol (PEG)
potassium chloride (KCl) potassiwn nitrate (KN03) potassium phoshate (K3P04)
magnesium sulphate (MgS04) sodiwn chloride (NaCl) glycerol and mannitol also can be
used to immerse the seed in hydration process The benefit of such salts is to supply the
seed with nitrogen and other nutrients essential for protein synthesis during germination
8
t I amp
26 Seed Storage
A basic requirement in seed production is a good seed storage The purpose of seed storage
is to preserve planting stocks from one season to the next (Larry amp McDonald 1995)
Generally we stored seeds for the commercial purpose for carry - over seeds or for genn
plasm seeds Maintaining the seed quality for the longest duration possible is the main
objective in seed storage Loss of gennination during storage cannot be stopped but it
could be reduce by giving good storage conditions According to Agrawal (1980) the
seeds are considered to be in storage from the moment they reach physiological maturity
until they genninate or until they are thrown away because they are dead or otherwise
worthless
The seed storage must be properly handled because it can affect the seed quality There are
some factors that can affect the seed longevity in storage such as the variety of the seed
initial seed quality moisture content relative humidity and storage temperature It is
necessary for the seed storage condition to be dry and cool effective in pest control and
have a proper sanitation Other than than storing only high quality of seed also can help in
better production From all of the factors temperature is one of the most important element
which influence seed viability and vigour during storage Agrawal (1980) mentioned that
the lower the temperature the longer the seeds maintain gennination capacity so
decreasing temperature and seed moisture is an effective means of maintaining seed quality
in storage Every seed lot should be gennination - tested when it is received for storage to
record the seed quality Subsequent periodic checks also should be made to detect
deterioration and if it occurs storage condition should be checked to discover the reasons
and to take corrective measures
9
middot
27 Seed Specific Density
Density of seed refers to it mass per unit volume The seed size and density will affect the
yield One of the method for grouping seeds into some specific density is the liquid density
separation system It is one of the easier and cheaper method compared to using a machine
When seed is loaded into a liquid some will float and some will sink That mean the
floating seeds density is lower than the liquids density Vice versa the sinking seeds
density is higher than the liquids density Wang and Alyarez (2008) explained that this
method of separating seeds works by adjusting the specific gravity of the liquid in the
reservoir so that a first portion of the seeds float near the surface of the liquid and a second
portion of the seeds sink near the bottom of the reservoir and providing an outlet flow of
the liquid from a location proximal to the top of the reservoir above the location of the
second portion of seeds By this way seeds of lower density can be separated from higher
density Kwong Sellman Jalink amp Schoor (2005) illustrated that separation by density is
commonly incorporated as part of the routine cleaning and grading process in flower seed
production procedures Other than grouping seeds according to their specific density this
density separation also can be used to remove broken partially filled empty or immature
seeds
28 Seed Deterioration
McDonald (2005) defined flower seed deterioration as deteriorative changes occuring with
time that increase the seeds vulnerability to external challenges and decrease the ability of
the seed to survive Important factors contribute to seed deterioration are genetics seed
structure seed chemistry physicalphysiological quality seed treatments relative humidity
10
and temperature Relative humidity is important because it will influence the moisture
content of seeds in storage while temperature determines the amount of moisture in the air
and influences the rate of deteriorative reactions in seeds According to Agrawal (1980)
the rate of deterioration increases as the seed moisture content increases This could be due
to the mold growth in and on the seed
11
~
30 MATERIAL AND METHOD
31 Material
Fresh matured fruits of Hibiscus sabdarifJa L (Plate I) were obtained from local fanns in
Kota Samarahan Seeds were extracted from these fruits cleaned and dried under shade
Dried seeds were dusted with Captan 80 and then placed in air-tight bottle to keep it in a
safe place before use in the subsequent experiment
Plate 1 Fresh fruit ofH sabdarifJa Plate 2 Fresh fruit of H sabdarifJa with seeds
32 Method
321 Preliminary Evaluation
Moisture content and germination was conducted as an initial evaluation to assess the
quality of seeds used in this research The procedures were those prescribed by Association
ofOfficial Seeds Analysts (AOSA 1985)
12
middot
322 Moisture Content Test
Four replicates of 10 seeds each were placed into saucers (Plate 5) and calibrated to get the
wet weight The seeds then were placed in an oven at 60degC for 48 hours After that the
seeds were weighed again to get the dry weight The percentage of moisture content of the
seeds was calculated based on the formula below (AOSA 1985)
Moisture content () = b - c x 100
b-a
where a = weight of empty saucer
b = weight of (a) + weight of seeds before drying
c = weight of (a) + weight of seeds after drying in the oven
323 Germination Test
Germination test was conducted base on the AOSA (1985) where 25 seeds with four
replications were used For every replicates seeds were put into a petry dish on 3 layers of
moist Whatman filter paper (Plate 6) The seeds were left to germinate by incubating it in a
Plant Growth Chamber (28OC) and was checked daily until 10 days Seed was considered
as germinated if the radicles were about 5mm length emerged from the seed coat The
germination percentage was calculated base on AOSA 1985 formula below
13
-----------------------------------1
Gennination () a x 100
b
where a = total number of germinated seeds
b = total number of seeds used
324 Seed Characteristic
The size of the H sabdariffa seeds was calibrated for its length width and thickness Ten
replicates of seeds were used for calibrating the physical characteristics Then the seed
weights of 10 100 and 1000 seeds were calibrated Four replicates of 10 100 and 1000
seed-weight classes were used
325 Hydration of Seeds
Seed of H sabdariffa was placed in distilled water for 0 2 4 6 8 10 and 12 hours at
temperatures of 28 30 35 and 40degC Then seeds were analyzed for the moisture content
and gennination
326 Data Analysis
The experimental units were arranged randomly based on Completely Randomized Design
(CRD) and data was analyzed by using Analysis of Variance (ANOV A) If there any
significant differences the mean were discriminated using Least Significant Different
(LSD)
14
r---~~------------------------------------------------------------------~
LIST OF FIGURES AND PLATES
Figure Page
Moisture content of Hsabdariffa seeds for different period of 15
hydration at 28degC
2 Moisture content of Hsabdariffa seeds for different period of 16
hydration at 30degC
3 Moisture content of Hsabdariffa seeds for different period of 17
hydration at 35degC
4 Moisture content of Hsabdariffa seeds for different period of 18
hydration at 40degC
5 Moisture content of Hsabdariffa seeds for different period of 19
hydration at 28 30 35 and 40degC
6 Gennination of Hsabdariffa seeds for different period of 20
hydration at 28degC
7 Gennination of Hsabdariffa seeds for different period of 21
hydration at 30degC
8 Gennination ofHsabdariffa seeds for different period of 22
hydration at 35degC
Gennination of Hsabdariffa seeds for different period of
hydration at 40degC
9 23
vii
10 24
Germination of HsabdarifJa seeds for different period of
hydration at 28 30 35 and 40degC
Plate
Fresh fruit of H sabdarifJa 11
2 Fresh fruit of H sabdarifJa with seeds 11
3 Fresh seeds of H sabdarifJa 16
4 Dried seeds of H sabdarifJa 16
viii
--_shy
I
I
Physical characteristic and germination of Hibiscus Sabdariffa L following hydration treatments
Syazwanie Binti Dzulbaimi
Plant Resource and Management Programme Faculty of Science and Technology
Universiti Malaysia Sarawak
ABSTRACT
A study was conducted to determine the physical characteristics and to assess and evaluate the effectiveness of hydration techniques in improving the performance of HibisclIs sabdariffa L seeds The average length width and thickness obtained for H sabdariffa seeds were 539 mm 455 mm and 234 mm respectively The average weight for a single seed was 002 g and it was estimated that one kg weight contained approximately 50000 seeds This indicated that the seeds used in this experiment were small In hydration treatment seeds of H sabdariffa were soaked in water at temperature 28 30 35 and 40degC for 0246 8 10 and 12 hours and were evaluated for moisture content and germination Results indicated that hydration treatment was not suitable for H sabdariffa seeds as it did not improved the germination The highest germination percentage of H sabdarifJa seeds following hydration treatment was 29 which was obtained for 2 hours hydration at 30degC
Key words Hibiscus sabdarifJa calibration hydration temperature
ABSTRAK
Satu kajian telah dijalankan IInt1lk menentukan ciri-ciri jizikal dall mengkaji serta menentukan keberkesanan teknik rawatall hidrasi dalam meningkatkan kadar pertllmbllhan biji benih Hibiscus sabdariffa Dalam penentlkuran biji benih pIrata panjang lebar dan ketebalan biji benih H sabdariffa yang diperoleh adalah 539 mm 455 mm dan 234 mm Purata berat sebiji benih adalah 002 g dan dianggarkan bahawa satll kilogram mengandungi lebih kurang 50000 biji benih Ini memmjukkan yang biji benih yang digllnakan unIlk kajian ini agak keci Dalam rawatan hidrasi biji benih H sabdariffa direndam di dalam air pada suhu 28degC 30degC 35degC dan 40degC Bagi setiap suhu yang digunakan biji benih direndam selama 0 2 4 6 8 10 dan 12 jam dan selepas itu kandungan kelembapan dan percambahan biji benih dinilai Keputlsan yang diperoleh menllnjukkan rawatan hidrasi adalah tidak sesuai bagi biji benih H sabdariffa memalldangkan peratls percambahan tidak meningkat berbanding sebelum rawatan Peratlls percambahan tertinggi bagi biji benih H sabdariffa selepas rawatan hidrasi adalah 29 iaitll bagi hidrasi selama 2 jam pada slhu 30degC
Kata kllnciHibisclS sabdarifJa kalibrasi hidrasi suhl
ix
I bull
10 INTRODUCTION
11 Background
Hibiscus sabdarifJa Linn or also known as Roselle or red sorrel belongs to the family
Malvaceae It is also known as asam paya asam susur and asam kumbang locally It is a
new commercial crop in Malaysia where it was brought from India (Amin et al 2008)
Since Roselle plantation only had started in early 1990s it is considered as a new crop in
Malaysia Two introduced varieties in Malaysia are Arab and Terengganu variety
However in 2009 Universiti Kebangsaan Malaysia had launched three new varieties
named UKMR-l UKMR-2 and UKMR-3 to help promote the Roselle industry Recently
Department of Agriculture had collaborated with F AMA entrepreneurs and growers in
order to strengthen the H sabdarifJa industry in Malaysia by expanding and marketing this
product locally and in foreign market
According to Qi et al (2005) H sabdarifJa is an annual mostly brannched and erect
shrub Its reddish stem can achieve 35m tall The leaves are dark green to red alternate
glabrous long - petiolate palmately divided into 3 - 7 lobes with serrate margins Having
both male and female organs the flower are red to yellow in color with a dark centre
containing short peduncles
H sabdarifJa can survive in a wann and humid tropical and subtropical climate It can be
grown in a greenhouse with litle shade but the best condition for growing it is under the
full sunlight (Qi et aI 2005) Roselle plants are suitable for tropical climates with wellshy
distributed rain - fall of 1500 - 2000 mm per year from sea level to about 600 m in
altitude (Amin et aI 2008) It can adapt to variety of soils but prefered a friable sandy
loam with humus
1
Current production of H sabdarifJa in Malaysia is about 240 tonnes annually Reported in
unpublished result from Agricultural Department of Terengganu in the process of
removing the calyces out of 3 tonnes of raw materials about 15 tonnes of the velvety
capsules containing the seeds are being disposed as a by - product and unexploited
(Halimatul Amin Mohd Esa Nawalyah amp Siti 2007) Amin et at (2008) mentioned that
the plant takes about three to four months to reach the commercial stage of maturity before
the flowers are harvested
12 Commercial Production
Many parts of H sabdariffa including seeds leaves fruits and roots are very useful
especially the fleshy red calyces They are used fresh for making wine juice jam jelly
syrup gelatin pudding cakes ice cream and flavors and also dried and brewed into tea
spice and used for butter pies sauces tarts and other desserts (Qi et al 2005) In a study
about commercial practice of Roselle beverage production Bolade et at (2009) reported
that the dried calyceswater ratios involved in the commercial practice ranged between
157 and 171 (wv) while the sweetness level of the beverage ranged between 112 and
133 degBrix The red calyces which contains high amount of vitamin C was used to make
those healthy beverages In Malaysia we also consume the leaves as vegetables There are
previous study shows that H sabdarifJa seeds can be used as a protein source Instead of
using protein from conventional sources protein isolates or concentrates from Roselle
seeds might be useful as low cost source of protein substitute in dietary supplement or food
ingredient in food industry thus alleviating the problem of protein scarcity (Halimatul et
al 2007)
2
middot
13 Medicinal Uses
H sabdariffa is said to be useful in traditional medicine as a digestive agent purgative and
diuretic among others Having two diuretic ingredients ascorbic acid and glycolic acid it
has the ability to increase urination and also valued for its mild laxative effect Containing
citric acid it also can be used as cooling herb The tonic tea produce from the leaves and
flowers is good for digestive and kidney function The heated leaves are applied to cracks
in the feet and on boils and ulcers to speed maturation (Qi et aI 2005) Other roselle s part
have also been reported to be folk remedy for cancer obesity diabetes and hypertension
(Bolad et aI 2009) Amin et al (2008) also added that the flower and fleshy fruits are
used in pharmaceutical industry to relieve symptoms of bronchitis and coughs Roselle s
lotion products are useful for treating sores and wounds
Another nutritional value of Roselle discovered by UKM was the high content of
hydroxycitric acid (HCA) which is widely used as the main ingredients in many
commercial weight loss and slimming products Recent studies by local and international
scientist have proven the effectiveness of HCA in reducing body weight This
characteristic makes Roselle a very valuable crop
14 Problem Statement
The demand for H sabdariffa production nowadays had increase due to its commercial and
medicinal values To fulfill these demands more good quality seeds are needed
Furthermore not many research had been done on improving the H sabdarifJa seeds as a
planting material eventhough the best propagation method for H sabdariffa is by seeds
Other than that disease has been reported as a limiting factor to the production of Roselle
worldwide (Sie R S et aI 2011)
3
1S Objectives
1 To detennine the physical characteristics of H sabdariffa seeds for 10 100 and
1000 seeds weight and for length width and thickness of the seed
2 To evaluate the germination of H sabdariffa seeds following hydration at different
temperatures
3 To determine the effectiveness of hydration technique in improving the gennination
of H sabdariffa seeds
4
-
Pusat Khidmat Maklumat Akadtmik VNIVERSm MALAYSIA SARAWAK
I J bull bull
20 LITERATURE REVIEW
21 Characteristic of Seed
Seeds quality can be affected by changes in environmental temperature and relative
humidity The characteristic of seed must be determine because it is important to ensure
high quality yield in field planting Seed germination can be affected by number of factors
such as level of oxygen temperature and moisture content Growth performance of seed
also can be related to seed size Therefore grading is important to obtain seed that can
grows better Some useful criteria for visual separation of seed includes the seed size
shape colour weight thickness density and surface texture
According to Jain and Bal (1997) roselle seeds are having average principal dimensions of
298-336 186-224 and 170-201 mm Omobuwajo et al (2000) found that the average of
roselle seeds length width and thickness were 558 521 and 281 mm respectively The
seed was reported to be important for its oil in some parts of Africa its native origin From
previous research by scientist roselle seed was among the highest protein-containing seeds
when compared with other seeds like passion fruit (Passiflora edulis) Amaranthus seeds
and Pisum sativum seeds Latest findings by Hainida et al (2008) found that the seed from
Malaysia are composed of 99 moisture 335 protein 221 lipids 130 available
carbohydrate 183 total dietary fibers and 75 ash
H sabdarifJa can be grown easily in most of the tropical country Like other hibiscus
species it is quite hard for roselle to be affected by pest and disease and maintaining this
plant in the field is not so difficult The seeds were sown in a suitable container and the
seedlings will be ready for planting at the age of 10-15 days However it is also important
to ensure that the seeds to be sown were in a good condition and quality to avoid losses
5
22 Seed Germination
Germination is the emergence and development from the seed embryo of those essential
structures which are indicative of the ability to produce a normal plant under favorable
conditions (AOSA 1991) According to Smith (2011) the sum of germination events
beginning with hydration and culminating with root emergence Several stages involved in
germination are imbibition of water activation of enzyme systems metabolism of storage
products and their transport and finally the emergence of the radicle and growth of the
seedling Seed germination is described as the emergence and development from the seed
embryo of those essential structures which for the kind of seed in question are indicative
of the ability to produce a normal plant under favorable conditions (AOSA 1978) In a
simpler contcxt germination is the emergence of the radicle through the seed coat
Depending on the species seeds can germinate as quick as a few days or for as long as
several years Due to the high respiration rate and some exudation and leakage through the
seed coat the germinating seedling will undergoes a net loss in dry weight before the
germination For a particular kind of species the germination condition is mostly affected
by temperature type of substrate on which to germinate the seed light condition and
moisture level
The germination test is commonly used to determine seed viability It is an analytical
procedure to evaluate seed germination under standardized favorable conditions that are
seldom if ever encountered in the field The germination test is merely an estimate and
has certain limitations as a universal estimate of seed quality However if these limitations
are recognized the germination test is a useful viability index The need for germination
test is to estimate how the seed will perform in the field
6
I bull bull
23 Seed Moisture Content
Amount of water in the seed that usually expressed in a percentage fonn is the seed
moisture content Agrawal (1980) found that the amount of moisture in the seed is
probably the most important factor influencing seed viability during storage By
determining the seeds moisture content the possible storage life for the seeds can be
predicted According to McDonald (2005) selecting the appropriate salt to achieve a seed
moisture content of 5-6 is the recommended method for achieving optimum seed
storage and minimizing seed deterioration It is necessary to dry seeds to the safe moisture
contents since the life of a seed and its span largely revolves around its moisture content
and the suitable moisture content for starchy seeds are less than 14 while for oily seeds
are less than 11 (Agrawal 1980)
24 Seed Vigour
According to Geneve (2005) seed vigour is defined by the Asociation of official Seed
analysts as those seed properties which detennine the potential for rapid unifonn
emergence and development of nonnal seedlings under the wide range of field conditions
Seed vigour is closely related with seed storage environment and duration During seed
storage the vigour of a seed lot is reduced prior to seed viability as indicated by standard
germination (Hampton amp TeKrony 1995) Major factors influencing the seed vigour in
stored seed lots are the environmental conditions during seed storage and the length of time
in storage Tetrazolium test is one of the method for detennining the seeds vigour To
minimize the decline in vigour and germination seeds should be undamaged
7
bull t I I
25 Seed Hydration
A process of supplying water to seed to maintain the fluid or moisture content in the seed
is called hydration According to Copeland and McDonald (1995) seed hydration is a
process whereby seeds are hydrated using various protocols and then redried to permit
routine handling This process results in increased germination rate more uniform
emergence germination under a broader range of environments and improved sedling
vigour and growth It is one of the priming treatments of seed which usually applied to
strengthen or increase vitality of seed Priming process can increase a rate of germination
as the treatment improves the seed vigor Hydration is one component of priming which
can increase germination with more range of environment condition The objective of seed
hydration technology is to increase the percentage and rate of germination expand the
range of temperatures over which the seed will germinate and increase the uniformity of
stand establishment To accomplish these objectives seeds must be hydrated in some way
at a moisture level sufficient to initiate the early events of germination but not sufficient to
permit radicle protrusion (Akers and Holley 1986) However the performance of the seed
might also being influence by other factors like the species type of seed and the seeds
quality
Water can be used for the hydration treatment because it had proved to enhance
germination emergence growth and yield of seed However other mediwns such as the
solution of potassium hydrophosphate (KH2P04) monobasic polyethylene glycol (PEG)
potassium chloride (KCl) potassiwn nitrate (KN03) potassium phoshate (K3P04)
magnesium sulphate (MgS04) sodiwn chloride (NaCl) glycerol and mannitol also can be
used to immerse the seed in hydration process The benefit of such salts is to supply the
seed with nitrogen and other nutrients essential for protein synthesis during germination
8
t I amp
26 Seed Storage
A basic requirement in seed production is a good seed storage The purpose of seed storage
is to preserve planting stocks from one season to the next (Larry amp McDonald 1995)
Generally we stored seeds for the commercial purpose for carry - over seeds or for genn
plasm seeds Maintaining the seed quality for the longest duration possible is the main
objective in seed storage Loss of gennination during storage cannot be stopped but it
could be reduce by giving good storage conditions According to Agrawal (1980) the
seeds are considered to be in storage from the moment they reach physiological maturity
until they genninate or until they are thrown away because they are dead or otherwise
worthless
The seed storage must be properly handled because it can affect the seed quality There are
some factors that can affect the seed longevity in storage such as the variety of the seed
initial seed quality moisture content relative humidity and storage temperature It is
necessary for the seed storage condition to be dry and cool effective in pest control and
have a proper sanitation Other than than storing only high quality of seed also can help in
better production From all of the factors temperature is one of the most important element
which influence seed viability and vigour during storage Agrawal (1980) mentioned that
the lower the temperature the longer the seeds maintain gennination capacity so
decreasing temperature and seed moisture is an effective means of maintaining seed quality
in storage Every seed lot should be gennination - tested when it is received for storage to
record the seed quality Subsequent periodic checks also should be made to detect
deterioration and if it occurs storage condition should be checked to discover the reasons
and to take corrective measures
9
middot
27 Seed Specific Density
Density of seed refers to it mass per unit volume The seed size and density will affect the
yield One of the method for grouping seeds into some specific density is the liquid density
separation system It is one of the easier and cheaper method compared to using a machine
When seed is loaded into a liquid some will float and some will sink That mean the
floating seeds density is lower than the liquids density Vice versa the sinking seeds
density is higher than the liquids density Wang and Alyarez (2008) explained that this
method of separating seeds works by adjusting the specific gravity of the liquid in the
reservoir so that a first portion of the seeds float near the surface of the liquid and a second
portion of the seeds sink near the bottom of the reservoir and providing an outlet flow of
the liquid from a location proximal to the top of the reservoir above the location of the
second portion of seeds By this way seeds of lower density can be separated from higher
density Kwong Sellman Jalink amp Schoor (2005) illustrated that separation by density is
commonly incorporated as part of the routine cleaning and grading process in flower seed
production procedures Other than grouping seeds according to their specific density this
density separation also can be used to remove broken partially filled empty or immature
seeds
28 Seed Deterioration
McDonald (2005) defined flower seed deterioration as deteriorative changes occuring with
time that increase the seeds vulnerability to external challenges and decrease the ability of
the seed to survive Important factors contribute to seed deterioration are genetics seed
structure seed chemistry physicalphysiological quality seed treatments relative humidity
10
and temperature Relative humidity is important because it will influence the moisture
content of seeds in storage while temperature determines the amount of moisture in the air
and influences the rate of deteriorative reactions in seeds According to Agrawal (1980)
the rate of deterioration increases as the seed moisture content increases This could be due
to the mold growth in and on the seed
11
~
30 MATERIAL AND METHOD
31 Material
Fresh matured fruits of Hibiscus sabdarifJa L (Plate I) were obtained from local fanns in
Kota Samarahan Seeds were extracted from these fruits cleaned and dried under shade
Dried seeds were dusted with Captan 80 and then placed in air-tight bottle to keep it in a
safe place before use in the subsequent experiment
Plate 1 Fresh fruit ofH sabdarifJa Plate 2 Fresh fruit of H sabdarifJa with seeds
32 Method
321 Preliminary Evaluation
Moisture content and germination was conducted as an initial evaluation to assess the
quality of seeds used in this research The procedures were those prescribed by Association
ofOfficial Seeds Analysts (AOSA 1985)
12
middot
322 Moisture Content Test
Four replicates of 10 seeds each were placed into saucers (Plate 5) and calibrated to get the
wet weight The seeds then were placed in an oven at 60degC for 48 hours After that the
seeds were weighed again to get the dry weight The percentage of moisture content of the
seeds was calculated based on the formula below (AOSA 1985)
Moisture content () = b - c x 100
b-a
where a = weight of empty saucer
b = weight of (a) + weight of seeds before drying
c = weight of (a) + weight of seeds after drying in the oven
323 Germination Test
Germination test was conducted base on the AOSA (1985) where 25 seeds with four
replications were used For every replicates seeds were put into a petry dish on 3 layers of
moist Whatman filter paper (Plate 6) The seeds were left to germinate by incubating it in a
Plant Growth Chamber (28OC) and was checked daily until 10 days Seed was considered
as germinated if the radicles were about 5mm length emerged from the seed coat The
germination percentage was calculated base on AOSA 1985 formula below
13
-----------------------------------1
Gennination () a x 100
b
where a = total number of germinated seeds
b = total number of seeds used
324 Seed Characteristic
The size of the H sabdariffa seeds was calibrated for its length width and thickness Ten
replicates of seeds were used for calibrating the physical characteristics Then the seed
weights of 10 100 and 1000 seeds were calibrated Four replicates of 10 100 and 1000
seed-weight classes were used
325 Hydration of Seeds
Seed of H sabdariffa was placed in distilled water for 0 2 4 6 8 10 and 12 hours at
temperatures of 28 30 35 and 40degC Then seeds were analyzed for the moisture content
and gennination
326 Data Analysis
The experimental units were arranged randomly based on Completely Randomized Design
(CRD) and data was analyzed by using Analysis of Variance (ANOV A) If there any
significant differences the mean were discriminated using Least Significant Different
(LSD)
14
10 24
Germination of HsabdarifJa seeds for different period of
hydration at 28 30 35 and 40degC
Plate
Fresh fruit of H sabdarifJa 11
2 Fresh fruit of H sabdarifJa with seeds 11
3 Fresh seeds of H sabdarifJa 16
4 Dried seeds of H sabdarifJa 16
viii
--_shy
I
I
Physical characteristic and germination of Hibiscus Sabdariffa L following hydration treatments
Syazwanie Binti Dzulbaimi
Plant Resource and Management Programme Faculty of Science and Technology
Universiti Malaysia Sarawak
ABSTRACT
A study was conducted to determine the physical characteristics and to assess and evaluate the effectiveness of hydration techniques in improving the performance of HibisclIs sabdariffa L seeds The average length width and thickness obtained for H sabdariffa seeds were 539 mm 455 mm and 234 mm respectively The average weight for a single seed was 002 g and it was estimated that one kg weight contained approximately 50000 seeds This indicated that the seeds used in this experiment were small In hydration treatment seeds of H sabdariffa were soaked in water at temperature 28 30 35 and 40degC for 0246 8 10 and 12 hours and were evaluated for moisture content and germination Results indicated that hydration treatment was not suitable for H sabdariffa seeds as it did not improved the germination The highest germination percentage of H sabdarifJa seeds following hydration treatment was 29 which was obtained for 2 hours hydration at 30degC
Key words Hibiscus sabdarifJa calibration hydration temperature
ABSTRAK
Satu kajian telah dijalankan IInt1lk menentukan ciri-ciri jizikal dall mengkaji serta menentukan keberkesanan teknik rawatall hidrasi dalam meningkatkan kadar pertllmbllhan biji benih Hibiscus sabdariffa Dalam penentlkuran biji benih pIrata panjang lebar dan ketebalan biji benih H sabdariffa yang diperoleh adalah 539 mm 455 mm dan 234 mm Purata berat sebiji benih adalah 002 g dan dianggarkan bahawa satll kilogram mengandungi lebih kurang 50000 biji benih Ini memmjukkan yang biji benih yang digllnakan unIlk kajian ini agak keci Dalam rawatan hidrasi biji benih H sabdariffa direndam di dalam air pada suhu 28degC 30degC 35degC dan 40degC Bagi setiap suhu yang digunakan biji benih direndam selama 0 2 4 6 8 10 dan 12 jam dan selepas itu kandungan kelembapan dan percambahan biji benih dinilai Keputlsan yang diperoleh menllnjukkan rawatan hidrasi adalah tidak sesuai bagi biji benih H sabdariffa memalldangkan peratls percambahan tidak meningkat berbanding sebelum rawatan Peratlls percambahan tertinggi bagi biji benih H sabdariffa selepas rawatan hidrasi adalah 29 iaitll bagi hidrasi selama 2 jam pada slhu 30degC
Kata kllnciHibisclS sabdarifJa kalibrasi hidrasi suhl
ix
I bull
10 INTRODUCTION
11 Background
Hibiscus sabdarifJa Linn or also known as Roselle or red sorrel belongs to the family
Malvaceae It is also known as asam paya asam susur and asam kumbang locally It is a
new commercial crop in Malaysia where it was brought from India (Amin et al 2008)
Since Roselle plantation only had started in early 1990s it is considered as a new crop in
Malaysia Two introduced varieties in Malaysia are Arab and Terengganu variety
However in 2009 Universiti Kebangsaan Malaysia had launched three new varieties
named UKMR-l UKMR-2 and UKMR-3 to help promote the Roselle industry Recently
Department of Agriculture had collaborated with F AMA entrepreneurs and growers in
order to strengthen the H sabdarifJa industry in Malaysia by expanding and marketing this
product locally and in foreign market
According to Qi et al (2005) H sabdarifJa is an annual mostly brannched and erect
shrub Its reddish stem can achieve 35m tall The leaves are dark green to red alternate
glabrous long - petiolate palmately divided into 3 - 7 lobes with serrate margins Having
both male and female organs the flower are red to yellow in color with a dark centre
containing short peduncles
H sabdarifJa can survive in a wann and humid tropical and subtropical climate It can be
grown in a greenhouse with litle shade but the best condition for growing it is under the
full sunlight (Qi et aI 2005) Roselle plants are suitable for tropical climates with wellshy
distributed rain - fall of 1500 - 2000 mm per year from sea level to about 600 m in
altitude (Amin et aI 2008) It can adapt to variety of soils but prefered a friable sandy
loam with humus
1
Current production of H sabdarifJa in Malaysia is about 240 tonnes annually Reported in
unpublished result from Agricultural Department of Terengganu in the process of
removing the calyces out of 3 tonnes of raw materials about 15 tonnes of the velvety
capsules containing the seeds are being disposed as a by - product and unexploited
(Halimatul Amin Mohd Esa Nawalyah amp Siti 2007) Amin et at (2008) mentioned that
the plant takes about three to four months to reach the commercial stage of maturity before
the flowers are harvested
12 Commercial Production
Many parts of H sabdariffa including seeds leaves fruits and roots are very useful
especially the fleshy red calyces They are used fresh for making wine juice jam jelly
syrup gelatin pudding cakes ice cream and flavors and also dried and brewed into tea
spice and used for butter pies sauces tarts and other desserts (Qi et al 2005) In a study
about commercial practice of Roselle beverage production Bolade et at (2009) reported
that the dried calyceswater ratios involved in the commercial practice ranged between
157 and 171 (wv) while the sweetness level of the beverage ranged between 112 and
133 degBrix The red calyces which contains high amount of vitamin C was used to make
those healthy beverages In Malaysia we also consume the leaves as vegetables There are
previous study shows that H sabdarifJa seeds can be used as a protein source Instead of
using protein from conventional sources protein isolates or concentrates from Roselle
seeds might be useful as low cost source of protein substitute in dietary supplement or food
ingredient in food industry thus alleviating the problem of protein scarcity (Halimatul et
al 2007)
2
middot
13 Medicinal Uses
H sabdariffa is said to be useful in traditional medicine as a digestive agent purgative and
diuretic among others Having two diuretic ingredients ascorbic acid and glycolic acid it
has the ability to increase urination and also valued for its mild laxative effect Containing
citric acid it also can be used as cooling herb The tonic tea produce from the leaves and
flowers is good for digestive and kidney function The heated leaves are applied to cracks
in the feet and on boils and ulcers to speed maturation (Qi et aI 2005) Other roselle s part
have also been reported to be folk remedy for cancer obesity diabetes and hypertension
(Bolad et aI 2009) Amin et al (2008) also added that the flower and fleshy fruits are
used in pharmaceutical industry to relieve symptoms of bronchitis and coughs Roselle s
lotion products are useful for treating sores and wounds
Another nutritional value of Roselle discovered by UKM was the high content of
hydroxycitric acid (HCA) which is widely used as the main ingredients in many
commercial weight loss and slimming products Recent studies by local and international
scientist have proven the effectiveness of HCA in reducing body weight This
characteristic makes Roselle a very valuable crop
14 Problem Statement
The demand for H sabdariffa production nowadays had increase due to its commercial and
medicinal values To fulfill these demands more good quality seeds are needed
Furthermore not many research had been done on improving the H sabdarifJa seeds as a
planting material eventhough the best propagation method for H sabdariffa is by seeds
Other than that disease has been reported as a limiting factor to the production of Roselle
worldwide (Sie R S et aI 2011)
3
1S Objectives
1 To detennine the physical characteristics of H sabdariffa seeds for 10 100 and
1000 seeds weight and for length width and thickness of the seed
2 To evaluate the germination of H sabdariffa seeds following hydration at different
temperatures
3 To determine the effectiveness of hydration technique in improving the gennination
of H sabdariffa seeds
4
-
Pusat Khidmat Maklumat Akadtmik VNIVERSm MALAYSIA SARAWAK
I J bull bull
20 LITERATURE REVIEW
21 Characteristic of Seed
Seeds quality can be affected by changes in environmental temperature and relative
humidity The characteristic of seed must be determine because it is important to ensure
high quality yield in field planting Seed germination can be affected by number of factors
such as level of oxygen temperature and moisture content Growth performance of seed
also can be related to seed size Therefore grading is important to obtain seed that can
grows better Some useful criteria for visual separation of seed includes the seed size
shape colour weight thickness density and surface texture
According to Jain and Bal (1997) roselle seeds are having average principal dimensions of
298-336 186-224 and 170-201 mm Omobuwajo et al (2000) found that the average of
roselle seeds length width and thickness were 558 521 and 281 mm respectively The
seed was reported to be important for its oil in some parts of Africa its native origin From
previous research by scientist roselle seed was among the highest protein-containing seeds
when compared with other seeds like passion fruit (Passiflora edulis) Amaranthus seeds
and Pisum sativum seeds Latest findings by Hainida et al (2008) found that the seed from
Malaysia are composed of 99 moisture 335 protein 221 lipids 130 available
carbohydrate 183 total dietary fibers and 75 ash
H sabdarifJa can be grown easily in most of the tropical country Like other hibiscus
species it is quite hard for roselle to be affected by pest and disease and maintaining this
plant in the field is not so difficult The seeds were sown in a suitable container and the
seedlings will be ready for planting at the age of 10-15 days However it is also important
to ensure that the seeds to be sown were in a good condition and quality to avoid losses
5
22 Seed Germination
Germination is the emergence and development from the seed embryo of those essential
structures which are indicative of the ability to produce a normal plant under favorable
conditions (AOSA 1991) According to Smith (2011) the sum of germination events
beginning with hydration and culminating with root emergence Several stages involved in
germination are imbibition of water activation of enzyme systems metabolism of storage
products and their transport and finally the emergence of the radicle and growth of the
seedling Seed germination is described as the emergence and development from the seed
embryo of those essential structures which for the kind of seed in question are indicative
of the ability to produce a normal plant under favorable conditions (AOSA 1978) In a
simpler contcxt germination is the emergence of the radicle through the seed coat
Depending on the species seeds can germinate as quick as a few days or for as long as
several years Due to the high respiration rate and some exudation and leakage through the
seed coat the germinating seedling will undergoes a net loss in dry weight before the
germination For a particular kind of species the germination condition is mostly affected
by temperature type of substrate on which to germinate the seed light condition and
moisture level
The germination test is commonly used to determine seed viability It is an analytical
procedure to evaluate seed germination under standardized favorable conditions that are
seldom if ever encountered in the field The germination test is merely an estimate and
has certain limitations as a universal estimate of seed quality However if these limitations
are recognized the germination test is a useful viability index The need for germination
test is to estimate how the seed will perform in the field
6
I bull bull
23 Seed Moisture Content
Amount of water in the seed that usually expressed in a percentage fonn is the seed
moisture content Agrawal (1980) found that the amount of moisture in the seed is
probably the most important factor influencing seed viability during storage By
determining the seeds moisture content the possible storage life for the seeds can be
predicted According to McDonald (2005) selecting the appropriate salt to achieve a seed
moisture content of 5-6 is the recommended method for achieving optimum seed
storage and minimizing seed deterioration It is necessary to dry seeds to the safe moisture
contents since the life of a seed and its span largely revolves around its moisture content
and the suitable moisture content for starchy seeds are less than 14 while for oily seeds
are less than 11 (Agrawal 1980)
24 Seed Vigour
According to Geneve (2005) seed vigour is defined by the Asociation of official Seed
analysts as those seed properties which detennine the potential for rapid unifonn
emergence and development of nonnal seedlings under the wide range of field conditions
Seed vigour is closely related with seed storage environment and duration During seed
storage the vigour of a seed lot is reduced prior to seed viability as indicated by standard
germination (Hampton amp TeKrony 1995) Major factors influencing the seed vigour in
stored seed lots are the environmental conditions during seed storage and the length of time
in storage Tetrazolium test is one of the method for detennining the seeds vigour To
minimize the decline in vigour and germination seeds should be undamaged
7
bull t I I
25 Seed Hydration
A process of supplying water to seed to maintain the fluid or moisture content in the seed
is called hydration According to Copeland and McDonald (1995) seed hydration is a
process whereby seeds are hydrated using various protocols and then redried to permit
routine handling This process results in increased germination rate more uniform
emergence germination under a broader range of environments and improved sedling
vigour and growth It is one of the priming treatments of seed which usually applied to
strengthen or increase vitality of seed Priming process can increase a rate of germination
as the treatment improves the seed vigor Hydration is one component of priming which
can increase germination with more range of environment condition The objective of seed
hydration technology is to increase the percentage and rate of germination expand the
range of temperatures over which the seed will germinate and increase the uniformity of
stand establishment To accomplish these objectives seeds must be hydrated in some way
at a moisture level sufficient to initiate the early events of germination but not sufficient to
permit radicle protrusion (Akers and Holley 1986) However the performance of the seed
might also being influence by other factors like the species type of seed and the seeds
quality
Water can be used for the hydration treatment because it had proved to enhance
germination emergence growth and yield of seed However other mediwns such as the
solution of potassium hydrophosphate (KH2P04) monobasic polyethylene glycol (PEG)
potassium chloride (KCl) potassiwn nitrate (KN03) potassium phoshate (K3P04)
magnesium sulphate (MgS04) sodiwn chloride (NaCl) glycerol and mannitol also can be
used to immerse the seed in hydration process The benefit of such salts is to supply the
seed with nitrogen and other nutrients essential for protein synthesis during germination
8
t I amp
26 Seed Storage
A basic requirement in seed production is a good seed storage The purpose of seed storage
is to preserve planting stocks from one season to the next (Larry amp McDonald 1995)
Generally we stored seeds for the commercial purpose for carry - over seeds or for genn
plasm seeds Maintaining the seed quality for the longest duration possible is the main
objective in seed storage Loss of gennination during storage cannot be stopped but it
could be reduce by giving good storage conditions According to Agrawal (1980) the
seeds are considered to be in storage from the moment they reach physiological maturity
until they genninate or until they are thrown away because they are dead or otherwise
worthless
The seed storage must be properly handled because it can affect the seed quality There are
some factors that can affect the seed longevity in storage such as the variety of the seed
initial seed quality moisture content relative humidity and storage temperature It is
necessary for the seed storage condition to be dry and cool effective in pest control and
have a proper sanitation Other than than storing only high quality of seed also can help in
better production From all of the factors temperature is one of the most important element
which influence seed viability and vigour during storage Agrawal (1980) mentioned that
the lower the temperature the longer the seeds maintain gennination capacity so
decreasing temperature and seed moisture is an effective means of maintaining seed quality
in storage Every seed lot should be gennination - tested when it is received for storage to
record the seed quality Subsequent periodic checks also should be made to detect
deterioration and if it occurs storage condition should be checked to discover the reasons
and to take corrective measures
9
middot
27 Seed Specific Density
Density of seed refers to it mass per unit volume The seed size and density will affect the
yield One of the method for grouping seeds into some specific density is the liquid density
separation system It is one of the easier and cheaper method compared to using a machine
When seed is loaded into a liquid some will float and some will sink That mean the
floating seeds density is lower than the liquids density Vice versa the sinking seeds
density is higher than the liquids density Wang and Alyarez (2008) explained that this
method of separating seeds works by adjusting the specific gravity of the liquid in the
reservoir so that a first portion of the seeds float near the surface of the liquid and a second
portion of the seeds sink near the bottom of the reservoir and providing an outlet flow of
the liquid from a location proximal to the top of the reservoir above the location of the
second portion of seeds By this way seeds of lower density can be separated from higher
density Kwong Sellman Jalink amp Schoor (2005) illustrated that separation by density is
commonly incorporated as part of the routine cleaning and grading process in flower seed
production procedures Other than grouping seeds according to their specific density this
density separation also can be used to remove broken partially filled empty or immature
seeds
28 Seed Deterioration
McDonald (2005) defined flower seed deterioration as deteriorative changes occuring with
time that increase the seeds vulnerability to external challenges and decrease the ability of
the seed to survive Important factors contribute to seed deterioration are genetics seed
structure seed chemistry physicalphysiological quality seed treatments relative humidity
10
and temperature Relative humidity is important because it will influence the moisture
content of seeds in storage while temperature determines the amount of moisture in the air
and influences the rate of deteriorative reactions in seeds According to Agrawal (1980)
the rate of deterioration increases as the seed moisture content increases This could be due
to the mold growth in and on the seed
11
~
30 MATERIAL AND METHOD
31 Material
Fresh matured fruits of Hibiscus sabdarifJa L (Plate I) were obtained from local fanns in
Kota Samarahan Seeds were extracted from these fruits cleaned and dried under shade
Dried seeds were dusted with Captan 80 and then placed in air-tight bottle to keep it in a
safe place before use in the subsequent experiment
Plate 1 Fresh fruit ofH sabdarifJa Plate 2 Fresh fruit of H sabdarifJa with seeds
32 Method
321 Preliminary Evaluation
Moisture content and germination was conducted as an initial evaluation to assess the
quality of seeds used in this research The procedures were those prescribed by Association
ofOfficial Seeds Analysts (AOSA 1985)
12
middot
322 Moisture Content Test
Four replicates of 10 seeds each were placed into saucers (Plate 5) and calibrated to get the
wet weight The seeds then were placed in an oven at 60degC for 48 hours After that the
seeds were weighed again to get the dry weight The percentage of moisture content of the
seeds was calculated based on the formula below (AOSA 1985)
Moisture content () = b - c x 100
b-a
where a = weight of empty saucer
b = weight of (a) + weight of seeds before drying
c = weight of (a) + weight of seeds after drying in the oven
323 Germination Test
Germination test was conducted base on the AOSA (1985) where 25 seeds with four
replications were used For every replicates seeds were put into a petry dish on 3 layers of
moist Whatman filter paper (Plate 6) The seeds were left to germinate by incubating it in a
Plant Growth Chamber (28OC) and was checked daily until 10 days Seed was considered
as germinated if the radicles were about 5mm length emerged from the seed coat The
germination percentage was calculated base on AOSA 1985 formula below
13
-----------------------------------1
Gennination () a x 100
b
where a = total number of germinated seeds
b = total number of seeds used
324 Seed Characteristic
The size of the H sabdariffa seeds was calibrated for its length width and thickness Ten
replicates of seeds were used for calibrating the physical characteristics Then the seed
weights of 10 100 and 1000 seeds were calibrated Four replicates of 10 100 and 1000
seed-weight classes were used
325 Hydration of Seeds
Seed of H sabdariffa was placed in distilled water for 0 2 4 6 8 10 and 12 hours at
temperatures of 28 30 35 and 40degC Then seeds were analyzed for the moisture content
and gennination
326 Data Analysis
The experimental units were arranged randomly based on Completely Randomized Design
(CRD) and data was analyzed by using Analysis of Variance (ANOV A) If there any
significant differences the mean were discriminated using Least Significant Different
(LSD)
14
I
Physical characteristic and germination of Hibiscus Sabdariffa L following hydration treatments
Syazwanie Binti Dzulbaimi
Plant Resource and Management Programme Faculty of Science and Technology
Universiti Malaysia Sarawak
ABSTRACT
A study was conducted to determine the physical characteristics and to assess and evaluate the effectiveness of hydration techniques in improving the performance of HibisclIs sabdariffa L seeds The average length width and thickness obtained for H sabdariffa seeds were 539 mm 455 mm and 234 mm respectively The average weight for a single seed was 002 g and it was estimated that one kg weight contained approximately 50000 seeds This indicated that the seeds used in this experiment were small In hydration treatment seeds of H sabdariffa were soaked in water at temperature 28 30 35 and 40degC for 0246 8 10 and 12 hours and were evaluated for moisture content and germination Results indicated that hydration treatment was not suitable for H sabdariffa seeds as it did not improved the germination The highest germination percentage of H sabdarifJa seeds following hydration treatment was 29 which was obtained for 2 hours hydration at 30degC
Key words Hibiscus sabdarifJa calibration hydration temperature
ABSTRAK
Satu kajian telah dijalankan IInt1lk menentukan ciri-ciri jizikal dall mengkaji serta menentukan keberkesanan teknik rawatall hidrasi dalam meningkatkan kadar pertllmbllhan biji benih Hibiscus sabdariffa Dalam penentlkuran biji benih pIrata panjang lebar dan ketebalan biji benih H sabdariffa yang diperoleh adalah 539 mm 455 mm dan 234 mm Purata berat sebiji benih adalah 002 g dan dianggarkan bahawa satll kilogram mengandungi lebih kurang 50000 biji benih Ini memmjukkan yang biji benih yang digllnakan unIlk kajian ini agak keci Dalam rawatan hidrasi biji benih H sabdariffa direndam di dalam air pada suhu 28degC 30degC 35degC dan 40degC Bagi setiap suhu yang digunakan biji benih direndam selama 0 2 4 6 8 10 dan 12 jam dan selepas itu kandungan kelembapan dan percambahan biji benih dinilai Keputlsan yang diperoleh menllnjukkan rawatan hidrasi adalah tidak sesuai bagi biji benih H sabdariffa memalldangkan peratls percambahan tidak meningkat berbanding sebelum rawatan Peratlls percambahan tertinggi bagi biji benih H sabdariffa selepas rawatan hidrasi adalah 29 iaitll bagi hidrasi selama 2 jam pada slhu 30degC
Kata kllnciHibisclS sabdarifJa kalibrasi hidrasi suhl
ix
I bull
10 INTRODUCTION
11 Background
Hibiscus sabdarifJa Linn or also known as Roselle or red sorrel belongs to the family
Malvaceae It is also known as asam paya asam susur and asam kumbang locally It is a
new commercial crop in Malaysia where it was brought from India (Amin et al 2008)
Since Roselle plantation only had started in early 1990s it is considered as a new crop in
Malaysia Two introduced varieties in Malaysia are Arab and Terengganu variety
However in 2009 Universiti Kebangsaan Malaysia had launched three new varieties
named UKMR-l UKMR-2 and UKMR-3 to help promote the Roselle industry Recently
Department of Agriculture had collaborated with F AMA entrepreneurs and growers in
order to strengthen the H sabdarifJa industry in Malaysia by expanding and marketing this
product locally and in foreign market
According to Qi et al (2005) H sabdarifJa is an annual mostly brannched and erect
shrub Its reddish stem can achieve 35m tall The leaves are dark green to red alternate
glabrous long - petiolate palmately divided into 3 - 7 lobes with serrate margins Having
both male and female organs the flower are red to yellow in color with a dark centre
containing short peduncles
H sabdarifJa can survive in a wann and humid tropical and subtropical climate It can be
grown in a greenhouse with litle shade but the best condition for growing it is under the
full sunlight (Qi et aI 2005) Roselle plants are suitable for tropical climates with wellshy
distributed rain - fall of 1500 - 2000 mm per year from sea level to about 600 m in
altitude (Amin et aI 2008) It can adapt to variety of soils but prefered a friable sandy
loam with humus
1
Current production of H sabdarifJa in Malaysia is about 240 tonnes annually Reported in
unpublished result from Agricultural Department of Terengganu in the process of
removing the calyces out of 3 tonnes of raw materials about 15 tonnes of the velvety
capsules containing the seeds are being disposed as a by - product and unexploited
(Halimatul Amin Mohd Esa Nawalyah amp Siti 2007) Amin et at (2008) mentioned that
the plant takes about three to four months to reach the commercial stage of maturity before
the flowers are harvested
12 Commercial Production
Many parts of H sabdariffa including seeds leaves fruits and roots are very useful
especially the fleshy red calyces They are used fresh for making wine juice jam jelly
syrup gelatin pudding cakes ice cream and flavors and also dried and brewed into tea
spice and used for butter pies sauces tarts and other desserts (Qi et al 2005) In a study
about commercial practice of Roselle beverage production Bolade et at (2009) reported
that the dried calyceswater ratios involved in the commercial practice ranged between
157 and 171 (wv) while the sweetness level of the beverage ranged between 112 and
133 degBrix The red calyces which contains high amount of vitamin C was used to make
those healthy beverages In Malaysia we also consume the leaves as vegetables There are
previous study shows that H sabdarifJa seeds can be used as a protein source Instead of
using protein from conventional sources protein isolates or concentrates from Roselle
seeds might be useful as low cost source of protein substitute in dietary supplement or food
ingredient in food industry thus alleviating the problem of protein scarcity (Halimatul et
al 2007)
2
middot
13 Medicinal Uses
H sabdariffa is said to be useful in traditional medicine as a digestive agent purgative and
diuretic among others Having two diuretic ingredients ascorbic acid and glycolic acid it
has the ability to increase urination and also valued for its mild laxative effect Containing
citric acid it also can be used as cooling herb The tonic tea produce from the leaves and
flowers is good for digestive and kidney function The heated leaves are applied to cracks
in the feet and on boils and ulcers to speed maturation (Qi et aI 2005) Other roselle s part
have also been reported to be folk remedy for cancer obesity diabetes and hypertension
(Bolad et aI 2009) Amin et al (2008) also added that the flower and fleshy fruits are
used in pharmaceutical industry to relieve symptoms of bronchitis and coughs Roselle s
lotion products are useful for treating sores and wounds
Another nutritional value of Roselle discovered by UKM was the high content of
hydroxycitric acid (HCA) which is widely used as the main ingredients in many
commercial weight loss and slimming products Recent studies by local and international
scientist have proven the effectiveness of HCA in reducing body weight This
characteristic makes Roselle a very valuable crop
14 Problem Statement
The demand for H sabdariffa production nowadays had increase due to its commercial and
medicinal values To fulfill these demands more good quality seeds are needed
Furthermore not many research had been done on improving the H sabdarifJa seeds as a
planting material eventhough the best propagation method for H sabdariffa is by seeds
Other than that disease has been reported as a limiting factor to the production of Roselle
worldwide (Sie R S et aI 2011)
3
1S Objectives
1 To detennine the physical characteristics of H sabdariffa seeds for 10 100 and
1000 seeds weight and for length width and thickness of the seed
2 To evaluate the germination of H sabdariffa seeds following hydration at different
temperatures
3 To determine the effectiveness of hydration technique in improving the gennination
of H sabdariffa seeds
4
-
Pusat Khidmat Maklumat Akadtmik VNIVERSm MALAYSIA SARAWAK
I J bull bull
20 LITERATURE REVIEW
21 Characteristic of Seed
Seeds quality can be affected by changes in environmental temperature and relative
humidity The characteristic of seed must be determine because it is important to ensure
high quality yield in field planting Seed germination can be affected by number of factors
such as level of oxygen temperature and moisture content Growth performance of seed
also can be related to seed size Therefore grading is important to obtain seed that can
grows better Some useful criteria for visual separation of seed includes the seed size
shape colour weight thickness density and surface texture
According to Jain and Bal (1997) roselle seeds are having average principal dimensions of
298-336 186-224 and 170-201 mm Omobuwajo et al (2000) found that the average of
roselle seeds length width and thickness were 558 521 and 281 mm respectively The
seed was reported to be important for its oil in some parts of Africa its native origin From
previous research by scientist roselle seed was among the highest protein-containing seeds
when compared with other seeds like passion fruit (Passiflora edulis) Amaranthus seeds
and Pisum sativum seeds Latest findings by Hainida et al (2008) found that the seed from
Malaysia are composed of 99 moisture 335 protein 221 lipids 130 available
carbohydrate 183 total dietary fibers and 75 ash
H sabdarifJa can be grown easily in most of the tropical country Like other hibiscus
species it is quite hard for roselle to be affected by pest and disease and maintaining this
plant in the field is not so difficult The seeds were sown in a suitable container and the
seedlings will be ready for planting at the age of 10-15 days However it is also important
to ensure that the seeds to be sown were in a good condition and quality to avoid losses
5
22 Seed Germination
Germination is the emergence and development from the seed embryo of those essential
structures which are indicative of the ability to produce a normal plant under favorable
conditions (AOSA 1991) According to Smith (2011) the sum of germination events
beginning with hydration and culminating with root emergence Several stages involved in
germination are imbibition of water activation of enzyme systems metabolism of storage
products and their transport and finally the emergence of the radicle and growth of the
seedling Seed germination is described as the emergence and development from the seed
embryo of those essential structures which for the kind of seed in question are indicative
of the ability to produce a normal plant under favorable conditions (AOSA 1978) In a
simpler contcxt germination is the emergence of the radicle through the seed coat
Depending on the species seeds can germinate as quick as a few days or for as long as
several years Due to the high respiration rate and some exudation and leakage through the
seed coat the germinating seedling will undergoes a net loss in dry weight before the
germination For a particular kind of species the germination condition is mostly affected
by temperature type of substrate on which to germinate the seed light condition and
moisture level
The germination test is commonly used to determine seed viability It is an analytical
procedure to evaluate seed germination under standardized favorable conditions that are
seldom if ever encountered in the field The germination test is merely an estimate and
has certain limitations as a universal estimate of seed quality However if these limitations
are recognized the germination test is a useful viability index The need for germination
test is to estimate how the seed will perform in the field
6
I bull bull
23 Seed Moisture Content
Amount of water in the seed that usually expressed in a percentage fonn is the seed
moisture content Agrawal (1980) found that the amount of moisture in the seed is
probably the most important factor influencing seed viability during storage By
determining the seeds moisture content the possible storage life for the seeds can be
predicted According to McDonald (2005) selecting the appropriate salt to achieve a seed
moisture content of 5-6 is the recommended method for achieving optimum seed
storage and minimizing seed deterioration It is necessary to dry seeds to the safe moisture
contents since the life of a seed and its span largely revolves around its moisture content
and the suitable moisture content for starchy seeds are less than 14 while for oily seeds
are less than 11 (Agrawal 1980)
24 Seed Vigour
According to Geneve (2005) seed vigour is defined by the Asociation of official Seed
analysts as those seed properties which detennine the potential for rapid unifonn
emergence and development of nonnal seedlings under the wide range of field conditions
Seed vigour is closely related with seed storage environment and duration During seed
storage the vigour of a seed lot is reduced prior to seed viability as indicated by standard
germination (Hampton amp TeKrony 1995) Major factors influencing the seed vigour in
stored seed lots are the environmental conditions during seed storage and the length of time
in storage Tetrazolium test is one of the method for detennining the seeds vigour To
minimize the decline in vigour and germination seeds should be undamaged
7
bull t I I
25 Seed Hydration
A process of supplying water to seed to maintain the fluid or moisture content in the seed
is called hydration According to Copeland and McDonald (1995) seed hydration is a
process whereby seeds are hydrated using various protocols and then redried to permit
routine handling This process results in increased germination rate more uniform
emergence germination under a broader range of environments and improved sedling
vigour and growth It is one of the priming treatments of seed which usually applied to
strengthen or increase vitality of seed Priming process can increase a rate of germination
as the treatment improves the seed vigor Hydration is one component of priming which
can increase germination with more range of environment condition The objective of seed
hydration technology is to increase the percentage and rate of germination expand the
range of temperatures over which the seed will germinate and increase the uniformity of
stand establishment To accomplish these objectives seeds must be hydrated in some way
at a moisture level sufficient to initiate the early events of germination but not sufficient to
permit radicle protrusion (Akers and Holley 1986) However the performance of the seed
might also being influence by other factors like the species type of seed and the seeds
quality
Water can be used for the hydration treatment because it had proved to enhance
germination emergence growth and yield of seed However other mediwns such as the
solution of potassium hydrophosphate (KH2P04) monobasic polyethylene glycol (PEG)
potassium chloride (KCl) potassiwn nitrate (KN03) potassium phoshate (K3P04)
magnesium sulphate (MgS04) sodiwn chloride (NaCl) glycerol and mannitol also can be
used to immerse the seed in hydration process The benefit of such salts is to supply the
seed with nitrogen and other nutrients essential for protein synthesis during germination
8
t I amp
26 Seed Storage
A basic requirement in seed production is a good seed storage The purpose of seed storage
is to preserve planting stocks from one season to the next (Larry amp McDonald 1995)
Generally we stored seeds for the commercial purpose for carry - over seeds or for genn
plasm seeds Maintaining the seed quality for the longest duration possible is the main
objective in seed storage Loss of gennination during storage cannot be stopped but it
could be reduce by giving good storage conditions According to Agrawal (1980) the
seeds are considered to be in storage from the moment they reach physiological maturity
until they genninate or until they are thrown away because they are dead or otherwise
worthless
The seed storage must be properly handled because it can affect the seed quality There are
some factors that can affect the seed longevity in storage such as the variety of the seed
initial seed quality moisture content relative humidity and storage temperature It is
necessary for the seed storage condition to be dry and cool effective in pest control and
have a proper sanitation Other than than storing only high quality of seed also can help in
better production From all of the factors temperature is one of the most important element
which influence seed viability and vigour during storage Agrawal (1980) mentioned that
the lower the temperature the longer the seeds maintain gennination capacity so
decreasing temperature and seed moisture is an effective means of maintaining seed quality
in storage Every seed lot should be gennination - tested when it is received for storage to
record the seed quality Subsequent periodic checks also should be made to detect
deterioration and if it occurs storage condition should be checked to discover the reasons
and to take corrective measures
9
middot
27 Seed Specific Density
Density of seed refers to it mass per unit volume The seed size and density will affect the
yield One of the method for grouping seeds into some specific density is the liquid density
separation system It is one of the easier and cheaper method compared to using a machine
When seed is loaded into a liquid some will float and some will sink That mean the
floating seeds density is lower than the liquids density Vice versa the sinking seeds
density is higher than the liquids density Wang and Alyarez (2008) explained that this
method of separating seeds works by adjusting the specific gravity of the liquid in the
reservoir so that a first portion of the seeds float near the surface of the liquid and a second
portion of the seeds sink near the bottom of the reservoir and providing an outlet flow of
the liquid from a location proximal to the top of the reservoir above the location of the
second portion of seeds By this way seeds of lower density can be separated from higher
density Kwong Sellman Jalink amp Schoor (2005) illustrated that separation by density is
commonly incorporated as part of the routine cleaning and grading process in flower seed
production procedures Other than grouping seeds according to their specific density this
density separation also can be used to remove broken partially filled empty or immature
seeds
28 Seed Deterioration
McDonald (2005) defined flower seed deterioration as deteriorative changes occuring with
time that increase the seeds vulnerability to external challenges and decrease the ability of
the seed to survive Important factors contribute to seed deterioration are genetics seed
structure seed chemistry physicalphysiological quality seed treatments relative humidity
10
and temperature Relative humidity is important because it will influence the moisture
content of seeds in storage while temperature determines the amount of moisture in the air
and influences the rate of deteriorative reactions in seeds According to Agrawal (1980)
the rate of deterioration increases as the seed moisture content increases This could be due
to the mold growth in and on the seed
11
~
30 MATERIAL AND METHOD
31 Material
Fresh matured fruits of Hibiscus sabdarifJa L (Plate I) were obtained from local fanns in
Kota Samarahan Seeds were extracted from these fruits cleaned and dried under shade
Dried seeds were dusted with Captan 80 and then placed in air-tight bottle to keep it in a
safe place before use in the subsequent experiment
Plate 1 Fresh fruit ofH sabdarifJa Plate 2 Fresh fruit of H sabdarifJa with seeds
32 Method
321 Preliminary Evaluation
Moisture content and germination was conducted as an initial evaluation to assess the
quality of seeds used in this research The procedures were those prescribed by Association
ofOfficial Seeds Analysts (AOSA 1985)
12
middot
322 Moisture Content Test
Four replicates of 10 seeds each were placed into saucers (Plate 5) and calibrated to get the
wet weight The seeds then were placed in an oven at 60degC for 48 hours After that the
seeds were weighed again to get the dry weight The percentage of moisture content of the
seeds was calculated based on the formula below (AOSA 1985)
Moisture content () = b - c x 100
b-a
where a = weight of empty saucer
b = weight of (a) + weight of seeds before drying
c = weight of (a) + weight of seeds after drying in the oven
323 Germination Test
Germination test was conducted base on the AOSA (1985) where 25 seeds with four
replications were used For every replicates seeds were put into a petry dish on 3 layers of
moist Whatman filter paper (Plate 6) The seeds were left to germinate by incubating it in a
Plant Growth Chamber (28OC) and was checked daily until 10 days Seed was considered
as germinated if the radicles were about 5mm length emerged from the seed coat The
germination percentage was calculated base on AOSA 1985 formula below
13
-----------------------------------1
Gennination () a x 100
b
where a = total number of germinated seeds
b = total number of seeds used
324 Seed Characteristic
The size of the H sabdariffa seeds was calibrated for its length width and thickness Ten
replicates of seeds were used for calibrating the physical characteristics Then the seed
weights of 10 100 and 1000 seeds were calibrated Four replicates of 10 100 and 1000
seed-weight classes were used
325 Hydration of Seeds
Seed of H sabdariffa was placed in distilled water for 0 2 4 6 8 10 and 12 hours at
temperatures of 28 30 35 and 40degC Then seeds were analyzed for the moisture content
and gennination
326 Data Analysis
The experimental units were arranged randomly based on Completely Randomized Design
(CRD) and data was analyzed by using Analysis of Variance (ANOV A) If there any
significant differences the mean were discriminated using Least Significant Different
(LSD)
14
I bull
10 INTRODUCTION
11 Background
Hibiscus sabdarifJa Linn or also known as Roselle or red sorrel belongs to the family
Malvaceae It is also known as asam paya asam susur and asam kumbang locally It is a
new commercial crop in Malaysia where it was brought from India (Amin et al 2008)
Since Roselle plantation only had started in early 1990s it is considered as a new crop in
Malaysia Two introduced varieties in Malaysia are Arab and Terengganu variety
However in 2009 Universiti Kebangsaan Malaysia had launched three new varieties
named UKMR-l UKMR-2 and UKMR-3 to help promote the Roselle industry Recently
Department of Agriculture had collaborated with F AMA entrepreneurs and growers in
order to strengthen the H sabdarifJa industry in Malaysia by expanding and marketing this
product locally and in foreign market
According to Qi et al (2005) H sabdarifJa is an annual mostly brannched and erect
shrub Its reddish stem can achieve 35m tall The leaves are dark green to red alternate
glabrous long - petiolate palmately divided into 3 - 7 lobes with serrate margins Having
both male and female organs the flower are red to yellow in color with a dark centre
containing short peduncles
H sabdarifJa can survive in a wann and humid tropical and subtropical climate It can be
grown in a greenhouse with litle shade but the best condition for growing it is under the
full sunlight (Qi et aI 2005) Roselle plants are suitable for tropical climates with wellshy
distributed rain - fall of 1500 - 2000 mm per year from sea level to about 600 m in
altitude (Amin et aI 2008) It can adapt to variety of soils but prefered a friable sandy
loam with humus
1
Current production of H sabdarifJa in Malaysia is about 240 tonnes annually Reported in
unpublished result from Agricultural Department of Terengganu in the process of
removing the calyces out of 3 tonnes of raw materials about 15 tonnes of the velvety
capsules containing the seeds are being disposed as a by - product and unexploited
(Halimatul Amin Mohd Esa Nawalyah amp Siti 2007) Amin et at (2008) mentioned that
the plant takes about three to four months to reach the commercial stage of maturity before
the flowers are harvested
12 Commercial Production
Many parts of H sabdariffa including seeds leaves fruits and roots are very useful
especially the fleshy red calyces They are used fresh for making wine juice jam jelly
syrup gelatin pudding cakes ice cream and flavors and also dried and brewed into tea
spice and used for butter pies sauces tarts and other desserts (Qi et al 2005) In a study
about commercial practice of Roselle beverage production Bolade et at (2009) reported
that the dried calyceswater ratios involved in the commercial practice ranged between
157 and 171 (wv) while the sweetness level of the beverage ranged between 112 and
133 degBrix The red calyces which contains high amount of vitamin C was used to make
those healthy beverages In Malaysia we also consume the leaves as vegetables There are
previous study shows that H sabdarifJa seeds can be used as a protein source Instead of
using protein from conventional sources protein isolates or concentrates from Roselle
seeds might be useful as low cost source of protein substitute in dietary supplement or food
ingredient in food industry thus alleviating the problem of protein scarcity (Halimatul et
al 2007)
2
middot
13 Medicinal Uses
H sabdariffa is said to be useful in traditional medicine as a digestive agent purgative and
diuretic among others Having two diuretic ingredients ascorbic acid and glycolic acid it
has the ability to increase urination and also valued for its mild laxative effect Containing
citric acid it also can be used as cooling herb The tonic tea produce from the leaves and
flowers is good for digestive and kidney function The heated leaves are applied to cracks
in the feet and on boils and ulcers to speed maturation (Qi et aI 2005) Other roselle s part
have also been reported to be folk remedy for cancer obesity diabetes and hypertension
(Bolad et aI 2009) Amin et al (2008) also added that the flower and fleshy fruits are
used in pharmaceutical industry to relieve symptoms of bronchitis and coughs Roselle s
lotion products are useful for treating sores and wounds
Another nutritional value of Roselle discovered by UKM was the high content of
hydroxycitric acid (HCA) which is widely used as the main ingredients in many
commercial weight loss and slimming products Recent studies by local and international
scientist have proven the effectiveness of HCA in reducing body weight This
characteristic makes Roselle a very valuable crop
14 Problem Statement
The demand for H sabdariffa production nowadays had increase due to its commercial and
medicinal values To fulfill these demands more good quality seeds are needed
Furthermore not many research had been done on improving the H sabdarifJa seeds as a
planting material eventhough the best propagation method for H sabdariffa is by seeds
Other than that disease has been reported as a limiting factor to the production of Roselle
worldwide (Sie R S et aI 2011)
3
1S Objectives
1 To detennine the physical characteristics of H sabdariffa seeds for 10 100 and
1000 seeds weight and for length width and thickness of the seed
2 To evaluate the germination of H sabdariffa seeds following hydration at different
temperatures
3 To determine the effectiveness of hydration technique in improving the gennination
of H sabdariffa seeds
4
-
Pusat Khidmat Maklumat Akadtmik VNIVERSm MALAYSIA SARAWAK
I J bull bull
20 LITERATURE REVIEW
21 Characteristic of Seed
Seeds quality can be affected by changes in environmental temperature and relative
humidity The characteristic of seed must be determine because it is important to ensure
high quality yield in field planting Seed germination can be affected by number of factors
such as level of oxygen temperature and moisture content Growth performance of seed
also can be related to seed size Therefore grading is important to obtain seed that can
grows better Some useful criteria for visual separation of seed includes the seed size
shape colour weight thickness density and surface texture
According to Jain and Bal (1997) roselle seeds are having average principal dimensions of
298-336 186-224 and 170-201 mm Omobuwajo et al (2000) found that the average of
roselle seeds length width and thickness were 558 521 and 281 mm respectively The
seed was reported to be important for its oil in some parts of Africa its native origin From
previous research by scientist roselle seed was among the highest protein-containing seeds
when compared with other seeds like passion fruit (Passiflora edulis) Amaranthus seeds
and Pisum sativum seeds Latest findings by Hainida et al (2008) found that the seed from
Malaysia are composed of 99 moisture 335 protein 221 lipids 130 available
carbohydrate 183 total dietary fibers and 75 ash
H sabdarifJa can be grown easily in most of the tropical country Like other hibiscus
species it is quite hard for roselle to be affected by pest and disease and maintaining this
plant in the field is not so difficult The seeds were sown in a suitable container and the
seedlings will be ready for planting at the age of 10-15 days However it is also important
to ensure that the seeds to be sown were in a good condition and quality to avoid losses
5
22 Seed Germination
Germination is the emergence and development from the seed embryo of those essential
structures which are indicative of the ability to produce a normal plant under favorable
conditions (AOSA 1991) According to Smith (2011) the sum of germination events
beginning with hydration and culminating with root emergence Several stages involved in
germination are imbibition of water activation of enzyme systems metabolism of storage
products and their transport and finally the emergence of the radicle and growth of the
seedling Seed germination is described as the emergence and development from the seed
embryo of those essential structures which for the kind of seed in question are indicative
of the ability to produce a normal plant under favorable conditions (AOSA 1978) In a
simpler contcxt germination is the emergence of the radicle through the seed coat
Depending on the species seeds can germinate as quick as a few days or for as long as
several years Due to the high respiration rate and some exudation and leakage through the
seed coat the germinating seedling will undergoes a net loss in dry weight before the
germination For a particular kind of species the germination condition is mostly affected
by temperature type of substrate on which to germinate the seed light condition and
moisture level
The germination test is commonly used to determine seed viability It is an analytical
procedure to evaluate seed germination under standardized favorable conditions that are
seldom if ever encountered in the field The germination test is merely an estimate and
has certain limitations as a universal estimate of seed quality However if these limitations
are recognized the germination test is a useful viability index The need for germination
test is to estimate how the seed will perform in the field
6
I bull bull
23 Seed Moisture Content
Amount of water in the seed that usually expressed in a percentage fonn is the seed
moisture content Agrawal (1980) found that the amount of moisture in the seed is
probably the most important factor influencing seed viability during storage By
determining the seeds moisture content the possible storage life for the seeds can be
predicted According to McDonald (2005) selecting the appropriate salt to achieve a seed
moisture content of 5-6 is the recommended method for achieving optimum seed
storage and minimizing seed deterioration It is necessary to dry seeds to the safe moisture
contents since the life of a seed and its span largely revolves around its moisture content
and the suitable moisture content for starchy seeds are less than 14 while for oily seeds
are less than 11 (Agrawal 1980)
24 Seed Vigour
According to Geneve (2005) seed vigour is defined by the Asociation of official Seed
analysts as those seed properties which detennine the potential for rapid unifonn
emergence and development of nonnal seedlings under the wide range of field conditions
Seed vigour is closely related with seed storage environment and duration During seed
storage the vigour of a seed lot is reduced prior to seed viability as indicated by standard
germination (Hampton amp TeKrony 1995) Major factors influencing the seed vigour in
stored seed lots are the environmental conditions during seed storage and the length of time
in storage Tetrazolium test is one of the method for detennining the seeds vigour To
minimize the decline in vigour and germination seeds should be undamaged
7
bull t I I
25 Seed Hydration
A process of supplying water to seed to maintain the fluid or moisture content in the seed
is called hydration According to Copeland and McDonald (1995) seed hydration is a
process whereby seeds are hydrated using various protocols and then redried to permit
routine handling This process results in increased germination rate more uniform
emergence germination under a broader range of environments and improved sedling
vigour and growth It is one of the priming treatments of seed which usually applied to
strengthen or increase vitality of seed Priming process can increase a rate of germination
as the treatment improves the seed vigor Hydration is one component of priming which
can increase germination with more range of environment condition The objective of seed
hydration technology is to increase the percentage and rate of germination expand the
range of temperatures over which the seed will germinate and increase the uniformity of
stand establishment To accomplish these objectives seeds must be hydrated in some way
at a moisture level sufficient to initiate the early events of germination but not sufficient to
permit radicle protrusion (Akers and Holley 1986) However the performance of the seed
might also being influence by other factors like the species type of seed and the seeds
quality
Water can be used for the hydration treatment because it had proved to enhance
germination emergence growth and yield of seed However other mediwns such as the
solution of potassium hydrophosphate (KH2P04) monobasic polyethylene glycol (PEG)
potassium chloride (KCl) potassiwn nitrate (KN03) potassium phoshate (K3P04)
magnesium sulphate (MgS04) sodiwn chloride (NaCl) glycerol and mannitol also can be
used to immerse the seed in hydration process The benefit of such salts is to supply the
seed with nitrogen and other nutrients essential for protein synthesis during germination
8
t I amp
26 Seed Storage
A basic requirement in seed production is a good seed storage The purpose of seed storage
is to preserve planting stocks from one season to the next (Larry amp McDonald 1995)
Generally we stored seeds for the commercial purpose for carry - over seeds or for genn
plasm seeds Maintaining the seed quality for the longest duration possible is the main
objective in seed storage Loss of gennination during storage cannot be stopped but it
could be reduce by giving good storage conditions According to Agrawal (1980) the
seeds are considered to be in storage from the moment they reach physiological maturity
until they genninate or until they are thrown away because they are dead or otherwise
worthless
The seed storage must be properly handled because it can affect the seed quality There are
some factors that can affect the seed longevity in storage such as the variety of the seed
initial seed quality moisture content relative humidity and storage temperature It is
necessary for the seed storage condition to be dry and cool effective in pest control and
have a proper sanitation Other than than storing only high quality of seed also can help in
better production From all of the factors temperature is one of the most important element
which influence seed viability and vigour during storage Agrawal (1980) mentioned that
the lower the temperature the longer the seeds maintain gennination capacity so
decreasing temperature and seed moisture is an effective means of maintaining seed quality
in storage Every seed lot should be gennination - tested when it is received for storage to
record the seed quality Subsequent periodic checks also should be made to detect
deterioration and if it occurs storage condition should be checked to discover the reasons
and to take corrective measures
9
middot
27 Seed Specific Density
Density of seed refers to it mass per unit volume The seed size and density will affect the
yield One of the method for grouping seeds into some specific density is the liquid density
separation system It is one of the easier and cheaper method compared to using a machine
When seed is loaded into a liquid some will float and some will sink That mean the
floating seeds density is lower than the liquids density Vice versa the sinking seeds
density is higher than the liquids density Wang and Alyarez (2008) explained that this
method of separating seeds works by adjusting the specific gravity of the liquid in the
reservoir so that a first portion of the seeds float near the surface of the liquid and a second
portion of the seeds sink near the bottom of the reservoir and providing an outlet flow of
the liquid from a location proximal to the top of the reservoir above the location of the
second portion of seeds By this way seeds of lower density can be separated from higher
density Kwong Sellman Jalink amp Schoor (2005) illustrated that separation by density is
commonly incorporated as part of the routine cleaning and grading process in flower seed
production procedures Other than grouping seeds according to their specific density this
density separation also can be used to remove broken partially filled empty or immature
seeds
28 Seed Deterioration
McDonald (2005) defined flower seed deterioration as deteriorative changes occuring with
time that increase the seeds vulnerability to external challenges and decrease the ability of
the seed to survive Important factors contribute to seed deterioration are genetics seed
structure seed chemistry physicalphysiological quality seed treatments relative humidity
10
and temperature Relative humidity is important because it will influence the moisture
content of seeds in storage while temperature determines the amount of moisture in the air
and influences the rate of deteriorative reactions in seeds According to Agrawal (1980)
the rate of deterioration increases as the seed moisture content increases This could be due
to the mold growth in and on the seed
11
~
30 MATERIAL AND METHOD
31 Material
Fresh matured fruits of Hibiscus sabdarifJa L (Plate I) were obtained from local fanns in
Kota Samarahan Seeds were extracted from these fruits cleaned and dried under shade
Dried seeds were dusted with Captan 80 and then placed in air-tight bottle to keep it in a
safe place before use in the subsequent experiment
Plate 1 Fresh fruit ofH sabdarifJa Plate 2 Fresh fruit of H sabdarifJa with seeds
32 Method
321 Preliminary Evaluation
Moisture content and germination was conducted as an initial evaluation to assess the
quality of seeds used in this research The procedures were those prescribed by Association
ofOfficial Seeds Analysts (AOSA 1985)
12
middot
322 Moisture Content Test
Four replicates of 10 seeds each were placed into saucers (Plate 5) and calibrated to get the
wet weight The seeds then were placed in an oven at 60degC for 48 hours After that the
seeds were weighed again to get the dry weight The percentage of moisture content of the
seeds was calculated based on the formula below (AOSA 1985)
Moisture content () = b - c x 100
b-a
where a = weight of empty saucer
b = weight of (a) + weight of seeds before drying
c = weight of (a) + weight of seeds after drying in the oven
323 Germination Test
Germination test was conducted base on the AOSA (1985) where 25 seeds with four
replications were used For every replicates seeds were put into a petry dish on 3 layers of
moist Whatman filter paper (Plate 6) The seeds were left to germinate by incubating it in a
Plant Growth Chamber (28OC) and was checked daily until 10 days Seed was considered
as germinated if the radicles were about 5mm length emerged from the seed coat The
germination percentage was calculated base on AOSA 1985 formula below
13
-----------------------------------1
Gennination () a x 100
b
where a = total number of germinated seeds
b = total number of seeds used
324 Seed Characteristic
The size of the H sabdariffa seeds was calibrated for its length width and thickness Ten
replicates of seeds were used for calibrating the physical characteristics Then the seed
weights of 10 100 and 1000 seeds were calibrated Four replicates of 10 100 and 1000
seed-weight classes were used
325 Hydration of Seeds
Seed of H sabdariffa was placed in distilled water for 0 2 4 6 8 10 and 12 hours at
temperatures of 28 30 35 and 40degC Then seeds were analyzed for the moisture content
and gennination
326 Data Analysis
The experimental units were arranged randomly based on Completely Randomized Design
(CRD) and data was analyzed by using Analysis of Variance (ANOV A) If there any
significant differences the mean were discriminated using Least Significant Different
(LSD)
14
Current production of H sabdarifJa in Malaysia is about 240 tonnes annually Reported in
unpublished result from Agricultural Department of Terengganu in the process of
removing the calyces out of 3 tonnes of raw materials about 15 tonnes of the velvety
capsules containing the seeds are being disposed as a by - product and unexploited
(Halimatul Amin Mohd Esa Nawalyah amp Siti 2007) Amin et at (2008) mentioned that
the plant takes about three to four months to reach the commercial stage of maturity before
the flowers are harvested
12 Commercial Production
Many parts of H sabdariffa including seeds leaves fruits and roots are very useful
especially the fleshy red calyces They are used fresh for making wine juice jam jelly
syrup gelatin pudding cakes ice cream and flavors and also dried and brewed into tea
spice and used for butter pies sauces tarts and other desserts (Qi et al 2005) In a study
about commercial practice of Roselle beverage production Bolade et at (2009) reported
that the dried calyceswater ratios involved in the commercial practice ranged between
157 and 171 (wv) while the sweetness level of the beverage ranged between 112 and
133 degBrix The red calyces which contains high amount of vitamin C was used to make
those healthy beverages In Malaysia we also consume the leaves as vegetables There are
previous study shows that H sabdarifJa seeds can be used as a protein source Instead of
using protein from conventional sources protein isolates or concentrates from Roselle
seeds might be useful as low cost source of protein substitute in dietary supplement or food
ingredient in food industry thus alleviating the problem of protein scarcity (Halimatul et
al 2007)
2
middot
13 Medicinal Uses
H sabdariffa is said to be useful in traditional medicine as a digestive agent purgative and
diuretic among others Having two diuretic ingredients ascorbic acid and glycolic acid it
has the ability to increase urination and also valued for its mild laxative effect Containing
citric acid it also can be used as cooling herb The tonic tea produce from the leaves and
flowers is good for digestive and kidney function The heated leaves are applied to cracks
in the feet and on boils and ulcers to speed maturation (Qi et aI 2005) Other roselle s part
have also been reported to be folk remedy for cancer obesity diabetes and hypertension
(Bolad et aI 2009) Amin et al (2008) also added that the flower and fleshy fruits are
used in pharmaceutical industry to relieve symptoms of bronchitis and coughs Roselle s
lotion products are useful for treating sores and wounds
Another nutritional value of Roselle discovered by UKM was the high content of
hydroxycitric acid (HCA) which is widely used as the main ingredients in many
commercial weight loss and slimming products Recent studies by local and international
scientist have proven the effectiveness of HCA in reducing body weight This
characteristic makes Roselle a very valuable crop
14 Problem Statement
The demand for H sabdariffa production nowadays had increase due to its commercial and
medicinal values To fulfill these demands more good quality seeds are needed
Furthermore not many research had been done on improving the H sabdarifJa seeds as a
planting material eventhough the best propagation method for H sabdariffa is by seeds
Other than that disease has been reported as a limiting factor to the production of Roselle
worldwide (Sie R S et aI 2011)
3
1S Objectives
1 To detennine the physical characteristics of H sabdariffa seeds for 10 100 and
1000 seeds weight and for length width and thickness of the seed
2 To evaluate the germination of H sabdariffa seeds following hydration at different
temperatures
3 To determine the effectiveness of hydration technique in improving the gennination
of H sabdariffa seeds
4
-
Pusat Khidmat Maklumat Akadtmik VNIVERSm MALAYSIA SARAWAK
I J bull bull
20 LITERATURE REVIEW
21 Characteristic of Seed
Seeds quality can be affected by changes in environmental temperature and relative
humidity The characteristic of seed must be determine because it is important to ensure
high quality yield in field planting Seed germination can be affected by number of factors
such as level of oxygen temperature and moisture content Growth performance of seed
also can be related to seed size Therefore grading is important to obtain seed that can
grows better Some useful criteria for visual separation of seed includes the seed size
shape colour weight thickness density and surface texture
According to Jain and Bal (1997) roselle seeds are having average principal dimensions of
298-336 186-224 and 170-201 mm Omobuwajo et al (2000) found that the average of
roselle seeds length width and thickness were 558 521 and 281 mm respectively The
seed was reported to be important for its oil in some parts of Africa its native origin From
previous research by scientist roselle seed was among the highest protein-containing seeds
when compared with other seeds like passion fruit (Passiflora edulis) Amaranthus seeds
and Pisum sativum seeds Latest findings by Hainida et al (2008) found that the seed from
Malaysia are composed of 99 moisture 335 protein 221 lipids 130 available
carbohydrate 183 total dietary fibers and 75 ash
H sabdarifJa can be grown easily in most of the tropical country Like other hibiscus
species it is quite hard for roselle to be affected by pest and disease and maintaining this
plant in the field is not so difficult The seeds were sown in a suitable container and the
seedlings will be ready for planting at the age of 10-15 days However it is also important
to ensure that the seeds to be sown were in a good condition and quality to avoid losses
5
22 Seed Germination
Germination is the emergence and development from the seed embryo of those essential
structures which are indicative of the ability to produce a normal plant under favorable
conditions (AOSA 1991) According to Smith (2011) the sum of germination events
beginning with hydration and culminating with root emergence Several stages involved in
germination are imbibition of water activation of enzyme systems metabolism of storage
products and their transport and finally the emergence of the radicle and growth of the
seedling Seed germination is described as the emergence and development from the seed
embryo of those essential structures which for the kind of seed in question are indicative
of the ability to produce a normal plant under favorable conditions (AOSA 1978) In a
simpler contcxt germination is the emergence of the radicle through the seed coat
Depending on the species seeds can germinate as quick as a few days or for as long as
several years Due to the high respiration rate and some exudation and leakage through the
seed coat the germinating seedling will undergoes a net loss in dry weight before the
germination For a particular kind of species the germination condition is mostly affected
by temperature type of substrate on which to germinate the seed light condition and
moisture level
The germination test is commonly used to determine seed viability It is an analytical
procedure to evaluate seed germination under standardized favorable conditions that are
seldom if ever encountered in the field The germination test is merely an estimate and
has certain limitations as a universal estimate of seed quality However if these limitations
are recognized the germination test is a useful viability index The need for germination
test is to estimate how the seed will perform in the field
6
I bull bull
23 Seed Moisture Content
Amount of water in the seed that usually expressed in a percentage fonn is the seed
moisture content Agrawal (1980) found that the amount of moisture in the seed is
probably the most important factor influencing seed viability during storage By
determining the seeds moisture content the possible storage life for the seeds can be
predicted According to McDonald (2005) selecting the appropriate salt to achieve a seed
moisture content of 5-6 is the recommended method for achieving optimum seed
storage and minimizing seed deterioration It is necessary to dry seeds to the safe moisture
contents since the life of a seed and its span largely revolves around its moisture content
and the suitable moisture content for starchy seeds are less than 14 while for oily seeds
are less than 11 (Agrawal 1980)
24 Seed Vigour
According to Geneve (2005) seed vigour is defined by the Asociation of official Seed
analysts as those seed properties which detennine the potential for rapid unifonn
emergence and development of nonnal seedlings under the wide range of field conditions
Seed vigour is closely related with seed storage environment and duration During seed
storage the vigour of a seed lot is reduced prior to seed viability as indicated by standard
germination (Hampton amp TeKrony 1995) Major factors influencing the seed vigour in
stored seed lots are the environmental conditions during seed storage and the length of time
in storage Tetrazolium test is one of the method for detennining the seeds vigour To
minimize the decline in vigour and germination seeds should be undamaged
7
bull t I I
25 Seed Hydration
A process of supplying water to seed to maintain the fluid or moisture content in the seed
is called hydration According to Copeland and McDonald (1995) seed hydration is a
process whereby seeds are hydrated using various protocols and then redried to permit
routine handling This process results in increased germination rate more uniform
emergence germination under a broader range of environments and improved sedling
vigour and growth It is one of the priming treatments of seed which usually applied to
strengthen or increase vitality of seed Priming process can increase a rate of germination
as the treatment improves the seed vigor Hydration is one component of priming which
can increase germination with more range of environment condition The objective of seed
hydration technology is to increase the percentage and rate of germination expand the
range of temperatures over which the seed will germinate and increase the uniformity of
stand establishment To accomplish these objectives seeds must be hydrated in some way
at a moisture level sufficient to initiate the early events of germination but not sufficient to
permit radicle protrusion (Akers and Holley 1986) However the performance of the seed
might also being influence by other factors like the species type of seed and the seeds
quality
Water can be used for the hydration treatment because it had proved to enhance
germination emergence growth and yield of seed However other mediwns such as the
solution of potassium hydrophosphate (KH2P04) monobasic polyethylene glycol (PEG)
potassium chloride (KCl) potassiwn nitrate (KN03) potassium phoshate (K3P04)
magnesium sulphate (MgS04) sodiwn chloride (NaCl) glycerol and mannitol also can be
used to immerse the seed in hydration process The benefit of such salts is to supply the
seed with nitrogen and other nutrients essential for protein synthesis during germination
8
t I amp
26 Seed Storage
A basic requirement in seed production is a good seed storage The purpose of seed storage
is to preserve planting stocks from one season to the next (Larry amp McDonald 1995)
Generally we stored seeds for the commercial purpose for carry - over seeds or for genn
plasm seeds Maintaining the seed quality for the longest duration possible is the main
objective in seed storage Loss of gennination during storage cannot be stopped but it
could be reduce by giving good storage conditions According to Agrawal (1980) the
seeds are considered to be in storage from the moment they reach physiological maturity
until they genninate or until they are thrown away because they are dead or otherwise
worthless
The seed storage must be properly handled because it can affect the seed quality There are
some factors that can affect the seed longevity in storage such as the variety of the seed
initial seed quality moisture content relative humidity and storage temperature It is
necessary for the seed storage condition to be dry and cool effective in pest control and
have a proper sanitation Other than than storing only high quality of seed also can help in
better production From all of the factors temperature is one of the most important element
which influence seed viability and vigour during storage Agrawal (1980) mentioned that
the lower the temperature the longer the seeds maintain gennination capacity so
decreasing temperature and seed moisture is an effective means of maintaining seed quality
in storage Every seed lot should be gennination - tested when it is received for storage to
record the seed quality Subsequent periodic checks also should be made to detect
deterioration and if it occurs storage condition should be checked to discover the reasons
and to take corrective measures
9
middot
27 Seed Specific Density
Density of seed refers to it mass per unit volume The seed size and density will affect the
yield One of the method for grouping seeds into some specific density is the liquid density
separation system It is one of the easier and cheaper method compared to using a machine
When seed is loaded into a liquid some will float and some will sink That mean the
floating seeds density is lower than the liquids density Vice versa the sinking seeds
density is higher than the liquids density Wang and Alyarez (2008) explained that this
method of separating seeds works by adjusting the specific gravity of the liquid in the
reservoir so that a first portion of the seeds float near the surface of the liquid and a second
portion of the seeds sink near the bottom of the reservoir and providing an outlet flow of
the liquid from a location proximal to the top of the reservoir above the location of the
second portion of seeds By this way seeds of lower density can be separated from higher
density Kwong Sellman Jalink amp Schoor (2005) illustrated that separation by density is
commonly incorporated as part of the routine cleaning and grading process in flower seed
production procedures Other than grouping seeds according to their specific density this
density separation also can be used to remove broken partially filled empty or immature
seeds
28 Seed Deterioration
McDonald (2005) defined flower seed deterioration as deteriorative changes occuring with
time that increase the seeds vulnerability to external challenges and decrease the ability of
the seed to survive Important factors contribute to seed deterioration are genetics seed
structure seed chemistry physicalphysiological quality seed treatments relative humidity
10
and temperature Relative humidity is important because it will influence the moisture
content of seeds in storage while temperature determines the amount of moisture in the air
and influences the rate of deteriorative reactions in seeds According to Agrawal (1980)
the rate of deterioration increases as the seed moisture content increases This could be due
to the mold growth in and on the seed
11
~
30 MATERIAL AND METHOD
31 Material
Fresh matured fruits of Hibiscus sabdarifJa L (Plate I) were obtained from local fanns in
Kota Samarahan Seeds were extracted from these fruits cleaned and dried under shade
Dried seeds were dusted with Captan 80 and then placed in air-tight bottle to keep it in a
safe place before use in the subsequent experiment
Plate 1 Fresh fruit ofH sabdarifJa Plate 2 Fresh fruit of H sabdarifJa with seeds
32 Method
321 Preliminary Evaluation
Moisture content and germination was conducted as an initial evaluation to assess the
quality of seeds used in this research The procedures were those prescribed by Association
ofOfficial Seeds Analysts (AOSA 1985)
12
middot
322 Moisture Content Test
Four replicates of 10 seeds each were placed into saucers (Plate 5) and calibrated to get the
wet weight The seeds then were placed in an oven at 60degC for 48 hours After that the
seeds were weighed again to get the dry weight The percentage of moisture content of the
seeds was calculated based on the formula below (AOSA 1985)
Moisture content () = b - c x 100
b-a
where a = weight of empty saucer
b = weight of (a) + weight of seeds before drying
c = weight of (a) + weight of seeds after drying in the oven
323 Germination Test
Germination test was conducted base on the AOSA (1985) where 25 seeds with four
replications were used For every replicates seeds were put into a petry dish on 3 layers of
moist Whatman filter paper (Plate 6) The seeds were left to germinate by incubating it in a
Plant Growth Chamber (28OC) and was checked daily until 10 days Seed was considered
as germinated if the radicles were about 5mm length emerged from the seed coat The
germination percentage was calculated base on AOSA 1985 formula below
13
-----------------------------------1
Gennination () a x 100
b
where a = total number of germinated seeds
b = total number of seeds used
324 Seed Characteristic
The size of the H sabdariffa seeds was calibrated for its length width and thickness Ten
replicates of seeds were used for calibrating the physical characteristics Then the seed
weights of 10 100 and 1000 seeds were calibrated Four replicates of 10 100 and 1000
seed-weight classes were used
325 Hydration of Seeds
Seed of H sabdariffa was placed in distilled water for 0 2 4 6 8 10 and 12 hours at
temperatures of 28 30 35 and 40degC Then seeds were analyzed for the moisture content
and gennination
326 Data Analysis
The experimental units were arranged randomly based on Completely Randomized Design
(CRD) and data was analyzed by using Analysis of Variance (ANOV A) If there any
significant differences the mean were discriminated using Least Significant Different
(LSD)
14
middot
13 Medicinal Uses
H sabdariffa is said to be useful in traditional medicine as a digestive agent purgative and
diuretic among others Having two diuretic ingredients ascorbic acid and glycolic acid it
has the ability to increase urination and also valued for its mild laxative effect Containing
citric acid it also can be used as cooling herb The tonic tea produce from the leaves and
flowers is good for digestive and kidney function The heated leaves are applied to cracks
in the feet and on boils and ulcers to speed maturation (Qi et aI 2005) Other roselle s part
have also been reported to be folk remedy for cancer obesity diabetes and hypertension
(Bolad et aI 2009) Amin et al (2008) also added that the flower and fleshy fruits are
used in pharmaceutical industry to relieve symptoms of bronchitis and coughs Roselle s
lotion products are useful for treating sores and wounds
Another nutritional value of Roselle discovered by UKM was the high content of
hydroxycitric acid (HCA) which is widely used as the main ingredients in many
commercial weight loss and slimming products Recent studies by local and international
scientist have proven the effectiveness of HCA in reducing body weight This
characteristic makes Roselle a very valuable crop
14 Problem Statement
The demand for H sabdariffa production nowadays had increase due to its commercial and
medicinal values To fulfill these demands more good quality seeds are needed
Furthermore not many research had been done on improving the H sabdarifJa seeds as a
planting material eventhough the best propagation method for H sabdariffa is by seeds
Other than that disease has been reported as a limiting factor to the production of Roselle
worldwide (Sie R S et aI 2011)
3
1S Objectives
1 To detennine the physical characteristics of H sabdariffa seeds for 10 100 and
1000 seeds weight and for length width and thickness of the seed
2 To evaluate the germination of H sabdariffa seeds following hydration at different
temperatures
3 To determine the effectiveness of hydration technique in improving the gennination
of H sabdariffa seeds
4
-
Pusat Khidmat Maklumat Akadtmik VNIVERSm MALAYSIA SARAWAK
I J bull bull
20 LITERATURE REVIEW
21 Characteristic of Seed
Seeds quality can be affected by changes in environmental temperature and relative
humidity The characteristic of seed must be determine because it is important to ensure
high quality yield in field planting Seed germination can be affected by number of factors
such as level of oxygen temperature and moisture content Growth performance of seed
also can be related to seed size Therefore grading is important to obtain seed that can
grows better Some useful criteria for visual separation of seed includes the seed size
shape colour weight thickness density and surface texture
According to Jain and Bal (1997) roselle seeds are having average principal dimensions of
298-336 186-224 and 170-201 mm Omobuwajo et al (2000) found that the average of
roselle seeds length width and thickness were 558 521 and 281 mm respectively The
seed was reported to be important for its oil in some parts of Africa its native origin From
previous research by scientist roselle seed was among the highest protein-containing seeds
when compared with other seeds like passion fruit (Passiflora edulis) Amaranthus seeds
and Pisum sativum seeds Latest findings by Hainida et al (2008) found that the seed from
Malaysia are composed of 99 moisture 335 protein 221 lipids 130 available
carbohydrate 183 total dietary fibers and 75 ash
H sabdarifJa can be grown easily in most of the tropical country Like other hibiscus
species it is quite hard for roselle to be affected by pest and disease and maintaining this
plant in the field is not so difficult The seeds were sown in a suitable container and the
seedlings will be ready for planting at the age of 10-15 days However it is also important
to ensure that the seeds to be sown were in a good condition and quality to avoid losses
5
22 Seed Germination
Germination is the emergence and development from the seed embryo of those essential
structures which are indicative of the ability to produce a normal plant under favorable
conditions (AOSA 1991) According to Smith (2011) the sum of germination events
beginning with hydration and culminating with root emergence Several stages involved in
germination are imbibition of water activation of enzyme systems metabolism of storage
products and their transport and finally the emergence of the radicle and growth of the
seedling Seed germination is described as the emergence and development from the seed
embryo of those essential structures which for the kind of seed in question are indicative
of the ability to produce a normal plant under favorable conditions (AOSA 1978) In a
simpler contcxt germination is the emergence of the radicle through the seed coat
Depending on the species seeds can germinate as quick as a few days or for as long as
several years Due to the high respiration rate and some exudation and leakage through the
seed coat the germinating seedling will undergoes a net loss in dry weight before the
germination For a particular kind of species the germination condition is mostly affected
by temperature type of substrate on which to germinate the seed light condition and
moisture level
The germination test is commonly used to determine seed viability It is an analytical
procedure to evaluate seed germination under standardized favorable conditions that are
seldom if ever encountered in the field The germination test is merely an estimate and
has certain limitations as a universal estimate of seed quality However if these limitations
are recognized the germination test is a useful viability index The need for germination
test is to estimate how the seed will perform in the field
6
I bull bull
23 Seed Moisture Content
Amount of water in the seed that usually expressed in a percentage fonn is the seed
moisture content Agrawal (1980) found that the amount of moisture in the seed is
probably the most important factor influencing seed viability during storage By
determining the seeds moisture content the possible storage life for the seeds can be
predicted According to McDonald (2005) selecting the appropriate salt to achieve a seed
moisture content of 5-6 is the recommended method for achieving optimum seed
storage and minimizing seed deterioration It is necessary to dry seeds to the safe moisture
contents since the life of a seed and its span largely revolves around its moisture content
and the suitable moisture content for starchy seeds are less than 14 while for oily seeds
are less than 11 (Agrawal 1980)
24 Seed Vigour
According to Geneve (2005) seed vigour is defined by the Asociation of official Seed
analysts as those seed properties which detennine the potential for rapid unifonn
emergence and development of nonnal seedlings under the wide range of field conditions
Seed vigour is closely related with seed storage environment and duration During seed
storage the vigour of a seed lot is reduced prior to seed viability as indicated by standard
germination (Hampton amp TeKrony 1995) Major factors influencing the seed vigour in
stored seed lots are the environmental conditions during seed storage and the length of time
in storage Tetrazolium test is one of the method for detennining the seeds vigour To
minimize the decline in vigour and germination seeds should be undamaged
7
bull t I I
25 Seed Hydration
A process of supplying water to seed to maintain the fluid or moisture content in the seed
is called hydration According to Copeland and McDonald (1995) seed hydration is a
process whereby seeds are hydrated using various protocols and then redried to permit
routine handling This process results in increased germination rate more uniform
emergence germination under a broader range of environments and improved sedling
vigour and growth It is one of the priming treatments of seed which usually applied to
strengthen or increase vitality of seed Priming process can increase a rate of germination
as the treatment improves the seed vigor Hydration is one component of priming which
can increase germination with more range of environment condition The objective of seed
hydration technology is to increase the percentage and rate of germination expand the
range of temperatures over which the seed will germinate and increase the uniformity of
stand establishment To accomplish these objectives seeds must be hydrated in some way
at a moisture level sufficient to initiate the early events of germination but not sufficient to
permit radicle protrusion (Akers and Holley 1986) However the performance of the seed
might also being influence by other factors like the species type of seed and the seeds
quality
Water can be used for the hydration treatment because it had proved to enhance
germination emergence growth and yield of seed However other mediwns such as the
solution of potassium hydrophosphate (KH2P04) monobasic polyethylene glycol (PEG)
potassium chloride (KCl) potassiwn nitrate (KN03) potassium phoshate (K3P04)
magnesium sulphate (MgS04) sodiwn chloride (NaCl) glycerol and mannitol also can be
used to immerse the seed in hydration process The benefit of such salts is to supply the
seed with nitrogen and other nutrients essential for protein synthesis during germination
8
t I amp
26 Seed Storage
A basic requirement in seed production is a good seed storage The purpose of seed storage
is to preserve planting stocks from one season to the next (Larry amp McDonald 1995)
Generally we stored seeds for the commercial purpose for carry - over seeds or for genn
plasm seeds Maintaining the seed quality for the longest duration possible is the main
objective in seed storage Loss of gennination during storage cannot be stopped but it
could be reduce by giving good storage conditions According to Agrawal (1980) the
seeds are considered to be in storage from the moment they reach physiological maturity
until they genninate or until they are thrown away because they are dead or otherwise
worthless
The seed storage must be properly handled because it can affect the seed quality There are
some factors that can affect the seed longevity in storage such as the variety of the seed
initial seed quality moisture content relative humidity and storage temperature It is
necessary for the seed storage condition to be dry and cool effective in pest control and
have a proper sanitation Other than than storing only high quality of seed also can help in
better production From all of the factors temperature is one of the most important element
which influence seed viability and vigour during storage Agrawal (1980) mentioned that
the lower the temperature the longer the seeds maintain gennination capacity so
decreasing temperature and seed moisture is an effective means of maintaining seed quality
in storage Every seed lot should be gennination - tested when it is received for storage to
record the seed quality Subsequent periodic checks also should be made to detect
deterioration and if it occurs storage condition should be checked to discover the reasons
and to take corrective measures
9
middot
27 Seed Specific Density
Density of seed refers to it mass per unit volume The seed size and density will affect the
yield One of the method for grouping seeds into some specific density is the liquid density
separation system It is one of the easier and cheaper method compared to using a machine
When seed is loaded into a liquid some will float and some will sink That mean the
floating seeds density is lower than the liquids density Vice versa the sinking seeds
density is higher than the liquids density Wang and Alyarez (2008) explained that this
method of separating seeds works by adjusting the specific gravity of the liquid in the
reservoir so that a first portion of the seeds float near the surface of the liquid and a second
portion of the seeds sink near the bottom of the reservoir and providing an outlet flow of
the liquid from a location proximal to the top of the reservoir above the location of the
second portion of seeds By this way seeds of lower density can be separated from higher
density Kwong Sellman Jalink amp Schoor (2005) illustrated that separation by density is
commonly incorporated as part of the routine cleaning and grading process in flower seed
production procedures Other than grouping seeds according to their specific density this
density separation also can be used to remove broken partially filled empty or immature
seeds
28 Seed Deterioration
McDonald (2005) defined flower seed deterioration as deteriorative changes occuring with
time that increase the seeds vulnerability to external challenges and decrease the ability of
the seed to survive Important factors contribute to seed deterioration are genetics seed
structure seed chemistry physicalphysiological quality seed treatments relative humidity
10
and temperature Relative humidity is important because it will influence the moisture
content of seeds in storage while temperature determines the amount of moisture in the air
and influences the rate of deteriorative reactions in seeds According to Agrawal (1980)
the rate of deterioration increases as the seed moisture content increases This could be due
to the mold growth in and on the seed
11
~
30 MATERIAL AND METHOD
31 Material
Fresh matured fruits of Hibiscus sabdarifJa L (Plate I) were obtained from local fanns in
Kota Samarahan Seeds were extracted from these fruits cleaned and dried under shade
Dried seeds were dusted with Captan 80 and then placed in air-tight bottle to keep it in a
safe place before use in the subsequent experiment
Plate 1 Fresh fruit ofH sabdarifJa Plate 2 Fresh fruit of H sabdarifJa with seeds
32 Method
321 Preliminary Evaluation
Moisture content and germination was conducted as an initial evaluation to assess the
quality of seeds used in this research The procedures were those prescribed by Association
ofOfficial Seeds Analysts (AOSA 1985)
12
middot
322 Moisture Content Test
Four replicates of 10 seeds each were placed into saucers (Plate 5) and calibrated to get the
wet weight The seeds then were placed in an oven at 60degC for 48 hours After that the
seeds were weighed again to get the dry weight The percentage of moisture content of the
seeds was calculated based on the formula below (AOSA 1985)
Moisture content () = b - c x 100
b-a
where a = weight of empty saucer
b = weight of (a) + weight of seeds before drying
c = weight of (a) + weight of seeds after drying in the oven
323 Germination Test
Germination test was conducted base on the AOSA (1985) where 25 seeds with four
replications were used For every replicates seeds were put into a petry dish on 3 layers of
moist Whatman filter paper (Plate 6) The seeds were left to germinate by incubating it in a
Plant Growth Chamber (28OC) and was checked daily until 10 days Seed was considered
as germinated if the radicles were about 5mm length emerged from the seed coat The
germination percentage was calculated base on AOSA 1985 formula below
13
-----------------------------------1
Gennination () a x 100
b
where a = total number of germinated seeds
b = total number of seeds used
324 Seed Characteristic
The size of the H sabdariffa seeds was calibrated for its length width and thickness Ten
replicates of seeds were used for calibrating the physical characteristics Then the seed
weights of 10 100 and 1000 seeds were calibrated Four replicates of 10 100 and 1000
seed-weight classes were used
325 Hydration of Seeds
Seed of H sabdariffa was placed in distilled water for 0 2 4 6 8 10 and 12 hours at
temperatures of 28 30 35 and 40degC Then seeds were analyzed for the moisture content
and gennination
326 Data Analysis
The experimental units were arranged randomly based on Completely Randomized Design
(CRD) and data was analyzed by using Analysis of Variance (ANOV A) If there any
significant differences the mean were discriminated using Least Significant Different
(LSD)
14
1S Objectives
1 To detennine the physical characteristics of H sabdariffa seeds for 10 100 and
1000 seeds weight and for length width and thickness of the seed
2 To evaluate the germination of H sabdariffa seeds following hydration at different
temperatures
3 To determine the effectiveness of hydration technique in improving the gennination
of H sabdariffa seeds
4
-
Pusat Khidmat Maklumat Akadtmik VNIVERSm MALAYSIA SARAWAK
I J bull bull
20 LITERATURE REVIEW
21 Characteristic of Seed
Seeds quality can be affected by changes in environmental temperature and relative
humidity The characteristic of seed must be determine because it is important to ensure
high quality yield in field planting Seed germination can be affected by number of factors
such as level of oxygen temperature and moisture content Growth performance of seed
also can be related to seed size Therefore grading is important to obtain seed that can
grows better Some useful criteria for visual separation of seed includes the seed size
shape colour weight thickness density and surface texture
According to Jain and Bal (1997) roselle seeds are having average principal dimensions of
298-336 186-224 and 170-201 mm Omobuwajo et al (2000) found that the average of
roselle seeds length width and thickness were 558 521 and 281 mm respectively The
seed was reported to be important for its oil in some parts of Africa its native origin From
previous research by scientist roselle seed was among the highest protein-containing seeds
when compared with other seeds like passion fruit (Passiflora edulis) Amaranthus seeds
and Pisum sativum seeds Latest findings by Hainida et al (2008) found that the seed from
Malaysia are composed of 99 moisture 335 protein 221 lipids 130 available
carbohydrate 183 total dietary fibers and 75 ash
H sabdarifJa can be grown easily in most of the tropical country Like other hibiscus
species it is quite hard for roselle to be affected by pest and disease and maintaining this
plant in the field is not so difficult The seeds were sown in a suitable container and the
seedlings will be ready for planting at the age of 10-15 days However it is also important
to ensure that the seeds to be sown were in a good condition and quality to avoid losses
5
22 Seed Germination
Germination is the emergence and development from the seed embryo of those essential
structures which are indicative of the ability to produce a normal plant under favorable
conditions (AOSA 1991) According to Smith (2011) the sum of germination events
beginning with hydration and culminating with root emergence Several stages involved in
germination are imbibition of water activation of enzyme systems metabolism of storage
products and their transport and finally the emergence of the radicle and growth of the
seedling Seed germination is described as the emergence and development from the seed
embryo of those essential structures which for the kind of seed in question are indicative
of the ability to produce a normal plant under favorable conditions (AOSA 1978) In a
simpler contcxt germination is the emergence of the radicle through the seed coat
Depending on the species seeds can germinate as quick as a few days or for as long as
several years Due to the high respiration rate and some exudation and leakage through the
seed coat the germinating seedling will undergoes a net loss in dry weight before the
germination For a particular kind of species the germination condition is mostly affected
by temperature type of substrate on which to germinate the seed light condition and
moisture level
The germination test is commonly used to determine seed viability It is an analytical
procedure to evaluate seed germination under standardized favorable conditions that are
seldom if ever encountered in the field The germination test is merely an estimate and
has certain limitations as a universal estimate of seed quality However if these limitations
are recognized the germination test is a useful viability index The need for germination
test is to estimate how the seed will perform in the field
6
I bull bull
23 Seed Moisture Content
Amount of water in the seed that usually expressed in a percentage fonn is the seed
moisture content Agrawal (1980) found that the amount of moisture in the seed is
probably the most important factor influencing seed viability during storage By
determining the seeds moisture content the possible storage life for the seeds can be
predicted According to McDonald (2005) selecting the appropriate salt to achieve a seed
moisture content of 5-6 is the recommended method for achieving optimum seed
storage and minimizing seed deterioration It is necessary to dry seeds to the safe moisture
contents since the life of a seed and its span largely revolves around its moisture content
and the suitable moisture content for starchy seeds are less than 14 while for oily seeds
are less than 11 (Agrawal 1980)
24 Seed Vigour
According to Geneve (2005) seed vigour is defined by the Asociation of official Seed
analysts as those seed properties which detennine the potential for rapid unifonn
emergence and development of nonnal seedlings under the wide range of field conditions
Seed vigour is closely related with seed storage environment and duration During seed
storage the vigour of a seed lot is reduced prior to seed viability as indicated by standard
germination (Hampton amp TeKrony 1995) Major factors influencing the seed vigour in
stored seed lots are the environmental conditions during seed storage and the length of time
in storage Tetrazolium test is one of the method for detennining the seeds vigour To
minimize the decline in vigour and germination seeds should be undamaged
7
bull t I I
25 Seed Hydration
A process of supplying water to seed to maintain the fluid or moisture content in the seed
is called hydration According to Copeland and McDonald (1995) seed hydration is a
process whereby seeds are hydrated using various protocols and then redried to permit
routine handling This process results in increased germination rate more uniform
emergence germination under a broader range of environments and improved sedling
vigour and growth It is one of the priming treatments of seed which usually applied to
strengthen or increase vitality of seed Priming process can increase a rate of germination
as the treatment improves the seed vigor Hydration is one component of priming which
can increase germination with more range of environment condition The objective of seed
hydration technology is to increase the percentage and rate of germination expand the
range of temperatures over which the seed will germinate and increase the uniformity of
stand establishment To accomplish these objectives seeds must be hydrated in some way
at a moisture level sufficient to initiate the early events of germination but not sufficient to
permit radicle protrusion (Akers and Holley 1986) However the performance of the seed
might also being influence by other factors like the species type of seed and the seeds
quality
Water can be used for the hydration treatment because it had proved to enhance
germination emergence growth and yield of seed However other mediwns such as the
solution of potassium hydrophosphate (KH2P04) monobasic polyethylene glycol (PEG)
potassium chloride (KCl) potassiwn nitrate (KN03) potassium phoshate (K3P04)
magnesium sulphate (MgS04) sodiwn chloride (NaCl) glycerol and mannitol also can be
used to immerse the seed in hydration process The benefit of such salts is to supply the
seed with nitrogen and other nutrients essential for protein synthesis during germination
8
t I amp
26 Seed Storage
A basic requirement in seed production is a good seed storage The purpose of seed storage
is to preserve planting stocks from one season to the next (Larry amp McDonald 1995)
Generally we stored seeds for the commercial purpose for carry - over seeds or for genn
plasm seeds Maintaining the seed quality for the longest duration possible is the main
objective in seed storage Loss of gennination during storage cannot be stopped but it
could be reduce by giving good storage conditions According to Agrawal (1980) the
seeds are considered to be in storage from the moment they reach physiological maturity
until they genninate or until they are thrown away because they are dead or otherwise
worthless
The seed storage must be properly handled because it can affect the seed quality There are
some factors that can affect the seed longevity in storage such as the variety of the seed
initial seed quality moisture content relative humidity and storage temperature It is
necessary for the seed storage condition to be dry and cool effective in pest control and
have a proper sanitation Other than than storing only high quality of seed also can help in
better production From all of the factors temperature is one of the most important element
which influence seed viability and vigour during storage Agrawal (1980) mentioned that
the lower the temperature the longer the seeds maintain gennination capacity so
decreasing temperature and seed moisture is an effective means of maintaining seed quality
in storage Every seed lot should be gennination - tested when it is received for storage to
record the seed quality Subsequent periodic checks also should be made to detect
deterioration and if it occurs storage condition should be checked to discover the reasons
and to take corrective measures
9
middot
27 Seed Specific Density
Density of seed refers to it mass per unit volume The seed size and density will affect the
yield One of the method for grouping seeds into some specific density is the liquid density
separation system It is one of the easier and cheaper method compared to using a machine
When seed is loaded into a liquid some will float and some will sink That mean the
floating seeds density is lower than the liquids density Vice versa the sinking seeds
density is higher than the liquids density Wang and Alyarez (2008) explained that this
method of separating seeds works by adjusting the specific gravity of the liquid in the
reservoir so that a first portion of the seeds float near the surface of the liquid and a second
portion of the seeds sink near the bottom of the reservoir and providing an outlet flow of
the liquid from a location proximal to the top of the reservoir above the location of the
second portion of seeds By this way seeds of lower density can be separated from higher
density Kwong Sellman Jalink amp Schoor (2005) illustrated that separation by density is
commonly incorporated as part of the routine cleaning and grading process in flower seed
production procedures Other than grouping seeds according to their specific density this
density separation also can be used to remove broken partially filled empty or immature
seeds
28 Seed Deterioration
McDonald (2005) defined flower seed deterioration as deteriorative changes occuring with
time that increase the seeds vulnerability to external challenges and decrease the ability of
the seed to survive Important factors contribute to seed deterioration are genetics seed
structure seed chemistry physicalphysiological quality seed treatments relative humidity
10
and temperature Relative humidity is important because it will influence the moisture
content of seeds in storage while temperature determines the amount of moisture in the air
and influences the rate of deteriorative reactions in seeds According to Agrawal (1980)
the rate of deterioration increases as the seed moisture content increases This could be due
to the mold growth in and on the seed
11
~
30 MATERIAL AND METHOD
31 Material
Fresh matured fruits of Hibiscus sabdarifJa L (Plate I) were obtained from local fanns in
Kota Samarahan Seeds were extracted from these fruits cleaned and dried under shade
Dried seeds were dusted with Captan 80 and then placed in air-tight bottle to keep it in a
safe place before use in the subsequent experiment
Plate 1 Fresh fruit ofH sabdarifJa Plate 2 Fresh fruit of H sabdarifJa with seeds
32 Method
321 Preliminary Evaluation
Moisture content and germination was conducted as an initial evaluation to assess the
quality of seeds used in this research The procedures were those prescribed by Association
ofOfficial Seeds Analysts (AOSA 1985)
12
middot
322 Moisture Content Test
Four replicates of 10 seeds each were placed into saucers (Plate 5) and calibrated to get the
wet weight The seeds then were placed in an oven at 60degC for 48 hours After that the
seeds were weighed again to get the dry weight The percentage of moisture content of the
seeds was calculated based on the formula below (AOSA 1985)
Moisture content () = b - c x 100
b-a
where a = weight of empty saucer
b = weight of (a) + weight of seeds before drying
c = weight of (a) + weight of seeds after drying in the oven
323 Germination Test
Germination test was conducted base on the AOSA (1985) where 25 seeds with four
replications were used For every replicates seeds were put into a petry dish on 3 layers of
moist Whatman filter paper (Plate 6) The seeds were left to germinate by incubating it in a
Plant Growth Chamber (28OC) and was checked daily until 10 days Seed was considered
as germinated if the radicles were about 5mm length emerged from the seed coat The
germination percentage was calculated base on AOSA 1985 formula below
13
-----------------------------------1
Gennination () a x 100
b
where a = total number of germinated seeds
b = total number of seeds used
324 Seed Characteristic
The size of the H sabdariffa seeds was calibrated for its length width and thickness Ten
replicates of seeds were used for calibrating the physical characteristics Then the seed
weights of 10 100 and 1000 seeds were calibrated Four replicates of 10 100 and 1000
seed-weight classes were used
325 Hydration of Seeds
Seed of H sabdariffa was placed in distilled water for 0 2 4 6 8 10 and 12 hours at
temperatures of 28 30 35 and 40degC Then seeds were analyzed for the moisture content
and gennination
326 Data Analysis
The experimental units were arranged randomly based on Completely Randomized Design
(CRD) and data was analyzed by using Analysis of Variance (ANOV A) If there any
significant differences the mean were discriminated using Least Significant Different
(LSD)
14
Pusat Khidmat Maklumat Akadtmik VNIVERSm MALAYSIA SARAWAK
I J bull bull
20 LITERATURE REVIEW
21 Characteristic of Seed
Seeds quality can be affected by changes in environmental temperature and relative
humidity The characteristic of seed must be determine because it is important to ensure
high quality yield in field planting Seed germination can be affected by number of factors
such as level of oxygen temperature and moisture content Growth performance of seed
also can be related to seed size Therefore grading is important to obtain seed that can
grows better Some useful criteria for visual separation of seed includes the seed size
shape colour weight thickness density and surface texture
According to Jain and Bal (1997) roselle seeds are having average principal dimensions of
298-336 186-224 and 170-201 mm Omobuwajo et al (2000) found that the average of
roselle seeds length width and thickness were 558 521 and 281 mm respectively The
seed was reported to be important for its oil in some parts of Africa its native origin From
previous research by scientist roselle seed was among the highest protein-containing seeds
when compared with other seeds like passion fruit (Passiflora edulis) Amaranthus seeds
and Pisum sativum seeds Latest findings by Hainida et al (2008) found that the seed from
Malaysia are composed of 99 moisture 335 protein 221 lipids 130 available
carbohydrate 183 total dietary fibers and 75 ash
H sabdarifJa can be grown easily in most of the tropical country Like other hibiscus
species it is quite hard for roselle to be affected by pest and disease and maintaining this
plant in the field is not so difficult The seeds were sown in a suitable container and the
seedlings will be ready for planting at the age of 10-15 days However it is also important
to ensure that the seeds to be sown were in a good condition and quality to avoid losses
5
22 Seed Germination
Germination is the emergence and development from the seed embryo of those essential
structures which are indicative of the ability to produce a normal plant under favorable
conditions (AOSA 1991) According to Smith (2011) the sum of germination events
beginning with hydration and culminating with root emergence Several stages involved in
germination are imbibition of water activation of enzyme systems metabolism of storage
products and their transport and finally the emergence of the radicle and growth of the
seedling Seed germination is described as the emergence and development from the seed
embryo of those essential structures which for the kind of seed in question are indicative
of the ability to produce a normal plant under favorable conditions (AOSA 1978) In a
simpler contcxt germination is the emergence of the radicle through the seed coat
Depending on the species seeds can germinate as quick as a few days or for as long as
several years Due to the high respiration rate and some exudation and leakage through the
seed coat the germinating seedling will undergoes a net loss in dry weight before the
germination For a particular kind of species the germination condition is mostly affected
by temperature type of substrate on which to germinate the seed light condition and
moisture level
The germination test is commonly used to determine seed viability It is an analytical
procedure to evaluate seed germination under standardized favorable conditions that are
seldom if ever encountered in the field The germination test is merely an estimate and
has certain limitations as a universal estimate of seed quality However if these limitations
are recognized the germination test is a useful viability index The need for germination
test is to estimate how the seed will perform in the field
6
I bull bull
23 Seed Moisture Content
Amount of water in the seed that usually expressed in a percentage fonn is the seed
moisture content Agrawal (1980) found that the amount of moisture in the seed is
probably the most important factor influencing seed viability during storage By
determining the seeds moisture content the possible storage life for the seeds can be
predicted According to McDonald (2005) selecting the appropriate salt to achieve a seed
moisture content of 5-6 is the recommended method for achieving optimum seed
storage and minimizing seed deterioration It is necessary to dry seeds to the safe moisture
contents since the life of a seed and its span largely revolves around its moisture content
and the suitable moisture content for starchy seeds are less than 14 while for oily seeds
are less than 11 (Agrawal 1980)
24 Seed Vigour
According to Geneve (2005) seed vigour is defined by the Asociation of official Seed
analysts as those seed properties which detennine the potential for rapid unifonn
emergence and development of nonnal seedlings under the wide range of field conditions
Seed vigour is closely related with seed storage environment and duration During seed
storage the vigour of a seed lot is reduced prior to seed viability as indicated by standard
germination (Hampton amp TeKrony 1995) Major factors influencing the seed vigour in
stored seed lots are the environmental conditions during seed storage and the length of time
in storage Tetrazolium test is one of the method for detennining the seeds vigour To
minimize the decline in vigour and germination seeds should be undamaged
7
bull t I I
25 Seed Hydration
A process of supplying water to seed to maintain the fluid or moisture content in the seed
is called hydration According to Copeland and McDonald (1995) seed hydration is a
process whereby seeds are hydrated using various protocols and then redried to permit
routine handling This process results in increased germination rate more uniform
emergence germination under a broader range of environments and improved sedling
vigour and growth It is one of the priming treatments of seed which usually applied to
strengthen or increase vitality of seed Priming process can increase a rate of germination
as the treatment improves the seed vigor Hydration is one component of priming which
can increase germination with more range of environment condition The objective of seed
hydration technology is to increase the percentage and rate of germination expand the
range of temperatures over which the seed will germinate and increase the uniformity of
stand establishment To accomplish these objectives seeds must be hydrated in some way
at a moisture level sufficient to initiate the early events of germination but not sufficient to
permit radicle protrusion (Akers and Holley 1986) However the performance of the seed
might also being influence by other factors like the species type of seed and the seeds
quality
Water can be used for the hydration treatment because it had proved to enhance
germination emergence growth and yield of seed However other mediwns such as the
solution of potassium hydrophosphate (KH2P04) monobasic polyethylene glycol (PEG)
potassium chloride (KCl) potassiwn nitrate (KN03) potassium phoshate (K3P04)
magnesium sulphate (MgS04) sodiwn chloride (NaCl) glycerol and mannitol also can be
used to immerse the seed in hydration process The benefit of such salts is to supply the
seed with nitrogen and other nutrients essential for protein synthesis during germination
8
t I amp
26 Seed Storage
A basic requirement in seed production is a good seed storage The purpose of seed storage
is to preserve planting stocks from one season to the next (Larry amp McDonald 1995)
Generally we stored seeds for the commercial purpose for carry - over seeds or for genn
plasm seeds Maintaining the seed quality for the longest duration possible is the main
objective in seed storage Loss of gennination during storage cannot be stopped but it
could be reduce by giving good storage conditions According to Agrawal (1980) the
seeds are considered to be in storage from the moment they reach physiological maturity
until they genninate or until they are thrown away because they are dead or otherwise
worthless
The seed storage must be properly handled because it can affect the seed quality There are
some factors that can affect the seed longevity in storage such as the variety of the seed
initial seed quality moisture content relative humidity and storage temperature It is
necessary for the seed storage condition to be dry and cool effective in pest control and
have a proper sanitation Other than than storing only high quality of seed also can help in
better production From all of the factors temperature is one of the most important element
which influence seed viability and vigour during storage Agrawal (1980) mentioned that
the lower the temperature the longer the seeds maintain gennination capacity so
decreasing temperature and seed moisture is an effective means of maintaining seed quality
in storage Every seed lot should be gennination - tested when it is received for storage to
record the seed quality Subsequent periodic checks also should be made to detect
deterioration and if it occurs storage condition should be checked to discover the reasons
and to take corrective measures
9
middot
27 Seed Specific Density
Density of seed refers to it mass per unit volume The seed size and density will affect the
yield One of the method for grouping seeds into some specific density is the liquid density
separation system It is one of the easier and cheaper method compared to using a machine
When seed is loaded into a liquid some will float and some will sink That mean the
floating seeds density is lower than the liquids density Vice versa the sinking seeds
density is higher than the liquids density Wang and Alyarez (2008) explained that this
method of separating seeds works by adjusting the specific gravity of the liquid in the
reservoir so that a first portion of the seeds float near the surface of the liquid and a second
portion of the seeds sink near the bottom of the reservoir and providing an outlet flow of
the liquid from a location proximal to the top of the reservoir above the location of the
second portion of seeds By this way seeds of lower density can be separated from higher
density Kwong Sellman Jalink amp Schoor (2005) illustrated that separation by density is
commonly incorporated as part of the routine cleaning and grading process in flower seed
production procedures Other than grouping seeds according to their specific density this
density separation also can be used to remove broken partially filled empty or immature
seeds
28 Seed Deterioration
McDonald (2005) defined flower seed deterioration as deteriorative changes occuring with
time that increase the seeds vulnerability to external challenges and decrease the ability of
the seed to survive Important factors contribute to seed deterioration are genetics seed
structure seed chemistry physicalphysiological quality seed treatments relative humidity
10
and temperature Relative humidity is important because it will influence the moisture
content of seeds in storage while temperature determines the amount of moisture in the air
and influences the rate of deteriorative reactions in seeds According to Agrawal (1980)
the rate of deterioration increases as the seed moisture content increases This could be due
to the mold growth in and on the seed
11
~
30 MATERIAL AND METHOD
31 Material
Fresh matured fruits of Hibiscus sabdarifJa L (Plate I) were obtained from local fanns in
Kota Samarahan Seeds were extracted from these fruits cleaned and dried under shade
Dried seeds were dusted with Captan 80 and then placed in air-tight bottle to keep it in a
safe place before use in the subsequent experiment
Plate 1 Fresh fruit ofH sabdarifJa Plate 2 Fresh fruit of H sabdarifJa with seeds
32 Method
321 Preliminary Evaluation
Moisture content and germination was conducted as an initial evaluation to assess the
quality of seeds used in this research The procedures were those prescribed by Association
ofOfficial Seeds Analysts (AOSA 1985)
12
middot
322 Moisture Content Test
Four replicates of 10 seeds each were placed into saucers (Plate 5) and calibrated to get the
wet weight The seeds then were placed in an oven at 60degC for 48 hours After that the
seeds were weighed again to get the dry weight The percentage of moisture content of the
seeds was calculated based on the formula below (AOSA 1985)
Moisture content () = b - c x 100
b-a
where a = weight of empty saucer
b = weight of (a) + weight of seeds before drying
c = weight of (a) + weight of seeds after drying in the oven
323 Germination Test
Germination test was conducted base on the AOSA (1985) where 25 seeds with four
replications were used For every replicates seeds were put into a petry dish on 3 layers of
moist Whatman filter paper (Plate 6) The seeds were left to germinate by incubating it in a
Plant Growth Chamber (28OC) and was checked daily until 10 days Seed was considered
as germinated if the radicles were about 5mm length emerged from the seed coat The
germination percentage was calculated base on AOSA 1985 formula below
13
-----------------------------------1
Gennination () a x 100
b
where a = total number of germinated seeds
b = total number of seeds used
324 Seed Characteristic
The size of the H sabdariffa seeds was calibrated for its length width and thickness Ten
replicates of seeds were used for calibrating the physical characteristics Then the seed
weights of 10 100 and 1000 seeds were calibrated Four replicates of 10 100 and 1000
seed-weight classes were used
325 Hydration of Seeds
Seed of H sabdariffa was placed in distilled water for 0 2 4 6 8 10 and 12 hours at
temperatures of 28 30 35 and 40degC Then seeds were analyzed for the moisture content
and gennination
326 Data Analysis
The experimental units were arranged randomly based on Completely Randomized Design
(CRD) and data was analyzed by using Analysis of Variance (ANOV A) If there any
significant differences the mean were discriminated using Least Significant Different
(LSD)
14
22 Seed Germination
Germination is the emergence and development from the seed embryo of those essential
structures which are indicative of the ability to produce a normal plant under favorable
conditions (AOSA 1991) According to Smith (2011) the sum of germination events
beginning with hydration and culminating with root emergence Several stages involved in
germination are imbibition of water activation of enzyme systems metabolism of storage
products and their transport and finally the emergence of the radicle and growth of the
seedling Seed germination is described as the emergence and development from the seed
embryo of those essential structures which for the kind of seed in question are indicative
of the ability to produce a normal plant under favorable conditions (AOSA 1978) In a
simpler contcxt germination is the emergence of the radicle through the seed coat
Depending on the species seeds can germinate as quick as a few days or for as long as
several years Due to the high respiration rate and some exudation and leakage through the
seed coat the germinating seedling will undergoes a net loss in dry weight before the
germination For a particular kind of species the germination condition is mostly affected
by temperature type of substrate on which to germinate the seed light condition and
moisture level
The germination test is commonly used to determine seed viability It is an analytical
procedure to evaluate seed germination under standardized favorable conditions that are
seldom if ever encountered in the field The germination test is merely an estimate and
has certain limitations as a universal estimate of seed quality However if these limitations
are recognized the germination test is a useful viability index The need for germination
test is to estimate how the seed will perform in the field
6
I bull bull
23 Seed Moisture Content
Amount of water in the seed that usually expressed in a percentage fonn is the seed
moisture content Agrawal (1980) found that the amount of moisture in the seed is
probably the most important factor influencing seed viability during storage By
determining the seeds moisture content the possible storage life for the seeds can be
predicted According to McDonald (2005) selecting the appropriate salt to achieve a seed
moisture content of 5-6 is the recommended method for achieving optimum seed
storage and minimizing seed deterioration It is necessary to dry seeds to the safe moisture
contents since the life of a seed and its span largely revolves around its moisture content
and the suitable moisture content for starchy seeds are less than 14 while for oily seeds
are less than 11 (Agrawal 1980)
24 Seed Vigour
According to Geneve (2005) seed vigour is defined by the Asociation of official Seed
analysts as those seed properties which detennine the potential for rapid unifonn
emergence and development of nonnal seedlings under the wide range of field conditions
Seed vigour is closely related with seed storage environment and duration During seed
storage the vigour of a seed lot is reduced prior to seed viability as indicated by standard
germination (Hampton amp TeKrony 1995) Major factors influencing the seed vigour in
stored seed lots are the environmental conditions during seed storage and the length of time
in storage Tetrazolium test is one of the method for detennining the seeds vigour To
minimize the decline in vigour and germination seeds should be undamaged
7
bull t I I
25 Seed Hydration
A process of supplying water to seed to maintain the fluid or moisture content in the seed
is called hydration According to Copeland and McDonald (1995) seed hydration is a
process whereby seeds are hydrated using various protocols and then redried to permit
routine handling This process results in increased germination rate more uniform
emergence germination under a broader range of environments and improved sedling
vigour and growth It is one of the priming treatments of seed which usually applied to
strengthen or increase vitality of seed Priming process can increase a rate of germination
as the treatment improves the seed vigor Hydration is one component of priming which
can increase germination with more range of environment condition The objective of seed
hydration technology is to increase the percentage and rate of germination expand the
range of temperatures over which the seed will germinate and increase the uniformity of
stand establishment To accomplish these objectives seeds must be hydrated in some way
at a moisture level sufficient to initiate the early events of germination but not sufficient to
permit radicle protrusion (Akers and Holley 1986) However the performance of the seed
might also being influence by other factors like the species type of seed and the seeds
quality
Water can be used for the hydration treatment because it had proved to enhance
germination emergence growth and yield of seed However other mediwns such as the
solution of potassium hydrophosphate (KH2P04) monobasic polyethylene glycol (PEG)
potassium chloride (KCl) potassiwn nitrate (KN03) potassium phoshate (K3P04)
magnesium sulphate (MgS04) sodiwn chloride (NaCl) glycerol and mannitol also can be
used to immerse the seed in hydration process The benefit of such salts is to supply the
seed with nitrogen and other nutrients essential for protein synthesis during germination
8
t I amp
26 Seed Storage
A basic requirement in seed production is a good seed storage The purpose of seed storage
is to preserve planting stocks from one season to the next (Larry amp McDonald 1995)
Generally we stored seeds for the commercial purpose for carry - over seeds or for genn
plasm seeds Maintaining the seed quality for the longest duration possible is the main
objective in seed storage Loss of gennination during storage cannot be stopped but it
could be reduce by giving good storage conditions According to Agrawal (1980) the
seeds are considered to be in storage from the moment they reach physiological maturity
until they genninate or until they are thrown away because they are dead or otherwise
worthless
The seed storage must be properly handled because it can affect the seed quality There are
some factors that can affect the seed longevity in storage such as the variety of the seed
initial seed quality moisture content relative humidity and storage temperature It is
necessary for the seed storage condition to be dry and cool effective in pest control and
have a proper sanitation Other than than storing only high quality of seed also can help in
better production From all of the factors temperature is one of the most important element
which influence seed viability and vigour during storage Agrawal (1980) mentioned that
the lower the temperature the longer the seeds maintain gennination capacity so
decreasing temperature and seed moisture is an effective means of maintaining seed quality
in storage Every seed lot should be gennination - tested when it is received for storage to
record the seed quality Subsequent periodic checks also should be made to detect
deterioration and if it occurs storage condition should be checked to discover the reasons
and to take corrective measures
9
middot
27 Seed Specific Density
Density of seed refers to it mass per unit volume The seed size and density will affect the
yield One of the method for grouping seeds into some specific density is the liquid density
separation system It is one of the easier and cheaper method compared to using a machine
When seed is loaded into a liquid some will float and some will sink That mean the
floating seeds density is lower than the liquids density Vice versa the sinking seeds
density is higher than the liquids density Wang and Alyarez (2008) explained that this
method of separating seeds works by adjusting the specific gravity of the liquid in the
reservoir so that a first portion of the seeds float near the surface of the liquid and a second
portion of the seeds sink near the bottom of the reservoir and providing an outlet flow of
the liquid from a location proximal to the top of the reservoir above the location of the
second portion of seeds By this way seeds of lower density can be separated from higher
density Kwong Sellman Jalink amp Schoor (2005) illustrated that separation by density is
commonly incorporated as part of the routine cleaning and grading process in flower seed
production procedures Other than grouping seeds according to their specific density this
density separation also can be used to remove broken partially filled empty or immature
seeds
28 Seed Deterioration
McDonald (2005) defined flower seed deterioration as deteriorative changes occuring with
time that increase the seeds vulnerability to external challenges and decrease the ability of
the seed to survive Important factors contribute to seed deterioration are genetics seed
structure seed chemistry physicalphysiological quality seed treatments relative humidity
10
and temperature Relative humidity is important because it will influence the moisture
content of seeds in storage while temperature determines the amount of moisture in the air
and influences the rate of deteriorative reactions in seeds According to Agrawal (1980)
the rate of deterioration increases as the seed moisture content increases This could be due
to the mold growth in and on the seed
11
~
30 MATERIAL AND METHOD
31 Material
Fresh matured fruits of Hibiscus sabdarifJa L (Plate I) were obtained from local fanns in
Kota Samarahan Seeds were extracted from these fruits cleaned and dried under shade
Dried seeds were dusted with Captan 80 and then placed in air-tight bottle to keep it in a
safe place before use in the subsequent experiment
Plate 1 Fresh fruit ofH sabdarifJa Plate 2 Fresh fruit of H sabdarifJa with seeds
32 Method
321 Preliminary Evaluation
Moisture content and germination was conducted as an initial evaluation to assess the
quality of seeds used in this research The procedures were those prescribed by Association
ofOfficial Seeds Analysts (AOSA 1985)
12
middot
322 Moisture Content Test
Four replicates of 10 seeds each were placed into saucers (Plate 5) and calibrated to get the
wet weight The seeds then were placed in an oven at 60degC for 48 hours After that the
seeds were weighed again to get the dry weight The percentage of moisture content of the
seeds was calculated based on the formula below (AOSA 1985)
Moisture content () = b - c x 100
b-a
where a = weight of empty saucer
b = weight of (a) + weight of seeds before drying
c = weight of (a) + weight of seeds after drying in the oven
323 Germination Test
Germination test was conducted base on the AOSA (1985) where 25 seeds with four
replications were used For every replicates seeds were put into a petry dish on 3 layers of
moist Whatman filter paper (Plate 6) The seeds were left to germinate by incubating it in a
Plant Growth Chamber (28OC) and was checked daily until 10 days Seed was considered
as germinated if the radicles were about 5mm length emerged from the seed coat The
germination percentage was calculated base on AOSA 1985 formula below
13
-----------------------------------1
Gennination () a x 100
b
where a = total number of germinated seeds
b = total number of seeds used
324 Seed Characteristic
The size of the H sabdariffa seeds was calibrated for its length width and thickness Ten
replicates of seeds were used for calibrating the physical characteristics Then the seed
weights of 10 100 and 1000 seeds were calibrated Four replicates of 10 100 and 1000
seed-weight classes were used
325 Hydration of Seeds
Seed of H sabdariffa was placed in distilled water for 0 2 4 6 8 10 and 12 hours at
temperatures of 28 30 35 and 40degC Then seeds were analyzed for the moisture content
and gennination
326 Data Analysis
The experimental units were arranged randomly based on Completely Randomized Design
(CRD) and data was analyzed by using Analysis of Variance (ANOV A) If there any
significant differences the mean were discriminated using Least Significant Different
(LSD)
14
I bull bull
23 Seed Moisture Content
Amount of water in the seed that usually expressed in a percentage fonn is the seed
moisture content Agrawal (1980) found that the amount of moisture in the seed is
probably the most important factor influencing seed viability during storage By
determining the seeds moisture content the possible storage life for the seeds can be
predicted According to McDonald (2005) selecting the appropriate salt to achieve a seed
moisture content of 5-6 is the recommended method for achieving optimum seed
storage and minimizing seed deterioration It is necessary to dry seeds to the safe moisture
contents since the life of a seed and its span largely revolves around its moisture content
and the suitable moisture content for starchy seeds are less than 14 while for oily seeds
are less than 11 (Agrawal 1980)
24 Seed Vigour
According to Geneve (2005) seed vigour is defined by the Asociation of official Seed
analysts as those seed properties which detennine the potential for rapid unifonn
emergence and development of nonnal seedlings under the wide range of field conditions
Seed vigour is closely related with seed storage environment and duration During seed
storage the vigour of a seed lot is reduced prior to seed viability as indicated by standard
germination (Hampton amp TeKrony 1995) Major factors influencing the seed vigour in
stored seed lots are the environmental conditions during seed storage and the length of time
in storage Tetrazolium test is one of the method for detennining the seeds vigour To
minimize the decline in vigour and germination seeds should be undamaged
7
bull t I I
25 Seed Hydration
A process of supplying water to seed to maintain the fluid or moisture content in the seed
is called hydration According to Copeland and McDonald (1995) seed hydration is a
process whereby seeds are hydrated using various protocols and then redried to permit
routine handling This process results in increased germination rate more uniform
emergence germination under a broader range of environments and improved sedling
vigour and growth It is one of the priming treatments of seed which usually applied to
strengthen or increase vitality of seed Priming process can increase a rate of germination
as the treatment improves the seed vigor Hydration is one component of priming which
can increase germination with more range of environment condition The objective of seed
hydration technology is to increase the percentage and rate of germination expand the
range of temperatures over which the seed will germinate and increase the uniformity of
stand establishment To accomplish these objectives seeds must be hydrated in some way
at a moisture level sufficient to initiate the early events of germination but not sufficient to
permit radicle protrusion (Akers and Holley 1986) However the performance of the seed
might also being influence by other factors like the species type of seed and the seeds
quality
Water can be used for the hydration treatment because it had proved to enhance
germination emergence growth and yield of seed However other mediwns such as the
solution of potassium hydrophosphate (KH2P04) monobasic polyethylene glycol (PEG)
potassium chloride (KCl) potassiwn nitrate (KN03) potassium phoshate (K3P04)
magnesium sulphate (MgS04) sodiwn chloride (NaCl) glycerol and mannitol also can be
used to immerse the seed in hydration process The benefit of such salts is to supply the
seed with nitrogen and other nutrients essential for protein synthesis during germination
8
t I amp
26 Seed Storage
A basic requirement in seed production is a good seed storage The purpose of seed storage
is to preserve planting stocks from one season to the next (Larry amp McDonald 1995)
Generally we stored seeds for the commercial purpose for carry - over seeds or for genn
plasm seeds Maintaining the seed quality for the longest duration possible is the main
objective in seed storage Loss of gennination during storage cannot be stopped but it
could be reduce by giving good storage conditions According to Agrawal (1980) the
seeds are considered to be in storage from the moment they reach physiological maturity
until they genninate or until they are thrown away because they are dead or otherwise
worthless
The seed storage must be properly handled because it can affect the seed quality There are
some factors that can affect the seed longevity in storage such as the variety of the seed
initial seed quality moisture content relative humidity and storage temperature It is
necessary for the seed storage condition to be dry and cool effective in pest control and
have a proper sanitation Other than than storing only high quality of seed also can help in
better production From all of the factors temperature is one of the most important element
which influence seed viability and vigour during storage Agrawal (1980) mentioned that
the lower the temperature the longer the seeds maintain gennination capacity so
decreasing temperature and seed moisture is an effective means of maintaining seed quality
in storage Every seed lot should be gennination - tested when it is received for storage to
record the seed quality Subsequent periodic checks also should be made to detect
deterioration and if it occurs storage condition should be checked to discover the reasons
and to take corrective measures
9
middot
27 Seed Specific Density
Density of seed refers to it mass per unit volume The seed size and density will affect the
yield One of the method for grouping seeds into some specific density is the liquid density
separation system It is one of the easier and cheaper method compared to using a machine
When seed is loaded into a liquid some will float and some will sink That mean the
floating seeds density is lower than the liquids density Vice versa the sinking seeds
density is higher than the liquids density Wang and Alyarez (2008) explained that this
method of separating seeds works by adjusting the specific gravity of the liquid in the
reservoir so that a first portion of the seeds float near the surface of the liquid and a second
portion of the seeds sink near the bottom of the reservoir and providing an outlet flow of
the liquid from a location proximal to the top of the reservoir above the location of the
second portion of seeds By this way seeds of lower density can be separated from higher
density Kwong Sellman Jalink amp Schoor (2005) illustrated that separation by density is
commonly incorporated as part of the routine cleaning and grading process in flower seed
production procedures Other than grouping seeds according to their specific density this
density separation also can be used to remove broken partially filled empty or immature
seeds
28 Seed Deterioration
McDonald (2005) defined flower seed deterioration as deteriorative changes occuring with
time that increase the seeds vulnerability to external challenges and decrease the ability of
the seed to survive Important factors contribute to seed deterioration are genetics seed
structure seed chemistry physicalphysiological quality seed treatments relative humidity
10
and temperature Relative humidity is important because it will influence the moisture
content of seeds in storage while temperature determines the amount of moisture in the air
and influences the rate of deteriorative reactions in seeds According to Agrawal (1980)
the rate of deterioration increases as the seed moisture content increases This could be due
to the mold growth in and on the seed
11
~
30 MATERIAL AND METHOD
31 Material
Fresh matured fruits of Hibiscus sabdarifJa L (Plate I) were obtained from local fanns in
Kota Samarahan Seeds were extracted from these fruits cleaned and dried under shade
Dried seeds were dusted with Captan 80 and then placed in air-tight bottle to keep it in a
safe place before use in the subsequent experiment
Plate 1 Fresh fruit ofH sabdarifJa Plate 2 Fresh fruit of H sabdarifJa with seeds
32 Method
321 Preliminary Evaluation
Moisture content and germination was conducted as an initial evaluation to assess the
quality of seeds used in this research The procedures were those prescribed by Association
ofOfficial Seeds Analysts (AOSA 1985)
12
middot
322 Moisture Content Test
Four replicates of 10 seeds each were placed into saucers (Plate 5) and calibrated to get the
wet weight The seeds then were placed in an oven at 60degC for 48 hours After that the
seeds were weighed again to get the dry weight The percentage of moisture content of the
seeds was calculated based on the formula below (AOSA 1985)
Moisture content () = b - c x 100
b-a
where a = weight of empty saucer
b = weight of (a) + weight of seeds before drying
c = weight of (a) + weight of seeds after drying in the oven
323 Germination Test
Germination test was conducted base on the AOSA (1985) where 25 seeds with four
replications were used For every replicates seeds were put into a petry dish on 3 layers of
moist Whatman filter paper (Plate 6) The seeds were left to germinate by incubating it in a
Plant Growth Chamber (28OC) and was checked daily until 10 days Seed was considered
as germinated if the radicles were about 5mm length emerged from the seed coat The
germination percentage was calculated base on AOSA 1985 formula below
13
-----------------------------------1
Gennination () a x 100
b
where a = total number of germinated seeds
b = total number of seeds used
324 Seed Characteristic
The size of the H sabdariffa seeds was calibrated for its length width and thickness Ten
replicates of seeds were used for calibrating the physical characteristics Then the seed
weights of 10 100 and 1000 seeds were calibrated Four replicates of 10 100 and 1000
seed-weight classes were used
325 Hydration of Seeds
Seed of H sabdariffa was placed in distilled water for 0 2 4 6 8 10 and 12 hours at
temperatures of 28 30 35 and 40degC Then seeds were analyzed for the moisture content
and gennination
326 Data Analysis
The experimental units were arranged randomly based on Completely Randomized Design
(CRD) and data was analyzed by using Analysis of Variance (ANOV A) If there any
significant differences the mean were discriminated using Least Significant Different
(LSD)
14
bull t I I
25 Seed Hydration
A process of supplying water to seed to maintain the fluid or moisture content in the seed
is called hydration According to Copeland and McDonald (1995) seed hydration is a
process whereby seeds are hydrated using various protocols and then redried to permit
routine handling This process results in increased germination rate more uniform
emergence germination under a broader range of environments and improved sedling
vigour and growth It is one of the priming treatments of seed which usually applied to
strengthen or increase vitality of seed Priming process can increase a rate of germination
as the treatment improves the seed vigor Hydration is one component of priming which
can increase germination with more range of environment condition The objective of seed
hydration technology is to increase the percentage and rate of germination expand the
range of temperatures over which the seed will germinate and increase the uniformity of
stand establishment To accomplish these objectives seeds must be hydrated in some way
at a moisture level sufficient to initiate the early events of germination but not sufficient to
permit radicle protrusion (Akers and Holley 1986) However the performance of the seed
might also being influence by other factors like the species type of seed and the seeds
quality
Water can be used for the hydration treatment because it had proved to enhance
germination emergence growth and yield of seed However other mediwns such as the
solution of potassium hydrophosphate (KH2P04) monobasic polyethylene glycol (PEG)
potassium chloride (KCl) potassiwn nitrate (KN03) potassium phoshate (K3P04)
magnesium sulphate (MgS04) sodiwn chloride (NaCl) glycerol and mannitol also can be
used to immerse the seed in hydration process The benefit of such salts is to supply the
seed with nitrogen and other nutrients essential for protein synthesis during germination
8
t I amp
26 Seed Storage
A basic requirement in seed production is a good seed storage The purpose of seed storage
is to preserve planting stocks from one season to the next (Larry amp McDonald 1995)
Generally we stored seeds for the commercial purpose for carry - over seeds or for genn
plasm seeds Maintaining the seed quality for the longest duration possible is the main
objective in seed storage Loss of gennination during storage cannot be stopped but it
could be reduce by giving good storage conditions According to Agrawal (1980) the
seeds are considered to be in storage from the moment they reach physiological maturity
until they genninate or until they are thrown away because they are dead or otherwise
worthless
The seed storage must be properly handled because it can affect the seed quality There are
some factors that can affect the seed longevity in storage such as the variety of the seed
initial seed quality moisture content relative humidity and storage temperature It is
necessary for the seed storage condition to be dry and cool effective in pest control and
have a proper sanitation Other than than storing only high quality of seed also can help in
better production From all of the factors temperature is one of the most important element
which influence seed viability and vigour during storage Agrawal (1980) mentioned that
the lower the temperature the longer the seeds maintain gennination capacity so
decreasing temperature and seed moisture is an effective means of maintaining seed quality
in storage Every seed lot should be gennination - tested when it is received for storage to
record the seed quality Subsequent periodic checks also should be made to detect
deterioration and if it occurs storage condition should be checked to discover the reasons
and to take corrective measures
9
middot
27 Seed Specific Density
Density of seed refers to it mass per unit volume The seed size and density will affect the
yield One of the method for grouping seeds into some specific density is the liquid density
separation system It is one of the easier and cheaper method compared to using a machine
When seed is loaded into a liquid some will float and some will sink That mean the
floating seeds density is lower than the liquids density Vice versa the sinking seeds
density is higher than the liquids density Wang and Alyarez (2008) explained that this
method of separating seeds works by adjusting the specific gravity of the liquid in the
reservoir so that a first portion of the seeds float near the surface of the liquid and a second
portion of the seeds sink near the bottom of the reservoir and providing an outlet flow of
the liquid from a location proximal to the top of the reservoir above the location of the
second portion of seeds By this way seeds of lower density can be separated from higher
density Kwong Sellman Jalink amp Schoor (2005) illustrated that separation by density is
commonly incorporated as part of the routine cleaning and grading process in flower seed
production procedures Other than grouping seeds according to their specific density this
density separation also can be used to remove broken partially filled empty or immature
seeds
28 Seed Deterioration
McDonald (2005) defined flower seed deterioration as deteriorative changes occuring with
time that increase the seeds vulnerability to external challenges and decrease the ability of
the seed to survive Important factors contribute to seed deterioration are genetics seed
structure seed chemistry physicalphysiological quality seed treatments relative humidity
10
and temperature Relative humidity is important because it will influence the moisture
content of seeds in storage while temperature determines the amount of moisture in the air
and influences the rate of deteriorative reactions in seeds According to Agrawal (1980)
the rate of deterioration increases as the seed moisture content increases This could be due
to the mold growth in and on the seed
11
~
30 MATERIAL AND METHOD
31 Material
Fresh matured fruits of Hibiscus sabdarifJa L (Plate I) were obtained from local fanns in
Kota Samarahan Seeds were extracted from these fruits cleaned and dried under shade
Dried seeds were dusted with Captan 80 and then placed in air-tight bottle to keep it in a
safe place before use in the subsequent experiment
Plate 1 Fresh fruit ofH sabdarifJa Plate 2 Fresh fruit of H sabdarifJa with seeds
32 Method
321 Preliminary Evaluation
Moisture content and germination was conducted as an initial evaluation to assess the
quality of seeds used in this research The procedures were those prescribed by Association
ofOfficial Seeds Analysts (AOSA 1985)
12
middot
322 Moisture Content Test
Four replicates of 10 seeds each were placed into saucers (Plate 5) and calibrated to get the
wet weight The seeds then were placed in an oven at 60degC for 48 hours After that the
seeds were weighed again to get the dry weight The percentage of moisture content of the
seeds was calculated based on the formula below (AOSA 1985)
Moisture content () = b - c x 100
b-a
where a = weight of empty saucer
b = weight of (a) + weight of seeds before drying
c = weight of (a) + weight of seeds after drying in the oven
323 Germination Test
Germination test was conducted base on the AOSA (1985) where 25 seeds with four
replications were used For every replicates seeds were put into a petry dish on 3 layers of
moist Whatman filter paper (Plate 6) The seeds were left to germinate by incubating it in a
Plant Growth Chamber (28OC) and was checked daily until 10 days Seed was considered
as germinated if the radicles were about 5mm length emerged from the seed coat The
germination percentage was calculated base on AOSA 1985 formula below
13
-----------------------------------1
Gennination () a x 100
b
where a = total number of germinated seeds
b = total number of seeds used
324 Seed Characteristic
The size of the H sabdariffa seeds was calibrated for its length width and thickness Ten
replicates of seeds were used for calibrating the physical characteristics Then the seed
weights of 10 100 and 1000 seeds were calibrated Four replicates of 10 100 and 1000
seed-weight classes were used
325 Hydration of Seeds
Seed of H sabdariffa was placed in distilled water for 0 2 4 6 8 10 and 12 hours at
temperatures of 28 30 35 and 40degC Then seeds were analyzed for the moisture content
and gennination
326 Data Analysis
The experimental units were arranged randomly based on Completely Randomized Design
(CRD) and data was analyzed by using Analysis of Variance (ANOV A) If there any
significant differences the mean were discriminated using Least Significant Different
(LSD)
14
t I amp
26 Seed Storage
A basic requirement in seed production is a good seed storage The purpose of seed storage
is to preserve planting stocks from one season to the next (Larry amp McDonald 1995)
Generally we stored seeds for the commercial purpose for carry - over seeds or for genn
plasm seeds Maintaining the seed quality for the longest duration possible is the main
objective in seed storage Loss of gennination during storage cannot be stopped but it
could be reduce by giving good storage conditions According to Agrawal (1980) the
seeds are considered to be in storage from the moment they reach physiological maturity
until they genninate or until they are thrown away because they are dead or otherwise
worthless
The seed storage must be properly handled because it can affect the seed quality There are
some factors that can affect the seed longevity in storage such as the variety of the seed
initial seed quality moisture content relative humidity and storage temperature It is
necessary for the seed storage condition to be dry and cool effective in pest control and
have a proper sanitation Other than than storing only high quality of seed also can help in
better production From all of the factors temperature is one of the most important element
which influence seed viability and vigour during storage Agrawal (1980) mentioned that
the lower the temperature the longer the seeds maintain gennination capacity so
decreasing temperature and seed moisture is an effective means of maintaining seed quality
in storage Every seed lot should be gennination - tested when it is received for storage to
record the seed quality Subsequent periodic checks also should be made to detect
deterioration and if it occurs storage condition should be checked to discover the reasons
and to take corrective measures
9
middot
27 Seed Specific Density
Density of seed refers to it mass per unit volume The seed size and density will affect the
yield One of the method for grouping seeds into some specific density is the liquid density
separation system It is one of the easier and cheaper method compared to using a machine
When seed is loaded into a liquid some will float and some will sink That mean the
floating seeds density is lower than the liquids density Vice versa the sinking seeds
density is higher than the liquids density Wang and Alyarez (2008) explained that this
method of separating seeds works by adjusting the specific gravity of the liquid in the
reservoir so that a first portion of the seeds float near the surface of the liquid and a second
portion of the seeds sink near the bottom of the reservoir and providing an outlet flow of
the liquid from a location proximal to the top of the reservoir above the location of the
second portion of seeds By this way seeds of lower density can be separated from higher
density Kwong Sellman Jalink amp Schoor (2005) illustrated that separation by density is
commonly incorporated as part of the routine cleaning and grading process in flower seed
production procedures Other than grouping seeds according to their specific density this
density separation also can be used to remove broken partially filled empty or immature
seeds
28 Seed Deterioration
McDonald (2005) defined flower seed deterioration as deteriorative changes occuring with
time that increase the seeds vulnerability to external challenges and decrease the ability of
the seed to survive Important factors contribute to seed deterioration are genetics seed
structure seed chemistry physicalphysiological quality seed treatments relative humidity
10
and temperature Relative humidity is important because it will influence the moisture
content of seeds in storage while temperature determines the amount of moisture in the air
and influences the rate of deteriorative reactions in seeds According to Agrawal (1980)
the rate of deterioration increases as the seed moisture content increases This could be due
to the mold growth in and on the seed
11
~
30 MATERIAL AND METHOD
31 Material
Fresh matured fruits of Hibiscus sabdarifJa L (Plate I) were obtained from local fanns in
Kota Samarahan Seeds were extracted from these fruits cleaned and dried under shade
Dried seeds were dusted with Captan 80 and then placed in air-tight bottle to keep it in a
safe place before use in the subsequent experiment
Plate 1 Fresh fruit ofH sabdarifJa Plate 2 Fresh fruit of H sabdarifJa with seeds
32 Method
321 Preliminary Evaluation
Moisture content and germination was conducted as an initial evaluation to assess the
quality of seeds used in this research The procedures were those prescribed by Association
ofOfficial Seeds Analysts (AOSA 1985)
12
middot
322 Moisture Content Test
Four replicates of 10 seeds each were placed into saucers (Plate 5) and calibrated to get the
wet weight The seeds then were placed in an oven at 60degC for 48 hours After that the
seeds were weighed again to get the dry weight The percentage of moisture content of the
seeds was calculated based on the formula below (AOSA 1985)
Moisture content () = b - c x 100
b-a
where a = weight of empty saucer
b = weight of (a) + weight of seeds before drying
c = weight of (a) + weight of seeds after drying in the oven
323 Germination Test
Germination test was conducted base on the AOSA (1985) where 25 seeds with four
replications were used For every replicates seeds were put into a petry dish on 3 layers of
moist Whatman filter paper (Plate 6) The seeds were left to germinate by incubating it in a
Plant Growth Chamber (28OC) and was checked daily until 10 days Seed was considered
as germinated if the radicles were about 5mm length emerged from the seed coat The
germination percentage was calculated base on AOSA 1985 formula below
13
-----------------------------------1
Gennination () a x 100
b
where a = total number of germinated seeds
b = total number of seeds used
324 Seed Characteristic
The size of the H sabdariffa seeds was calibrated for its length width and thickness Ten
replicates of seeds were used for calibrating the physical characteristics Then the seed
weights of 10 100 and 1000 seeds were calibrated Four replicates of 10 100 and 1000
seed-weight classes were used
325 Hydration of Seeds
Seed of H sabdariffa was placed in distilled water for 0 2 4 6 8 10 and 12 hours at
temperatures of 28 30 35 and 40degC Then seeds were analyzed for the moisture content
and gennination
326 Data Analysis
The experimental units were arranged randomly based on Completely Randomized Design
(CRD) and data was analyzed by using Analysis of Variance (ANOV A) If there any
significant differences the mean were discriminated using Least Significant Different
(LSD)
14
middot
27 Seed Specific Density
Density of seed refers to it mass per unit volume The seed size and density will affect the
yield One of the method for grouping seeds into some specific density is the liquid density
separation system It is one of the easier and cheaper method compared to using a machine
When seed is loaded into a liquid some will float and some will sink That mean the
floating seeds density is lower than the liquids density Vice versa the sinking seeds
density is higher than the liquids density Wang and Alyarez (2008) explained that this
method of separating seeds works by adjusting the specific gravity of the liquid in the
reservoir so that a first portion of the seeds float near the surface of the liquid and a second
portion of the seeds sink near the bottom of the reservoir and providing an outlet flow of
the liquid from a location proximal to the top of the reservoir above the location of the
second portion of seeds By this way seeds of lower density can be separated from higher
density Kwong Sellman Jalink amp Schoor (2005) illustrated that separation by density is
commonly incorporated as part of the routine cleaning and grading process in flower seed
production procedures Other than grouping seeds according to their specific density this
density separation also can be used to remove broken partially filled empty or immature
seeds
28 Seed Deterioration
McDonald (2005) defined flower seed deterioration as deteriorative changes occuring with
time that increase the seeds vulnerability to external challenges and decrease the ability of
the seed to survive Important factors contribute to seed deterioration are genetics seed
structure seed chemistry physicalphysiological quality seed treatments relative humidity
10
and temperature Relative humidity is important because it will influence the moisture
content of seeds in storage while temperature determines the amount of moisture in the air
and influences the rate of deteriorative reactions in seeds According to Agrawal (1980)
the rate of deterioration increases as the seed moisture content increases This could be due
to the mold growth in and on the seed
11
~
30 MATERIAL AND METHOD
31 Material
Fresh matured fruits of Hibiscus sabdarifJa L (Plate I) were obtained from local fanns in
Kota Samarahan Seeds were extracted from these fruits cleaned and dried under shade
Dried seeds were dusted with Captan 80 and then placed in air-tight bottle to keep it in a
safe place before use in the subsequent experiment
Plate 1 Fresh fruit ofH sabdarifJa Plate 2 Fresh fruit of H sabdarifJa with seeds
32 Method
321 Preliminary Evaluation
Moisture content and germination was conducted as an initial evaluation to assess the
quality of seeds used in this research The procedures were those prescribed by Association
ofOfficial Seeds Analysts (AOSA 1985)
12
middot
322 Moisture Content Test
Four replicates of 10 seeds each were placed into saucers (Plate 5) and calibrated to get the
wet weight The seeds then were placed in an oven at 60degC for 48 hours After that the
seeds were weighed again to get the dry weight The percentage of moisture content of the
seeds was calculated based on the formula below (AOSA 1985)
Moisture content () = b - c x 100
b-a
where a = weight of empty saucer
b = weight of (a) + weight of seeds before drying
c = weight of (a) + weight of seeds after drying in the oven
323 Germination Test
Germination test was conducted base on the AOSA (1985) where 25 seeds with four
replications were used For every replicates seeds were put into a petry dish on 3 layers of
moist Whatman filter paper (Plate 6) The seeds were left to germinate by incubating it in a
Plant Growth Chamber (28OC) and was checked daily until 10 days Seed was considered
as germinated if the radicles were about 5mm length emerged from the seed coat The
germination percentage was calculated base on AOSA 1985 formula below
13
-----------------------------------1
Gennination () a x 100
b
where a = total number of germinated seeds
b = total number of seeds used
324 Seed Characteristic
The size of the H sabdariffa seeds was calibrated for its length width and thickness Ten
replicates of seeds were used for calibrating the physical characteristics Then the seed
weights of 10 100 and 1000 seeds were calibrated Four replicates of 10 100 and 1000
seed-weight classes were used
325 Hydration of Seeds
Seed of H sabdariffa was placed in distilled water for 0 2 4 6 8 10 and 12 hours at
temperatures of 28 30 35 and 40degC Then seeds were analyzed for the moisture content
and gennination
326 Data Analysis
The experimental units were arranged randomly based on Completely Randomized Design
(CRD) and data was analyzed by using Analysis of Variance (ANOV A) If there any
significant differences the mean were discriminated using Least Significant Different
(LSD)
14
and temperature Relative humidity is important because it will influence the moisture
content of seeds in storage while temperature determines the amount of moisture in the air
and influences the rate of deteriorative reactions in seeds According to Agrawal (1980)
the rate of deterioration increases as the seed moisture content increases This could be due
to the mold growth in and on the seed
11
~
30 MATERIAL AND METHOD
31 Material
Fresh matured fruits of Hibiscus sabdarifJa L (Plate I) were obtained from local fanns in
Kota Samarahan Seeds were extracted from these fruits cleaned and dried under shade
Dried seeds were dusted with Captan 80 and then placed in air-tight bottle to keep it in a
safe place before use in the subsequent experiment
Plate 1 Fresh fruit ofH sabdarifJa Plate 2 Fresh fruit of H sabdarifJa with seeds
32 Method
321 Preliminary Evaluation
Moisture content and germination was conducted as an initial evaluation to assess the
quality of seeds used in this research The procedures were those prescribed by Association
ofOfficial Seeds Analysts (AOSA 1985)
12
middot
322 Moisture Content Test
Four replicates of 10 seeds each were placed into saucers (Plate 5) and calibrated to get the
wet weight The seeds then were placed in an oven at 60degC for 48 hours After that the
seeds were weighed again to get the dry weight The percentage of moisture content of the
seeds was calculated based on the formula below (AOSA 1985)
Moisture content () = b - c x 100
b-a
where a = weight of empty saucer
b = weight of (a) + weight of seeds before drying
c = weight of (a) + weight of seeds after drying in the oven
323 Germination Test
Germination test was conducted base on the AOSA (1985) where 25 seeds with four
replications were used For every replicates seeds were put into a petry dish on 3 layers of
moist Whatman filter paper (Plate 6) The seeds were left to germinate by incubating it in a
Plant Growth Chamber (28OC) and was checked daily until 10 days Seed was considered
as germinated if the radicles were about 5mm length emerged from the seed coat The
germination percentage was calculated base on AOSA 1985 formula below
13
-----------------------------------1
Gennination () a x 100
b
where a = total number of germinated seeds
b = total number of seeds used
324 Seed Characteristic
The size of the H sabdariffa seeds was calibrated for its length width and thickness Ten
replicates of seeds were used for calibrating the physical characteristics Then the seed
weights of 10 100 and 1000 seeds were calibrated Four replicates of 10 100 and 1000
seed-weight classes were used
325 Hydration of Seeds
Seed of H sabdariffa was placed in distilled water for 0 2 4 6 8 10 and 12 hours at
temperatures of 28 30 35 and 40degC Then seeds were analyzed for the moisture content
and gennination
326 Data Analysis
The experimental units were arranged randomly based on Completely Randomized Design
(CRD) and data was analyzed by using Analysis of Variance (ANOV A) If there any
significant differences the mean were discriminated using Least Significant Different
(LSD)
14
30 MATERIAL AND METHOD
31 Material
Fresh matured fruits of Hibiscus sabdarifJa L (Plate I) were obtained from local fanns in
Kota Samarahan Seeds were extracted from these fruits cleaned and dried under shade
Dried seeds were dusted with Captan 80 and then placed in air-tight bottle to keep it in a
safe place before use in the subsequent experiment
Plate 1 Fresh fruit ofH sabdarifJa Plate 2 Fresh fruit of H sabdarifJa with seeds
32 Method
321 Preliminary Evaluation
Moisture content and germination was conducted as an initial evaluation to assess the
quality of seeds used in this research The procedures were those prescribed by Association
ofOfficial Seeds Analysts (AOSA 1985)
12
middot
322 Moisture Content Test
Four replicates of 10 seeds each were placed into saucers (Plate 5) and calibrated to get the
wet weight The seeds then were placed in an oven at 60degC for 48 hours After that the
seeds were weighed again to get the dry weight The percentage of moisture content of the
seeds was calculated based on the formula below (AOSA 1985)
Moisture content () = b - c x 100
b-a
where a = weight of empty saucer
b = weight of (a) + weight of seeds before drying
c = weight of (a) + weight of seeds after drying in the oven
323 Germination Test
Germination test was conducted base on the AOSA (1985) where 25 seeds with four
replications were used For every replicates seeds were put into a petry dish on 3 layers of
moist Whatman filter paper (Plate 6) The seeds were left to germinate by incubating it in a
Plant Growth Chamber (28OC) and was checked daily until 10 days Seed was considered
as germinated if the radicles were about 5mm length emerged from the seed coat The
germination percentage was calculated base on AOSA 1985 formula below
13
-----------------------------------1
Gennination () a x 100
b
where a = total number of germinated seeds
b = total number of seeds used
324 Seed Characteristic
The size of the H sabdariffa seeds was calibrated for its length width and thickness Ten
replicates of seeds were used for calibrating the physical characteristics Then the seed
weights of 10 100 and 1000 seeds were calibrated Four replicates of 10 100 and 1000
seed-weight classes were used
325 Hydration of Seeds
Seed of H sabdariffa was placed in distilled water for 0 2 4 6 8 10 and 12 hours at
temperatures of 28 30 35 and 40degC Then seeds were analyzed for the moisture content
and gennination
326 Data Analysis
The experimental units were arranged randomly based on Completely Randomized Design
(CRD) and data was analyzed by using Analysis of Variance (ANOV A) If there any
significant differences the mean were discriminated using Least Significant Different
(LSD)
14
middot
322 Moisture Content Test
Four replicates of 10 seeds each were placed into saucers (Plate 5) and calibrated to get the
wet weight The seeds then were placed in an oven at 60degC for 48 hours After that the
seeds were weighed again to get the dry weight The percentage of moisture content of the
seeds was calculated based on the formula below (AOSA 1985)
Moisture content () = b - c x 100
b-a
where a = weight of empty saucer
b = weight of (a) + weight of seeds before drying
c = weight of (a) + weight of seeds after drying in the oven
323 Germination Test
Germination test was conducted base on the AOSA (1985) where 25 seeds with four
replications were used For every replicates seeds were put into a petry dish on 3 layers of
moist Whatman filter paper (Plate 6) The seeds were left to germinate by incubating it in a
Plant Growth Chamber (28OC) and was checked daily until 10 days Seed was considered
as germinated if the radicles were about 5mm length emerged from the seed coat The
germination percentage was calculated base on AOSA 1985 formula below
13
-----------------------------------1
Gennination () a x 100
b
where a = total number of germinated seeds
b = total number of seeds used
324 Seed Characteristic
The size of the H sabdariffa seeds was calibrated for its length width and thickness Ten
replicates of seeds were used for calibrating the physical characteristics Then the seed
weights of 10 100 and 1000 seeds were calibrated Four replicates of 10 100 and 1000
seed-weight classes were used
325 Hydration of Seeds
Seed of H sabdariffa was placed in distilled water for 0 2 4 6 8 10 and 12 hours at
temperatures of 28 30 35 and 40degC Then seeds were analyzed for the moisture content
and gennination
326 Data Analysis
The experimental units were arranged randomly based on Completely Randomized Design
(CRD) and data was analyzed by using Analysis of Variance (ANOV A) If there any
significant differences the mean were discriminated using Least Significant Different
(LSD)
14
-----------------------------------1
Gennination () a x 100
b
where a = total number of germinated seeds
b = total number of seeds used
324 Seed Characteristic
The size of the H sabdariffa seeds was calibrated for its length width and thickness Ten
replicates of seeds were used for calibrating the physical characteristics Then the seed
weights of 10 100 and 1000 seeds were calibrated Four replicates of 10 100 and 1000
seed-weight classes were used
325 Hydration of Seeds
Seed of H sabdariffa was placed in distilled water for 0 2 4 6 8 10 and 12 hours at
temperatures of 28 30 35 and 40degC Then seeds were analyzed for the moisture content
and gennination
326 Data Analysis
The experimental units were arranged randomly based on Completely Randomized Design
(CRD) and data was analyzed by using Analysis of Variance (ANOV A) If there any
significant differences the mean were discriminated using Least Significant Different
(LSD)
14