IDENTIFICATION AND DISTRIBUTION OF ARBUSCULAR MYCORRHIZAL SPORES IN SELECTED PLANT

ROOTS AND THEIR SURROUNDING SOIL

Tiang Bi Ren

(39159)

QK Bachelor of Science with Honours604.2

(Resource Biotechnology) M92 2015T55l

2015

Pusat Khidmat Maklumat Akaclem ik UN IVER ITI MA LAYSIA SARAWA '

Identification and Distribution of Arbuscular Mycorrhizal Spores in Selected Plant Roots and Their Surrounding Soil

Tiang Bi Ren (39159)

This project is submitted in fulfillment of the requirement for the Degree of Bachelor of

Science with Honours

(Resource Biotechnology)

Supervisor: Dr. Samuel Lihan Co-supervisor: Dr. Rebicca Edward

Resource Biotechnology Department of Molecular Biology

Faculty of Resource Science and Technology Universiti Malaysia Sarawak

I.

i

ACKNOWLEDGEMENT

First and foremost, I would like to express my fullest gratitude to my research supervisor,

Dr. Samuel Lihan, in the process of completion in this project. I am extremely grateful to

my supervisor for sharing his expertise, valuable guidance and encouragements which are

vital for me to complete my research. Without his valuable time and patience, I am

certainly that this research work could not be carried out smoothly and effectively.

Next, I would also like to express my sincere appreciation to Rakiya Abdullahi, the

postgraduate student of Bacteriology Laboratory. I am thankful for her full assistance,

valuable advices, and concerns during my research work in the Bacteriology Laboratory.

Without her fullest help, it will be hard for me to accomplish my experiment well. Thus, I

sincerely felt grateful for her help in my work .

I would also like to take this opportunity to show my gratitude to my family

members especially my parents for their constant support, encouragement and attention

towards me for me to complete my research work. With an unceasing support from my

family, I was able to complete my research project smoothly.

On top of that, I would like to extend my gratefulness to all my colleagues in

Bacteriology Laboratory. I am thankful for having colleagues who always share their

knowledge and ~dvice to me in my project work.

Last but not least, I would like to record my sincere gratitude to everyone, who either

directly or indirectly has lent their hands in guiding and making my research work a

success .

'.

UNIVERSITI MALAYSIA SARA WAK

Grade:

Please tick (.J) Fina f Year Project Report [Z] Masters D PhD D

DECLARATION OF ORIGINAL WORK

This declaration is made on the .... JQ~~......day of.....~~.~.~ ....2015.

Student's Declaration:

I, Tiang Bi Ren, 39159, ofFaculty of Resource Science and Technology hereby declare that the work entitled

"Identifiction and Distribution of Arbuscular Mycorrhizal Spores in Selected Plant Roots and Their

Surrounding Soil" is my original work. r have not copied from any other students' work or from any other

sources except where due reference or acknowledgement is made explicitly in the text, nor has any part been

written for me by another person.

TI~ 81 RW (3~\r~) Date submitted Name of the student (Matric No.)

Supervisor's Declaration:

I, Samuel Lihan hereby certities that the work entitled "Identifiction and Distribution of Arbuscular

Mycorrhizal Spore~ in Selected Plant Roots and Their Surrounding Soil" was prepared by the above named

student, and was submitted to the "FACULTY" as a * partiaVfull fultillment for the conferment of Bachelor

of Science with Honours, and the aforementioned work, to the best of my knowledge, is the said student's

work.

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I declare that Project/Thesis is classi fied as (Please tick (~)):

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research was done)*

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• This Project/Thesis is the sole legal property of Universiti Malaysia Sarawak (UNl MAS).

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III

I

Pusat hicfmat Makfumat Akadem ik UNIVER. IT I . A AYS A ARAWAK

TABLE OF CONTENTS

ACKNOWLEDGEMENT

DECLARATION II

T ABLE OF CONTENTS

ABSTRACT

IV

LIST OF ABBREVIATIONS VII

LIST OF FIGURES VIII

LIST OF TABLES IX

1.0 INTRODUCTION 2.

2.0 LITERATURE REVIEW 4

2.1 Mycorrhizae 4

2.2 Arbuscular Mycorrhizae Fungi (AMF) 4

2.2.1 History of Arbuscular Mycorrhiza Fungi 4

2.2.2 Morphological Features of AM fungi 5

2.3 Arbuscular Mycorrhiza Fungi Symbiosis 7

2.4 Benefits of Arbuscular Mycorrhiza Fungi 8 ..

2.4.1 Increased Nutrient Uptake in Plants 8

2.4.2 Bio-control Agent 8

2.4.3 Potential as Bio-fertilizer 9

2.4.4 Drought Tolerance 9

2.4.5 Salt Tolerance 10

IV

,.......

2.5 Studies on the AM Fungi Diversity 12

2.6 Studies on the Soil Physicochemical Properties on Colonization of AM fungi 14

3.0 MATERIALS AND METHODS 15

3.1 Collection of Soil and Root Samples 15

3.2 Observation of AM Fungi Spores in Plant Root Samples 16

3.3 Isolation of AM Fungi Spores 16

3.3.1 Wet Sieving and Decanting Technique 16

3.3.2 Sucrose Density Gradient Centrifugation Technique 18

3.4 Quantification of AM fungi Spores 19

3.5 Staining of AM Fungi Roots 19

3.6 Assessment of AM Fungi Root Colonization 20

3.7 Identification of AM fungi Spores 21

3.8 Soil Physicochemical Analysis 22

3.8.1 Physical Analysis of Soil Samples 22

3.8.2 Chemical Analysis of Soil Samples 25

3.9 Statistical Analysis 25

4.0 RESULTS 26

4.1 Observation of AM Fungi Spores 26

4.2 Quantification of AM Fungi Spore 27

4.3 Estimation of AM Fungi Root Colonization 28

4.4 Identification of AM Fungi Spores 29

v

4.5 Physicochemical AnaLysis of Soil Samples 33

4.5.1 Physical Analysis of Soil Samples 33

4.5.2 Chemical Analysis of Soil Samples 34

4.6 Statistical Analysis 35

5.0 DISCUSSION 36

CONCLUSION 43

REFERENCES 45

APPENDICES 50

"

VI

I

LIST OF ABBREVIATIONS

AM Arbuscular Mycorrhiza

AMF Arbuscular Mycorrhiza Fungi

INVAM International Culture Collection of Vesicular and Arbuscular Mycorrhizal

Fungi

PVP Polyvinyl Pyrrolidone

SPSS Statistical Package for Social Sciences

J.1m micrometer

VAM Vesicular Arbuscular M ycorrhiza

."

VII

j

LIST OF FIGURES

Figure 1 Main structures of AM fungi 6

Figure 2 Sampling sites for plants 15

Figure 3 Sieves used for wet sieving and decanting method 17

Figure 4 Sievings collected in different Petri dishes 17

Figure 5 Flow diagram for "Feel Method" 24

Figure 6 AM spores observed under microscope at magnification of lOOX 26

Figure 7 Glomus spp. 30

Figure 8 Rhizophagus spp. 30

Figure 9 Acaulospora spp. 31

Figure 10 Funnel({ormis spp. 31

Figure 11 Isolated types of AM spores 32

VIII

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I:

....

Table I

Table 2

Table 3

Table 4

Table 5

Table 6

I

LIST OF TABLES

Enumeration of spores of AM fungi per gram of soil samples 27

Percentage of root colonization of AM fungi in soil samples 28

Identification of isolated AM fungi spores 29

Characteristics of soil texture analysis of plant samples 33

Reading of pH meter from soil samples 34

Pearson correlation analysis of soil pH and colonization rate of AM fungi 35

IX

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Identification and Distribution of Arbuscular Mycorrhizal Spores in Selected Plant Roots and Their Surrounding Soil

Tiang Bi Ren (39159)

Resource Biotechnology Programme Department of Molecular Biology

Faculty of Resource Science and Technology University Malaysia Sarawak

ABSTRACT

Arbuscular mycorrhizas fungi (AMF) possess great potential in ecosystem especially in agricultural system as they can contribute towards the sustainability of the system. AMF have the ability to increase nutrient uptake, alleviate environmental stresses, and potential as bio-fertilizer. Thus, the aim of this project was to isolate, characterize and identify morphologically types of AMF present in selected plants sampled around Kota Samarahan which consists of Cassava, Carica papaya, Cymbopogon, and Pandanus ammyllifolius, quantification of AMF spores, and analysis of soil physicochemical properties towards colonization of AMF. Isolation process of AMF spores was carried out using wet sieving and decanting method and spores were characterized morphologically. From result obtained, genera of AMF isolated consisted of Acaulospora, Funneliformis, Glomus, and Rhizophagus whereby genus Glomus was found dominant among all plant samples. It also showed that soil texture and pH affect the colonization rate of AMF. Cassava recorded the highest colonization rate of AMF with 20% whereas Cymbopogon recorded lowest AMF colonization rate of 10% only. From this study, AMF spores were successfully isolated from different plant roots and characterized morphologically. Soil texture and soil pH were found to affect the colonization rate of AMF in plants.

Key words: Arbuscular mycorrhiza fungi (AM F), isolation, physicochemical, colonization

ABSTRAK

Kulat Arbuskel Mikoriza (KAM) memiliki potensi yang besar dalam ekosistem terutamanya dalam bidang pertanian kerana KAM dapat meyumbang kepada kemampanan ekosistem. KAM mempunyai kemampuan untuk meningkatkan pengambilan nutrisi,penguranagan tekanan alam sekitar, serta potensi sebagai bio-baja. Justeru, objektif projek ini adalah un/uk mengasingkan, mengcirikan dan mengenalpasti spora KAM secara m01jologi dalam tumbuhan yang disampel sekilar Kota Samarahan yang terdiri daripada Cassava, Carica papaya, Cymbopogon, dan Pandanus amaryllifolius, pengkuantifikasian spora KAM dan analisis faktor jizikokimia /anah terlradap kolonisasi KAM. Proses isolasi spora KAM dijalankan dengan menggunakan teknik pengayakan basah dan dekantasi dan spora KAM diidentifikasi secm'a morfologi. Daripada hasil kepulusan, genus KAM yang diperoleh terdiri daripada genus Acaulospora, Funneliformis, Glomus dan Rhizophagus dan genus Glomus mendominasi dalam semua tumbuhan. Kajian ini juga menunjukkan tekstltr tanah dan pH mempengaruhi kadar kolonisasi KAM. Cassava merekodkan kadar kolonisasi yang tertinggi dengan 20 peratus manakala Cymbopogon merekodkan kadar kolonisasi yang terendah iaitu sebanyak J0 peratus sahaja. Daripada kajian ini, spora KAM Berjaya diisolasi daripada pelbagai akar tumbuhan, dan dikenalpasti secara morfologi. Tekstur tanah dan pH tanah didapati mempengaruhi kadar kolonisasi AMF dalam tumbuhan.

Kata kunci: Kulat Arbuskel Mikoriza (KAM), isolasi, fizikokimia, kolonisasi

1.0 INTRODUCTION

Various kinds of microorganisms are present in the soil whereby each of these

microorganisms plays an important role in diverse physiological events in the soil

environment. One of these microorganisms that are involved in these activities IS

mycorrhiza fungi. Mycorrhiza is a special interaction between two eukaryotes which

comprises of an obligate biotrophic fungus and a host plant whereby both associates obtain

benefits from the mutual relationship (Bonfante & Genre, 2008). These mycorrhiza fungi

form a symbiotic relationship whereby the plant supply sugars to the fungi and in return

the fungi supply phosphorus to the plant cells. There are six major groups of mycorrhizas

which comprises of ectomycorrhizae, endomycorrhizae, ectendomycorrhizae, arbutoid

mycorrhizae, monotropoid mycorrhizae and orchid mycorrhizae (Harley & Smith, 1983).

Amongst diverse types of mycorrhizaes, Arbuscular Mycorrhiza (AM) fungi is one

of the commonly and recurring endomycorrhiza around the world (Shanna & Yadav,

20 13). The most distinct char acteristic of AM fungi is the growth of arbuscules which are

highly branched inside the root cortical cells. The fungi will grow between cortical cells

originally and then penetrate the host cell and subsequently grows in the cells fonning new

sections which contain high molecular complexity materials that are enveloped by the host

cell.

Presently, AM fungi is one of the enctomycorrhiza which are being studied

intensively by researchers as AM fungi exists in most natural ecosystem and agricultural

field and play pivotal role in the growth, health and productivity of plants (Kanchana &

Gupta, 2012). The ability of AM fungi to fonn close associations with most species of

plants and significantly improving the uptake of mineral nutrients in exchange of

photosynthetates, decrease stresses caused by biotic and abiotic factors and huge potential

2

as bio-fertilizer have greatly drawn attention of researchers to study it intensively for a

productive agricultural practices.

AM fungi have diversity of species and genera and their composition are greatly

different between every plant species. According to Bever et al. (200 1), the understanding

of roles of each individual AM fungi species is comparatively inadequate even though

records of ecological benefits by AM fungi in overall are plentiful. Various researches

indicated that the growth of plant is influenced by the corresponding fungal species and

particular plant involved (Bever et al., 200 1).

However, there were not much researches conducted on the status of AMF in

Malaysia. Therefore, this study is conducted to investigate the type of indigenous species

of AM fungi that are present in various plants which are tapioca plant, papaya, lemongrass

and pandan leaves that sampled around Kota Samarahan, Sarawak.

The objectives of this study being conducted are:

• To isolate and quantify Arbuscular Mycorrhiza (AM) fungi spores from different

plant roots and their surrounding soil;

• To characterize morphologically isolated Arbuscular Mycorrhiza (AM) fungi

spores present in the sample plants;

• To study the effect of soil physicochemical properties towards the colonization of

Arbuscular Mycorrhiza (AM) fungi.

3

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2.0 LITERATURE REVIEW

2.1 Mycorrhizae

Mycorrhizae fungi are found to be occurring abundantly in all natural ecosystems and play

a vital role in these ecosystems. According to a study done by Peyronel et al. (1969) ,

earliest classification of mycorrhizae was based on the comparative location .of fungi in the

roots whereby these mycorrhizaes was divided into endomycorrhizae, ectomycorrhizae,

and ectendomycorrhizae. However, these classifications were not sufficient to determine

the variety of associations involving mycorrhiza. Thus, from the study done by Harley and

Smith (1983), commonly accepted classifications of mycorrhiza were proposed by them

which divided the mycorrhiza into six groups that consists of endomycorrhiza,

ectomycorrhiza, ectendomycorrhiza, arbutoid mycorrhiza, monotropoid mycorrhiza and

orchid mycorrhiza.

2.2 Arbuscular Mycorrhizae Fungi (AMF)

2.2.1 History of Arbuscular Mycorrhiza Fungi

Arbuscular mycorrhiza fungi are known as obligate symbiotic fungi which can be found in

most species of terrestrial plants. AM fungi belongs to the phylum Glomeromycota which

includes eleven families and twenty-five genera of species are a key component in the soil

microbiota and Signify the most significant terrestri~l symbiosis (Bumi et al., 2011).

According to Brundrett (2004), AM fungi were found to evolve from 460 million

years ago from the records of fossils found from Ordovician Age. From these records, it

proved that AM fungi play an important role in the colonization of most terrestrial plants in

the natural ecosystems. According to Strack et al. (2003), the term "mycorrhiza" originated

from Greek word whereby ' myces ' means fungus and 'rhiza' means root and this term was

4

usat Khidmpt Makhmlat Akad mil< UN1VERSITI MALAY IA SARAWAK

jproposed in 1885 by Bernhardt Frank. Arbuscular mycorrhiza was named so due to a

prevalent characteristic whereby they have a unique branching arrangement called

arbuscules growing inside the cortical cells of host plant roots. The formation of arbuscules

greatly increases the contact area between the plant and the fungus . The exchange of

nutrients such as phosphorus, zinc and copper from the soil to the plant took place in the

arbuscules with the aid of AM fungi and in return, the plant provides carbohydrate to the

fungi.

2.2.2 Morphological Features of AM fungi

AM fungi is made up of three major constituents which comprises of the root itself where

the root supplied carbon in the form of sugars to the fungi; the formation of arbuscules

within the cortical cells of the plant root that enables the contact between the plant

cytoplasm and the fungus and also the structure of extraradical hyphae which help in

nutrients uptake (Smith & Read, 2008) . •

Spores of AM fungi formed on the subtending hyphae as swellings which occurred

either in the soil or in the roots of host plant. These spores contain storage lipids which

function as energy source for pre-symbiotic growth. According to Hosny et al. (1998), AM

fungi spores are found to comprise of cytoplasm and contain abundant nuclei which vary

from 576 nuclei and can reach up to 35000 nuclei in different species of plants.

Blaszkowski et al. (2002) stated that nearly all sp@res of AM fungi devetoped thick walls

which can have more than one layer. However, the color, morphology and the composition

of the spores differ dependent on the species of the AM fungi . Variation in these

characteristics caused the size of the spores to be varied greatly between the species which

can be from 20 J!m to 100 J!m (Dalpe et aI., 2005).

5

According to Bago et al. (1998), arbuscules are structures that are tree-shape like

and fonned by repeated dichotomous branching inside the cell wall of root cortical cell of

host plant. Arbuscules act as an indicator of active mycorrhiza in the soil. Arbuscules

function as the main site for exchange of nutrients and signaling exchange between a host

plant and a fungus (Parniske, 2008).

Dalpe et al. (2005) reported that vesicles of AM fungi were formed from the

swellings of intraradical hyphae and it is filled with lipids and glycolipids which are

necessary for growth of host plant. Vesicles are fonned to collect storage products in many

vesicular arbuscular mycorrhizal (V AM) relationships. On top of that, extensively fonned

extraradical mycelium in the fungi helps in the exchange of nutrients between the roots and

the fungi (Aggangan et al., 2011). Figure 1 showed the main structures that were

associated with AM fungi in plant root.

{ {

matrical' pore

lr

r t: cort x

t: cpidcrJ:'T1 i

h bae

Figure 1: Main structures of AM fungi (Moore, 2011).

6

')

2.3 Arbuscular Mycorrhiza Fungi Symbiosis

Smith and Read (2008) reported that various researches conducted by scientists indicated

that many plants do not interact directly with the soil environment; instead, these plants

interact indirectly with the soil environment through the arbuscular mycorrhiza fungi .

Klironomos (2003) stated that AM fungi possessed the capability of forming symbiosis

with plants making the Glomeromycota being characterized as a generalist organism in

most plants.

The mechanism of converting phosphate that passes through fungal membrane into

polyphosphate which are bounded within the vesicles and transported back through the

hyphae to the host plant by the AM fungi enabled the host plant's phosphate necessity to

be filled up to ninety percent and thus increases the phosphate uptake significantly (van der

Heijden et al., 2008). Extraradical myceliums that are present in AM fungi functions as an

extension of roots for the host plant which efficiently exploit available resources for the

plant by extending its mycelium to the surface of the soil (Ollson et ai, 2002). Aggangan et

al. (2011) stated that extraradical myceliums which are extensively formed in the fungi

help in the exchange of nutrients between the plant roots and the fungi.

Finlay (2008) stated that the hyphae of AM fungi were able to access and release

nutrients that were physically or physiologically inaccessible to the roots of the plant

" which increases the uptake of nutrients in plants. The hyphae of AM fungi are able to

obtain nutrients in the soil through the accelerated process of decomposition and lysing of

soil microbes (Ollson et al., 2002).

7

)

2.4 Benefits of Arbuscular Mycorrhiza Fungi

2.4.1 Increased Nutrient Uptake in Plants

In a study done by Naher et al. (2013) on inoculated AM plants, AM fungi are being

reported to help to increase the uptake of nutrients such as phosphorus and zinc compared

to non-inoculated plants. AM fungi are ubiquitous and fonn a symbiotic relationship with

the roots of most terrestrial plant on the Earth. The association between AM fungi and

plant benefits the plant nutrition, growth and survival due to their enhanced exploitation of

soil nutrients.

From a study done by Al-Karaki and Al-Raddad (as cited in Naher et al., 2013),

mineral contents in wheat plants that are inoculated with AM fungi are found to be higher

compared to non-inoculated wheat plants. Besides, Wu et al. (2011) conducted a study on

the nutrient uptake in peach plants that were inoculated with AM fungi with non­

inoculated peach plants. From the result 'obtained, it showed that mycorrhizal inoculation

increased the nutrient uptake and growth perfonnance of peach seedlings. It also showed

that Glomus mossea exhibited best mycorrhizal efficiency on the growth and nutrient

acquisition ofpeach seedlings.

2.4.2 Bio-control Agent

AM fungi are found to be able to activate and increase the defense level of plants against

soil-borne pathogens. Al-Askar and Rashad reported that AM fungi playa pivotal role in

protecting bean plant against Fusarium solani (as cited in Naher et al., 2013). According to

Naher et al. (2013), a number of findings showed that mycorrhizal inoculation directly or

indirectly increased the plant defense mechanism against pathogens.

8

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In another study done by Akhtar and Siddiqui (2007), they reported that the

establishments of AM fungi in plant roots have been shown to reduce damages caused by

soil-borne plant pathogens with enhancement of resistance in mycorrhizal plants. In a

study done by Vailleau et al. (2007), they conducted experiment on tomato plants which

was inoculated with AM fungi . Their result showed positive result whereby the inoculated

tomato plants reduce the infestation of Ralstonia solanacearum which is a causal of

bacteria wilt disease in crop plant.

2.4.3 Potential as Bio-fertilizer

According to Sadhana (2014), bio-fertilizer is organic substances whereby it makes use of

the microorganisms to increase the fertility of soil and AM fungi are among these

microorganisms which play important role in the application ofbio-fertilizer. AM fungi are

found to enhance the growth and productivity of crops when applied as bio-fertilizer to the

crops. Sadhana (2014) stated that the use of AM fungi as bio-fertilizer do not cause any

pollution and are environment-friendly for agricultural practice.

Bhat et al. (20 I 0) studied the effect of Rhizobium and V AM fungi on green grain

(Vigna radiate L. wilczek) under temperate conditions. From the result obtained in the

study, there was a significant effect of Rhizobium and V AM on the level of nitrogen levels

in grain. The nodulation, nitrogen, phosphorus and calcium content in the grain and also

crude protein content increases significantly.

2.4.4 Drought Tolerance

According to various researches done, AM fungi were able to protect plants from drought

stress and its damaging results. Drought stress was thought as one of the most essential

abiotic factor that will limit the growth of plants (Porcel & Luiz-Lozano, 2004). From the

9

study done by Porcel and Luiz-Lozano (2004) on the tolerance of drought in soybean

plants, they found out that soybean plants that were inoculated with AM fungi were

protected against drought compared to non-inoculated AM fungi soybean plants. Their

result showed that AM-inoculated plants have significantly higher shoot-biomass

production and leaf water potential than non-inoculated plants.

According to Jeffries and Barea (2012), AM fungi symbiosis often resulted in the

altered rates of the water movement into, through and also out of the host plant, with

positive consequences for the physiology of the plant and tissue hydration. Accordingly,

AM fungi symbiosis able to protect crop plants against the damaging effects of the water

deficit in the plant and that the AM fungi contribution to plant drought tolerance resulting

from a combination of physical, nutritional and also cellular effect of the plant.

2.4.5 Salt Tolerance

According to Abdel Latef and Miransan (2014), salt stress was a huge threat that

unfavorably hanned the productivity and growth of plant. Evelin et al. (2009) stated that

there were various researches done by scientists on the mechanisms that are involved in the

alleviation of salt stress by AM fungi and the significant role played by AM fungi in these

stress. There are various mechanisms employed by AM fungi which comprises of

biochemical mechanism, physiological and morecular mechanism to reduce salt stress in

plant (Evelin et al.. 2009). Abdel Latef and Miransari (2014) stated that AM fungi help in

the tolerance of salt in plant by various mechanisms such as increasing nutrient uptake in

plant, manufacturing growth honnone in plant, and improving the activity of

photosynthesis and usage of water efficiency in the plant.

Estrada et al. (2013) conducted a study on the salt tolerance efficiency in AM fungi

inoculated maize plant. In the study, Estrada et al. (2013) researched whether indigenous

10

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)

AM fungi that were isolated from location with salinity problem would help the maize

plant to reduce effects of salt stress better than non-inoculated AM fungi maize plants.

From their findings, they concluded that maize plant that were inoculated with native AM

fungi showed higher shoot dry biomass at all levels of salinity. They also stated that

inoculated maize plant displayed an increasing potassium ion (K+) and reduced sodium ion

(Na+) buildup compared to non-mycorrhizal plants.

In another study done by Yang et al. (2014), they researched on the salt tolerance

response in AM fungi inoculated apple seedlings. They conducted this study to investigate

the reaction of apple seedlings which were inoculated with AM fungi under various levels

of salinity stress. From the result obtained, it showed that apple seedlings that were

inoculated with AM fungi displayed a better tolerance towards salt stress and had a better

growth compared to non-inoculated apple seedlings.

11

)

2.5 tudies on the AM Fungi Diversity

Symanczik et al. (2014) conducted a research on the occurrence of AM fungi in Arabian

Peninsula. According to the study, they reported on the desert habituated AM fungi found

in the Arabian Peninsula. Symanczik et al. (2014) were able to successfully isolate and

recorded few species of AM fungi from the whole Arabian Peninsula for the very first time.

As these isolates were from extremely dry region, these isolated AM fungi were thought as

an asset to conserve the biodiversity in the desert area especially in the region of Arabian

Penin ula.

In another study done by Muthuraj et al. (2014), they researched on the diversity

existence of AM fungi in forty-six medicinal plants in India. From ~he study, Muthuraj et

al. (2014) were able to isolate 30 AM fungal spores from four genera which are

AClllospora, Gigaspora, Glomus, and Scutellospora and characterized them

morphologically. The study conducted showed that AM fungal symbiosis were found in

various medicinal plants and had great influence on the host plant. This made AM fungi an

important microorganism in the natural ecosystem environment.

Shanna and Yadav (2013) studied on the occurrence of AM fungi in the local

barley field. From the study done, they perfoIm isolation of AM fungi spores from the

local barley field and able to isolate various species of AM fungi from few genera which

mainly comprised of Glomus, Gigaspora, and SClitellospora fOIm the barley field . Based

on the result obtained, ShaIma and Yadav (2013) found out that the local barley fields

which contain AM fungi had a better growth compared to fields that lacked of AM fungi

associations.

On top of that, from another study conducted by Nasrullah et al. (2010), they

studied on the occurrence of AM fungi in the wheat-maize cropping system in Pakistan.

12

Various species of AM fungi were successfully isolated from wheat and being

characterized morphologically to identify the species of AM fungi present. From the study

done, they also conclude that the rate of the root infection and density of infection by AM

fungi were influenced by type of soil and locations of plants under different agrological

conditions.

Khades and Rodrigues (2009) researched on the existence of AM fungi in six

varietie of Carica papaya L. in a tropical agro-based ecosystem. From the result obtained,

Khades and Rodrigues managed to document important association of AM fungi with six

varieties of Carica papaya L. They concluded that AM fungi symbiosis between the host

plant and the fungi were controlled by various edaphic factors whereby these factors

directly influenced the enumeration of species associated with papaya varieties from the

agro-based ecosystem in various places .

13

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