PROCEEDINGS OF THE 2 d INTERNATIONAL SYMPOSIUM ON TEMULAWA.K AND THE 40th MEETING OF NATIONAL WORKING GROUP ON INDONESIA MEDICINAL PLANT

LAY-OUT

Titis Arifiana, SSi

December, 2011

ISBN No. 978-979-25-1209-0

Address

Biopharmaca Research Center Institute of Research and Community Services - Bogor Agricultural University

Kampus IPB Taman Kencana

Jln. Taman Kencana No. 3, Bogor 16128, INDONESIA

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Email: bfarmaka@gmail.com Website: http://biofarmaka.ipb.ac.id

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RECTOR SPEECH

The 2nd International Symposium on Temulawak the 40th Meeting of National Working Group on Indonesian Medicinal Plant

Assalamualaikum wr. wb. Salam sejahtera bagi kita semua Good morning.

It is our pleasure today that we are all here to attend the important event so called 'Globalization of Jamu Brand Indonesia' which consists of several agenda such as the 2 nd International Symposium on Temulawak (Curcuma xanthorrhiza); the 40th Meeting of National Working Group on Indonesian Medicinal Plant; Workshops, Business Meeting, Jamu Festival, 'Jamu' Batik Design Competition and Temulawak 'Welcome Drink' Formula Competition here in Bogor, Indonesia.

As most of us may still remember that 3 years ago, in 2008, I attended 'Gelar Kebangkitan Jamu Brand Indonesia' and the First International Symposium on Temulawak. The event was officially opened by the President of the Republic of Indonesia, in Istana Negara, Jakarta; while the symposium was held here in the same place, IPB International Conference Center. Now, we are here again to attend the Globalization of Jamu Brand Indonesia. For the two important events, held in 2008 and 2011, multisectors and international stakeholders are involved, and IPB is significantly taking parts. This clearly indicates that continuous improvement to achieve our goal, to make Jamu for the Word Quality of Live, is really our concern.

We all know that Indonesia is the second largest countries in the world regarding biodiversity. The Indonesian people are used to their natural resources with knowledge they inherited from their ancestors. For example, medicinal plants, animals, and microbes are applied as preventive, promotive and curative alternatives. Jamu has long been known and applied by the society; and nowadays, there is also an increasing tendency of using herbal medicine that is popularly known as 'back to nature' for curing diseases worldwide. Jamu business keeps increasing, I heard that almost reach to 10 trillion IDR in 2010; and the support from Indonesian government becomes stronger and real actions have also been conducted, such as : Scientification of Jamu, and Roadmap of Jamu Development will be launched and used as a national guidance for Jamu development. So I believe that through the strong commitment from Jamu stakeholders, our vision that Jamu for the World Quality of Live can be achieved in the near future .

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Distinguish Guest, Ladies, and Gentleman,

Bogor Agricultural University (IPB) with its vision to becoming a world class research university with core competences in tropical agriculture and bioscience with entrepreneur character, and one of our mission is to improve the welfare of human beings through the application of developed science and technology are clearly in line with our effort to improve Jamu development.

Within this context, Biopharmaca Research Center IPB keeps focusing its research development and optimizing its efforts to develop qualified biopharmaca products with the support of strong networking with international and national partners. IPB also contributes to the national policy development of Indonesian biopharmaca and committed its existence in education and research development in order to achieve national and international reputation. Currently, IPB is proposing the establishment of Indonesian Biopharmaca Center (IBC) through the support of Japan International Corporation Agency (JICA), and the visibility study of the project is now being conducted. The IBC is expected to be a center of excellence in Biopharmaca Research Development within the country and will contribute to international reputation.

Today as part of Globalization of Jamu Brand Indonesia, we also have an important international event, namely "The 2"d International Symposium on Temulawak (Curcuma xanthorriza)" which is conducted by Biopharmaca Research Center, IPB in collaboration with Indonesian government institutions, private sectors, and foreign partners. The theme of the symposium is utilization and application of Curcuma xanthorrhiza through scientific and technological approach toward better and healthy life.

We know that temulawak is known as one of the Indonesian indigenous herbals, which mostly used as the main ingredient for traditional medicine or 'Jamu'. The popularity of temulawak is increasing along with its commercial use and research result applications. Many scientists have conducted research to reveal the secret of temulawak. Temulawak can be used for various purposes such as for maintaining human health, animal health, and supplement beverages to increase appetite and to keep fresh our stamina. As a continuation of our commitment, IPB is conducting various aspects of research based on temulawak, e.g. brain tonic, cardiovascular diseases, diabetic; further, our research output on avian flu has been registered for patent.

Nowadays, in the middle of modern lifestyle, temulawak is occupying place in our society's heart and with its various benefits, thus temulawak deserves to be Indonesian "ginseng" herbal. IPB fully support the three days activities involving researchers and scientists all over the world to share their experience and experties which will be conducted in IPB International Convention Center.

We do hope that through Globalization of Jamu Brand Indonesia which are involving biopharmaca stakeholders within the country and abroad, modernization of Indonesian medicine/Jamu will be accelerated and generated benefits to increase our health and welfare.

We would like to ask to the Minister of Coordinator of People's Welfare, Republic of Indonesia to officially open The Globalization of Jamu Brand Indonesia.

Finally, thank you to all of ypu who make this precious event possible.

Billa hi taufik wal hidayah, wassalamualaikum wr. wb.

Bogor, May 26, 2011 Rector of IPB,

Prof Dr Herry Suhardiyanto, MSc

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~ ISBN No. 978-979-25-1209-0

Proceedings of The Second International Symposium on Temulawak -- • • ~ ~~ ,-..;.. ~ •' -·-. I • '''' • .-.~r -'~:~,· .'·:•

Page

Preface iii Remark from Chairman of Organizing Committee iv Acknowledgement from Rector of Bogor Agricultural University v

Botany 1. The Genetic Variation Analysis of Temulawak (Curcuma xanthorrhiza Roxb.) In Java Island Using Random 3

Amplification of Polymorphic DNA (RAPD) Method Budi Setiadi Daryono, Hastuti, Tara Puri Ducha Rahmani

2. The Effectivity of Indoi-3-Butyric Acid (IBA), 6-Benzyl Amino Purin (BAP), and Sucrose to Improve In 7 Vitro Multiplication of Temulawak (Curcuma xanthorrizha Roxb.) Darda Efendi, Cecep S. Darma, Yustika, De/vi Maretta, Churiyah

3. In Vitro Microrhizome Formation of Temulawak (Curcuma xanthorrhiza Roxb.) 11 De/vi Maretta, Darda Efendi, Sandra Arifin Aziz, Dodo R. Sastra

4. Identification of Genetic Diversity on (Curcuma xanthorriza) Collection Clones In Java and Madura 16 Nur Azizah

5. Responses of temulawak (Curcuma xanthorrhiza) explant in vitro growth in photoautotrof condition 22 Pertamawati

6. Substitution of Macro Nutrient Using Composite Fertilizer on Micropropagia and Conservaton of Java 26 Trumeric (Curcuma xanthorhiza) In Vitro Sitti Fatimah Syahid and Natalini Nova Kristina

7. The Ethnobotanical Aspects of Curcuma spp. In Javanese Traditional Medicine 29 Mu/yati Rahayu, Harini M. Sangat and Vera Budi Lestari

8. Effect of Fertilization by the Local Farmers on Fiber and Curcumin Ingredients of Temulawak (Curcuma 34 xanthorriza Roxb.) in Kulon Progo, D.I. Yogyakarta Fibrianty and Retno Utami Hatmi

9. Micropropagation of Curcuma Xanthorhyza 37 Tatik Wardiyati, Evi Muakhiroh, Moch.Dawam, Sukendar

10. Feasibility Study Farming Management of Temulawak Curcuma Xhanthorhiza By Using Different Seed 40 Rhizome Size Ermiati and Sukarman

11. Yield Stability Evaluation on Six UB Clones of Temulawak (Curcuma xanthorriza Robx) 45 Kuswanto and Nur Azizah

12. The Use of Biopesticide for Increasing Ginger 48 Noorwitri Utami, Jeni Hariyanti, Lukita Devy, Arief Arianto

13. Induction of Flowering in Large White Ginger (Zingiber officinale Rose.) using Paclobutrazol 52 Melati, Palupi E R, Bermawie N

14. Seven Species of Endangered Medicinal Plants (Akar Kuning, Gaharu, Kapung-Kapung, Kempas, Kulim, 56 Pasak Bumi, and Pulai) at Bukit Tigapuluh National Parks Francisca Murti Setyowati and Wardah

15. Growth and Bioactive Compound of Guava (Psidium guajava) on Different Frequencies of Leaf Harvesting 62 Sandra Arifin Aziz

16. Seeds Corypha: Medicine Properties and Distribution in Purwodadi Botanic Garden 67 Rony Irawanto

17. Plant Damaging Organisms Associated With King's Bitter Plant (Andrographis paniculata) in West Java 72 and Their Recommended Control Measures Setyowati Retno Djiwanti

18. Medicinal Plants Species in Mount Merapi National Park before the Eruption in 2010 and Its Cultivated in 79 Bogar Botanical Garden Sudarmono, R. 5 . Purwantoro

19. Daemonorops draco as Superior Medicinal Plants : Opportunities and Challenges 83 Tati Rostiwati, Rachman Effendi, Tuti Herawati

20. Diversity of Ahidoidea and Coccoidea (Hemiptera) on Some Medicinal Plants 87 Tri L. Mardiningsih and Dewi Sartiami

21. Growth and Production of Portulaca oleracea L. at Different Plant Ages Affected By Poultry Manure 91 Maya Melati , Sandra Arifin Aziz, and Mohammad Tarih Suseno

ISBN No. 978-979-25-1209-0

Proceedings ofThe Second International Symposium on Temulawak ...... -- ./'. \ •• -~- ' - ~··~ :.....~ ·:- •• - ....... ).•' c ~.- 1 .. , .-·-.,.~.-

22. Reduction of Macronutrient Concentrations on The In Vitro Growth of Red Ginger (Zingiber officina/e) 95 Betalini Widhi Hapsari, Tri Muji Ermayanti, Deritha E. Rantau, Rudiyanto

23 . Morphological Variation of Echinacea purpurea (L.) Moench Accessions In Medicinal Plant and Traditional 100 Medicine Research and Development Office Dyah Subositi and Fauzi

24. Graptophyllum pictum (L.) Griff.: The Collection of Purwodad i Botanic Garden and Its Potential Uses 104 Esti Endah Ariyanti

25. The Effect of 2,4D to The Formulation and Growth of Saga Callus (Abrus precatorius L) Developed from 108 Seeds by In Vitro Propagation Deliah Seswita, Octivia Trisilawati

Efficacy 26. Efficacy of Temulawak (Curcuma xanthorrhiza Roxb.) Extract to Increase Blood T Lymphocyte Count in 113

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Obese Adults Mira Dewi, Cesilia Meti Dwiriani, M. Januwati,Muhammad Aries Supplementation of Concentrate With Different Levels of Temulawak (C. xanthorrhiza, Roxb) on Milk Production of Lactating Frisien Holland Cows E. Sulistyowati, U. Santoso, and I. Badarina

Effect of Curcuma xanthorrhiza Roxb. Decoct on Glucose Absorption Level in Intestine of Male Rat of Wistar Strain Diah Dhianawaty D, Andreanus A. Soemardji, Samsudin Surialaga, Anna Martiana S., Ruslin Antiplasmodial Activity Assay on the Combination of Curcumin and Mangosteen in Vitro against Plasmodium falciparum Strain 3D7 Culture Syamsudin, Mathilde Gonsali Djong Potency of Curcuma xanthorrhiza Extract as a Natural Growth Promoter in Pregnant Ewes with Superovulation Andriyanto, Wasmen Manalu

Inhibitory Activity of Curcuma domestica, Curcuma xanthorrhiza and Zingiber zerumbet Mixed Extract in Proliferation of Colon Cancer Cells HCT (ATCC-CCL 116) Latifah K Darusman, Wulan Tri Wahyuni, Nuriman Zebua Neuroprotective Effect of Temulawak (Curcuma xanthorhiza) on Braine Nerve Cell Damage Induced by Lipopolysaccharide (LPS) Min Rahminiwati, Ita Djuwita, Latifah K Darusman, Siti Sa 'diah Tekokak (Solanum torvum Sw.) Extract Potency as Tyrosinase Inhibitor Irmanida Batubara, Latifah K. Darusman, Irma H. Suparto, Trivadila Antioxidant Activity of N-Hexane Extracts of The Leaves of Soursop (Annona muricata L.) Eka Prasasti Nur Rachmani, Tuti Sri Suhesti Toxicity and Antibacterial Effect of Solanum torvum (Tekokak) Leaves and Fruit Extracts. M Agung Zaim A, Yusrida Hasibuan, Nunuk Kurniati N, Irmanida Batubara Potency of Keji Beling (Stachytarpheta mutabilis) and Soursop (Annona muricata) Extracts as HMGCoA Reductase Inhibitor Irmanida Batubara, Salina Febriany, Latifah K Darusman, Tohru Mitsunaga Capacity of Soursop Leaves as Antioxidant, Antibacterial, and Tyrosinase Inhibitor Susi Indariani, Irmanida Batubara, Latifah K Darusman Cytotoxic Activ ity of Zingiber offlcinale, Piper retrofractum, and Their Combinations on Breast Cancer Cell Line (T47D) Hendri Wasito, Trisnowati, Heny Ekowati The Effect of Yellow Pumpkin Juice on Serum LDL/HDL Ratio in Male White Rat (Rattus Norvegicus) Fed with High Cholesterol Diet Harliady Dany Prabowo,Ismawati, Sri Wahyuni

Effects of Olfactory Stimulation with Scent of Cypress (Callitris g/aucophylla) Essential Oil on Br~wn Adipose Tissue Sympathetic Nerve Activity of Rat J () p ft W Ryunosuke Matsuoka and Tohru Mitsunaga f"l Biological Activities of Two Diterpenes from Jatropha Plants Sahidin, I ; Ardiansyah; Muhammad Taher; Ajeng Diantini; and Marianti A. Manggau

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ISBN No. 978-979-25-1209-0 .

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Proceedings of The Second International Symposium on Temulawak

In Vitro Microrhizome Formation of Temulawak (Curcuma xanthorrhiza Roxb.)

Delvi Maretta1a, Dania Efendi2a, Sandra Arifin Aziz2 b, Dodo R. Sastra 1b

1 Perekayasa/Peneliti Badan Pengkajian dan Penerapan Teknologi (BPPT Gd 2 Lt 17 Jl MH. Thamrin No 8 Jakarta Pusat 10340; delvi2001ina@yahoo.com 1•)

2a,b Staf Pengajar Departemen Agronomi dan Hortikultura, Institut Pertanian Bogor (Kampus IPB Darmaga, Jl Raya Darmaga 16680; (dardaefendi @yahoo .com 2•; sandraaziz@yahoo. com 2b)

ABSTRACT

The aims of this research were to find the influence of the physical form of media (liquid and solid), composition of media (full and half strenght concentration Murashige and Skoog (MS) media), benzylaminopurine (BAP) and sucrose concentration to the growth of explants and formation of micro rhizome. Research consists of three experiments that used factorial design . Cultures of all experiments were incubated in the dark for 16 hours day · '. The results showed that microrhizomes of Temulawak (Curcuma xanthorrhiza Roxb.) was formed in liquid half and full strength MS. Interaction BAP and sucrose had significant effect on rhizomes formation at 18 weeks . The rhizomes formation frequency was higher on media enhanced with BAP 1 and 2 gl-1 and higher sucrose concentration. More and larger micro rhizomes produced obtain from small size shoots.

Keywords: temulawak, l iquid MS media, BAP, sucrose, microrhizome

INTRODUCTION

Temulawak (Curcuma xanthorrhiza Roxb.) is a medicinal

plant native to Indonesia . Various studies have revealed

the efficacy of this plant for human health. Peculiar

efficacy of it's rhizomes are mainly caused by the yellow

compound, curcuminoids and volatile oil, xanthorrizol.

The efficacy of curcuminoids are to neutralize toxins,

relieve pain, increases bile secretion, lowering blood

levels of cholesterol and triglycerides, while xanthorrizol

used to treat breast cancer, lung, ovarian, and

antibacterial prevention of tooth enamel damage (Sidik

2006).

Nowdays, seed propagation of temulawak still apply in

conventional technique by using quite old rhizome . The

higher weight of seed rhizomes planting will increase the

yield (Djakamihardja eta/. 1985) . Per plant production of

temulawak from seed weighing 40 g rhizome higher than

plant from the lower-weighted rhizome seedling (Kasiran

2008) . The using of large rhizome as seed to obtain high

yields causes tendly large amount of seed need, thus

reducing the amount of rhizome for consumption or for

processing .

In vitro technique is a technology that can be applied to

overcome the seed needs in the form of micro rhizomes.

Tyagi et at. (2006) and Anisuzzaman et at. (2008) stated

that micro rhizome can be planted directly without

acclimatization process before . Storage and transport of

micro rhizome as seed will be easier. Micro rhizome can

be used for germplasm conservations also so that more

efficient and longer time storage.

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Induction micro rhizome process is called organogenesis .

This process occurs when cells of explant divide and then

differentiate to form an organ. Cells plant regeneration is

determined by genotype, type and age of explants, plant

growth regulators, medium, temperature and radiation .

In the plant, formation of tuber or rhizome organs will

take place on certain conditions (Wattimena 2006).

Salisbury and Ross (1992) stated that in vitro formation

of plant storage organs would be induced by sucrose and

plant growth regulators that were added to the media .

Several studies have been conducted to induce micro

rhizome formation such on ginger plant (Rahmawati et

a/. 2003, Tyagi eta/. 2006), temuputih (Anisuzzaman et

a/. 2008), turmeric (Shirgurkar eta/. 2001), but has not

been done on temulawak. So, it is necessary to conduct

in vitro micro rhizome formation studies on temulawak.

The purposes of this research were to find the influence

of the physical form of media (liquid and solid),

composition of media (full and half strenght

concentration MS media), BAP and sucrose concentration

to the growth of explants and formation of micro

rhizome .

MATERIAL AND METHODS

These researches were conducted at Laboratorium

Pengembangan Teknologi Industri Agro dan Biomedika

(LAPTIAB) BPPT, Puspiptek Area, Serpong lasted from

October 2008 until May 2010. This study consists of

three series of experiments :

a) Experiment I: Effect of physical forms of media and

sucrose to growth and

micro rhizome

formation of temulawak

ISBN No. 978-979-25-1209-0

Proceedings of The Second International Symposium on Temulawak

b) Experiment II : Effect of the composition of MS media

and sucrose to growth and format ion of temulawak

micro rhizome

c) Experiment III : Effect of BAP and sucrose to growth

and formation of temulawak micro rhizome

Experiment I: Effect of Physical Form of Media and Sucrose Concentration

The media used in the experiments was MS composition

media. This experiment used factorial design . Factor A

was the physical form of medium that cons ists of 2

levels : solid and liquid . Factor B was the sucrose

concentrat ion with 4 levels of concentration were 30, 60,

90 and 120 gL-1 • There were 8 treatment combinations

(in a complete randomized design) with 10 replications

per treatment so that there were 80 units experiments.

An exper imental unit was a bottle containing one

explants . Solid medium was made by adding agar

powder (8 gl-1 medium) into solution. Cultures

maintained in the dark for 16 hours every day.

Experiment II: Effect of MS Media Composition and Sucrose Concentration

The physical condition of the media (solid or liquid) used

was determined based on the results of experiment I.

This experiment used factorial design . Factor A was the

composition of MS media consisting 2 levels: half and full

strenght MS media . Factor B was the sucrose

concentration with 4 levels of concentrations : 30, 60, 90

and 120 gL-1 . There were 8 treatment combinations (in a

complete randomized design) with 20 replications per

t reatment so that there were 160 experimental units (a

bottle contains! explants) . During the experiments, the

culture maintained in the dark for 16 hours every day .

Experiment III: Effect of BAP and sucrose concentration

The physical condition and composition of MS media

used were determined based on the results of

experiment I and II. This experiment also used factorial

design . Factor A was BAP concentrations, consisting 4

levels : 0, 1, 2, and 3 mgL-1. While factor B was sucrose

concentration, consisting 4 levels : 30, 60, 90 & 120 gl-1 .

There were 16 treatment combinations (in a completely

randomized design) and 10 replications in each

treatment, so there were 160 units experiment. In all

experiments, the parameter of data that was observed :

number of rhizome induction, swelling diameter, plantlet

height, leaf length, leaf width, and root length .

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Statistics Model

The design of experiments I , II and III were completely

randomized factorial design . Data analyzing with ANOVA

and followed with Duncan Multiple Range Test (DMRT) at

level 5% error if the result was significantly different.

RESULTS AND DISCUSSION

Experiment I: Effect of Physical Form of Media and Sucrose Concentration

Physical forms of medium and sucrose concentration

interactions hadn 't significant effect to micro rhizome

formation until the end of the experiment for 16 weeks .

Micro rhizome formation induction was influenced by

physical forms medium but not influenced by sucrose

concentration . In the liqu id medium t he number of micro

rhizome induction higher than in the solid medium (Table

1) .

Plantlets with higher size, broader leaves and longer

roots on solid medium produced swell-induction of micro

rhizome fewer and smaller than the plantlets in liquid

medium. Micro rhizome formations in liquid medium

have been successfully conducted to Curcuma aromatica

Salisb. (Nayak eta/. 2000) and Curcuma /onga L. (Islam

2004 ) . In liquid medium higher plantlet size produced

less number of m icro rhizome (correlation -0 .99* ) . Leaf

length and leaf width addition were inversely

proporsional to the addition of the amount of micro

rhizomes that was produced .

Table 1 . The number of micro rh izome induction and plantl et size (1 5 WAP*)

Varia bel

Number of Micro Rhizome I nduct ion Swelli ng Diameter (em) Plantlet Height (em) Leaf Length (em) Leaf Width (em) Root Length (em)

* WAP : Weeks After Planted

Treatments Solid Medium Liquid Medium

0. 33 1.22

0.20 0. 53 6.79 4.17 3.47 1.99 1.17 0.89 1.78 1.02

Experiment II: Effect of MS Media Composition and Sucrose Concentration

In the first experiment number of micro

rhizome induct ion occur higher in the liquid media

than in the solid media . Because of it , media t ha t

was used in the second experiment was liquid media .

Until 18 WAP micro rhizome formation was not

influenced by single factors and by interaction of both

treatments . Although it wasn 't signif icantly d iffe rent,

amount of micro rhizome showed higher on strength half

MS medium than on full concentration MS medium .

ISBN No. 978-979-25-1209-0

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Proceedings of The Second International Symposium on Temulawak

Formation of turmeric micro rhizome (Curcuma tonga

Linn .) according to Sh irgurkar et at. (2001) optimum on

half strength MS media, whereas according to Islam

(2004) the number and size optimum on % MS media.

Unlike to that statements, Kenya et at. (2002) stated

that the number of in -vitro micro bulblet of two cultivars

lily (Avignon and Bergamo) higher on full concentration

MS media than on lower concentration media.

0.15 § .c y (full MS)= -0.001x + 0.13 1:1) ~ 0.10

' • R2 = 0.7543

"= G> • , . E 0 ' N ~ :e

0.05

E 112 MS) = -0.0005x + 0.073 • ... y

'E R2 = 0.2873 ' 0.00

0 30 60 90 120 150 sucrose (gil)

Figure 1 . Micro rh izome weight on half strenght and full concentration MS media.

Note : - - - linear ( full MS) ---- linear (1/2 MS)

Micro rhizome that formed on half strength MS media

had lower weight than micro rhizome formed on full

concentration MS media (Fig . 1). Rhizome d iameter was

almost same on both media (Fig . 2) . Thus, number of

micro rhizome on half strenght MS media higher with

lower weight and smaller size than micro rhizome formed

on full concentration MS media .

Experiment III: Effect of BAP and sucrose concentration

Based on the results of two previous trials, the media

used was half strenght liquid MS media . In this

experiment, interaction between BAP and sucrose

affected significantly to the number of micro rhizome at

18 WAP. The DMRT results demonstrate that interaction

BAP 2 gl-1 and sucrose 120 g l -1 formed the most number

of micro rhizome.

1.00

y 112 MS)= -0.0054x + 0.7292 • R2 = 0.604

0 30 60 90 120 150 sucrose (gil)

Figure 2 . Diameter of the micro rhizome on half and full strength MS medi um

Note : - - - linear (full MS) ----- linear (1/2 MS)

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Along with the increasing of sucrose, tota l micro rhizome

formation also increased on media without BAP and with

1, 2 gl-1 BAP (Fig 3a, b, c) . Otherwise, the number of

micro rhizome progressively decreasing on media with 3

gl-1 BAP (Fig . 4) . According to Rahmawati et at. (2003)

the amount of ginger Uahe emprit) micro rh izome

increased along with increasing of BAP and sucrose

concentration . Explants tend to store excess nutrients

from the media in the rhizomes form. In vitro culture of

Curcuma aromatica Sal isb L on medium with 60- 90 gL-1

sucrose (Nayak 2000) , Curcuma tonga on medium with

40-80 gL-1 sucrose (Shirgurkar et at. 2001) and 60 -100

gl-1 (Islam, 2004) were the optimum sucrose

concentration for increasing the number and size of

micro rhizome. Anisuzzaman et at. (2008) reported that

70% of in-vitro shoots of Curcuma zedoaria (Christm.)

Roscoe forming micro rhizome occurred on medium

containing 60 gl-1 sucrose.

Figure 3 . Micro rhizome from treatment 90 g/1 sucrose and BAP 0 gL-1(a), 120 g/1 sucrose and BAP 1 gL-1 (b), 120 g/1 sucrose dan BAP 2 gL-1

ISBN No. 978-979-25-1209-0

Proceedings of The Second International Symposium on Temulawak

..

y(BAP 2 mgL-') = 0.0292x -1 .38 R2 =060"

I

I I

~ ~

I

y (B,t> 1 mgL-1 )j,D.0142x - 0 67 I . R2 = (T74"

,' / ,' /

I ./ • -.... I • '~ (tanpa BAP) = 0 0108x- 0.33

I R' = 0.49" , y(BAP;,;L-'~33x +0 63

O +----;~--~~--~.--~R'-=~0~.00~---.

0 30 60 150

Figure 4. Number of micro rhizome on medium without BAP and with BAP 1, 2, 3 mgL-1

---- - linear (without BAP) -- -- linear (BAP 1 mgL-1) - - - linear (BAP 2 mgl-1 )

linear (BAP 3 mgL-1) * : significant a a = 5%

Sucrose as carbon source can be used for food storage

filling. One of food storage organ is micro rhizome.

Salisbury and Ross (1992) stated that in the storage

organs, carbohydrates accumulate as aminoplas by

result of sucrose translocation . Trigiano and Gray (2005)

stated that sucrose was the most often used as carbon

and energy sources in the in-vitro culture because it was

naturally synthesized and transported by plants.

On 18 WAP, with the presence of BAP in the medium,

plantlet height and number of shoots significantly

correlated with the number and diameter of micro

rhizome. In the medium with BAP 1 mgl-1, more shoot

would be produced more micro rhizome. The increasing

of plantlet height followed by the decreasing of number

and diameter of rhizomes. Such was the case, more

micro rhizome with large-sized was harvested from small

size shoot (Tables 2 and 3).

Table 2. Correlation between leaf length, number of senescene leaves, plantlet height, root length, and number of shoots to micro rhizome number at 18 WAP

Varia bel

Leaf Length Number of Senescene Leaves Plantlet Height Root Length Number of Shoots

Number of Rhizomes BAPO BAPl BAP 2 mgl-1 mgL-1 mgL-1

0.09 -0.09 -0.41

-0.24 -0.05 -0.13

0.05 -0.58* -0 .82** -0.23 -0 .27 -0 .35 -0.23 0.64* 0.32

*: significant at o = 5% ,••: highly significant at o = 1%

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Table 3. Correlation between leaf length, number of senescene leaves, plantlet height, root length, number of shoots to rh izome diameter micro at 18 WAP

Micro Rhizome Diameter Variabel BAPO BAP 1 BAP 2

mgl-1 mgL-1 mgL-1

Leaf Length 0.21 -0.10 -0.07 Number of -0 .24 -0.15 -0.13 Senescene Leaves Plantlet Height 0.08 -0.54* -0.52* Root Length -0.30 -0.28 -0.16 Number of Shoot -0.31 0.58* 0.52* .. significant at o = 5%

Micro rhizome formations in all three experiments were

closely related with the concentration of sucrose in the

medium. The high sucrose concentration causes medium

osmotic pressure higher than in the plant cells so that

the water becomes unavailable for plants. These

conditions can be interpreted that water stress happen in

the plant. Haryati (2003) stated that water stress during

vegetative growth would affect the source intensity such

as size of canopy, while at the generative growth would

affect the size of sinks such as part of plant that were

usually harvested (fruit or rhizome).

CONCLUSI ON AND SUGGESTION

Temulawak micro rhizome formed in liquid half and full

strength MS medium. Interaction of BAP and sucrose

significantly affected to micro rhizome number that found

on 18 MSP. The presence of BAP 1 and 2 mgl-1 in the

medium will increase the number of micro rhizome

during the increasing of sucrose concentration . More and

larger micro rhizome produced obtain from small size

shoots .

REFERENCES

Anisuzzaman M, Sharmin SA, Mandai SC, Sultana R, Khalekuzzaman M, Alam I , Alam MF. 2008. In vitro Microrhizome Induction in Curcuma zedoaria (Christm.) Roscoe-A Conserbation Prioritized Medicinal Plant. Journal of Biological Sciences p 1-5.

Djakamihardja S, Setyadiredja P, Sudjono I. 1985. Budidaya Tanaman Temulawak (Curcuma xanthorrhiza Roxb.) dan Prospek Pengembangannya di Indonesia. Di dalam : Prosiding Simposium Nasional Temulawak; Bandung, 17 Sept 1985. Bandung : Lembaga Peneli tian Universitas Padjadjaran . him 149-160.

Haryati. 2003. Pengeruh Cekaman Air terhadap Pertumbuhan dan Hasil Tanaman. Program Studi Hasil Pertanian. Medan : Universitas Sumatera Utara .

Islam MA, Kloppstech K, Jacobsen J. 2004. Efficient procedure for in-vitro microrhizome induction in Curcuma tonga L. (Zingiberaceae)-A medicinal plant of tropical Asia . Plant Tissue Cult. 14(2) : 123-134.

Kasiran . Penelitian Produksi Temulawak (Curcuma xanthorriza Roxb.) melalui Penggunaan Ukuran Bibit yang Berbeda. Buletin Penelitian Sistem Kesehatan Vol. 11 No 3 Juli 2008: 270-275

Kenya AP, Murdaningsih HK, Herawati T, Darsa JS. 2002. Taggap dua kultivar lili terhadap kombinasi medium MS dan gula pasir untuk konservasi in-vitro. Zuriat 13 (2) :87-96.

ISBN No. 978-979-25-1209-0