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Jour. Myan.Acad. Arts & Se. 2005 Vol. III. Nb.f(ii)

Meristem Culture Gf Aloe vera L;

Kay Thi Mya', Thida Myint-^, Myo •Han-', and Khin Maung

Shoot tips,of Aloe vera L were excised and inoculated in basal and modified MS (Murashige and Skoog, 1962) medium. Growth regulators and growth factors were supplemented to the basal medium. Development of explants, initiation of and multiplication of slibots in culture were recorded.

Introduction

• •. ' • #>.'•-• . • Aloes is a genus of 160 species ofBkerophytic native of Africa. It spreaded into East and West Indies, jfndia, China and other countries. (Chopra, 1982). They range in size from little one-inch miniatures to massive colonies consisting of hundreds of 2-ft diameter plants. Although most Aloes have some medicinal or comnierciaEvalue, the most commonly known is Aloe vera L. It is a large succulent growing up to 1.5 metres in height, with a strong fibrous and a-large stem supporting a of narrow lanceolate . The leaves are whitish green on both sides and bear spiny teeth on the margins. The yellow to drooping grows in a long at the top of the stalk. The is a triangular containing numerous . All Aloes are semitropical succulent plants, and may only be grown outdoors. Aloe vera is a succulent, and^as such,, stores a large quantity of water within its leaves and root system. Works in the applied sciences using in vitro technology were carried out on plants important for agriculture or other spheres of human activity. In vitro technology is a combination of methods Resigned for super effective vegetative propagation of plants. This technology allows to restore whole plants from very small individual fragments of any plant organ or , (i.e. stems, leaves, root tips, flowers, , , grains and so on). The technology is based on the idea that il^s possible in principle to regenerate a whole organism from an individual cell. The problem here is to provide necessary conditions for such regeneration;

1,2 Lecturers, Department of Botany, University of Yangon 3 Associate, Professor, Department of Botany, University of Yangon 4 Adviser, Department of Botany, University of Yangon 250 Jour. Myan. Acad. Ar&& Sc, 2G05 Vol. HI. No. 4(ii) Botany

Cells and tissue culture is. widely used for vegetative propagation called micropropagatioo of the plants. Using in vitro technology some plants, e.g. Aloe, are carried out in micropropasation.

Material aud Methods

(1) Stem tips including apical rneristei.fi.,of Aloe vera L. (2) base from aseptic plant of A. vera L.

Table 1. Composition of basal.MSimedismi (Miirashiage-.&'Skoog, 1962)

CompoiflMd .: '••' .•Amount p;®r.lit<^r of medium .^:.v '• ••; : : NH4O3. • ••< ' / ^ J./1650 : : •nig . .••••.••• ••• ;' :#X • •• .'. .-.V*i.\ • . . : KNQ3. r .•""•* -1900- mg .V : . • .. \ " , • ->? ' • '. KH2PO4 t • 170

$ !V •• •

MgSO4. 7H2O •••' ' 1 37°

.•CaCl2.4H2O.'- • '• ' 1 .-440 mg .

FeSO4. 7H2O | 6.2' mg : • .

m • MnSO4:.4H2O- , •• 1 '."••22. mg

NaMO4. 7H2O ""I '0.25 mg \ • i'A KI\- ,, ' , . •••.• 1 °-83: mg ; '• -.,.. ,.-.•.• V

C11SQ4; 7H2O ' '§0.025 rn'g- . •••••. ; . "

CoCl2. 4H2O .. • 2 0.025. mg

ZnSO4. 7H2O • • '1 8.6 mg

• ':• • •'% - •.

FeSO4. 7H2O • - .• §. ' 27.8 mg ;

Na2EDTA • • -M3 • 37.3 mg Sucrose • p;. 30 g • ;' " •• Jour. Myari. Acad.Arts & Sc. 2005 Vol. III. No. 4(i8) Botany 251

Table 2. Composition of Modified MS Medium

Compound Amount per liter of medium NH4O3 1650^ mg

;KNO3 1900fJmg KH2PO4 170f-mg -L ir-'

M0S04. 7H2O :'.370p.mg j

Ca Cl2. 4H2O 440£mg

FeSO4. 7H2O 6.2|ymg •-.'•'' '^i

MnSO4.4H2O . • .'• . 7 :22i,;.mg- • • . • ."...•-• t' ,,'-• , •' .

NaMO4. 7H2O .. '. • -• 0-25|[mg' ••••;• :..• • 0.8^; mg KI ; { : " -•: '

CuSO4. 7H2O 0.025A mg .

CoCl2:4H2O 0.025« mg

1 •- • . '••. • -f- :- • •'•'• "•• •• ......

Z11SO4. 7H2O 8.6f mg

FeSO4. 7H2O 27.8J mg

Na2EDTA 37-.3V mg

2;4-Dr.'••-•• • ' 0.5| mg Kinetin 0.2f mg . : 1 :...••• ThiaminHCl 0.1? mg

: ' •' •.•• ' . J- ••••.. . •• Pyridoxine. HG1 .0-.5i. mg : Sucrose 252 Jour. Myan.Acad.Arts & Sc.2005 Vol. Hl.vNo; 4(ii) Botany

Culture media: Basal 'MS' medium (Murashige and Skoog,1962 -as shown in Table 1) and modified MS medium (as shown in Table 2).

Surface sterilization: Leafy tips of Aloe vera L.I were washed with water thoroughly under tap. Then, leaves were removed |and leafless tips were swapped with ;70% ethanql. After that they were clean with sterile water and dipped in (1,5;% Cpcbrex solution for 20 minutes. IFinally dissected explants were rinsed with sterile distilled water and inoculated in culture media.

Procedure: After surface sterilization,|stem tips including apical meristem of Aloe vera L. were inoculated in basalt and modified MS medium. The synthetic growing media balanced in minerals was basal MS medium and that was compared to modified MS containing to help the cells grow and callus formation. All the work is; done in totally sterile cabinet in order to avoid the unwelcome growth of fungi and bacteria. Explants are kept in closed vessels. Each vessel contains afgrowing medium with agar. Cultures are placed in specially equipped] rooms with adjustable microclimate: temperature, humidity, and illumination.

^Results ••H ",'"'"•'- . . The explants in both media;developed into . However, growth in basal medium was relatively slow. Growth regulators supported healthy growth of shoots in modified MS>medium. (Table 3) Shoots cultured in vitro:^ere cut and the base was sprouted in the above media. Multiple shoots sprouted in modified MS medium, while basal MS medium induced only a few small snoots. (Table 3)

- • • •" i '•'"'' V . . Segments of leaf-base q^tained from aseptically grown plants in culture were placed on both basal and modified MS media. Further growth and development of explants in basal amedium were not observed until three months. They turned to brown ini colour and died within the culture period. Jour.Myan. Acad. Arts & Sc.2005 Vol. III. No. 4(ii) Botany 253

Gallus formation on explantswas observed only in modified MS medium. It initiated insecond month but it slightly developed in the third month.. Most of them turned brown :in< the fourth month. However,: few culture were survived and well growth of callus mass was observed after subculture. (Table 3)

Table 3. Growth and development of explant in different treatments. Explants Treatments Observation Stem tips from intact plant Basal MS ,; Very slow growth of shoots Modified MS Healthy growth of shoots from in vitro culture Basal MS, Few shoot development from the base in some culture ModifiedMS Development of multiple shoots in culture • . • • ' '• r if

Leaf base from aseptic plant Basal MS v ; Failed to survive ModifiedMS An initiative of callus formation

Discussion Merist^em culture technique is very effective in propagation of rare arid difficult in cultivation plants. This technology also allow propagating unique mutations and hybrid plants existing only in single specimen. In vitro technology allows obtaining about a million specimens from only one meristem. Most, of the plants are obtained using in vitro technology, after that they were cultivated as ordinary plants growing in the soil. Alternatively they may be adapted to the greenhouse conditions.:; The most useful technique for in vitro propagation of Aloe is auxiliary shoot induction. Plants dormant axils are activated and grown to new plants. Some iiloes can be propagated this way while other may be propagated through . Another useful technique is the induction of callus and regeneration of this callus into new shoots. . Although they are different ways: to produce clonal propagatiosn of Aloe, only meristem culture technique has been attempted in the present work. In comparison of basal and modified MS medium, the latter showed better results (Table 3). It mav be 254 Jour. Myatu Acad. Arts & Set2005 Vol. Ill, No. 4(ii) Botany

due to the growth regulators and growth factors (Table 1,2); Many workers have applied combinations of growth regulators ( and ) \ to auxiliary shoot produced in. meristem culture of differentjplant species. IAA and BA in Dianthus caryophyllus; NAA and Kinetin in Chrysanthemum morifolium (Tisserat, 1925). In the present study, combination of 2,4 -D and kinetin has also promoted growth and multiplication of shoot in shoot tiprmeristem culture of Aloe vera L. (Table 3)

References Backer; C: A. 1968. of Java. Vol. lif. N.V.P. NoordhofrGroningen. The Netherland. Bailey, L.H. 1939. The standard Cyclopedia of . Vol. I, A - £.' The Macmillan Co.Ltd., New York. • 'i ' Chopra, R N., I. Q. Chopra, K. L. Handa and L. D. Kapur. 1982, Chopra's Indigenous Drugs of India. 2nd ed, U. N. Dhur& Sons (P) Ltd., Calcutta. Cronquist, 1981, An Integrated System of Classification of, Flowering. Plants. Columbia University Press. New Yqi;kj! ; De, K.K. 1997. An Introduction to Plant-Tissue Culture (Repr.). New Central Book Agency (p.) Ltd., Calcutta. \- .

Hartmann, H.T. and D.E. Kester, 1968. Plant Propagation: Principles and Praties (2nd ed.). Prentice-Hall, Inc. Englewpod Cliffs. New Jassy. 'Hooker, J. D. 1890. Flpra of British, India. 'Vol. VI L, Reeye^ &,.Co. Ltd., London. Street,H.E.1977, Plant Tissue and Cell Culture. (2nd ed.) University, of California Press. California. . Lawrence, H.M. 1964. of Vascular Plants. The Macmillan Company, New York.

• \ ' ' ... Thorpe, T;A- 1981. :. Method and Application in Agriculture. Academic - . Press. Inc. New York. >';. >} •'? ..•• .; .;.'.;: , Tisserat, B. 1985. Embryogenesis, Orgarioge. nesis and Plant Regeneration in R;A Dixon: Edit: Culture A Practical Approach:. IRL press. Washington DC.

' . " .. "'•••. ''• •w-i'J-' '. ' .' ..*; • th •.' •••:. • Trease, G.E and W.C. Evans. 1978, A Text, Book of'Pharmacognosy. (11 ed.) Bailliere; Tindall Cassel. London. ' J * -

••• '•-"'•- '" ' '' ' f, , ' th •'':'1 ••••'•'•.•s":- '-. '- '. Trease, G.E. and W.C. Evans. 2002. Pharmdcognosy. (15 ; ed.) Bailliere;-; Tindall Cassell: London.