ISSN: 0975-766X CODEN: IJPTFI Available Online through Research Article www.ijptonline.com INVITRO REGENERATION OF BANANA (MUSA SPP.) M. Guru Prasad 1, SK. Jaffar* 2, Ch. Manjula 3 Regional Agriculture research station, Tirupathi -517 507. Dept of Biochemistry, Acharya nagarjuna university, Guntur -522 510. Dept of Foods& Nutritional Science, Acharya Nagarjuna University, Guntur- 522 510. Email:shaikjaffar2008@yahoo.com Received on 03-10-2011 Accepted on 18-10-2011 Abstract This study describes a method of propagation for one of the cultural of banana developed by the Indian research agricultural research station. The principle character of this good in its sweetness, good smell and high yield, which has made this particular popular in India. Invitro shoot tip were formed with in 2-3 weeks when meristems were carefully isolated from field grown plants and after proper sterilization implanted in semi solid medium with 2.0 mg/l 6-Benzyl amino purine (BAP). Rate of shoot proliferation increased with BAP and kinetin. Regeneration of cornets was cleanup and shoot multiplications occurred when MS was enriched with 2mg/L BAP and 1.0mg/L Kinetin with the increase of subculture frequency of shoot proliferation was enhanced. Addition of 0.1mg/L IAA, increased shoot elongation and stimulated growth of the shoots, respectively. The micro shoots rooted well within 2 weeks in ½ MS supplemented with 0.5mg/L indol-3-butyric acid. Rooting percentage and growth were much better in high medium in comparison to semi solid medium. Elimination of agar from the root induction media reduced the cost of production significantly. After proper acclimatization, rooted plantlets were transferred to polythene bags containing garden soil and humus (1:1). Two weeks after transplantation, 98% plants survived and flashed new leaves. No morphological variants were observed during the passing of micro propagation. Introduction: Banana (Musa spp.) is the 4 th largest food crop in the world, a staple food for nearly 400 million people and an essential source of income for many natural economics. It is the nutrients fruit, rich in carbohydrates IJPT Dec-2011 Vol. 3 Issue No.4 3440-3445 Page 3440
and minerals as well as good of vitamins. There are few fruit crops that contain as much vitamins as banana and from the aint of view banana appears to enea out compete apple, the most ranked fruit is temperate countries. Banana is one of the most important and most handly grows fruit crops in India. As banana is parts of its traditional propagation is done by small shoots or snekers (4 to 5 sneakers plant) from the parent by multiplication through this method. The major reason is believed to be unavailability of healthy and virms. A free sneaker as traditional propagation is inadequate to restore the affected farmers. Use of plant tissue culture is necessary for germplasm exchange. Conservation and rapid multiplication, which invitro seed germination plays a critical role in generating hybrid plants. Sensual researchers have reported the regeneration of Musa spp via micro propagation. To meet the demand of the farmers to commercialized a good culture like and to immense total banana production in the country a large number of plantlets are needed mithum a specific tree pecoid thus invitro clonal propagation could play a role to reduce this scarcity of plantlets. Materials and Methods Two weeks old sword suckers attached to fruiting mother plants where used as a source of shoot tips and served as the primary explants. Mother plants were carefully inspected. After separating the suckers from the mother plant, the pseudo stem at the lower part containing the meristem was selected. It was washed under running tap water for 15 20min. The unsheathing leaf bases and corn tissues where removed to obtained a block measuring 10cm long, containing the shoot apex. The explants where soaked in tween 80 solution for 7 min and then washed thrice with distilled water. The meristematic tissue were surface sterilized under laminar air flow by immersing in 0.1% mercuric chloride solution for 15min and washed thoroughly 4 times with distilled autoclaved water to remove any traces of mercuric chloride. Then the shoot tip were decapitated and a block of tissue was excised and transferred to MS medium containing different phytoharmones supplemented with 3% sucrose and 0.8% difco bacto agar in test tubes and culture vesicles. The ph of the medium was adjusted to 5.8 before autoclaving and the culture conditions was maintained a 24± 2 0 C under 16hr cool white fluorescent tube light (4000 lux). IJPT Dec-2011 Vol. 3 Issue No.4 3440-3445 Page 3441
Shoot proliferation SK. Jaffar * et al. /International Journal Of Pharmacy&Technology The excised cornlets were transferred aseptically, in a MS medium supplemented with different concentrations (0.5, 0.8, 1.0, 1.5, 1.8, 2.0, 2.2 and 3.0mg/L) of 6-BAP for single shoot regeneration. Multiplication phase The regenerated shoot lets obtained from the earlier experiment, were cut in to small picies and were used as source of explants. The explants were cultured on regeneration medium (multiplication) with different concentrations of BAP and Kinetin to observe the response of cytokines and organic supplements. Root induction phase For root induction, healthy shoot lets were transferred on to half MS medium with IBA of various concentrations (0.1-0.8mg/L). Hardening After sufficient rooting, good and healthy plantlets were properly hardened through gradual exposure to sun light for one week and then transferred to a shade house. Rooted plantlet were carefully was with tap water and transferred to soil, consisting of mixture of vermiculate, coconut peat (1:1) plantlets were kept in the shade house at a relative humidity of 80-90%. Data assembly and scrutiny. Table-1: Effect of benzyl amino purine on shoot multiplication in banana. S.no Growth regulation No of No of shoots % of shoot BAP kinetin explants frequency 1 0.5 0.2 8 - - 2 0.8 0.4 8 - - 3 1.0 0.6 8 2-3 20 4 1.5 0.8 8 3-4 30 5 1.8 0.9 8 4-7 50 6 2.0 1.0 8 7-10 80 7 2.2 1.2 8 5-6 40 8 3.0 1.5 8 5-6 40 IJPT Dec-2011 Vol. 3 Issue No.4 3440-3445 Page 3442
SK. Jaffar * et al. /International Journal Of Pharmacy&Technology Table-2: Effect of indole butyric acid on root initiation on banana. S.no 1 2 3 4 5 6 7 Growth regulator Indole butyric acid 0.1 0.2 0.3 0.4 0.5 0.6 0.8 No of days Rooting frequency poor poor good good good Best Results and Discussion The effect of BAP on shoot proliferation and elongation was tested by adding different concentrations of BAP to a basal medium. Results are presented in (Table-1). A morphogenic response of the cultured shoot tips was visible as a swelling and greenish colour after 10-12 days of inoculation. With shoot length of 1.4 2.5 cm shoot proliferation and elongation response was strongest in the MS medium enriched with BAP at 2mg/L. it is to be noted that in the presence study, shoot culture was examined for six weeks. Frequency of shoot proliferation was increased with increase of subculture in the same medium containing MS + BAP at 2mg/L. Shoot proliferation In this experiment, it has shown good response with the regenerated shoots and then cultured in a MS medium containing different concentrations of BAP and KN. It has shown multiple shoots 2-6 and multiplication frequency (62.9%) and highest number of multiple shoots was achieved in a MS medium with BAP and KN. Explants IJPT Dec-2011 Vol. 3 Issue No.4 3440-3445 Page 3443
showed a reduced rate of multiplication with profuse bulbs and globular structure, stunned clumps and decreased shoot growth at al higher concentrations of BAP and KN. Root induction stage The healthier micro shoots were recovered and sub cultured on half MS with 1mg/L IBA to induce roots and to raise plantlets. The efficiency of auxins for root initiation was evaluated to optimize the medium composition. The rooting ability of recorded and data are presented in (Table-2). When liquid medium was used, plantlets showed expended use and healthy rasomatous bases. Rooting can be stimulated when individual shoots are transferred to a basal medium. Acclimatization and field observations The invitro raised plantlets where transferred to a hardening for 7 days and then acclimatized in to the prepared soil media contain vormiculate and coconut peat and no morphological variations were observed from mother plant. Conclusion The developed clonal propagation method can serves as a convenient method for large scale plant let s to meet the disease free, healthy plants and ultimately it can enhance the economic benefit of the cultivars. Acknowledgement Author thankful to Regional Agriculture research station, Tirupati, providing laboratory facilities to carry out the experiment. References 1. Escalant, J.V. and Jeisson, C.1989.Somatic embryogenesis and plants from immature zygotic embryos of species Musa acuminate and Musa barbisiana.plant cell report 7665-668. 2. Escalant, J.V., Teisson, c. and cote, F.X.1984.Amplified somatic embryogenesis from male flowers of triploid banana and plantain cultivars (Musa spp).in vitro cell development biology 30p:181-186 October 1994.societ for invitro biology. IJPT Dec-2011 Vol. 3 Issue No.4 3440-3445 Page 3444
3. Grapin., A., Schwendimen, J and Teisson, C.1996.Somatic embryogenesis in plantain banana.invitro cellular developmental biolog plant 32:66-71. 4. Jorden, M. and Velizo, J.1996.improvement of somatic embryogenesis in highland papaya cell suspension.plant cell, Tissue and Organ culture 44:189-194. 5. R.A. De fossard, Tissue culture for plant propagator, university of New England, Armidale, 1976. 6. P.Debergh, and R.Zimmermen Ed, Micropropagation technology and application, Kluwer Academic, Dordrecht, 1990, 484pp. 7. T.murashinge and F.skoog. Plant propagation through tissue culture.annu.rev.plant phsiol.25:135-197(1973) 8. O.L.Gamborg, Media preparation, plant tissue culture manual (K.Lindse ed.), Kluwer academic, Dordrecht 1991.ppI-24. 9. R.Levin, V.Gaba, B.Tal, S.Hirsch, D.oenola and I.k.Vasil.Automated plant tissue culture for mass propagation, Biotechnolog 6:1035-1040(1988) 10.Paris,B.,Strosse,H.;Van Den Hende,s.,Swennen,R.Sucrose pre culture to simplify crop reservation of banana meristem cultures.croletters 2002,23,375-384. 11.Zhu,G.;Genius,J.M.C.,Dussert.,S;Swenner,R,Panis,B.Change in sugar, sterol and fatty acid composition in banana meristem caused b sucrose induced acclimation and its effects on crop reservation.plant phsiol.2006,128-80-94. 12. Loescher, W.H.Phsiolog and metabolism of sugar alcohols in higher plants.phsiol.plant 1984, 75,102-109. Corresponding Author: SK. Jaffar*, Email:shaikjaffar2008@yahoo.com IJPT Dec-2011 Vol. 3 Issue No.4 3440-3445 Page 3445