shoots with a length of (3 - 5 cm) were excised and transferred to MS medium supplemented with different concentrations of auxins such as IBA, IAA and NAA (0.1 - 2.0 mg/l) alone and in combination with KIN. In all the concentrations tried, highest number of roots was observed on MS medium with IBA 1 mg/L + 0.2 mg/L KN. Highest rooting frequency (85%), with a maximum number of roots (12.2 ± 0.53) was observed in this combination, results are tabulated in Table 3.

4. Conclusion

From the results it is noticeable that fungicide bavistin, ethylene inhibitor silver thiosulphate does not shows any harmful effect on shoot regeneration of Asclepias curassavica. Thus these promissory compounds may be useful as a media supplement to develop efficient protocols for in vitro propagation as it favors the shoot formation. Casein hydrolysate can be used as an important growth factor that promotes shoot regeneration frequency of Asclepias curassavica in the present study. Activated charcoal

Table 3. Effect of different growth adjuvants on multiple shoot regeneration from nodal explants of A. curassavica in MS media supplemented with different growth regulators. Observations: After 4 weeks (The results are mean (SE±) of 20 independent determinations).

removed the phenolic exudates completely and prevented the browning of media and thus enhanced the frequency of regeneration.

5. Acknowledgements

S Hemadri Reddy would like to thank Department of Biotechnology, S.V. University, Tirupati, Andhra Pradesh for providing Doctoral fellowship to complete this work.


  1. M. E. Endress and P. V. Bruyns, “A Revised Classification of the Apocynaceae s.l.,” Botanical Review, Vol. 66, No. 1, 2000, pp. 1-56. doi:10.1007/BF02857781
  2. S. S. Samant, U. Dhar and L. M. S. Palni, “Medicinal Plants of Indian Himalaya: Diversity, Distribution Potential Values,” Himavikas Publication, No. 13, Gyan Prakash, Nainital, 1998.
  3. N. Komalavalli and M. V. Rao, “In Vitro Micropropagation of Gymnema elegans—A Rare Medicinal Plant,” Indian Journal of Experimental Biology, Vol. 35, 1997, pp. 1088-1092.
  4. P. Knogstrup, S. Baldursson and J. V. Norgard, “Ex Situ Genetic Conservation by Use of Tissue Culture,” Operational Botany, Vol. 113, 1992, pp. 49-53.
  5. M. F. Fay, “In What Is in Vitro Culture Appropriate to Plant Conservation?” Biodiversity and Conservation, Vol. 3, No. 2, 1994, pp. 176-183. doi:10.1007/BF02291887
  6. I. K. Vasil and T. A. Thorpe, “Plant Cell and Tissue Culture,” Kluwer Academic Publishers, Dordrecht, 1994.
  7. S. Govil and S. C. Gupta, “Commercialisation of Plant Tissue Culture in India,” Plant Cell, Tissue and Organ Culture, Vol. 51, No. 1, 1997, pp. 65-73. doi:10.1023/A:1005873221559
  8. R. Shields, S. J. Robinson and P. A. Anslow, “Use of Fungicides in Plant Tissue Culture,” Plant Cell Reports, Vol. 3, No. 1, 1984, pp. 33-36. doi:10.1007/BF00270226
  9. L. Burgos and N. Alburquerque, “Ethylene Inhibitors and Low Kanamycin Concentrations Improve Adventitious Regeneration from Apricot Leaves,” Plant Cell Reports, Vol. 21, No. 12, 2003, pp. 1167-1174. doi:10.1007/s00299-003-0625-6
  10. E. M. Beyer, “Effect of Silver Ion, Carbon Dioxide, and Oxygen on Ethylene Action and Metabolism,” Plant Physiology, Vol. 63, No. 1, 1979, pp. 169-173. doi:10.1104/pp.63.1.169
  11. G. L. Chi, D. G. Barfield, G. E. Sim and E. C. Pua, “Effect of AgNO3 and Aminoethoxyvinylglycine on in Vitro Shoot and Root Organogenesis from Seedling Explants of Recalcitrant Brassica genotypes,” Plant Cell Reports, Vol. 9, No. 4, 1990, pp. 195-198. doi:10.1007/BF00232178
  12. D. D. Songstad, D. R. Duncan and J. M. Widholm, “Effect of 1-Aminocyclopropane-1-carboxylic Acid, Silver Nitrate, and Noborn-Adiene on Plant Regeneration from Maize Callus Cultures,” Plant Cell Reports, Vol. 7, No. 4, 1988, pp. 262-265. doi:10.1007/BF00272538
  13. K. M. Charaibi, A. Latche, J. P. Roustan and J. Fallot, “Stimulation of Shoot Regeneration from Cotyledons of Helianthus annuus by the Ethylene Inhibitors, Silver and Cobalt,” Plant Cell Reports, Vol. 10, 1991, pp. 204-207.
  14. H. P. Bais, G. Sudha and G. A. Ravishankar, “Influence of Putrescine, Silver Nitrate and Polyamine Inhibitors on the Morphogenetic Response in Untransformed and Transformed Tissues of Cichorium intybus and Their Regenerants,” Plant Cell Reports, Vol. 20, No. 6, 2001, pp. 547-555. doi:10.1007/s002990100367
  15. P. Sujana and C. V. Naidu, “Influence of Bavistin, Cefotaxime, Kanamycin and Silver Thiosulphate on Plant Regeneration of Mentha piperita (L.)—An Important Multipurpose Medicinal Plant,” Journal of Phytology, Vol. 3, 2011, pp. 36-40.
  16. D. Preethi, T. M. Sridhar and C. V. Naidu, “Effect of Bavistin and Silver Thiosulphate on in Vitro Plant Regeneration of Stevia rebaudiana,” Journal of Phytology, Vol. 3, 2011, pp. 74-77.
  17. P. Bantawa, O. S. Roy, P. Ghosh and T. K. Mondal, “Effect of Bavistin and Adenine Sulphate on in Vitro Shoot Multiplication of Picrorhiza scrophulariiflora Pennell.: An Endangered Medicinal Plant of Indo-China Himalayan Regions,” Plant Tissue Culture and Biotechnology, Vol. 19, No. 2, 2009, pp. 237-245.
  18. N. Komalavalli and M. V. Rao, “In Vitro Micropropagation of Gymnema sylvestre—A Multipurpose Medicinal Plant,” Plant Cell, Tissue and Organ Culture, Vol. 61, No. 2, 2000, pp. 97-105. doi:10.1023/A:1006421228598
  19. C. J. Delp, “Benzimidazole and Related Fungicides,” In: H. Lyr, Ed., Modern Selective Fungicides, 2nd Edition, Gustav Fisher Verlag, Jena, 1987, pp. 291-303.
  20. R. K. Tripathi and S. Ram, “Induction of Growth and Differentiation of Carrot Callus Cultures by Carbendazim and Benzimidazole,” Indian Journal of Experimental Biology, Vol. 20, 1982, pp. 674-677.
  21. P. C. Garcia, R. M. Rivero, J. M. Ruiz and L. Romero, “The Role of Fungicides in the Physiology of Higher Plants: Implications for Defense Responses,” Botanical Review, Vol. 69, No. 2, 2003, pp. 162-172. doi:10.1663/0006-8101(2003)069[0162:TROFIT]2.0.CO;2
  22. K. G. M. Skene, “Cytokinin-Like Properties of the Systemic Fungicide Benomyl,” Journal of Horticultural Science, Vol. 47, 1972, pp. 179-182.
  23. T. H. Thomas, “Investigations into the Cytokinin-Like Properties of Benzimidazole Derived Fungicides,” Annals of Applied Biology, Vol. 76, No. 2, 1974, pp. 237-241. doi:10.1111/j.1744-7348.1974.tb07977.x
  24. D. Raghuramulu, “In Vitro Morphogenetic Studies of Hemidesmus indicus and Cynanchum callialatum. (Asclepiadaceae),” Ph.D. Thesis, Sri Krishnadevaraya University, Ananthapur, 2001.
  25. R. W. Joy-Iv, K. R. Patel and T. A. Thorpe, “Ascorbic Acid Enhancement of Organogenesis in Tobacco Callus,” Plant Cell, Tissue and Organ Culture, Vol. 13, No. 3, 1988, pp. 219-228. doi:10.1007/BF00043670
  26. A. Ahuja, M. Verma and S. Grewal, “Clonal Propagation of Ocomum Species by Tissue Culture,” Indian Journal of Experimental Biology, Vol. 20, 1982, pp. 455-458.
  27. N. Sharma and K. P. S. Chandel, “Effect of Ascorbic Acid on Axillary Shoot Induction in Tylophoro indica (Burm. f.) Merril,” Plant Cell, Tissue and Organ Culture, Vol. 29, No. 2, 1992, pp. 109-113. doi:10.1007/BF00033615
  28. V. Sarasan, E. Y. Sonia and G. M. Nair, “Regeneration of Indian Sarasapilla, Hemidesmus indicus R. Br., through Organogenesis and Somatic Embryogenesis,” Indian Journal of Experimental Biology, Vol. 32, 1994, pp. 284- 287.
  29. E. F. George and P. D. Sherrington, “Handbook and Directory of Commercial Laboratories,” In: Plant Propagation by Tissue Culture, Exegetics Ltd., Eversley, 1984, p. 709.
  30. C. G. Sudha, “Tissue Culture Studies on Three Rare Medicinal Plants of India, Holostemma anulare, Janakia arayalpathra and Rauwolfia micrantha,” Ph.D. Thesis, University of Kerala, India, 1996.
  31. T. Chakradhar, “In Vitro Culture, Physiological, Phytochemical and Antimicrobial Studies of a Medicinal Plant. Wattakaka volubilis (L.f.) stapf (Asclepiadaceae),” Ph.D. Thesis Submitted to S.K. University, Ananthapur, 2004.


*Corresponding author.

Journal Menu >>