In Vitro Plant Regeneration of Morus indica L. cv. V1 Using Leaf Explant


Adventitious bud induction and plantlet regeneration were studied in a popular mulberry variety, V1 using leaf as an explant. Fully expanded leaf explants were cultured on Murashige and Skoog’s (MS) medium supplemented with thidiazuron (TDZ) (0.5-4.0 mg/l), 6-benzylaminopurine (BAP) (0.5-2.0 mg/l), indole acetic acid (IAA) (2.0 mg/l), gibberlic acid (GA3) (1.0-2.0 mg/l) silver nitrate (AgNO3) (2.0 mg/l) and different carbon sources such as sucrose, fructose and glucose (10%-30%) either individually or in combination to induce adventitious buds and regeneration. The highest percentage (63%) of adventitious bud formation and regeneration (68%) was achieved in the medium containing MS with TDZ (1.0 mg/l), IAA (2.0 mg/l) and AgNO3 (2.0 mg/l). For subsequent regeneration and shoot elongation the MS medium having BAP (1.0 mg/l), GA3 (2.0 mg/l) and AgNO3 (2.0 mg/l) was found to be suitable. Amongst the carbon sources tested, the most suitable carbon source was found to be sucrose (3%) followed by fructose (2%) for adventitious bud formation. Excised in vitro shoots were rooted (60%-80%) in half strength MS medium supplemented with indole-3-butyric acid (1.0 mg/l). The well rooted plantlets were hardened in soil + sand + farm yard manure (FYM) mixture with a success rate of 70%-90%. Since in vitro regeneration is highly genotype-dependent in mulberry, the standardized protocol can be effectively used for further improvement of this leading genotype using biotechnological approaches.

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M. Raghunath, K. Nataraja, J. Meghana, R. Sajeevan, M. Rajan and S. Hussaine Qadri, "In Vitro Plant Regeneration of Morus indica L. cv. V1 Using Leaf Explant," American Journal of Plant Sciences, Vol. 4 No. 10, 2013, pp. 2001-2005. doi: 10.4236/ajps.2013.410249.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] B. C. Das, “Mulberry Taxonomy, Cytogenetics and Breeding,” National Seminar on Silk Research and Development, Bangalore, 10-13 March 1983.
[2] S. Oka and K. Ohyama, “In Vitro Initiation of Adventitious Buds and Its Modification by High Concentration of Benzyladenine in Leaf Tissue of Mulberry (Morus alba),” Canadian Journal of Botany, Vol. 59, No. 1, 1981, pp. 68-74.
[3] P. Narayan, S. Chakraborthy and G. Subba Rao, “Regeneration of Plantlets from Callus of Stem Segments of Mature Plant Morus alba L.,” Proceedings of the Indian National Science Academy, Vol. B5, 1989, p. 469.
[4] A. K. Jain and R. K. Datta, “Shoot Organogenesis and Plant Regeneration Mulberry (Morus bombycis koidz): Factors Influencing Morphogenetic Potential in Callus Cultures,” Plant Cell, Tissue and Organ Culture, Vol. 29, No. 1, 1992, pp. 43-50.
[5] H. Machii, “Organogenesis from Immature Leaf Cultures in Mulberry, Morus alba L.,” Journal of Sericultural Science of Japan, Vol. 61, No. 6, 1992, pp. 512-519.
[6] Sreenathrao and M. K. Raghunath, “Callus Initiation, Growth and Plant Regeneration in Mulberry (Morus alba L.) Cultivars,” Journal of the Swamy Botanical Club, Vol. 10, 1993, pp. 17-21.
[7] T. Sawaguchi, H. Yamunouchi, A. Koyama and H. Machii, “Effect of Culture Conditions on Adventitious Bud Formation from Cotyledons and Primary Leaves of Mulberry,” Journal of Sericultural Science of Japan, Vol. 68, No. 50, 1992, pp. 362-363.
[8] K. A. Kathiravan, Ganapathi and A. Shajahan, “Adventitious Shoot Formation and Plant Regeneration from Callus Cultures of Mulberry (Morus alba L.),” Sericologia, Vol. 37, 1997, pp. 727-733.
[9] K. Vijayan, S. P. Chakraborthy and B. N. Roy, “Plant Regeneration from Leaf Explants of Mulberry: Influence of Sugar, Genotype and 6-Benzyladenine,” Indian Journal of Experimental Biology, Vol. 38, No. 5, 2000, pp. 504-508.
[10] A. Kapur, S. Bhatnagar and P. Khurana, “Efficient Regeneration from Mature Leaf Explants of Indian Mulberry via Organogenesis,” Sericiologia, Vol. 41, 2001, pp. 207-214.
[11] S. Bhatnagar, A. Kapur and P. Khurana, “TDZ-Mediated Differentiation in Commercially Valuable Indian Mulberry M. indica Cultivars K2 and DD,” Plant Biotechnology, Vol. 18, No. 1, 2001, pp. 61-65.
[12] R. S. Sajeevan, S. Jeba Singh, Karaba N. Nataraja and M. B. Shivanna, “An Efficient in Vitro Protocol for Multiple Shoot Induction in Mulberry, Morus alba L. Variety V1,” International Research Journal of Plant Sciences, Vol. 2, No. 8, 2011, pp. 254-261.
[13] T. Murashige and F. Skoog, “A Revised Medium for Rapid Growth and Bioassay with Tobacco Tissue Cultures,” Plant Physiology, Vol. 15, No. 3, 1962, pp. 473-497.
[14] P. Khurana, S. Bhatnagar and S. Kumari, “Thidiazuron and Woody Plant Tissue Culture,” Journal of Plant Biology, Vol. 32, 2005, pp. 1-12.
[15] M. K. Raghunath and S. Ravindran, “High Frequency Adventitious Bud Induction and Plant Regeneration from Leaf Explants of Mulberry (Morus spp.) Genotypes,” National Conference on Sericulture Innovations Before and Beyond, CSR&TI, Mysore, 28-29 January 2011, p. 4.
[16] M. K. Raghunath, S. Lal and P. Khurana, “In Vitro Plant Regeneration from Different Explants of Elite Mulberry (Morus Sp.) Genotypes: AR-12, DD and S-13,” Bangladesh Journal of Sericology, Vol. 2 & 3, 2009, pp. 31-40.
[17] M. Hossain, S. M. Rahman, A. Zaman, Jorader and R. Islam, “Micropropagation of Morus laevigata Wall. From Mature Trees,” Plant Cell Reports, Vol. 11, No. 10, 1992, pp. 522-524.
[18] S. Bhatnagar, M. Das and P. Khurana, “Rapid in Vitro TDZ-Mediated Micropropagation of M. Indica C176 and C776 through Axillary Buds,” Indian Journal of Sericology, Vol. 41, 2002, pp. 80-83.

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