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Callus induction and plantlet regeneration in Withania somnifera (L.) dunal

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Summary

Callus induction was observed from hypocotyl, root, and cotyledonary leaf segments, grown on Murashige and Skoog (MS) medium supplemented with various concentrations and combinations of 2,4-dichlorophenoxyacetic acid (2,4-D) and kinetin (KN). Maximum callusing (100%) was obtained from root and cotyledonary leaf segments grown on MS medium supplemented with a combination of 2 mg l−1 (9.1 μM) 2,4-D and 0.2 mg l−1 (0.9 μM) KN. The calluses, when subcultured in the same medium, showed profuse callusing. However, these calluses remained recalcitrant to regenerate regardless of the quality and combinations of plant growth regulators in the nutrient pool. When hypocotyl segments were used as explants, callus induction was noticed in 91% of cultures which showed shoot regeneration on MS medium supplemented with 2 mg l−1 2,4-D and 0.2 mg l−1 KN. These shoots were transferred to fresh medium containing various concentrations and combinations of 6-benzyladenine (BA) and N6-(2-isopentenyl)adenosine (2-iP). Maximum shoot multiplication was observed after 60 d of the second subculture on MS medium containing 2 mg l−1 (8.9 μM) BA. These shoots were rooted best (87%) on MS medium containing 2 mg l−1 (9.9 μM) indole-3-butyric acid (IBA). The plantlets were transferred to the field after acclimatization and showed 60% survival.

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References

  • Abbas, A. M.; Jones, J. K.; Caligari, P. D. S. Clonal propagation by in vitro culture of Corchorus (jute). Plant Cell Tiss. Organ Cult. 47:231–238; 1996.

    Article  CAS  Google Scholar 

  • Abhyankar, G. A.; Chinchanikar, G. S. Response of Withania somnifera Dunal leaf explants in vitro. Phytomorphology 46:249–252; 1996.

    Google Scholar 

  • Baburaj, S.; Gunasekaran, K. In vitro differentiation of shoots from leaf callus cultures of Withania somnifera (L.) Dunal. J. Ind. Bot. Soc. 74:323–324; 1995.

    Google Scholar 

  • Banerjee, S.; Schwemmin, A. M.; Schwemmin, D. J. In vitro propagation of Solanum sarrachoides Sendt. J. Plant Physiol. 119:461–466; 1985.

    Google Scholar 

  • Basu, P.; Chand, S. Anthocyanin accumulation in Hyoscyamus muticus L. tissue cultures. J. Biotechnol. 52:151–159; 1996.

    Article  CAS  Google Scholar 

  • Burger, D. W. In vitro micropropagation of Eucalyptus sideroxylon. HortScience 22:496–497; 1987.

    CAS  Google Scholar 

  • Campbell, C. T.; Tomes, D. T. Establishment and multiplication of red clover plants by in vitro shoot tip culture. Plant Cell Tiss. Organ Cult. 3:49–57; 1984.

    Article  CAS  Google Scholar 

  • Chandler, S. F.; Dodds, J. H.; Henshaw, G. G. Factors affecting adventitious shoot formation in Solanum laciniatum Ait. callus cultures. In: 5th Meeting on Plant Tissue Culture; 1982:133–134.

  • Emke, A.; Eilert, U. Steroidal alkaloids in tissue cultures and regenerated plants of Solanum dulcamara. Plant Cell Rep. 5:31–34; 1986.

    Article  CAS  Google Scholar 

  • Filippone, E.; Lurquin, P. F. Stable transformation of egg plant (Solanum melongena L.) by cocultivation of tissues with Agrobacterium tumefaciens carrying a binary plasmid vector. Plant Cell Rep. 8:370–373; 1989.

    Article  CAS  Google Scholar 

  • Furmanowa, M.; Gajdzis-Kuls, D.; Ruszkowska, J.; Czarnocki, Z.; Obidoska, G.; Sadowska, A.; Rani, R.; Upadhyay, S. N. In vitro propagation of Withania somnifera and isolation of withanolides with immunosuppressive activity. Planta Med. 67:146–149; 2001.

    Article  PubMed  CAS  Google Scholar 

  • Gomes-da-Cunha, A. C.; Fernandes-Ferreira, M. Somatic embryogenesis, organogenesis and callus growth kinetics of flax. Plant Cell Tiss. Organ Cult. 47:1–8; 1996.

    Article  Google Scholar 

  • Hammerschlag, F. A.; Bauchan, G. R.; Scorza, R. Factors influencing in vitro multiplication and rooting of peach cultivars. Plant Cell Tiss. Organ Cult. 8:235–242; 1987.

    Article  CAS  Google Scholar 

  • Iriondo, J. M.; de la Iglesia, M.; Perez, C. Micropropagation of Elaeagnus angustifolia from mature trees. Tree Physiol. 15:691–693; 1995.

    PubMed  Google Scholar 

  • Kulkarni, A. A.; Thengane, S. R.; Krishnamurthy, K. V. Direct in vitro regeneration of leaf explants of Withania somnifera (L.). Dunal. Plant Sci. 119:163–168; 1996.

    Article  CAS  Google Scholar 

  • Linington, I. M.; Kew, R. B. G. Micropropagation of Dipterocarpus alatus and D. intricatus. In Vitro 25:Pt. 2, 63A; 1989.

    Google Scholar 

  • Malhotra, C. L.; Das, P. K.; Dhalla, N. S.; Prasad, K. Studies on Withania ashwagandha Kaul. Part III. The effect of total alkaloids on the cardiovascular system and respiration. Ind. J. Med. Res. 49:448–460; 1961.

    CAS  Google Scholar 

  • Mathews, H. Morphogenetic responses from in vitro cultured seedling explants of mung bean (Vigna radiata L. Wilczek). Plant Cell Tiss. Organ Cult. 11:233–240; 1987.

    Article  Google Scholar 

  • Meyers, L. S.; Grossen, N. E. Analysis of independent group designs. In: Behavioral research, theory, procedure and design. San Francisco: W.H. Freeman and Co.; 1974:237–252.

    Google Scholar 

  • Myers, J. M.; Simon, P. W. Continuous callus production and regeneration of garlic (Allium sativum L.) using root segments from shoot tip-derived plantlets. Plant Cell Rep. 17:726–730; 1998.

    Article  CAS  Google Scholar 

  • Murashige, T.; Skoog, F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15:473–497; 1962.

    Article  CAS  Google Scholar 

  • Ozias-Akins, P.; Perera, S. Organogenesis in cultured adventitious root segments and in protoplast derived callus of sweet potato. HortScience 25:121; 1990.

    Google Scholar 

  • Pattnaik, S. K.; Chand, P. K. Rapid clonal propagation of three mulberries, Morus cathayana Hemsl., M. ihou Koiz. and M. serrata Roxb., through in vitro culture of apical shoot buds and nodal explants from mature trees. Plant Cell Rep. 16:503–508; 1997.

    CAS  Google Scholar 

  • Rani, G.; Grover, I. S. In vitro callus induction and regeneration studies in Withania somnifera. Plant Cell Tiss. Organ Cult. 57:23–27; 1999.

    Article  Google Scholar 

  • Rani, K. J.; Rao, T. N.; Raghunatham, G.; Rao, P. V. Studies on callus growth and differentiation in safflower. Ind. J. Genet. 56:458–461; 1996.

    Google Scholar 

  • Rathore, K. S.; Goldsworthy, A. Electrical control of shoot regeneration in plant tissue cultures. Bio/Technology 3:1107–1109; 1985.

    Article  Google Scholar 

  • Ray, S.; Jha, S. Production of withaferin A in shoot cultures of Withania somnifera. Planta Med. 67:432–436; 2001.

    Article  PubMed  CAS  Google Scholar 

  • Roja, G.; Heble, M. R.; Sipahimalani, A. T. Tissue cultures of Withania somnifera: morphogenesis and withanolide synthesis. Phytother. Res. 5:185–187; 1991.

    Article  CAS  Google Scholar 

  • Romano, L. A.; Noronha, C.; Martins-Loucao, M. A. Factors affecting in vitro rooting of Quercus suber. In Vitro 28:Pt. 2, 116A; 1992.

    Google Scholar 

  • Roy, S. C.; Sarkar, A. In vitro regeneration and micropropagation of Aloe vera L. Sci. Hort. 47:107–113; 1991.

    Article  Google Scholar 

  • Sen, J.; Sharma, A. K. Micropropagation of Withania somnifera from germinating seeds and shoot tips. Plant Cell Tiss. Organ Cult. 26:71–73; 1991.

    Article  CAS  Google Scholar 

  • Shahzad, A.; Hasan, H.; Siddiqui, S. A. Callus induction and regeneration in Solanum nigrum L. in vitro. Phytomorphology 49:215–220; 1999.

    Google Scholar 

  • Sharma, K. K.; Thorpe, T. A. In vitro propagation of mulberry (Morus alba L.) through nodal segments. Sci. Hort. 42:307–320; 1990.

    Article  Google Scholar 

  • Sharma, S. K.; Ramamurthy, V. Micropropagation of 4-year-old elite Eucalyptus tereticornis trees. Plant Cell Rep. 19:511–518; 2000.

    Article  CAS  Google Scholar 

  • Subhash, K.; Christopher, T. Direct plantlet formation in cotyledon cultures of Capsicum frutescens. Curr. Sci. 57:99–100; 1988.

    Google Scholar 

  • Sumana, K. R.; Kaveriappa, K. M.; Bhat, H. S. In vitro micropropagation of Holarrhena pubescens. J. Med. Arom. Plant Sci. 21:299–303; 1999.

    CAS  Google Scholar 

  • Suzuki, S.; Fujino, H.; Tatsuo, Y.; Yamazaki, N.; Yoshizaki, M. Rapid propagation of Stephania cephalantha Hayata by tissue culture. Jap. J. Breed. 42:769–777; 1992.

    Google Scholar 

  • Tandon, P.; Rathore, T. S. Regeneration of plantlets from hypocotyl-derived callus of Coptis teeta. Plant Cell Tiss. Organ Cult. 28:115–117; 1992.

    Article  Google Scholar 

  • Thakur, R.; Rao, P. S.; Bapat, V. A. In vitro plant regeneration in Media azedarach. Plant Cell Rep. 18:127–131; 1998.

    Article  CAS  Google Scholar 

  • Tiwari, S. K.; Singh, S. P. Micropropagation of Withania somnifera by tissue culture. Van. Sandesh 15:19–20; 1991.

    Google Scholar 

  • Tiwari, V.; Singh, B. D.; Tiwari, K. N. Shoot regeneration and somatic embryogenesis from different explants of Brahmi [Bacopa monnieri (L.) Wettst.]. Plant Cell Rep. 17:538–543; 1998.

    Article  CAS  Google Scholar 

  • Tyagi, P.; Kothari, S. L. Micropropagation of Capparis decidua in vitro shoot proliferation on nodal explants of mature tree and seedling explants. J. Plant Biochem. Biotechnol. 6:19–23; 1997.

    Google Scholar 

  • Uma Devi, P.; Sharada, A. C.; Solomon, F. E. Antitumor and radiosensitizing effects of Withania somnifera (Ashwagandha) on a transplantable mouse tumor sarcoma 180. Ind. J. Exp. Biol. 31:607–611; 1993.

    Google Scholar 

  • Uma Devi, P.; Sharada, A. C.; Solomon, F. E.; Kamath, M. S. In vivo growth inhibitory effect of Withania somnifera (Ashwagandha) on a transplantable mouse tumor sarcoma 180. Ind. J. Exp. Biol. 30:169–172; 1992.

    Google Scholar 

  • Varghese, S. K.; Inamdar, J. A.; Kalia, K.; Subramanian, R. B.; Nataraj, M. Micropropagation of Aegle marmelos (L.) Cor. Phytomorphology 43:87–92; 1993.

    Google Scholar 

  • Zhao, Y. X.; Yao, D. Y.; Harris, P. J. C. Plant regeneration from callus and explants of Sesbania spp. Plant Cell Tiss. Organ Cult. 34:253–260; 1993.

    Article  CAS  Google Scholar 

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  1. Department of Botanical and Environmental Sciences, Guru Nanak Dev University, 143 005, Amritsar, India

    Gita Rani, G. S. Virk & Avinash Nagpal

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  1. Gita Rani
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  2. G. S. Virk
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  3. Avinash Nagpal
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Correspondence to Avinash Nagpal.

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Rani, G., Virk, G.S. & Nagpal, A. Callus induction and plantlet regeneration in Withania somnifera (L.) dunal. In Vitro Cell.Dev.Biol.-Plant 39, 468–474 (2003). https://doi.org/10.1079/IVP2003449

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