Micropropagation and tissue culture of the endangered medicinal plant (Withania somnifera L.) in Bangladesh

Paper Details

Research Paper 01/04/2019
Views (252) Download (14)
current_issue_feature_image
publication_file

Micropropagation and tissue culture of the endangered medicinal plant (Withania somnifera L.) in Bangladesh

Md. Forhad Hossain, Md. Naimul Hassan, Sumon Karmakar, Mutasim Billah, Md Abu Reza, Khandaker Md. Khalid-Bin- Ferdaus
Int. J. Biosci.14( 4), 292-304, April 2019.
Certificate: IJB 2019 [Generate Certificate]

Abstract

The present study was under taken to establish an efficient protocol considering various cultural aspects for plant regeneration, multiplication and rooting in vitro of Withania somnifera L. because of its medicinal value and to its survivability. Shoot tip and nodal segment were cultured aseptically on MS medium containing different concentrations and combinations of cytokinins (BAP and KIN) and auxins (NAA, IBA, and 2, 4-D) for direct and indirect regeneration. The explants were inoculated on MS medium supplemented with 8g/l Agar and 30g/l sucrose, different combination and concentration of plant growth regulators (auxin and cytokinins) was used for plant regeneration. The optimum pH was adjacent to 5.8 and autoclaved at 121±2°C for 20 minutes and incubated at 25±2°C under 16/8 hours photoperiod. In this study, the best shoot multiplication for direct regeneration was observed in 2.0mg/l BAP+0.2mg/l NAA, the longest shoot was observed 5.20cm and 5.00cm respectively. In case of indirect regeneration, highest response of callus was obtain by shoot tip (80.2%) and nodal segment (84.0%) culture on MS medium supplemented with 2mg/l 2, 4-D+0.2mg/l KIN. The best proliferation (6.5%) of adventitious bud was observed in BAP 2mg/l maximum rooting of the micro cuttings of regenerated shoots were successfully achieved with 2.0mg/l IBA and 1/2 MS+IBA2mg/l, the result was obtained 85% and 80% respectively. The plantlets were successfully transferred to soil and the percentage of survivability in such condition was obtains 87%. Micropropagation technique is the most effective way to rescue this medicinally endangers plant.

VIEWS 9

Abhyankar G, Chinchanikar G. 1996. Response of Withania somnifera Dunal leaf explants in vitro. Phytomorphology 46(3), 249-252.

Ahmad M, Saleem S, Ahmad AS, Ansari MA, Yousuf S, Hoda MN, Islam F. 2005. Neuroprotective effects of Withania somnifera on 6-hydroxydopamine induced Parkinsonism in rats. Human & experimental toxicology 24(3), 137-147.

Al-Khayri JM, Huang FH, Morelock TE. 1991. Regeneration of spinach from leaf callus. HortScience 26(7), 913-914.

Andallu B, Radhika B. 2000. Hypoglycemic, diuretic and hypocholesterolemic effect of winter cherry (Withania somnifera, Dunal) root.

Antonisamy R, Manickam V. 1999. Conservation through micropropagration and restoration of selected rare and endangered medicinal plants of South India, XVI International Botanical Congress. held in, 1-7.

Bhattacharya SK, SatyanK, Ghosal S. 1997. Antioxidant activity of glycowithanolides from Withania somnifera. Indian Journal of Experimental Biology 35(3), 236-239.

Bornman CH. 1983. Possibilities and constraints in the regeneration of trees from cotyledonary needles of Picea abies in vitro. Physiologia Plantarum 57(1), 5-16.

Chopra R, Chopra I, Handa K, Kapur L. 1958. Indigenous Drugs of India (p 436–437). UN Dhur and Sons, Calcutta.

Datta K, Datta S. 1984. Rapid clonal multiplication of Angelonia salicariefolia through tissue culture. Plant cell, tissue and organ culture 3(3), 215-220.

Fanizza G, Ricciardi L. 1988. The response of a range of genotypes of Vitis vinifera to sequential shoot tip cultures at high temperatures. Euphytica 39(1), 19-23.

Furmanowa M, Gajdzis-Kuls D, Ruszkowska J, Czarnocki Z, Obidoska G, Sadowska, A. Upadhyay SN. 2001. In vitro propagation of Withania somnifera and isolation of withanolides with immuno suppressive activity. Planta Medica 67(02), 146-149.

Henselová M, Slováková Ľ, Martinka M, Zahoranová A. 2012. Growth, anatomy and enzyme activity changes in maize roots induced by treatment of seeds with low-temperature plasma. Biologia 67(3), 490-497.

Kattimani K, Reddy Y, Rao BR.1999. Effect of pre-sowing seed treatment on germination, seedling emergence, seedling vigour and root yield of Ashwagandha (Withania somnifera Daunal.). Seed Science and technology 27(2), 483-488.

Kirtikar K, Basu B. 1918. Indian Medicinal Plants Vol-3: Bishen Singh Mahendra Pal Singh And Periodical Experts.

Kulkarni AA, Thengane S, Krishnamurthy K. 1996. Direct in vitro regeneration of leaf explants of Withania somnifera (L.) Dunal. Plant Science 119(1-2), 163-168.

Kulkarni AA, Thengane S, Krishnamurthy K. 2000. Direct shoot regeneration from node, internode, hypocotyl and embryo explants of Withania somnifera. Plant cell, tissue and organ culture 62(3), 203-209.

Kulkarni S, Dhir A. 2008. Withania somnifera: an Indian ginseng Progress in neuro-psychopharmacology and biological psychiatry 32(5), 1093-1105.

Majumdar D. 1955. Withania somnifera Dunal. Part II: alkaloidal constituents and their chemical characterizationInd. Journal of Pharmacology 17, 158-161.

Mohanty I, Arya DS, Dinda A, Talwar KK, Joshi S, Gupta SK. 2004. Mechanisms of cardioprotective effect of Withania somnifera in experimentally induced myocardial infarction. Basic & Clinical Pharmacology & Toxicology 94(4), 184-190.

Murashige T, Skoog F. 1962. A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiologia Plantarum 15(3), 473-497.

Ohyama K, Oka S. 1980. Bud and root formation in hypocotyl segments of Broussonetia kazinoki Sieb. in vitro. Plant and Cell Physiology 21(3), 487-492.

Pramanik TK, Datta S. 1986. Plant regeneration and ploidy variation in culture derived plants of Asclepias curassavica L. Plant Cell Reports 5(3), 219-222.

Rani G, Grover I. 1999. In vitro callus induction and regeneration studies in Withania somnifera. Plant cell, tissue and organ culture 57(1), 23-27.

Rani G, Virk G, Nagpal A. 2003. Callus induction and plantlet regeneration in Withania somnifera (L.) Dunal. In Vitro Cellular & Developmental Biology-Plant 39(5), 468-474.

Ray S, Jha S. 2001. Production of withaferin A in shoot cultures of Withania somnifera. Planta Medica 67(05), 432-436.

Roja G, Heble M, Sipahimalani A. 1991. Tissue cultures of Withania somnifera: morphogenesis and withanolide synthesis. Phytotherapy Research 5(4), 185-187.

Saritha K, Naidu C. 2007. In vitro flowering of Withania somnifera Dunal.—an important antitumor medicinal plant. Plant Science 172(4), 847-851.

Selva E, Stouffs B, Briquet M. 1989. In vitro propagation of Vicia faba L. by micro-cutting and multiple shoot induction. Plant cell, tissue and organ culture 18(2), 167-179.

Sen J, Sharma A. 1991. Micropropagation of Withania somnifera from germinating seeds and shoot tips. Plant cell, tissue and organ culture 26(2), 71-73.

Sharma KK, Thorpe TA. 1990. In vitro propagation of mulberry (Morus alba L.) through nodal segments. Scientia Horticulturae 42(4), 307-320.

Sharma M, Ali D, Batra A. 2010. Plant regeneration through in vitro somatic embryogenesis in Ashwagandha: Researcher.

Siddique N, Bari M, Shahnewaz S, Rahman M, Hasan M, Khan M, Islam M. 2004. Plant Regeneration of Withania somnífera (L.) Diurni (Ashwagandha) from Nodal Segments Derived Callus an Endangered Medicinal Plant in Bangladesh. Journal of Biological Sciences 4(2), 219-223.

Singh B, Saxena A, Chandan B, Gupta D, BhutaniK, Anand K. 2001. Adaptogenic activity of a novel, withanolide free aqueous fraction from the roots of Withania somnifera Dun. Phytotherapy Research 15(4), 311-318.

Sivanesan I. 2007. Direct regeneration from apical bud explants of Withania somnifera Dunal.

Sivanesan I, Murugesan K. 2008. An Efficient Regeneration from Nodal Expiants of Withania somnífera Dun.  Asian Journal of Plant Sciences 7(6), 551-556.

Tohda C, Kuboyama T, Komatsu K. 2005. Search for natural products related to regeneration of the neuronal network Neurosignals 14(1-2), 34-45.

Vakeswaran V, Krishnasamy V. 2003. Improvement in storability of Ashwagandha (Withania somnifera Dunal) seeds through pre-storage treatments by triggering their physiological and biochemical properties. Seed Technology 203-203.