Shoot regeneration from leaf explants of Withania coagulans (Stocks) Dunal and genetic stability evaluation of regenerates with RAPD and ISSR markers

Abstract

Edited by J Van StadenKeywords: Endangered Ex vitro Leaf regeneration Vegetable rennet and withanolides Withania coagulans (Stocks) Dunal (Solanaceae) is a critically endangered medicinal plant known for its multiple medicinal properties. We report here an efficient and high frequency plantlet regeneration system through direct organogenesis in W. coagulans using in vitro-derived leaves as explants. On Murashige and Skoog's (MS) medium containing 4.44 μM 6-benzylaminopurine (BAP) 73.7 ± 4.3% explants responded and produced 11.4 ± 0.9 shoot buds per explant. The regenerated shoot buds were elongated (6.7 ± 0.22 cm) on MS medium with 1.11 μM BAP and 0.57 μM indole-3-acetic acid (IAA). The elongated shoots were rooted both in vitro and ex vitro. The regenerates were acclimatized by slow and gradual exposure to different regimes of temperature and relative humidity. The leaf-regenerates were assessed for genetic stability using RAPD and ISSR molecular markers and confirmed true-to-type. The in vitro regeneration system developed would be useful for genetic restoration program as the overexploitation and reproductive failure forced W. coagulans towards the verge of complete extinction. The regeneration method would also be useful for genetic transformation for genetic improvement and improved phyto-pharming.

RAPD and ISSR Marker-Based Comparative Evaluation of Genetic Diversity Among Indian Germplasms of Euryale ferox: an Aquatic Food Plant

“…The RAPD marker has been utilized in detecting genetic variability in several species of plants [8][9][10][11][12]. The ISSR marker has been also proven to provide a strong, reproducible, easy, fast and economical means to evaluate genetic variability and identify variations between narrowly associated genotypes in several species of plants [11][12][13][14][15][16].…”

An Efficient Method for Direct Shoot Regeneration from Leaf Explants of Solanum nigrum Induced by Thidiazuron

“…Similar results of regeneration from leaf explants using TDZ has been reported in primrose (Gyves et al, 2001), mulberry (Chitra and Padmaja, 2005), Paulownia tomentosa (Corredoira et al, 2008) and Jatropha curcus (KhuranaKaul et al, 2010), which are in agreement with our results. Similarly, In vitro shoot regeneration has also been reported in plants of family Solanaceae (Magioli et al, 1998;Mallaya and Ravishankar, 2013;Rathore et al, 2016). While, Hussain et al (2005) reported that field grown explants of different varieties of S. tuberosum had high regeneration potential.…”

In vitro Shoot Multiplication of Withania coagulans (Stocks) Dunal

“…High concentration of TDZ resulted in hyperhydricity of shoots reported in Pluchea lanceolata (Kher et al 2014). TDZ at a higher concentration resulted in callus formation from leaf explants derived from in vitro raised shoots in W. coagulans is reported recently by Rathore et al (2016). Effectiveness of mT for axillary shoot multiplication and adventitious regeneration is reviewed by Aremu et al (2011).…”