An Efficient in vitro Leaf-based Regeneration and Evaluation of Genetic Fidelity Using ISSR Markers in Woodfordia fruticosa (L.) Kurz

Abstract

An efficient in vitro leaf regeneration protocol has been established in Woodfordia fruticosa L. (Lathyraceae), a threatened woody shrub. The leaf segments derived from in vitro-grown plants were cultured on Murashige and Skoog's medium supplemented with Thidiazuron (2.27, 4.54, 6.81, and 9.08 μM) and 6-Benzyladenine (4.4, 8.90, 13.30, and 17.70 μM) alone or in combination with Indole-3-acetic acid (1.14 and 2.28 μM). Maximum number of shoots (15.60 ± 1.15) with highest shoot length (2.90 ± 0.24) were regenerated directly from the leaf explants with a combination of TDZ (4.54 μM) and IAA (2.28 μM), whereas the intervening callus phase was observed in the media supplemented with TDZ or BA alone. The in vitro-regenerated shoots were rooted (100%) on half-strength MS salts fortified with 4.90 μM IBA. The regenerated plantlets hardened in the greenhouse showed an 85% survival rate. ISSR PCR of in vitro-regenerated plantlets reveals their genetic fidelity with the mother plant.

Keywords:

• Woodfordia fruticosa
• PCR
• ISSR
• genetic fidelity

Acknowledgments

Author Mallesham Bulle is thankful to the UGC for the financial assistance in the form of an SAP-DRS fellowship.