A rapid and efficient protocol for in vitro plant regeneration of Lathyrus sativus L. (Grass pea) through multiple shooting

Abstract

A rapid, simple and reproducible protocol for in vitro multiple shoot induction and whole plant regeneration was developed from two cultivars of Lathyrus sativus L., viz. Nirmal B1 and a locally available variety. Cotyledonary node, shoot tip of five-day-old germinated seeds and nodal explant of ten-day-old seedlings were used as experimental material. The explants were cultured in Murashige and Skoog (MS) media fortified with different concentrations (0.125-2.5 mg/L) of 6- Benzylaminopurine (BAP) and Thidiazuron (TDZ) for multiple shoot proliferation. The maximum number of multiple shoot induction and proliferation (12 – 14 shoots / explant) was observed in cotyledonary node explant within 30 days using 1.5 mg/L BAP. The micro shoots were subcultured in the same media containing low concentration of IAA (0.03 mg/L) for further elongation. Up to 85% of shoots developed roots following their transfer to half strength of Murashige and Skoog MS media containing 0.25 mg/L Indole-3-butyric acid (IBA) within 25 days. Rooted plantlets were successfully hardened under culture conditions and were subsequently established in the greenhouse with survival rate of 85%. The entire in vitro regeneration process took relatively short period of time (90-100 days). Thus, the present study remained successful in developing a rapid and efficient in vitro regeneration through direct organogenesis of Lathyrus sativus for subsequent development of efficient transformation systems.