In vitro Plant Regeneration from Cotyledonary Node Explants of Pigeon Pea (Cajanus cajan L.) Mill sp. cv. Vamban II via Direct Organogenesis

S Naresh

Abstract


A simple and efficient protocol was developed for in vitro propagation of Pigeon pea [Cajanus cajan (L.) Mill sp.], Vamban II. The seeds were removed from the pods and the cotyledon along with embryonal axes were cultured on MS medium, with 3% sucrose and supplemented with various concentrations of BAP and KIN (1 mg/L, 2 mg/L, 5 mg/L and 10 mg/L). The cotyledonary node along with the mass of shoot - initials originating from 12 day old seedling were excised by removing the epicoyl and cotyledons within 1 - 2 mm of the node and used as explants. Direct shoot initials were observed from cotyledonary node explants after 2 weeks of culture initiation. The maximum number of multiple shoots was observed at the concentration of 1 mg/L of BAP. Addition of IAA (0.5 mg/L) to BAP (1 mg/L, 2 mg/L, 5 mg/L and 10 mg/L) enhanced the callus with multiple shoots. The optimum level of BAP (with 0.5 IAA) that promoted the highest number of multiple shoot (6.4 shoots/explants) was 1 mg/L. Addition of NAA (0.5 mg/L) to BAP (1, 2, 5, 10 mg/L) enhances shoot bud with callus. Multiple shoots (4) were observed in the medium supplemented with NAA (0.5 mg/L) and BAP (1 mg/L). Well developed elongated shoots (2 to 3 cm) were excised and  cultured on ½ strength MS medium supplemented with different concentrations of IBA (0.1 - 0.5 mg/L) for root induction. Among these concentrations, 0.3 mg/L IBA produced the maximum number of roots. The highest percentage of plant regeneration via multiple shoots was noticed on MS medium supplemented with 1.0 mg/L. The mean number of shoots decreased with increasing in the concentration of BAP up to 1.0 mg/L. In addition cotyledonary node system has been efficiently used recently for Agrobacterium- mediated gene transfer technology in other legumes with high transformation frequencies.


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