Agrobacterium Mediated Transformation of Pigeonpea (Cajanus Cajan L Milllsp) var LRG-41 from Axillary Bud.

A reliable method of plant regeneration has been achieved from Axillary buds. Shoots appeared from explants when cultured on Murashige and skoog (MS) medium supplemented with BAP (Benzyl amino purine), Napthalene acetic acid (NAA) and Kinetin at various combinations. Elongated shoots were rooted with 70.6% rooting frequency in MS medium with indole buteric acid (IBA) at 1.0mg/l. The rooted plantlets were established well in soilrite mixture medium with 91% success and days taken for acclimatization were 12.8 days. This protocol was used in Agrobacterium mediated transformation. The transformation was carried out using the Agrobacterium strain LBA4404 containing the binary vector pCAMBIA2301 harboring npt II as selectable marker and GUS as reporter gene.

Pattern of Plant Regeneration from Shoot Tip Explants of Pigeonpea (Cajanus Cajan L Milllsp) var LRG-41.

An efficient direct shoot bud differentiation and multiple shoot induction from shoot tip explants of pigeon pea (Cajanus cajan L.) has been achieved. The frequency of shoot bud regeneration was influenced by the type of explants, genotype and concentrations of cytokinin. Explants viz. shoot tip isolated from 10 day old seedlings showed better explants response Explants were cultured on Murashige and skoog (MS) medium augmented with different concentrations of BAP and NAA. Among the various concentrations tested, 2.0mg/l BAP (Benzyl amino purine) and 0.1 mg/l Napthalene acetic acid (NAA) were found to be the best for maximum shoot bud differentiation. Percentage, as well as the number of shoots per explant showing differentiation of shoot buds was higher on MS media supplement with BAP and optimal BAP concentration for shoot regeneration was 2mg/l. The elongated shoots were successfully rooted on MS medium containing different concentrations of auxins. Among them indole buteric acid (IBA) at 1.0mg/l induced maximum frequency of rooting. Regenerated plants were successfully established in soil where 91% of them have been developed into morphologically normal and fertile plants. This method can thus be advantageously applied in the production of transgenic pigeon pea plants.