Organogenesis of Phaseolus angularis L.: high efficiency of adventitious shoot regeneration from etiolated seedlings in the presence of N6-benzylaminopurine and thidiazuron

摘要

A step-wise procedure for the regeneration of fertile plants by organogenesis from cultures of the economically important Phaseolus angularis L., cultivars: KS-6, KS-7 and KS-8 using etiolated seedlings was established. Pre-culture of 5-day old seedling explants with MS (Murashige and Skoog (1962) Physiol Plant 15:473-493) + B-5-vitamins (Gamborg et al. (1968) Exp Cell Res 50:151-158) liquid medium containing either 5.0 mu M TDZ or 5.0 mu M BAP under dark condition was essential for organogenesis. Bud growth and shoot multiplication were stimulated by reducing the BAP concentrations from 5.0 to 2.5 mu M after 3 weeks. The maximum frequency of shoot induction was 65.2% (33.8 +/- 2.54 shoots/explant) in cultivar KS-8 followed by KS-7 34.6% (23.4 +/- 1.91 shoots/explant) and KS-6 30.6% (21.2 +/- 2.28 shoots/explant). The multiplied buds elongated after transferring to solid MSB5 medium supplemented with 4.0 mu M GA(3), 12.5 mu M AgNO3 and 0.4 mu M IBA. Up to 98% rooting efficiency of was obtained when the shoots were pulse-treated with liquid medium containing 4.5 mu M IBA for 10 min. The rooted plantlets were transferred to pots in the greenhouse, where they grew, mature, flowered and bared pod normally. The efficient shoot bud induction capability was found to be cultivar dependent. All the three cultivars tested formed multiple shoots. This efficient and rapid regeneration system may also be helpful for Agrobacterium- or particle gun-mediated transformation for this important legume crop.