High frequency plant regeneration from cotyledonary node explants of Cucumis sativus L. cultivar ‘Green Long’ via adventitious shoot organogenesis and assessment of genetic fidelity by RAPD-PCR technology

Abstract

Influence of cytokinins, silver nitrate (AgNO3) and auxins on plant regeneration from cucumber was investigated. The cotyledonary node explants were cultured on MS medium augmented with various concentrations (0.5-2.5mg l-1) of 6-benzyl amino purine (BAP) and kinetin (KIN) for shoot bud induction. BAP at 1.5mg l-1 was found to be the best concentration for induction of high frequency of multiple shoots (98.4%). Interestingly, maximum percent of multiple shoot regeneration (100%) as well as number of shoot buds (54.6 shoots/culture) was recorded on MS medium containing the combination of 4.5mg l-1 AgNO3 and 1.5mg l-1 BAP. Multiple shoot bud regeneration frequency as well as the number of shoots was positively correlated with the concentrations of AgNO3. Addition of silver nitrate in the medium not only enhanced the rate of multiple shoot bud regeneration but also elongation of shoot buds was observed. The highest percent of rooting (96.2%) was noticed on a medium containing the combination of indole 3-butyric acid (IBA), 1.5mg l-1 and KIN 0.5mg l-1. Acclimatized plantlets were successfully established in the field where the survival rate observed was 72%. The RAPD profiles of in vitro regenerated plants were found to be highly monomorphic and identical banding pattern with mother plant. DNA fingerprinting results confirmed that the tissue culture plantlets were found to be true-to-type. The present study describes efficient protocol for high frequency plant regeneration via adventitious shoot organogenesis in cucumber.