Influence of adenine sulphate on multiple shoot induction in Clitoria ternatea L. and analysis of phyto-compounds in in vitro grown plants

Abstract

In the present study, we have proposed that the supplementation of adenine sulphate enhanced the multiple shoot induction and regeneration of Clitoria ternatea (L.) and it enhances the functional groups present in in vitro grown plants were confirmed through FTIR analysis. The juvenile nodal explants cultured on Murashige and Skoog (MS) medium supplemented with 1.0 mg L−1 BAP, 0.5 mg L−1 IBA, amino acids cysteine 150 mg L−1 and alanine 30 mg L−1 showed best shoot induction frequency (72.3%) after 2 weeks of incubation in light. Further, the influence of adenine sulphate was examined where 45 mg L−1 of adenine sulphate on MS medium produced 79.9% of multiple shoot induction after 15 days of incubation. Maximum percentage of root induction (76.1%) was observed on half-strength MS medium containing 0.1 mg L−1 IBA and NAA. The rooted plantlets were transferred to soil cups and maintained in the shade house for maturity and flowering. FTIR analysis clearly revealed the enhancement of important functional groups like alkanes, amines, carboxylic acids, alcohols, aldehydes, proteins, sulfonates, nitro compounds and halogenated compounds in in vitro regenerated plants through adenine sulphate. The genetic fidelity of in vitro propagated and control plants analyzed using random amplified polymorphic DNA and inter simple sequence repeat markers showed no dissimilarity between control and in vitro propagated plants. This protocol will be useful for large scale suspension culture and genetic transformation studies.