In vitro regeneration of the tropical multipurpose leguminous tree Sesbania grandiflora from cotyledon explants

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Abstract
A system using cotyledon pieces as explants and a BAP/NAA containing medium was developed for in vitro mass propagation of Sesbania grandiflora, a tropical nitrogen-fixing leguminous tree. The age and the lighting conditions of seedlings providing the explants were shown to be critical factors for both bud induction and bud elongation. Optimal choice for an efficient and reproducible bud induction process consisted of dark-grown seedlings, 24/36 h-old-post-imbibition, that yielded up to 96% of explants producing more than 30 buds each, after one week in culture. Bud development occurred throughout a direct organogenesis pathway, from the proximal and adaxial cut surface of the explants as proved by histological studies. Additional sites of regeneration were also obtained after wounding on the epidermal surface of explants, suggesting a large distribution of regenerative cells all along the explants. Bud elongation, i.e. stem differentiation and leaf growth, was improved by bud isolation from cotyledon explants and their further subculture in liquid bud elongation media for one week. Rooting was obtained on an auxin medium after 3 weeks and plants were established in soil with 92% success.