Benzyladenine-induced adventitious shoot regeneration from hypocotyls of adzukibean (Vigna angularis {Willd.} Ohwi and Ohashi)

Abstract

A complete in vitro plant regeneration system via adventitious shoot-bud formation from seedling hypocotyls of adzukibean was developed. Cotyledonary node (CN) and root explants from 4-d-old in vitro-germinated seedlings of 4 cultivars were cultured on agar-solidified R medium consisting of MS salts, B5 vitamins, 3.0% (w/v) sucrose and 4.4 μM 6-benzyladenine (BA). Shoot buds arose adventitiously at the basipetal cut of the hypocotyl in 40 to 85% of type 1 CN (8-10 mm) explants 6 weeks later with cvs Bansei aki azuki and Tanbadainagon azuki exhibiting significantly higher response. Shorter type 2 (4-5 mm) CN explants responded poorly (0-6%) in culture while intact roots were regenerative but at lower frequencies (5 to 31%). In cv Bansei aki azuki, optimum shoot initiation (78.7%) was achieved when type 1 CN explants from BA-preconditioned seedlings were re-cultured in R medium (SC0). BA supplied at 8.8 μM during SC0 significantly reduced the shoot initiation response to 46.7%. In the same manner, substituting equimolar concentration (4.4 μM) of kinetin (KN), zeatin (Z) or thidiazuron (TDZ) to BA during SC0 yielded no or significantly lower responses. Mean shoot production ranged from 6.2 to 9.8 shoots per explant 6 weeks after subculture (SC1) and varied significantly among the 4 cultivars used. Efficient rooting (93-96%) was achieved on excised shoots transferred to MS salts + B5 vitamins basal medium alone. The tissue culture system covered only 16 weeks to completion. Regenerated plantlets were transferred to soil with 86-100% survival rate and all surviving plants were morphologically normal and seed-fertile.

Source or Periodical Title

Plant Growth Regulation

ISSN

1676903

Page

147-153

Document Type

Article

Subject

BA-preconditioning, Cotyledonary node explant, Cytokinin, Plant regeneration system, Tissue culture, Vigna angularis

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