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Dry seeds from two lines of common bean (Phaseolus vulgaris L.) and one cultivar of faba bean (Vicia faba L.) were germinated on Murashige and Skoog (MS) medium containing B5 vitamins, 30 g sucrose/liter, and either 2.5, 5.0, or 7.5 µM benzyladenine (BA). Axenic seed cultures were grown at 22 to 24C in darkness and under continuous light from cool-white fluorescent tubes (40 µmol·m -2 ·s -1 ). Explant tissues were prepared from cotyledonary nodes (CN) and primary nodes (PN) of 14-day-old seedlings. Explants were cultured on corresponding seedling growth medium and maintained under continuous cool-white light (40 µmol·m -2 ·s -1 ). The percentages of CN and PN (in one line of common bean) explants that regenerated shoots and the number of shoots per explant (in all germplasm) were highest when nodal tissues were prepared from seedlings germinated in darkness. These responses were optimal on medium containing 5 µM BA during seedling growth and subsequent culture of explants. The number of shoots per explant was two to five times higher on explants cultured on medium with 0.25 to 1.0 µM forchlorfenuron (CPPU) or thidiazuron (TDZ) than on medium with 5 µM BA. Higher (2.5 and 5 µM) CPPU and TDZ concentrations inhibited shoot elongation and stimulated callus production. Histological analyses indicated that adventitious meristems formed 6 to 8 days after explant culture. Progenies from regenerated plants appeared similar to plants raised from the original seed stocks. Chemical names used: N- (phenylmethyl) -1 H- purin-6-amine (benzyladenine, BA); N- (2-chloro-4-pyridyl)- N'- phenylurea (forchlorfenuron, CPPU); N- phenyl -N' -1,2,3-thiadiazol-5-ylurea (thidiazuron, TDZ). Tissue culture of grain legumes using cotyledonary node ex- plants from in vitro-grown seedlings has been documented for organogenic competence (Jackson and Hobbs, 1990; McClean and Grafton, 19889; Wright et al., 1986). BA supplements (5 to 10 µM) in the medium during germination and subsequent explant culture were one of the most critical factors for organ- ogenic differentiation and shoot multiplication in this regener- ation system. N- (2-furanylmethyl)-1 H- purin-6-amine (kinetin) was ineffective (McClean and Grafton, 1989) for inducing mul- tiple-shoot organogenesis in common bean or in pea (Pisum sativum L.) (Jackson and Hobbs, 1990), and neither (E)-2-methyl- 4-(1H-purin-6-ylamino)-2-buten-1-ol (zeatin) nor N- (3-methyl- 2-butenyl)-1 H- purin-6-amine (2iP) could be substituted for BA in soybean (Glycine max (L.) Merr.) shoot regeneration studies

Dark Preconditioning, CPPU, and Thidiazuron Promote Shoot Organogenesis on Seedling Node Explants of Common and Faba Beans