Plant regeneration via direct shoot organogenesis from seedling explants of pole sitao (Vigna unguiculata L. Walp. var. Sesquipedalis L. Koern.)
2004
Avenido, R.A. | Dimaculangan, J.G. | Welgas, J.N. | del Rosario, E.E.,Philippines Univ. Los Banos, College, Laguna (Philippines). Inst. of Plant Breeding
Complete system for in vitro shoot induction, plantlet recovery and ex vitro establishment for pole sitao (Vigna unguiculata L. Walp. var sesquipedalis L. Koern.) was established for the first time using the protocol originally developed for mungbean and other Asiatic Vigna spp. as model. Cotyledon and cotyledonary node (CN) explants excised from aseptic seedlings were cultured using MS (Murashige and Skoog, 1962) salts and B5 vitamins (Gamborg et al, 1968) medium with 1.0 to 2.0 mg/L benzyladenine (BA). On the average, percent shoot formation in cultivars 'UPL PS 1' and 'UPL PS2' were higher using CN (67.6 and 80.4%) than that of cotyledons (25.2 and 17.8%). However, higher responses can be obtained from cotyledons if younger seedlings (i.e., 1 to 2-d-old) were used. A test on 8 pole sitao genotypes (cultivars and advanced breeding lines) for shoot regeneration efficiency revealed genotypic effects with a range of 43.3 to 100%. Individual shoots were excised and rooted at high frequencies (90-100%) using agar-solidified MS-B5 basal medium with 100 ml/L coconut water. Survival of acclimatized regenerants ranged from 47.4 to 88.9% when transplanted to a 1:1 mixture of garden soil and coir dust under greenhouse conditions. All tissue culture-derived seedlings are morphologically normal, flowered and produced pods filled with seeds under field conditions. The system may be applied in micropropagation of interspecific hybrids and other biotechnology-assisted manipulations (ex. in vitro selection and mutagenesis and genetic engineering) in pole sitao.
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