Somatic embryogenesis in a recalcitrant cultivar of alfalfa (Medicago sativa L.) in an improved medium
A simple culture protocol was developed in order to screen genotypes with a regenerating ability from a recalcitrant cultivar of alfalfa (Medicago sativa L.). Twenty six cultivars or germplasm samples were tested for their ability to form somatic embryos. The frequency of somatic embryogenesis of th...
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Veröffentlicht in: | Bulletin of the National Grassland Research Institute (Japan) 1991-03 (44) |
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Zusammenfassung: | A simple culture protocol was developed in order to screen genotypes with a regenerating ability from a recalcitrant cultivar of alfalfa (Medicago sativa L.). Twenty six cultivars or germplasm samples were tested for their ability to form somatic embryos. The frequency of somatic embryogenesis of the three Japanese cultivars was all low (0-2%) and they were considered to be recalcitrant cultivars. One Japanese cultivar, "Tachiwakaba" was used as a model plant for improving the media protocol. Four media (B 2, B 5h, UM and SH) were compared for their ability to induce somatic embryogenesis. Somatic embryogenesis occurred only in the UM medium. The effect of four cytokinins, e.g. Kinetin, Benzyladenine (BA), N-[2-isopentenyl] adenine (2 ip), and Zeatin was compared in a UM medium supplemented with 2mg/l 2,4-dichlorophenoxyacetic acid (2,4-D). Ba was more effective than any other cytokinins in terms of both callus growth and somatic embryogenesis. The effect of the combination of 2,4-D and BA at various concentrations was also examined. The number of somatic embryos formed was the largest at 4mg/l 2,4-D. Below 2 mg/l 2,4-D, somatic embryogenesis did not occur. The effect of concentration of BA was not appreciable in the range of 0.125-1.0 mg/l. The number of somatic embryos formed at 4 mg/l 2,4-D and 0.125 mg/l BA was four times as large as that at 2 mg/l 2,4-D and 0.25 mg/l Kinetin which corresponds to the original composition of the UM medium. It is concluded that the UM medium containing 4 mg/l 2,4-D supplemented with BA rather than Kinetin as a cytokinin is a suitable medium for the induction of somatic embryogenesis in hypocotyl-derived calluses of alfalfa "Tachiwakaba" among the media tested |
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ISSN: | 0385-0196 |