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以野生天蓝苜蓿种子发育5~7d无菌苗的子叶、下胚轴和种子苗叶片为外植体,对其愈伤组织诱导及其分化的基本培养与外源激素组合和浓度配比进行试验。结果表明:对子叶、下胚轴和叶片愈伤组织诱导最适宜的基础培养基分别是MS、B5和SH;激素的种类及其浓度配比对于愈伤组织的诱导因不同的外植体有很大的差异。其中,子叶、下胚轴以MS+NAA(0.5~1.0 mg/L)+6-BA(0.7mg/L)效果较好,叶片以SH+NAA(0.5~1.0mg/L)+2,4-D(1.5~5 mg/L)+6-BA(0.7mg/L)较好,少量下胚轴在MS+NAA(0.5mg/L)+6-BA(0.7mg/L)和MS+NAA(0.5mg/L)+KT(0.5 mg/L)的培养基中直接分化出胚状体,并成功发育成植株。在培养基Whb5上,子叶、下胚轴和叶片的分化都得到了很好的效果,分化率分别为43%、57%、40%,15~30d后都能再生植株。
The cotyledons, hypocotyls and seedling leaves of sterile seedlings of wild alfalfa cultivated for 5 ~ 7 days were used as explants, and their callus induction and differentiation were cultured with exogenous hormone combination and concentration ratio . The results showed that the most suitable basal medium for inducing cotyledon, hypocotyl and leaf callus were MS, B5 and SH, respectively. The types and concentrations of hormones were different for callus induction Great difference. Among them, the effects of MS + NAA (0.5-1.0 mg / L) + 6-BA (0.7 mg / L) on cotyledons and hypocotyls were better, (0.5mg / L) + 6-BA (0.7mg / L) and MS + NAA (0.5mg / L) NAA (0.5mg / L) + KT (0.5mg / L) medium directly differentiated into embryoid body, and successfully developed into plants. On the medium Whb5, the differentiation of cotyledons, hypocotyls and leaves all achieved good results. The rates of differentiation were 43%, 57% and 40% respectively, and the plants could be regenerated after 15-30 days.