甘蓝型油菜花药培养技术研究

Research on Anther Culture Technology of Brassica napus(Rape)

[目的]筛选甘蓝型油菜愈伤组织的最佳分化培养基,探讨甘蓝型油菜花药培养技术。[方法]在油菜始花期,取其主茎顶端蕾盘的中圈花蕾,将花药接种于B5+2,4 D 1.0mg/L+KT 1.5mg/L的培养基进行诱导,将诱导出的愈伤组织接种于不同配方的12种培养基进行分化培养。[结果]甘蓝型油菜花药培养取花粉处于单核后期至双核初期,对应于外部特征是蕾盘中圈10～15个花蕾,所产生的愈伤组织百分率最高(16%);12种不同配方的培养基中以B5+KT 2.0mg/L+BA 2.0mg/L+IAA 0.1mg/L或MS+KT 2.0mg/L+BA 2.0mg/L+IAA 0.1mg/L培养基上分化成苗最好,分化率分别为75%和70%,将分化的小苗置于1/2MS培养基上诱导生根,可长成完整幼苗。[结论]花药培养可以加速油菜育种进程。

[Objective] The aim was to screen the best differentiation medium of callus for Brassica napus and explore the anther culture technology of B. napus. [Method] The centre circle of flower bud in bud disk on top main stem of B. napus was taken at the initial time of flowering and the anthers were inoculated in culture medium of B5＋2,4 D 1.0 mg/L ＋ KT 1.5 mg/L for induction. The induced calli were inoculated to 12 t kinds of mediums with different formulas for differentiation eulture.[Result] In the anther culture of B. napus, the pollens were taken from the mononucleated anaphase stage to binucleated initial stage, the corresponding external characteristics was 10～15 buds in central circle of bud disk, from which the percentage of produced callus was the highest （16%）. Among 12 kinds of mediums with different formulas, the seedling differentiation was the best for the mediums of B5 ＋ KT 2.0 mg/L ＋ BA2.0 mg/L ＋ IAA 0.1 mg/L or MS ＋ KT 2.0mg/L ＋ BA 2,0 mg/L ＋ IAA 0.1 mg/L, with differentiation rate of 75% and 70% resp. When the differentiated young seedlings were transferred on 1/2 MS medium for inducing root formation, they could grow into the complete seedling. [Conclusion] Anther culture could acceleratethe breeding process of B. napus.