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为了优化小麦成熟胚培养体系,在前人报道的小麦成熟胚培养方法的基础上对小麦成熟种子处理及取胚方法进行了改进,发现胚切伤略带胚乳的取胚方法及浸泡过夜后4℃低温处理法可以提高小麦成熟胚的再生效率;从19个小麦品种(系)中筛选出5个在此培养体系下具有较高再生率的基因型,分别为农大408、农大3787、保丰104、农大3965和京花1号,其再生率分别为73.11%、68.30%、65.30%、63.40%和57.60%。进一步利用基因枪轰击法对小麦品种辽春18和保丰104成熟胚诱导的愈伤组织进行了转化,分别获得转GUS基因和转Gcn5RNAi基因的小麦T0代植株,经分子检测其转化率分别达到1.35%和1.80%。证明优化后的小麦成熟胚培养技术可以应用于小麦遗传转化研究。
In order to optimize the culture system of wheat mature embryos, we improved the method of maturing and taking embryos of wheat on the basis of the methods of wheat mature embryo culture previously reported. ℃ low temperature treatment can improve the regeneration efficiency of mature wheat embryos; 5 of 19 wheat cultivars (lines) screened in this culture system has a high regeneration rate of genotypes were Nongda 408, Nongda 3787, Paul Fung 104, Nongda 3965 and Jinghua 1, the regeneration rates were 73.11%, 68.30%, 65.30%, 63.40% and 57.60% respectively. The callus induced by the mature wheat germplasms of Liaochun 18 and Baofeng 104 was further transformed by gene gun bombardment method. T0 transgenic plants with GUS gene and Gcn5RNAi gene were obtained respectively. The transformation rate of the transgenic plants reached 1.35% and 1.80%. Proved optimized wheat mature embryo culture technology can be applied to wheat genetic transformation research.