基因型和外植体对小麦离体培养的影响

为建立适合小麦遗传转化的高效离体培养体系,本实验用14个优良小麦新品种(品系)为材料,对不同基因型的小麦进行幼胚离体培养研究,通过培养基激素的变化,寻找最适合遗传转化的基因型及培养基成分;结果表明:不同基因型间差异显著,其中09P200和SH905在小麦离体培养中反应较好,幼胚愈伤组织绿原基分化率和出苗率显著高于其他基因型,可作为幼胚培养和转基因受体材料的优良基因型,在分化培养基中添加100mg/L水解酪蛋白、100mg/L脯氨酸和2.0mg/LKT对愈伤组织诱导分化及成苗效果较好,所有供试基因绿原基分化率、成苗率平均分别为31.93%和14.42%;另外采用4个不同基因型的小麦幼胚和成熟胚为外植体,通过继代次数的改变,筛选各外植体最适合遗传转化的培养条件;结果表明幼胚继代两次、成熟胚继代三次可以获得相对理想的绿原基分化率,且此时的愈伤组织是遗传转化较理想的受体材料。

Effects of Genotypes and Explants on in vitro Culture of Wheat

To establish a high efficiency in vitro culture system suitable for genetic transformation of wheat,14 different wheat varieties(strains) were used for research on in vitro culture of embryonic wheat of different genotype.Through changing the hormone in the culture media,the most suitable genotype and the appropriate composition of culture media were found.The results show that the difference among genotypes is significant.The 09P200 and SH905 had grown better and had the higher frequency of green plantlet differentiation and plant generation of green primordia than the other genotypes in calluses of embryos,so they were the optimum genotype for immature embryos culture.The optimum differentiation media was with 100 mg/L hydrolysis casein,100 mg/L praline and 2.0 mg/L KT.The average rate of green plantlet differentiation and plant generation of green primordia were 31.93% and 14.42% respectively.In addition,using other 4 immature and mature embryos of wheat as explants,which has different genotypes,the most suitable culture conditions for genetic transformation were screened out through changing the subculture times.The results show that the relatively ideal differentiation rate of green primordia could be obtained when immature embryos of wheat are subcultured two times and the mature embryos are subcultured three times.Moreover at this time the calluses are the better recipientfor genetic transformation.