卡那霉素在小麦愈伤组织遗传转化中的应用

The Application of Kanamycin in the Genetic Transformation of Wheat Immature

为建立利用卡那霉素筛选的小麦遗传转化的有效体系,采用不同浓度的卡那霉素对小麦品种的幼胚愈伤组织进行处理,系统地研究了不同品种、不同愈伤组织诱导培养基、不同诱导培养时间下愈伤组织对卡那霉素筛选的反应。结果表明,不同小麦品种的愈伤组织对卡那霉素的抗性存在显著差异;同一品种不同培养条件下产生的愈伤组织对卡那霉素的反应存在显著差异;小麦品种河农827在N培养基上诱导培养14d得到的愈伤组织在含有15mg.L-1卡那霉素的分化培养基上筛选,效果较好。基因枪介导转化河农827的愈伤组织,用卡那霉素筛选得到的抗性再生苗经PCR检测,检测出12.5%的阳性株。

In order to establish the effective selecting system by using kanamycin as screening reagent in transgenic wheat, different concentrations of kanamycin added in regeneration media were tested for different wheat genotypes, different induction media and different induction duration. The results in- dicated that the sensitivity of the immature embryo callus to kanamycin differs significantly among the four genotypes studied, and among culture conditions, i.e. induction media and induction duration. For genotype Henong 827, culture condition combination with N medium induction under 14 days cul- ture and subsequently being screened with 15 mg · L-1 kanamycin was suggested. Using this method, a frequency （12.5G） of PCR-positive transgenic plants for Henong 827 was obtained. Furthermore, the feasibility of this method in transgenic wheat was discussed.