病毒复制酶基因Nib8和ERF转录因子W17基因枪法共转化小麦

Co-Transferring of Virus Replicase Gene Nib8 and ERF Gene W17 into Wheat Through Microprojectile Bombardment

将对小麦黄花叶病毒表现高抗的复制酶基因Nib8和具有广谱抗病性的ERF基因W17分别构建到单子叶高效组成型表达载体上,采用基因枪共转化法转化到小麦品种扬麦12和扬麦16中,PCR检测共获得Nib8基因的阳性转基因植株42株,W17基因的阳性转基因植株48株,及两个功能基因均为阳性的转基因植株6株。以扬麦12为受体的转化率分别为1.53%(Nib8)、4.87%(W17)和0.42%(Nib8+W17);以扬麦16为受体的转化率分别为2.05%(Nib8)、0.86%(W17)和0.20%(Nib8+W17)。对2个功能基因都呈阳性的6个植株进行Southern blotting分析,进一步证实功能基因已经整合到小麦基因组中。本研究为获得具有综合抗病性的小麦新材料奠定了基础。

Wheat yellow mosaic virus replicase gene Nib8 and broad disease-resistant ERF gene W17 were integrated into high-efficient constitutive expression vector for monocots and then co-transferred into wheat through microprojectile bombardment. Positive plants of To generation were identified by using PCR method, among which 42 plants were confirmed to be with Nib8, 48 plants with W17, and 6 plants with both Nib8 and W17. The transformation frequency for Yangmai 12 was 1.53% （Nib8）, 4.87% （W17）, and 0.42% （ Nib8 ＋ W17）, respectively, while for Yangmai 16 it was 2.05% （Nib8）, 0.86% （W17）, and 0.20% （Nib8 ＋ W17）. Six positive plants containing two target genes of W17 and Nib8 were analyzed by southern blotting, which further demonstrated that the functional genes were successfully integrated into wheat genome. This study provided a basis for obtaining new resource of comprehensive disease-resistant wheat materials.