转反义trxs基因小麦株系01TY18遗传分析及抗穗发芽特性

Inheriting Analysis of Anti-trxs Gene and Pre-harvest Sprouting Resistant Characteristic in Transgenic Line 01TY18

以豫麦18为受体导入反义trxs基因获得的株系01TY18(T0)为材料,运用PCR、实时荧光RT-PCR以及离体整穗发芽的方法,对反义trxs基因在转基因小麦中的遗传稳定性、基因表达和抗穗发芽特性进行了研究.结果表明,8个T1代转基因株系中有6个株系目的基因检测呈阳性,且在以后的世代中能够稳定遗传并呈典型的孟德尔单基因3∶1分离规律;反义trxs基因在6个转基因株系中能够正常表达且表现出显著的抗穗发芽特性.与非转基因对照相比,转基因株系穗粒发芽率和穗发芽度平均分别降低62%(P<0.01)和50.8%(P<0.01).

Using the transgenic line 01TY18 with anti-trxs gene and non-transgenic cultivar ‘Yumai18＇ as testing materials,we carried out the inheritance stability of anti-trxs gene,foreign gene expression and preharvest sprouting resistance in transgenic lines 01TY18 by PCR,real-time RT-PCR and intact spike sprou ring assay. The results indicated that there were 6 positive lines with the anti-trots gene among transgenic lines in T1 progeny. The segregation ratio of the anti-trxs gene in T2-T4 progeny of 6 transgenic lines was 3 ： 1,which was in accordance with Mendel＇s rule of dominant gene with one locus. The anti-trxs gene could express in 6 T1 transgenic lines by detection of real time PCR and 6 lines showed stronger resistance to pre harvest sprouting in 30 days after anthesis. Compared with non-transgenic plant, the average sprouting ratio of seeds on spikes and the spike sprouting degree were respectively reduced by 62% （P〈0.01） and 50.8%（P〈0.01）.