转反义trxs基因小麦分子鉴定及抗穗发芽株系的筛选  

Molecular Identification and Screening of Pre-harvest Sprouting Resistance Lines in Transgenic Wheat with Anti-sense trxs Gene

作　　者：陈新[1] 孟晓丹[1] 王亚英[1] 肖瑞霞[1] 张斌[1] 任江萍[1] 尹钧[1] 

机构地区：[1]河南农业大学国家小麦工程技术研究中心,河南郑州450002

出　　处：《河南农业科学》2013年第4期19-23,共5页Journal of Henan Agricultural Sciences

基　　金：国家转基因生物新品种培育重大专项(2011ZX08002-002)

摘　　要：为给小麦抗穗发芽育种提供基础材料和参考依据,分别以豫麦18、豫麦70和豫麦34为受体,对转反义trxs基因小麦01TY18、01TY70和01TY34高代株系进行PCR分子检测,并对阳性株系进行离体整穗发芽和α-淀粉酶活性测定。结果表明,经PCR检测T7代40个转基因株系中,阳性株系占55%。与非转基因对照相比,转基因株系的穗粒发芽率和α-淀粉酶活性都有所下降,其中有19个株系穗粒发芽率显著低于对照,占阳性转基因株系的86.4%;有16个株系α-淀粉酶活性较对照显著降低,占阳性转基因株系的72.7%。综合PCR检测、穗粒发芽率和α-淀粉酶活性测定结果,共筛选出转反义trxs基因的抗穗发芽小麦株系15个,其中01TY18 8个、01TY70 5个、01TY34 2个,占所有转基因株系的37.5%、阳性转基因株系的68.2%。To provide the basic materials and reference for pre-harvest sprouting resistance in wheat breeding,the high generation transgenic lines 01TY18,01TYT0 and 01TY34 with anti-trxs gene and their non-transgenic parents Yumai 18, Yumai 70 and Yumai 34 were used as testing materials to carry out the PCR detection,and the determination of intact spike sprouting and α-amylase activity in positive lines. The results of PCR detection showed that the positive lines with the target gene were 55 % of the 40 T7 transgenic lines. Compared with the non-transgenic control, the spike-seed sprouting rate and the a-amylase activity in positive transgenic lines decreased. Among them,19 lines were significantly lower than the control in spike-seed sprouting rates, accounting for 86.4 % of the positive transgenic lines,while 16 lines were significantly lower than the control in a-amylase activity,accounting for 72.7 % of the positive transgenic lines. Based on the results of PCR detection,spike spouting and a-amylase activity screening, 15 positive transgenic lines with pre-harvest sprouting resistance were selected, among which, 8 lines were from 01TY18,5 from 01TY70,and 2 from 01TY34,accounting for 68.2% of the positive transgenic lines and 37.5% of all transgenic lines,respectively.

关 键 词：小麦 反义TRXS基因 转基因株系 离体整穗发芽 Α-淀粉酶活性 

分 类 号：S512.1[农业科学—作物学]