反义TrxS基因在小麦中的表达及对小麦种子蛋白酶活性和蛋白组分的影响  

Expression of Anti-TrxS and Its Effect on Protease Activities and Protein Ingredients in Transgenic Wheat Seeds

作　　者：任江萍[1] 李磊[1] 王新国[1] 牛洪斌[1] 李永春[1] 孟凡荣[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：《麦类作物学报》2008年第6期941-945,共5页Journal of Triticeae Crops

基　　金：国家自然科学基金项目(30370877);河南省杰出青年基金项目(0512001600)

摘　　要：为了揭示反义硫氧还蛋白基因(anti-TrxS)在抗穗发芽小麦中的作用机制,探讨转基因小麦中蛋白酶活性和蛋白组分的变化规律,以转反义TrxS基因小麦株系为材料,用荧光定量RT-PCR、SDS-PAGE和生理指标测定的方法,对转基因株系和对照种子萌发过程中反义TrxS基因表达、蛋白酶活性和蛋白组分进行了检测。结果表明,反义TrxS基因在RNA水平上能够表达;转基因小麦籽粒蛋白酶活性明显下降,种子吸涨12、24、48、729、6和120 h后,转基因小麦种子蛋白酶活性比对照分别降低27.3%、30.7%、19.1%、19.4%、23.7%和13.5%;转基因小麦籽粒谷蛋白、醇溶蛋白的SDS-PAGE分析表明,反义TrxS基因的导入抑制了转基因小麦籽粒谷蛋白和醇溶蛋白大分子亚基二硫键(S-S)向巯基(-SH)的转化,从而使蛋白质的降解延缓。To clarify the function mechanism of antisense - thioredoxin s （anti-TrxS） gene in transgenic wheat in pre-harvest sprouting resistane, the anti-TrxS gene expression, protease activity and protein ingredients in transgenic line and non-transgenic cuhivar of Yumai 34 were detected By real-time RT-PCR, enzyme activity assay and SDS-PAGE during seed germination stage. The results indicated that anti-TrxS gene could express in transgenic wheat seeds. During 12-120 h of seed imbibing, the activities of protease were 27.3%, 30. 7%,19.1%,19.4%,23.7% and 13.5% lower than that of non- transgenie seeds, respectively. The results of analysis of SDS-PAGE of glutelin and gliadin indicated that anti-TrxS inhibited the tromsform of disulfide bonds（S-S） to the sulfhydryl（-SH） of transgenic line seeds and velocity.

关 键 词：小麦 反义TRXS基因 表达 蛋白酶 蛋白组分 

分 类 号：S512.1[农业科学—作物学] S312