Expression And Physiological And Bio-chemical Effects Of Exogenous Anti-TrxS Gene In Transgenic Wheat And Its Pre-Harvest Sprouting Resistance

Wheat pre-harvest sprouting is a worldwide climate disaster. It effected wheat yield, quality and its seed value severely. Pre-harvest sprouting is caused by a series of proteases, in which α-amylase plays a key role. Thioredoxin (Trx) plays an important role in regulating activity of enzymes by deoxidizing S-S to -SH in wheat plant. Therefore.we have transferred the anti-trxs gene into wheat (Triticum aestivum L.) cultivar 895004 and Yangmai 5. Transgenic wheat lines T1～T3 generations had been detected and analyzed by molecular method and obtained 00T89 and 00TY5 transgenic generations. More studies should be done for the sake of genetic rule of exogenous anti-trxs gene and its expression and effects. Therefore, by PCR, PCR-Southern blotting and nested-PCR, the present study continued to detecte the target gene at DNA level. The transcriptional changes of target gene and the abundance of mRNA were also identified. The changes of thioredoxin h redox activities and its related enzyme activities were evaluated in the process of forming and afterripening and germinating of seeds respectively, and so for the carbon and nitrogen metabolism levels in seeds by these active substances. Under artificial rainfall, we tested the seed germination and ear sprouting resistances. The agricultural and quality traits in the transgenic lines also were analysed. The main results were as following:1 The molecular detection, genetic analysis and resistance detection on herbicide (bialaphos) of transgenic anti-trx s gene lines00T89 and 00TY5, T4～T6 generations of the transgenic lines, were amplified for target gene in genome DNA by molecular methods. The PCR analysis showed positive results indicating exogenous anti-trxs gene had been successfully transferred and steadily inherited in different lines of 00T89 and 00TY5, and was proved to be specific and veritable by PCR-Southern blotting. The result also indicated that the ratio of target gene in 70-80% T4 transgenic lines followed by 7:1( x~ 2=3.816). But the ratio of negative plants in T5 progeny lines rose. The ratio of target gene inT6 progeny lines of 00T89 and 00TY5 followed by 12:1( x~2=3.816) and 10:1 ( x~2=3.61),...