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Rs-AFP2属于r-硫堇类抗菌肽,主要通过形成离子通道直接破坏细胞来杀灭病原菌。本研究通过基因枪介导法结合对目标基因的分子检测,证明已将外源Rs-AFP2基因转入小麦推广品种扬麦12中。通过逐株抗纹枯病接种鉴定、PCR、PCR-Southern blot、Southern blot和RT-PCR/荧光定量RT-PCR(Q-RT-PCR)分析,对转Rs-AFP2基因小麦T1至T4代植株跟踪检测。结果表明,Rs-AFP2在转基因小麦中能够稳定遗传,以单拷贝整合到小麦基因组中,遗传方式符合孟德尔遗传规律,并能在转录水平上表达。对转Rs-AFP2基因小麦的抗病性、主要农艺性状以及Rs-AFP2表达活性分析结果表明,与受体扬麦12相比,Rs-AFP2表达活性高的转基因小麦植株对纹枯病抗性有明显提高,其抗病性可以遗传,而主要农艺性状没有明显差异,证明可以利用Rs-AFP2基因和基因工程途径创制抗纹枯病小麦新种质。
Rs-AFP2 is an r-thionine antibacterial peptide, which kills pathogenic bacteria mainly by directly disrupting cells through the formation of ion channels. In this study, the molecular detection of the target gene by gene gun mediated method proved that the exogenous Rs-AFP2 gene was transferred into the wheat variety Yangmai 12. A Toxoplasma gondii pollination strain was obtained by inoculation identification of resistant strain of Rhizoctonia solani, PCR, PCR-Southern blot, Southern blot and RT-PCR / Q-RT-PCR analysis. Tracking test. The results showed that Rs-AFP2 could be stably inherited in transgenic wheat and integrated into the genome of wheat with a single copy. The genetic pattern of Rs-AFP2 accorded with Mendelian inheritance and could be expressed at transcriptional level. Analysis of disease resistance, major agronomic traits and Rs-AFP2 expression activity of Rs-AFP2 transgenic wheat showed that resistance to sheath blight resistance in transgenic wheat plants with high Rs-AFP2 expression activity There was no significant difference in the main agronomic traits, which proved that Rs-AFP2 gene and genetic engineering could be used to create new wheat germplasm with resistance to sheath blight.