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持绿基因(Stay Green gene)在植物衰老过程中起到了重要的作用.本研究利用同源序列PCR方法、 Gateway重组技术和子叶节遗传转化方法,对大豆持绿基因GmSGR进行RNAi载体的构建,并将其转入到大豆受体中,获得转基因阳性植株.结果显示,分别获得长度为786 bp的GmSGRl CDS序列,长度为816 bp的GmSGR2 CDS序列及355 bp的attB-GmSGRi干扰片段.依据RNAi引物设计原则和Gateway重组技术,将干扰片段依次导入到入门载体pDONR221和表达载体pB7GWIW2 ( Ⅱ )中,并转化农杆菌EHA105,通过子叶节法将其转入大豆受体中,最终获得阳性转基因植株.本研究在mRNA水平沉默同源靶基因,干扰大豆GmSGR基因功能,结果显示其叶片和种子有趋向持绿的特性.本研究为探究持绿基因的遗传效应,作用机理提供实践参考,为高光效大豆的遗传改良,探讨植物衰老延缓机制,提高产量和品质等提供理论支持.“,”Stay Green gene plays an important role in plant senescence. In this study, the homologous sequence PCR method, Gateway and the cotyledonary node transformation method were used to construct the RNAi vector of stay green GmSGR and transferred into soybean receptor to obtain transgenic plants. The results showed that GmSGR1 sequence of 786 bp in length, GmSGR2 sequence of 816 bp in length and attB-GmSGRi fragment of 355 bp in length were obtained respectively. According to the design principles of RNAi primers and Gateway recombination technology, the interference fragments were introduced into the intron vector pDONR221 and the inducible vector pB7GWIW2 (Ⅱ) in turn, and then transformed into Agrobacterium tumefaciens EHA105, transferred into the soybean receptor through the cotyledonary node method to obtain the positive transgenic Plant. Silencing the homologous target gene at the mRNA level knocked out the function of soybean GmSGR gene and observed that the leaves and seeds tended to be stay green. The research might provide a practical reference for exploring the genetic effect and mechanism of green gene, and provide theoretical support for the genetic improvement of high light efficiency soybean, exploring the mechanism of plant aging retardation, improving yield and quality.