RNA Interference Vector Construction and Genetic Transformation of Stay Green Gene GmSGR in Soybean
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.
SoybeanStay green geneGene cloneRNA interference vectorGenetic transformation
NETL
NSTL
万方数据
维普
