Construction and Transformation of Starch Synthases Gene RNAi Expression Vector Regulating by Potato Tuber Specificity Promoter

Abstract:

Using the fused gene-gbs3s2 composed of the gbss（1-261），ssⅢ（2164-2407） and ssⅡ（161-441） gene cDNA fragments as target，we constructed ihRNAi expression vector pART-GF regulated by GBSS of potato tuber specificity promoter. The inverted repeat construct was transformed to elite potato cultivars NHD3，NF and NC by Agrobacterium LBA4404-mediated transformation，PCR technology was used to detect transgenic plants，and the ratio of amylose eand amylopectin in transgenic mini-potato were determined by dual-wavelength spectrophotometry. Ten regenerated plants with kanamycin resistance were obtained through resistance screening and PCR detection. Among them，two was NC plants，three was NF plants，and five was NHD3 plants，respectively. The ratio of amylose and amylopectin in transgenic mini-potato were detected to decrease significantly. IhRNAi expression vector pART-GF regulated by GBSS of potato tuber specificity promoter was successfully constructed and breeding material of transgenic potato was obtained.