Molecular cloning of C4-specific Ppc gene of sorghum and its high level expression in transgenic rice  

作　　者：ZHANGFang CHIWei WANGQiang ZHANGQide WUNaihu 

机构地区：[1]InstituteofGeneticsandDevelopmentalBiology,ChineseAcademyofSciences,Beijing100080,China [2]InstituteofBotanyChineseAcademyofSciences,Beijing100093,China

出　　处：《Chinese Science Bulletin》2003年第17期1835-1840,共6页

摘　　要：In order to improve the carbon-assimilation ability of C3 plants, we isolated a C4-specific photosynthetic enzyme gene, Ppc (encode phosphoenolpyruvate carboxylase, PEPCase) from the genome of the C4 plant, sorghum, and transformed rice with it. As shown by sequence analysis, the gene is composed of 10 exons and 9 introns, and the full-length transcript is 5989 bp long. A recombinant expres-sion vector, p1301PEPC, was constructed by inserting the gene into a plasmid vector, pCAMBIA1301, which was then transformed into two japonica rice varieties, Nongken 58 and Zhonghua 10, using an Agrobacterium-mediated transforma-tion system. PCR analysis, activity measurement of PEPCase, and protein-, RNA- and DNA-based hybridization all con-firmed the successful integration of the C4-specific Ppc gene into the nuclear genome of rice and its high level expression. Physiological studies revealed the photosynthetic features characterizing C4 plants such as marked lowering of CO2 compensation point and photorespiration rate, and im-proved carboxylation efficiency. This study provides useful experimental materials and opens up new avenues for fur-ther studies on improving photosynthetic efficiency of elite varieties of rice.In order to improve the carbon-assimilation ability of C3 plants, we isolated a C4-specific photosynthetic enzyme gene, Ppc (encode phosphoenolpyruvate carboxylase, PEPCase) from the genome of the C4 plant, sorghum, and transformed rice with it. As shown by sequence analysis, the gene is composed of 10 exons and 9 introns, and the full-length transcript is 5989 bp long. A recombinant expres-sion vector, p1301PEPC, was constructed by inserting the gene into a plasmid vector, pCAMBIA1301, which was then transformed into two japonica rice varieties, Nongken 58 and Zhonghua 10, using an Agrobacterium-mediated transforma-tion system. PCR analysis, activity measurement of PEPCase, and protein-, RNA- and DNA-based hybridization all con-firmed the successful integration of the C4-specific Ppc gene into the nuclear genome of rice and its high level expression. Physiological studies revealed the photosynthetic features characterizing C4 plants such as marked lowering of CO2 compensation point and photorespiration rate, and im-proved carboxylation efficiency. This study provides useful experimental materials and opens up new avenues for fur-ther studies on improving photosynthetic efficiency of elite varieties of rice.

关 键 词：Ppc基因 光合酶基因 磷酸烯醇丙酮酸羧化酶 高粱 分子克隆 转基因水稻 基因表达 

分 类 号：Q943.2[生物学—植物学] S511[农业科学—作物学]