农业生物技术学报2012，Vol.20Issue(5)：481-488,8.DOI:10.3969/j.issn.1674-7968.2012.05.003
甘蔗尿苷二磷酸葡萄糖焦磷酸化酶基因(UGPase)转化拟南芥及转基因植株的生理特性分析
Transformation of UDP-glucose Pyrophosphorylase Gene (UGPase) from Saccharum officinarum into Arabidopsis thaliana and Analysis of Physiological Characteristics of Transgenic Plants
摘要
Abstract
UGPase (UDP-glucose pyrophosphorylase) is one of the enzymes which mainly involves in plant carbohydrate metabolism, and plays an important role in the process of plant growth and development. The plant expression vector was constructed by ligating the cDNA fragment of UGPase from Saccharum Officinanm into expression vector pBI121, then the recombinant plasmid pBI-UGP was confirmed by restriction enzyme digestion analysis with BamH I /Sac I , and further verified by DNA sequencing. The expression vector was introduced into Arabidopsis thaliana by floral-dip method, with Agrobacterium tumefaciens as media. Five To transgenic plants were selected by combining kanamycin screening with PCR detection. After selecting by kanamycin screening, T1 transgenic plants were identified by PCR and Southern blot, the results showed that the target gene was transformed into Arabidopsis thaliana successfully, and different transgenic plants contained various copy numbers of UGPase from Saccharum officinanm. T2 transgenic plants were identified by PCR and RT-PCR, the results indicated that the gene not only inherited stably in self-progeny, but also expressed at the RNA level. Meanwhile, content of soluble sugar, sucrose and starch were determinated in T2 transgenic plants and wild plants. Compared to wild plants, the soluble sugar content of transgenic plants had little change, however, sucrose content of transgenic plants improved obviously, increased by 50.85%~96.99%. Starch content of transgenic plants was lower than that of wild plants, decreased by 9.69%~36.76%. The research proved that UGPase plays a critical role in the process of converting sugar and starch, and the direction of its catalysis influneces distribution of these products in plant organization. It will lay foundation for further study of UGPase.关键词
尿苷二磷酸葡萄糖焦磷酸化酶(UGPase)/载体构建/转化/糖含量测定Key words
UGPase/ Vector construction/ Transformation/ Sugar content引用本文复制引用
连玲,叶冰莹,陈如凯,谢华安,张建福,陈由强..甘蔗尿苷二磷酸葡萄糖焦磷酸化酶基因(UGPase)转化拟南芥及转基因植株的生理特性分析[J].农业生物技术学报,2012,20(5):481-488,8.基金项目
国家自然科学基金项目(No.30871575) （No.30871575）
农业部现代农业产业技术体系建设专项资金(No.CARS-20-4-4) （No.CARS-20-4-4）
