麻疯树毒蛋白(curcin)基因转化水稻的研究

Study on Transformation of Rice for Curcin Gene of Jatropha curcas

以粳稻(Japonica)品种日本晴与中花11号为材料,对影响农杆菌介导水稻转化体系的几个因素进行了研究,建立了农杆菌介导的有效水稻转化体系.在该体系中,首次使用浓度为10-4g/mL嘌呤合成抑制剂acivicin在转化前对水稻愈伤组织进行预处理,结果表明此法可有效提高GUS基因的瞬时表达率.同时,利用插入了麻疯树毒蛋白(curcin)基因的植物表达质粒pBI121-curcin,通过农杆菌介导转化水稻,获得了多株转基因植株.通过PCR鉴定,证实curcin基因已整合到水稻基因组中.

Several important factors have been investigated on affecting Agrobacterium-mediated rice transformation system by using rice cultivars Nipponbare and Zhonghua No. 11, and an efficient Agrobacterium-mediated rice transformation system was developed. In this system, it was found that the GUS transient expression of transformed rice calli was improved by prior exposure to 10^-4g/mL acivicin, which is a kind of purine synthesis inhibitors, before infection with A. tumefaciens. At the same time, using the plasmid pBI121- curcin which carried curcin gene of Jatropha curcas, transgenicrice plants were obtained by the Agrobacterium-mediated rice transformation system. Most transgenic plants were identified by PCR ananlysis and the results indicated that erucin gene had been integrated into the genome of transgenic rice plants, which provide an efficient way for transferring useful foreign genes to rice plants and lay the basis for the further study on the function of curcin gene.