运用农杆菌介导的针刺法将外源基因转入水稻

Transforming gene into rice through agrobacterium-mediation assisted by needle puncture

选用含转化质粒p BI121-GUS的农杆菌为转化载体菌株,以水稻南粳45为转基因受体品种,用含有外源基因的农杆菌菌液浸蘸接种针,然后直接侵染受体水稻发芽种子的胚芽生长点,不进行愈伤组织培养,直接完成转基因操作。结果表明,生长成植株的48株中,16株经PCR检测和GUS组织化学检测均为阳性,说明外源基因不仅导入水稻基因组中,且已在蛋白质水平上表达。与传统的农杆菌介导法相比,本方法不需诱导愈伤组织以及之后的一系列组织培养过程,节省了大量的人力和财力,转化过程所需时间缩短,具有简单、快速、高效的优点,是一种改良的水稻转基因新技术。

The solution of agrobacterium EHA105 containing plasmid pBI121-GUS was used to infect the apical mer-istem of japonica rice variety, Nanjing45 through agrobacterium-mediation assisted by needle puncture to transform gus gene into rice without callus induction and a series of tissue culture. Among 48 survived seedlings, 16 positive plants were veri-fied by PCR detection and GUS histochemical assays, indicating that the GUS gene has been transformed into rice genome and expressed on protein level. Compared to the traditional method of agrobacterium-mediated transformation, the present method does not need to induce or culture callus, and can save a large amount of labor, time and costs. It is a simple, quick and efficient way for rice gene transformation.