根癌农杆菌介导反义蜡质基因获得水稻植株研究初报

Primary Studies on Transformation of Waxy Antisense Gene Mediated by Agrobacterium in Japonica Rice

采用根癌农杆菌介导法,将水稻反义蜡质基因导入寒地4个水稻品种(系)成熟胚的愈伤组织.这些愈伤组织经连续2次25～50 mg/L潮霉素抗性筛选,获得抗性愈伤组织的转化频率为12.15%～25.95%,经分化培养,不同品种的绿苗转化频率分别为2.49%～4.43%.分化成苗的植株经GUS检测,80%以上的植株叶片呈阳性反应,显示兰色.经PCR检测,也能扩增出外源基因相应的DNA片断,证明外源基因已整合到转基因植株中.

Waxy antisense gene was transferred into calli derived from immature embryos of four rice varieties by Agrobacterium tumefaciens.These calli were cultived on selective media containing 25~50 mg/L Hygromycin two times, 12.15%~25.95% resistant calli .These resistant calli were transferred to differentiation and regeneration media, 2.49%~4.43% calli grown up to plants. Assay of Gus activity showed that more than 80% of transgenic plants are Gus positive. A few transgenic plants which were negative for the Gus assay were analysed by PCR,foreign DNA fragments were detected in these plants. The results showed that foreign genes have been inserted into rice plants.