农杆菌介导萌动种胚转化法将抗草甘膦基因导入蓖麻

Introduction of glyphosat resistant gene into Ricinus communis L. by Agrobacterium mediated transformation using germinating embryos

为简化和提高转化效率,以蓖麻品种汾蓖10号为材料,采用非组培的农杆菌介导萌动种胚转化法,获得抗草甘膦植株T0。方法是在生长点附近针刺3针,超声波(强度800W)处理后,与农杆菌共培养16h。在随机选取的800株转化处理植株中,检测到5株PCR阳性植株,平均转化率为6.3‰;在喷施除草剂后存活的26株转化处理植株中,经PCR和Southern blot检测,获得了3株阳性抗草甘膦T0植株。本方法操作简单,成本低,避免了冗长繁琐的组织培养过程,为蓖麻遗传转化研究提供了新的思路和方法。

To improve the castor（ Ricinus communis L.） gene transformation,imbibed seeds（ germinating embryos） were transformed with herbicide resistant gene CP4- EPSPS by Agrobacterium- mediated transformation without tissue culture. Result showed that pricking 3 stitches at meristematic region（ apical point） of germinating seeds followed by ultra- sonication（ 800 W）,and co- cultured with Agrobacterium for 16 h were the optimum transformation conditions. 5 putative transgenic plants were detected from 800 randomly- selected treated seedlings,with average positive rate of 6. 3 ‰. After glyphosate screening,3 positive T0 seedlings were obtained out of26 survivals by PCR and Southern blot. This method was much simpler and more effective than the procedure of tedious tissue culture.