将抗真菌病基因导入马铃薯的研究

Studies on Introduction of Antifungal Genes into Potato

实验采用微束激光穿刺技术和农杆菌介导方法 ,将携带有 型烟草β 1 ,3葡聚糖酶基因和 型菜豆几丁质酶基因的 p BLGC质粒导入马铃薯 ( Solanum tuberosum L .)品种“津引 8号”。微束激光穿刺技术转化中 ,获得了 7株卡那霉素抗性再生苗 ,分子检测证明 ,其中有 1株为菜豆几丁质酶基因单基因转化植株。在农杆菌介导方法中得到了 1 3株卡那霉素抗性再生苗 ,其中有 3株为烟草β 1 ,3葡聚糖酶基因单基因转化植株。实验证明 ,同常规农杆菌介导方法相比 ,微束激光穿刺技术更为简便有效 ,细胞损伤小 ,为马铃薯遗传转化提供了一个外源基因直接导入的新途径。

The binary vector pBLGC, harboring tobacco class Ⅰ β1,3glucanase and bean class Ⅰ chitinase, were successfully introduced into the microtuber discs of potato (Solanum tuberosum L.) cultivars 'Jinyin 8' by using the Laser microbeam Puncture Techniques(LMPT) and Agrobacteriummediated methods. In the LMPT transformation, 7 kanamycinresistant regenerated plantlets have been obtained and the PCR and Southern blot analyses confirmed that 1 plant was bean class Ⅰ chitinasepositive. In the parallel experiment of Agrobacteriummediated methods, 3 of 13 kanamycinresistant regenerated plantlets was tobacco class Ⅰ β1,3glucanasepositive according to PCR and PCRSouthern analyses. It proved that both methods were applicable for potato transformation and that the LMPT was more effective and has less celldamage.