利用微束激光穿刺法将抗真菌基因导入棉花的研究初报

Studies on Introduction of Antifungal Genes into Cotton Plants by Laser Microbeam Puncture

以棉花感受态萌动种胚作为外源基因转化的受体,用激光微束穿刺法将β-1,3-葡聚糖酶及几丁质酶双价基因导入棉花,所构建的植物表达载体pB IBGC携带有筛选标记npt-Ⅱ(新霉素磷酸转移酶)基因。激光转化处理的种胚经卡那霉素筛选,已经获得抗性小植株。研究了微束激光转化法用于棉花感受态萌动胚转化预培养的时间、高渗液对材料的处理等。研究表明:用微束激光转化处理种胚是一种操作简便、重复性好的转化方法,用该法可将外源基因导入植物感受态萌动胚,避开植株离体再生的困难。

Using cotton susceptive seed-germinating embryos as transgenic receptor, the bivalent genes pBIBGC, which includes β-1,3-glucanase gene and chitinase gene, was introduced into cotton by laser microbeam Puncture Techniques （ LMPT）. There are npt- Ⅱ （ neomycin phosphotransferase Ⅱ ） gene as screening marker in the plant bivalent expression vector. From treated embryos by laser transformation, resistant plantlets have been obtained after kanamycin resistance screening. In this research, we have also studied some key factors that influenced LMPT, such as the duration of pretreatment of susceptive seed-germinating embryos and hypertonic treatment, and so on. The results indicated that the LMPT was easy to operate and repetitive. The foreign genes were introduced into susceptive seed-germinating embryos by this technique, which avoids difficulties of regeneration of plantlets.