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通过激光微束穿刺法 ,将 β 1,3 葡聚糖酶及几丁质酶基因双价植物表达载体 pBLGC导入棉花幼胚。转化一代棉花幼苗用蘸根法进行抗黄萎病筛选 ,将存活的苗移入病圃进行了卡那霉素(Kan 1% )抗性测定。结果表明 ,在移栽的 2 9株幼苗中 ,有 9株表现出明显的Kan抗性 ,对 9株抗Kan植株进行了PCR检测 ,结果显示 ,其中 7株表现为阳性。病圃中的T1代转基因植株在经历了黄萎病发病高峰期后 ,7株PCR阳性植株表现明显抗病 ,并已正常开花结铃 ,其他T1代植株及对照植株全部因后期发病死亡。这初步证明外源基因已整合到棉花基因组中 ,且使转基因植株对黄萎病表现出一定的抗性。
The bivalent plant expression vector pBLGC of β 1,3-glucanase and chitinase gene was introduced into the immature embryo of cotton by means of laser microbeam puncture. A new generation of cotton seedlings was transformed into resistant Verticillium dahliae by dip root method. Surviving seedlings were transferred to the nursery for kanamycin (Kan 1%) resistance assay. The results showed that 9 out of 29 transplanted seedlings showed obvious Kan resistance. Nine of the Kan plants were tested by PCR. The results showed that 7 of them were positive. The T1 transgenic plants in the nursery showed seven disease-resistant plants after the peak of the incidence of Verticillium wilt. The T1 plants had flowering and boll-forming. All the other T1 and all the control plants died due to the later stage. This preliminary evidence that foreign genes have been integrated into the cotton genome, and the transgenic plants showed resistance to Verticillium wilt.