论文部分内容阅读
利用基因枪法将含有潮霉素抗性基因ｈｐｔ，ｇｕｓＡ报告基因和雪花莲外源凝集素基因ｇｎａ的２个质 粒（ｐＷＲＧ１５１５和ｐＲＳＳＧＮＡ１）共转化粳稻品种鄂宜１０５号和鄂晚５号种子胚诱导的愈伤组织．从轰击的 ３２９块愈伤组织中共再生出６１株独立转基因植株．ＰＣＲ／Ｓｏｕｔｈｅｒｎ ｂｌｏｔ分析显示，７９％的转基因植株含有 所有３个外源基因．Ｗｅｓｔｅｒｎ ｂｌｏｔ分析发现，４８株含ｇｎａ基因的转基因植株中有３６株（７５％）在不同水平上 表达了ＧＮＡ，ＧＮＡ最高表达量占总可溶性蛋白的０．５％．遗传分析证实外源基因在转基因后代植株中大 多以孟德尔方式遗传．从其Ｒ１代为 ３：１孟德尔分离方式遗传的后代Ｒ２代植株中，鉴定出含有并表达 所有３个外源基因的５个独立转基因植株纯系．褐飞虱生物鉴定和喂养试验表明，转基因纯系对褐飞虱 具有显著的抑制作用，具体表现为降低褐飞虱存活率和繁殖力、延缓褐飞虱发育进度及减少褐飞虱进 食量，即表达ＧＮＡ的转基因水稻纯系对褐飞虱具有抗性作用．
Two plasmids containing the hygromycin resistance gene hpt, gusA reporter gene and the snowdrop lectin gene gna (pWRG1515 and pRSSGNA1) were co-transformed into the seed embryos of Oryi 105 and Erzhuang 5 by using the gene gun method Of callus. A total of 61 independent transgenic plants were regenerated from the bombarded 329 callus. PCR / Southern blot analysis showed that 79% of the transgenic plants contained all three foreign genes. Western blot analysis showed that 36 (75%) of 48 transgenic plants containing gna gene expressed GNA at different levels, and the highest expression level of GNA was 0.5% of total soluble protein. Genetic analysis confirmed that most of the exogenous genes are inherited in Mendelian fashion in transgenic progeny plants. Out of R2 progeny plants derived from the R1 generation 3: 1 Mendelian segregation, five independent transgenic lines that contained and expressed all three foreign genes were identified as pure lines. The brown planthopper biopsy and feeding experiments showed that the transgenic lines had a significant inhibitory effect on N. lugens, specifically reduced the survival rate and fertility of the brown planthopper, delayed the development of the brown planthopper and reduced the food intake of the brown planthopper, that is, With resistance.