农杆菌介导葡萄糖氧化酶基因转化油菜

以甘蓝型油菜带柄子叶为转化受体材料,采用农杆菌介导法进行葡萄糖氧化酶(Glucose oxidase,GO)基因的转化,经卡那霉素抗性筛选获得大量的不定芽.PCR检测结果显示其中22株呈阳性.淀粉-KI染色结果表明在转基因植株中检测到H2O2.在转基因植株×非转基因植株(F1代)中,卡那霉素抗性筛选显示86%的株系呈现典型的孟德尔单基因显性遗传.F1代盛花期茎秆菌核病活体接种结果证实,转基因植株后代菌核病抗性明显提高.证实GO基因已转入油菜植株中并得到有效表达.

Agrobacterium tumefaciens Mediated Introduction of Glucose Oxidase Gene into Brassica napus L.

To transform the glucose oxidase(GO) gene into rapeseed(Brassica napus L.) so as to enhance the resistance to Sclerotinia sclerotiorum,the cotyledon petiols of rapeseed were used as recipients of GO gene introduction mediated by Agrobacterium tumefaciens.Cotyledon petiols produced a lot of adventitious buds on selection medium supplemented with kanamycin.GO gene was confirmed by PCR on rapeseed genome.The hydrogen peroxide generated by the transgenic plants was detected by color reaction with starch-KI which proved that the glucose oxidase generated by the expression of GO gene had exerted its function on rapeseed.Kanamycin resistance in 86% of F1 of non-transgenic plants and transgenic plants presented typical Mendelian single dominance gene inheritance.By stem toothpick inoculation in F1 generation during fowereing,transgenic plants exhibited an increased resistance against Sclerotinia sclerotiorum than non transgenic plants did, which confirmed that GO gene had been transferred into and expressed in rapeseed.