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以抗大豆食心虫品系东农8004的胚尖为外植体,通过对芽诱导和芽伸长培养基进行优化,建立8004的胚尖转化体系,利用该体系进行cry1C*基因的转化,对获得的抗性再生植株进行分子鉴定和抗虫性鉴定。结果表明:在芽诱导培养基中附加1.67 mg·L-16-BA,丛生芽诱导率最高,达到80%,在芽伸长培养基中附加0.5 mg·L-1GA3和0.1mg·L-1IAA,芽伸长率最高为54%。共获得94株PPT(100μg·m L-1)抗性再生植株,PCR检测阳性14株,阳性率为14.89%。对4株T0代阳性植株进行大豆食心虫抗性鉴定,其中C-1植株的食心虫抗性明显高于野生型8004,说明在大豆中表达cry1C*基因能提高大豆对食心虫的抗性。
The embryo tip of Dongnong 8004 was used as explants to optimize the culture medium of buds and shoot elongation. The embryo tip transformation system of 8004 was established and the cry1C * gene was transformed by this system. Resistant regenerated plants for molecular identification and insect resistance identification. The results showed that 1.67 mg · L-16-BA was added into shoot induction medium, the induction rate of shoots was the highest, reached 80%, and 0.5 mg · L-1 GA3 and 0.1 mg · L-1 IAA , Bud elongation up to 54%. A total of 94 strains of PPT (100μg · m L-1) -resistant plants were obtained, of which 14 were positive by PCR. The positive rate was 14.89%. Four T0 generation positive plants were identified for their resistance to Siberian bean worms. The resistance of C-1 plants to Drosophila melanogaster was significantly higher than that of wild type 8004, indicating that expression of cry1C * gene in Soybean could increase the resistance of Soybean to Daucus.