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以河北省优良大豆品种冀豆15、五星2号和NF-58的子叶节为受体材料,利用农杆菌介导法进行遗传转化,探讨了影响农杆菌侵染后子叶节不定芽诱导的因素。结果表明:以萌发6 d、4℃处理24 h的子叶节为外植体,农杆菌侵染后经超声波处理30 s、共培养基中添加20 mg·L-1硝酸银,能够提高子叶节丛生芽诱导率,转化植株经草铵膦筛选以及PCR检测,T0代转化率达0.97%。利用该体系对大豆品种五星2号进行At NHX5基因的遗传转化,获得3株T0代RT-PCR检测阳性植株,且2-1号T1代阳性植株检测具有一定耐盐性,初步证明获得了转At NHX5基因的大豆新材料。
Agrobacterium-mediated genetic transformation was carried out using cotyledon nodes of excellent soybean varieties Jidou 15, Wuxing 2 and NF-58 in Hebei Province as the receptor materials, and the factors affecting the induction of adventitious buds of cotyledonary node after Agrobacterium tumefaciens infection were discussed . The results showed that the cotyledonary node which was treated for 6 days and 4 ℃ for 24 hours was used as explant and the Agrobacterium tumefaciens was sonicated for 30 seconds after the infection. The addition of 20 mg · L-1 silver nitrate in the co-culture medium could increase the cotyledonary node The induction rate of clustered buds, the transformation of plants by glufosinate screening and PCR detection, T0 generation conversion rate of 0.97%. The system was used to carry out genetic transformation of At NHX5 gene in soybean variety Wuxing No.2 to obtain 3 T0 generation RT-PCR positive plants, and the 2-1 generation T1 positive plants were tested for salt tolerance. At NHX5 gene of soybean new material.