农杆菌法转化茎尖获得转SNC1基因棉花及其T_1植株对棉花枯萎病的抗性

Agrobacterium-mediated Transformation of SNC1 Gene into Cotton Shoot Apex and Fusarium Wilt Resistance in T_1 Plants

以陆地棉中35和军棉1号的茎尖为外植体,利用农杆菌介导法将含有拟南芥抗病基因SNC1(sup-pressor of npr1-1,constitutive 1)转入棉花。对外植体培养时期、农杆菌侵染时间和共培养时间进行改良,实验结果表明,在外植体培养1d,菌液侵染20min,并且共培养保持在2d能够获得较高的遗传转化效率。PCR以及RT-PCR对再生植株T0代和T1代棉花的检测结果表明,SNC1基因已经整合到棉花的基因组中并得到表达。利用浸根法,对T1代转基因棉花接种棉花枯萎病菌强致病力菌株(Fusarium oxysporum f.sp.Vasinfectum),与对照比较,T1代转基因棉花的枯萎病抗性明显提高。

Using shoot apex from ＂Zhong 35＂ and ＂Junmian No. 1＂ as explant, we have successfully transferred the disease resistance gene SNC1 （suppressor ofnprl-1, constitutive） cloned from A rabidopsis thaliana into cotton by Agrobacterium-mediated transformation. The transformation conditions, including cultural stage of the explant, infection time, and co-culture time, were also optimized. The results showed that the relative high transformation rate was obtained following conditions as follows： one day of explant culture, 20 min of infection time, and two days of co-culture. PCR and RT-PCR test of To and T1 generations revealed that the SNC1 gene was successfully integrated into cotton and expressed. In addition, T1 generation infected with Fusarium oxysporum f. sp. Vasinfec- turn through root-dip method were discovered to own improved resistance to the disease compared with non-trans- genic plants.