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  被引量:7

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

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作  者:雷江荣[1] 王冬梅[1] 邵林[1] 危晓薇[1] 黄乐平[1] 

机构地区:[1]新疆农业科学院核技术生物技术研究所,乌鲁木齐830091

出  处:《分子植物育种》2010年第2期252-258,共7页Molecular Plant Breeding

基  金:农业部转基因专项(2009ZX08005-011B)资助

摘  要: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.

关 键 词:SNC1 农杆菌介导法 茎尖 陆地棉 枯萎病 

分 类 号:S562[农业科学—作物学]

 

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