黄瓜转新型抗菌蛋白基因GNK2-1及其抗枯萎病的研究  

Overexpression of a Novel Antifungal Protein Gene GNK2-1 Results in Elevated Resistance of Transgenic Cucumber to Fusarium oxysporum

作　　者：刘缙[1] 田花丽[1] 王亚红[1] 郭蔼光[1] 

机构地区：[1]西北农林科技大学生命科学学院,陕西省农业分子生物学重点实验室,杨凌712100

出　　处：《植物学报》2010年第4期411-418,共8页Chinese Bulletin of Botany

基　　金：国家转基因植物研究与开发专项(No.JY-03-A-11-01)

摘　　要：GNK2-1为一种来自银杏(Ginkgo biloba)种仁的新型抗真菌蛋白,具有较强的真菌抗性且性质稳定。序列分析表明,其结构与所有已知的抗真菌蛋白不同,而与富含半胱氨酸的植物类受体激酶的胞外结构域相似。为探索GNK2-1基因在黄瓜(Cucumis sativus)抗病反应中的作用,利用基因重组技术构建了GNK2-1的高效组成型表达载体,并利用根癌农杆菌(Agrobacterium tumefaciens)介导转入黄瓜栽培品种农城3号(Cucumis sativus' Nongcheng No.3')基因组中。通过对获得的抗性植株进行PCR、RT-PCR和Western blot检测分析,结果表明GNK2-1基因可在T0代转基因植株中转录表达,并能在T1代转基因黄瓜中稳定遗传。离体枯萎病抗性鉴定结果表明,转GNK2-1基因的黄瓜对枯萎病的抗性增强,GNK2-1可以作为黄瓜抗病性改良的潜在基因资源。A novel antifungal protein,ginkbilobin2-1（GNK2-1）,from Ginkgo biloba seed kernels,was proved to have a stable and significant inhibition effect on fungus growth.The protein showed no similarity to other pathogenesis-related proteins but did show homology to the extracellular domain of plant cysteine-rich receptor-like kinases.In order to improve resistance of transgenic plants to fungal infection,we used RT-PCR to amplify GNK2-1 from G.biloba seeds and transformed the gene into cucumber（Cucumis sativus） cultivar Nongcheng 3 via Agrobacterium-mediated method.Transgenic plants were obtained and the expression of the transgene was confirmed by PCR,RT-PCR and western blot analysis.Resistance tests against Fusarium oxysporum showed that expression of the GNK2-1 in transgenic cucumber plants conferred antifungal activity against this disease.The GNK2-1 gene may be a good candidate for breeding new cucumber varieties with resistance to fungal blight disease.

关 键 词：抗菌蛋白 黄瓜 枯萎病 转基因 

分 类 号：S642.2[农业科学—蔬菜学]