西瓜ClPDF2.1蛋白的原核表达及其对香蕉枯萎病菌的抑制活性测定  

Prokaryotic Expression and Antifungal Activity of Watermelon Cl PDF2.1 Against Fusarium oxysporum f. sp. cubense 4

作　　者：张继友[1] 景晓辉[2] 吴伦英[1] 刘国道[3] 

机构地区：[1]海南大学农学院,海南海口570228 [2]北京大学生命科学学院,北京100871 [3]中国热带农业科学院品种资源研究所,海南儋州571737

出　　处：《热带作物学报》2015年第6期1161-1165,共5页Chinese Journal of Tropical Crops

基　　金：国家自然科学基金项目(No.70963003);国家973计划项目(No.2007CB08903)

摘　　要：尖孢镰刀菌古巴专化型侵染香蕉引起的香蕉枯萎病严重危害中国香蕉产业。植物防卫素是一类小的富含半胱氨酸的抗真菌蛋白。本研究旨在了解西瓜(Citrullus lanatus)防卫素蛋白Cl PDF2.1对香蕉枯萎病菌的抑制活性。提取西瓜RNA后反转录为c DNA,以西瓜c DNA为模板,克隆Cl PDF2.1基因,将测序正确的Cl PDF2.1连接到载体p GEX-6P-1,随后转化Trans BL21(DE3)p Lys S,IPTG诱导GST-Cl PDF2.1融合蛋白表达16 h后,经SDS-PAGE电泳及Western blot鉴定GST-Cl PDF2.1融合蛋白的表达,进行GST-Cl PDF2.1融合蛋白体外抑制香蕉枯萎病菌生长的功能初探。本研究为进一步培育转基因香蕉抗病品种奠定基础。Banana wilt disease, caused by the pathogenic fungus Fusarium oxysporum f. sp. cubense 4 （Foc 4）, is regarded as one of the mostserious threat to banana production. Plant defensins are cysteine-rich innate anti- microbial proteins that play an important role in plant disease resistance, particularly to fungal diseases. In this paper, watermelon defensin C1PDF2.1 cDNA sequences were cloned and then inserted into vector pGEX-6P-1 with glutathione-S-transferase （GST） tag to construct the prokaryotic expression plasmid. The recombinant plasmids pGEX-6P-1-C1PDF2.1 were transformed into Trans BL21 （DE3） pLysS and expressed in the prokaryotic expression system. Results of SDS-PAGE and western blot after Glutathione Sepharose 4B affinity chromatography purification, showed that the specific fusion protein was successfully constructed and expressed after IPTG induction. The antifungal activity of watermelon defensins proteins C1PDF2.1 against Foc 4 growth indicated that C1PDF2.1 could decrease Foc 4 hypha growth by 81.7% at eighty micromolar concentration. This study showed that CIPDF2.1 could be a potential resistance gene for controlling banana Fusarium wilt disease.

关 键 词：西瓜防卫素基因Cl PDF2.1 原核表达 抗真菌活性 香蕉枯萎病菌 

分 类 号：S651[农业科学—果树学]