棉花尖孢镰刀菌绿色荧光蛋白的转化  

Transformation of Green Fluorescent Protein in Fusarium oxysporum of Cotton

作　　者：侯甲男 李武[2] 杨晓杰[2] 孙瑶[2] 谢德意[2] 汪蒙蒙 房卫平[2] 赵付安[2] Hou Jianan;Li Wu;Yang Xiaojie;Sun Yao;Xie Deyi;Wang Mengmeng;Fang Weiping;Zhaoruan(School of Life Sciences, Henan University, Kaifeng, 475001;Economic Research Institute, Henan Academy of Agricultural Sciences, Zhengzhou,450002)

机构地区：[1]河南大学生命科学学院,开封475001 [2]河南省农业科学院经济作物研究所,郑州450002

出　　处：《分子植物育种》2019年第10期3247-3252,共6页Molecular Plant Breeding

基　　金：国家棉花转基因重大专项子课题(2016ZX08005-001-005);中原学者项目(172101510001);2018河南省财政预算项目(20188101);河南省杰出人才项目(154200510026);河南省农业科学院优秀青年科技基金计划项目(2018YQ07)共同资助

摘　　要：棉花枯萎病是棉花的主要病害之一,严重威胁棉花产量和品质。绿色荧光蛋白(GFP)是发光水母(Aequorea victoria)体内的一种蛋白,作为报告基因拥有诸多优点,目前已被广泛应用于植物研究当中。为研究枯萎菌浸染棉花的过程及植病互作分子机理,本研究利用PEG介导的原生质体转化方法,将绿色荧光蛋白(GFP)基因转入到棉花枯萎病菌中,创制转报告基因株系。结果表明:转化子连续转接2代后仍能够稳定遗传,且荧光强度良好。经荧光共聚焦显微镜观察和PCR验证GFP基因已成功转入到菌株中,转化所得的菌株将为后续可视化检测和分析枯萎病菌对棉花的侵染等研究提供帮助。Cotton Fusarium wilt is one of the main diseases of cotton and seriously threatens cotton yield and quality. Green fluorescent protein(GFP) is a kind of protein in Aequorea victoria. As a reporter gene, it has many advantages and has been widely used in plant research. In order to study the process of cotton infected by Fusarium oxysporum and the molecular mechanism of plant-pathology interaction, the method of PEG mediated protoplast transformation was applied to transform the GFP genes into Fusarium oxysporum and transgenic reporter lines were created. The results showed that the transformants with GFP could be inherited steadily and their fluorescence signal was kept strongly after more than two successive generations. It was confirmed that the GFP genes had been successfully transferred into the strain of Fusarium oxysporum by observation of fluorescence confocal microscopy and PCR verification. The acquisition of the transformants with GFP would lay a solid foundation for subsequent study of the visual detection and analysis about the infection of cotton by Fusarium oxysporum.

关 键 词：棉花尖孢镰刀菌 绿色荧光蛋白 原生质体转化 

分 类 号：S435.621.2[农业科学—农业昆虫与害虫防治]