摘要
【目的】苦瓜枯萎病是由尖孢镰刀菌苦瓜专化型(Fusarium oxysporum f.sp.momodicae)侵染引起的一种重要土传病害。为有效防控该病害,对其病原菌进行绿色荧光蛋白基因(gfp)标记,为研究病原菌在苦瓜植株体内的侵染特性提供可视化跟踪检测手段。【方法】采用农杆菌介导的遗传转化方法对苦瓜枯萎病原菌强致病性野生型菌株FJAT-3018进行绿色荧光蛋白基因(gfp)标记,通过对转化子的菌落形态观察、生长速率和致病性测定,筛选出遗传稳定的转化子。【结果】野生型菌株FJAT-3018的转化效率约为14.5个转化子/10^6个孢子。经10次继代培养,筛选获得的转化子在菌落形态、生长速率和致病性方面与野生型菌株FJAT-3018无明显差异,gfp基因在转化子的菌丝体和分生孢子中均能强表达;通过激光共聚焦显微镜扫描跟踪发现,转化子能够在苦瓜植株根部和茎部侵染与定殖。【结论】绿色荧光蛋白基因已成功转入到苦瓜野生型菌株FJAT-3018中,其转化子具有良好的遗传稳定性且致病力不受影响。
【Objective】As a genetic marker,the green fluorescent protein gene(gfp)of Fusarium oxysporum f.sp.momodicae(Fom),one of the most serious fungal pathogens that caused the fusarium wilt on bitter gourd,was identified and transformed into bacterium to facilitate the study on the infection process and control of the disease.【Method】A highly pathogenic wild-type strain,FJAT-3018,isolated from diseased bitter gourds was transformed with the gene encoding gfp using an Agrobacterium-mediated method.The genetically stable transformants,FJAT-31290 and FJAT-31284,were verified by their colony morphology,growth rate,and pathogenicity.【Result】The efficiency of transforming FJAT-3018 was approximately 14.5 transformants per 10^6 spores.After 10 generations of subculture,the transformants did not significantly differ from the wild-type strain with respect to the colony morphology,growth rate,and pathogenicity.Strong constitutive expression of gfp could be seen in the fungal hyphae and conidia of the transformants.In addition,under a confocal laser scanning microscope,the transformant-infected bitter gourd plants showed growth of the fungal hyphae inside the roots and stem xylem.【Conclusion】The gfp gene had been successfully transformed into FJAT-31290 and FJAT-31284 with genetic stability and without apparent pathogenicity deviation from the highly pathogenic wild-type strain FJAT-3018.
作者
陈燕萍
刘欣
肖荣凤
朱育菁
林永胜
刘波
CHEN Yanping;LIU Xin;XIAO Rongfeng;ZHU Yujing;Lin Yongsheng;LIU Bo(Institute of Agricultural Biological Resource Research,Fujian Academy of Agricultural Sciences,Fuzhou,Fujian 350003,China)
出处
《福建农业学报》
CAS
CSCD
北大核心
2020年第11期1228-1233,共6页
Fujian Journal of Agricultural Sciences
基金
福建省科技计划公益类专项(2018R1017-9)
福建省农业科学院科技创新团队建设项目(STIT2017-2-8)。
关键词
苦瓜
枯萎病
绿色荧光蛋白
基因标记
Bitter gourd
Fusarium oxysporum
green fluorescent protein
genetic marker
