西瓜枯萎病菌和哈茨木霉的GFP标记及根部动态定殖比较  

Comparison of GFP Markers and Root Dynamic Colonization of Fusarium oxysporum f.sp.niveum and Trichoderma harzianum in Watermelon

作　　者：孟素玲 王媛[1] 顾欣[1] 王新谱[1] MENG Suling;WANG Yuan;GU Xin;WANG Xinpu(School of Agriculture,Ningxia University,Yinchuan 750021,China)

机构地区：[1]宁夏大学农学院,银川750021

出　　处：《西北农业学报》2024年第2期355-363,共9页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：国家自然科学基金(41761054);农业高质量发展和生态保护科技创新示范项目(NGSB-2021-10-01)。

摘　　要：为明确西瓜枯萎病菌和生防哈茨木霉在西瓜根部动态定殖的差异,采用PEG-CaCl2介导的原生质体转化法,将携带绿色荧光蛋白基因的pCT74质粒分别与尖镰孢菌西瓜专化型和哈茨木霉M3菌株的基因组整合获得相应的转化子;将转化子接种于土壤,利用激光共聚焦显微镜示踪并比较两菌株转化子对西瓜幼苗根部的侵染动态过程。结果表明,转化子连续传代6次仍能发出绿色荧光且荧光强度稳定,能够稳定遗传,经PCR验证绿色荧光蛋白基因转入成功。盆栽试验中,两种转化子均能成功定殖于西瓜幼苗根部,在定殖初期,尖镰孢菌西瓜专化型在根内的扩散速度快于哈茨木霉M3菌株。To clarify the differences in colonization dynamics of watermelon roots by Fusarium oxys-porum f.sp.niveum(FON)and Trichoderma harzianum,the protoplast mediated by PEG-CaCl2 method was used to integrate the pCT74 plasmid carrying the green fluorescent protein(GFP)gene,so as to obtain the corresponding transformants with the genomes of FON and T.harzianum M3 strains,respectively.The transformants were inoculated in soil,and laser scanning confocal microscope(LSCM)was used to trace and compare the dynamic of root infestation of watermelon seedlings by the two strains.The results showed that the transformants could still emit green fluorescence with stable fluorescence intensity for six consecutive passages and could be inherited stably,and the green fluorescent protein gene was successfully transferred by PCR.In this experiment,both transformants were successfully colonized in the roots of watermelon seedling.At the early stage of colonization,the FON spread faster in the roots than the T.harzianum M3 strain.The relationship between Tri-choderma,FON and host plants,environment provides a basis for clarifying the colonization pattern of Trichoderma.

关 键 词：哈茨木霉 尖镰孢菌西瓜专化型 绿色荧光蛋白 定殖 

分 类 号：S436.5[农业科学—农业昆虫与害虫防治]