棉花枯萎病菌新生理型菌株的分子鉴定  

Molecular identification of the new genotype strains of Fusarium oxysporum f.sp.vasinfectum in China

作　　者：郭庆港[1] 王培培[1] 鹿秀云[1] 董丽红[1] 赵卫松[1] 张晓云[1] 李社增[1] 马平[1] GUO Qing-gang, WANG Pei-pei, LU Xiu-yun, DONG Li-hong, ZHAO Wei-song, ZHANG Xiao-yun, LI She-zeng, MA Ping(Institute of Plant Protection, Hebei Academy of Agricultural and Forestry Sciences, Integrated Pest Management Center of Hebei Province, Key Laboratory of IPM on Crops in Northern Region of North China, Ministry of Agriculture, Baoding 071000, Chin)

机构地区：[1]河北省农林科学院植物保护研究所,河北省农业有害生物综合防治工程技术研究中心,农业部华北北部作物有害生物综合治理重点实验室,保定071000

出　　处：《植物病理学报》2018年第2期145-153,共9页Acta Phytopathologica Sinica

基　　金：国家现代棉花产业技术体系(CARS-18-15);河北省财政专项(F17C10007)

摘　　要：Fusarium oxysporum f.sp.vasinfectum(Fov)已报道有8个生理小种和一个澳大利亚生理型,在我国只报道有3、7、8号小种。前期研究发现,我国还有一个与3、7、8号小种具有较大遗传差异的Fov新生理型。本研究通过对3、7、8号小种和新生理型菌株的翻译延伸因子(EF-1α)基因序列比对分析,发现该新生理型菌株具有特异性碱基序列。根据这些特异性碱基序列,设计了一对针对新生理型菌株的特异性引物,并证明该引物可以特异性检测Fov新生理型菌株。利用该引物对采自河北省主要植棉区的77株Fov菌株进行筛选,鉴定出2株潜在的新生理型菌株。从这2个Fov菌株中克隆出EF-1α和β-tubulin基因序列,通过构建系统发育树,证明这2个Fov菌株为新生理型菌株,而且与澳大利亚生理型菌株具有较近的亲缘关系。本研究建立的分子检测方法可用于棉花枯萎病菌新生理型菌株的鉴定。Fusarium oxysporum f. sp. vasinfectum （Fov）, the pathogen of cotton fusarium wilt, was discrimi- nated into eight races and an Australian genotype in the world based on their pathogenicity to differential hosts. The races of 3, 7 and 8 had been reported in China, but a suspected novel genotype of Fov was frequently iden- tified in our lab. The purpose of this study was to develop a PCR-based technique for fast detection of the novel genotype strains from large number of samples collected in various cotton-growing areas. By performing gene se- quence alignment of elongation factor （EF-la） between races 3, 7, 8 and the novel genotype strain, several new genotype strain-specific bases were obtained. A pair of novel genotype strain-specific primers was designed based on those specific bases and used for PCR detection. It was confirmed that the primer set was specific for the novel genotype strains. Seventy seven Fov isolates collected from the main cotton-growing areas in Hebei province were tested with the novel genotype strain-specific primers, and two suspected isolates were obtained. The phylogenetic tree analysis revealed that these two suspected isolates formed a single clade with the novel genotype strain and showed a closer genetic relationship to the Australian genotype strain than the other races based on the combined EF-la and beta-tubulin genes sequence data. It was concluded that the new method could be used for rapid detection of the novel genotype strain of Fov.

关 键 词：尖孢镰孢菌 特异性 分子检测 遗传多样性 

分 类 号：S476[农业科学—农业昆虫与害虫防治]