甘肃省马铃薯镰刀菌干腐病优势病原的分离鉴定  

Isolation and identification of the dominant pathogens causing potato Fusarium dry rot in Gansu Province

作　　者：李金花[1,2] 王蒂[2] 柴兆祥[1] Lester W.Burgess 

机构地区：[1]甘肃农业大学草业学院,兰州730070 [2]甘肃省作物遗传改良与种质创新重点实验室,兰州730070 [3]澳大利亚悉尼大学农业食品与自然资源学院

出　　处：《植物病理学报》2011年第5期456-463,共8页Acta Phytopathologica Sinica

基　　金：甘肃省科技厅项目(2GS054-A41-005-01);甘肃省作物遗传改良与种质创新重点实验室开放基金项目(GKLAU0603);"农业部公益性行业科研专项(3-6-3)";甘肃省教育厅研究生导师科研资助项目(1002-05)

摘　　要：为明确甘肃马铃薯镰刀菌干腐病的优势病原,2006年12月～2007年3月由西至东从甘肃张掖、天祝、永登、临洮、渭源和西和等6县市的马铃薯贮藏窖中采集表现有镰刀菌干腐病症状的马铃薯薯块,以组织分离法分离病原,单孢纯化镰刀菌(Fusarium spp.)菌株后,以形态学为基础,参照Nelson镰刀菌分类系统进行鉴定。结果表明:6个采样区共分离到293株镰刀菌菌株,其中以接骨木镰刀菌(F.sambucinum)和茄病镰刀菌(F.solani)出现频率高,是优势种。分析发现第一优势种随采样区而异,张掖、天祝和渭源采集的样品中茄病镰刀菌分离频率分别为42.6%、42.1%和32.4%,接骨木镰刀菌分离频率分别为14.8%、5.3%和26.5%,茄病镰刀菌为第一优势种;永登、临洮和西和采集的样品中接骨木镰刀菌分离频率分别为52.1%、50.9%和55.2%,茄病镰刀菌的分离频率分别为23.3%、32.7%和20.7%,接骨木镰刀菌为第一优势种。本文进一步对其在PDA、CLA上的培养特征进行了观察和描述。按照柯赫氏法则用混合菌株接种法对大西洋(Atlantic)、夏波蒂(Shepody)以及一地方品种进行致病性测定,证实了优势菌种对块茎的致病性。利用EF-1α基因引物(EF-1H和EF-2T)对接骨木镰刀菌菌株GAUF-F12进行基因组DNA的PCR扩增,将PCR产物回收测序后在GenBank上比对,菌株GAUF-F12与GenBank登记的接骨木镰刀菌5个菌株的同源性均达99%;用DNASTAR分析软件将同源性较高的登记菌株的序列与GAUF-F12菌株构建同源性树,结果表明:该菌株与以上5个接骨木镰刀菌菌株均位于同源性树的同一分支,聚为一类,与形态学的鉴定结果一致。To know the dominant pathogens of potato Fusarium dry pot in Gansu,tubers showing symptoms of dry rot were sampled at farmers＇ storage caves in Zhangye city,Tianzhu,Yongdeng,Lintao,Weiyuan and Xihe county from Feb.2006 to Mar.2007.The pathogens were isolated by tissue isolation method,the Fusa-rium obtained were identified according to the taxonomic system of Nelson after being sub-cultured and single-spored.The results showed that the total 293 Fusarium isolates were obtained from the 12 storage caves in six sampling sites,and the dominant species were F.sambucinum and F.solani.The results also indicated that the dominant species were different in different ecological areas,for example,F.solani was the dominant pathogen in Zhangye,Tianzhu and Weiyuan,and where its isolation frequencies were 42.6%,42.1% and 32.4% respectively,and that of F.sambucinum were 14.8%、5.3% and 26.5% respectively.F.sambucinum was the dominant pathogen in Yongdeng,Lintao and Xihe,and where its isolation frequencies were 52.1%、50.9% and 55.2%,and that of F.solani were 23.3%、32.7% and 20.7%.The morphological characteristics of these two species on PDA and CLA were observed and described in details in this paper.According to Koch＇s Postulate,the pathogenicity of the dominant pathogen were tested with mixed isolates on potato varieties Atlantic,Shepody and a local variety from Tongwei county.It was verified that both of the two dominant Fusarium species were pathogenic to potato tubers.Genomic DNA was isolated from pure cultures of GAUF-F12 strain of F.sambucinum and amplified with primer pairs EF-1α（EF-1H and EF-2T）.The PCR product was then sequenced,and aligned in GenBank.It was indicated that there was a very close relationship between GAUF-F12 and the 5 F.sambucinum isolates,and the max similarity of their sequences was 99%.DNASTAR was used to draw the phylogenetic tree of isolate GAUF-F12.It was showed that strain GAUF-F12 and the above 5 F.sambucinum isolates were in the same cluster,which was consistent with the results

关 键 词：马铃薯镰刀菌干腐病 优势病原 F.sambucinum F.solani EF-1α基因 PCR 

分 类 号：S435.32[农业科学—农业昆虫与害虫防治]