3种瓜类枯萎病菌Fusarium oxysporum的分子检测  

Molecular detection of Fusarium oxysporum in cucumber,watermelon and melon

作　　者：张淑梅[1,2] 赵晓宇[1] 张先成[1] 孟利强[1] 李晶[1] 张云湖[1] 王玉霞[1] 

机构地区：[1]黑龙江省科学院微生物研究所,哈尔滨150010 [2]东北农业大学园艺学院,哈尔滨150030

出　　处：《植物病理学报》2010年第6期636-641,共6页Acta Phytopathologica Sinica

基　　金：黑龙江省自然科学基金项目(200839)

摘　　要：Fusarium oxysporum is one of the most important phytopathogens and cause Fusarium wilt disease in cucumber,watermelon and melon,etc.In this study,a pair of species-specific primers Fc-1 and Fc-2 was synthesized based on differences in internal transcribed spacer sequences of Fusarium genus.With the primers,a specific 315 bp PCR product was amplified from five F.oxysporum isolates isolated from cucumber,watermelon and melon,infected cucumber and watermelon tissues,while no product was obtained from other fourteen fungi,healthy cucumber and watermelon tissues.The detection sensitivity is 100 fg for genomic DNA of F.oxysporum and 1 000 spores/g soil for the soil pathogens.In contrast,the nested PCR with two pairs of primers(ITS1/ITS4 and Fc-1 /Fc-2) increased the sensitivity by 100-fold.In addition,one-step PCR could also detect F.oxysporum in symptomless cucumber root of 7 dpi(days post inoculation) and in infected cucumber and watermelon tissues at the early stage of disease development.Therefore,the developed PCR-based method enabled rapid,sensitive and reliable detection of F.oxysporum.It also provides the detection method for early monitoring and diagnosis of the pathogen as well as the plant disease management guidance.Fusarium oxysporum is one of the most important phytopathogens and cause Fusarium wilt disease in cucumber,watermelon and melon,etc.In this study,a pair of species-specific primers Fc-1 and Fc-2 was synthesized based on differences in internal transcribed spacer sequences of Fusarium genus.With the primers,a specific 315 bp PCR product was amplified from five F.oxysporum isolates isolated from cucumber,watermelon and melon,infected cucumber and watermelon tissues,while no product was obtained from other fourteen fungi,healthy cucumber and watermelon tissues.The detection sensitivity is 100 fg for genomic DNA of F.oxysporum and 1 000 spores/g soil for the soil pathogens.In contrast,the nested PCR with two pairs of primers（ITS1/ITS4 and Fc-1 /Fc-2） increased the sensitivity by 100-fold.In addition,one-step PCR could also detect F.oxysporum in symptomless cucumber root of 7 dpi（days post inoculation） and in infected cucumber and watermelon tissues at the early stage of disease development.Therefore,the developed PCR-based method enabled rapid,sensitive and reliable detection of F.oxysporum.It also provides the detection method for early monitoring and diagnosis of the pathogen as well as the plant disease management guidance.

关 键 词：瓜类枯萎病菌 分子检测 发病率 常见病害 瓜类作物 黄瓜 西瓜 甜瓜 

分 类 号：S436.421[农业科学—农业昆虫与害虫防治]