利用ITS1和ITS4通用引物扩增香蕉枯萎病菌核酸片段鉴定其生理小种  

Utility of Universal Primers, ITS1 and ITS4, to Amplify Sequences for Race Identification of Fusarium oxysporum f.sp. cubense

作　　者：曹永军[1,2] 程萍[2] 喻国辉[2] 黎永坚[2] 杨紫红[2] 周林[3] 

机构地区：[1]华南农业大学资源环境学院,广东广州510642 [2]珠海市农业科学研究中心,广东珠海519075 [3]华中农业大学生命科学技术学院,湖北武汉430070

出　　处：《热带作物学报》2010年第7期1098-1102,共5页Chinese Journal of Tropical Crops

基　　金：广东省科技厅教育部产学研结合项目(2007B090400083);珠海市科技计划项目(PC20081078;PC20082043)资助

摘　　要：利用真菌核糖体rDNA区通用引物ITS1和ITS4,扩增采集自香蕉的13株镰刀菌包括18S rDNA部分序列、ITS1-5.8S-ITS2全部序列和28S rDNA部分序列的片段,通过BLAST序列比对分析,确定13株菌为尖孢镰刀菌[Fusarium oxysporum]。采用最大简约法,以层出镰刀菌[F.proliferatum(EU151490)]和茄病镰刀菌[F.solani(GQ376116)]为外群,将13株菌的序列与BLAST检索获得的尖镰孢古巴专化型(F.oxysporum f.sp.Cubense)相应序列构建了系统发育树。系统发育分析结果显示,13株菌与NCBI登录的4个尖镰孢古巴专化型菌株一起被分成3个亚群,亚群聚类结果与回接鉴定结果及文献报道的生理小种结果完全一致。Thirteen different Fusarium strains were isolated from banana in Guangdong Province. Their ribosomal DNA internal transcribed spacer sequences, including 18S rDNA partial sequence, internal transcribed spacer 1, 5.8S rDNA, and internal transcribed spacer 2 complete sequence and 28S rDNA partial sequence, were amplified by using universal primers ITS1 and ITS4. The thirteen isolates were identified as F. oxysporum by the Basic Local Alignment Search Tool （BLAST） in NCBI. The amplified sequences of the thirteen strains and 4 F. oxysporum f. sp. cubense strains which had been registered in NCBI were used to phylogentic analysis taking F. proliferatum （EU151d90） and Fusarium. solani （GQ376116） from GenBank as outgroups. The results showed that all strains were classified into three subgroups, and the results of subgroups were identical to the strains reported in literature.

关 键 词：香蕉枯萎病菌 rDNA—ITS 系统发育分析 

分 类 号：S436.8[农业科学—农业昆虫与害虫防治]