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

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.