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从发病的番木瓜茎基部组织分离培养得到纯分离物,采用形态学和分子生物学方法进行分离物鉴定。经过致病性测定证实,分离物确定为番木瓜枯萎病的病原菌;通过形态学观察表明,与茄腐镰刀菌(Fusarium solani)形态一致。根据真菌的核糖体DNA(rDNA)内转录间隔区(ITS)的保守性和变异性,采用通用引物ITS1和ITS4进行PCR扩增,然后将PCR产物纯化后直接测序,对测试菌株与10株来自GenBank中的菌株ITS序列进行聚类分析,结果表明,测试菌株与GenBank上注册的茄腐镰刀菌GU134886、GQ451337、EU625405、EF534183和GQ376115聚在同一分枝上,分子鉴定结果与形态学观察结果一致,此为国内首次报道。
Pure papaya was isolated and cultured from the diseased papaya stalks, and the isolates were identified by morphological and molecular biological methods. The result of pathogenicity test confirmed that the isolate was the pathogen of Fusarium wilt. The morphological observation indicated that it was consistent with the morphology of Fusarium solani. According to the conservatism and variability of the internal transcribed spacer (ITS) region of ribosomal DNA (rDNA) in fungi, PCR was amplified by using universal primers ITS1 and ITS4, and then the PCR products were directly sequenced after purification. The results of cluster analysis of ITS sequences in GenBank showed that the test strains were clustered on the same branch with Fusarium solani GU134886, GQ451337, EU625405, EF534183 and GQ376115 registered on GenBank, and the molecular identification results were consistent with the morphological observations This is the first time in China.