摘要
炭疽菌属是一重要的植物病害真菌属,可引起多种植物病害,在农业生产上造成重大危害。黑痣菌属真菌引起的黑痣病也是一种常见病害,在刚竹属植物上普遍发生,还是许多牧草的病原菌。本试验在前人研究的基础上,对西北地区炭疽病和黑痣病病原菌进行鉴定,为《中国真菌志·炭疽菌属》、《中国真菌志·黑痣菌属》的撰写做基础工作。
鉴定炭疽病标本200余份,共鉴定出炭疽菌属的5个种,尖孢炭疽菌Colletotrichumacutatum,高粱炭疽菌C.subineolum,胶孢炭疽菌C.gloeosporioides,黑线炭疽菌C.dematium,球炭疽菌C.coccodes。对27株炭疽菌菌株进行ITS序列分析、构建系统发育树,研究表明形态学鉴定与ITS序列分析两种方法表现出很好的一致性和可靠性,形态学鉴定结果与分子结果相同。利用特异引物CaInt2/ITS4对分离自葡萄和枸杞上的炭疽菌进行PCR扩增,显示引起葡萄炭疽病的病原菌为胶孢炭疽菌(C.gloeosporioides)和尖孢炭疽菌(C.acutatum),引起枸杞炭疽病的病原菌为尖孢炭疽菌(C.acutatum),尖孢炭疽菌(C.acutatum)为我国葡萄和枸杞炭疽病的新病原。
鉴定黑痣菌标本100余份,鉴定出黑痣菌属3个种,禾草黑痣菌Phyllachoragraminis,虎尾草黑痣菌P.chloris-virgatae,禾草黑痣菌椭圆孢变种P.graminis var.ellipticta。对7个寄主上的黑痣菌的LSU片段进行克隆测序,通过比对分析设计出一个特异引物Phyl(5'-CRGCGCAAGTCTGGTCAMAGG-3')。利用特异引物ITS1/Phy1对24份标本上的黑痣菌进行PCR扩增、核苷酸序列分析,发现该引物对于禾本科禾亚科植物上的黑痣菌效果较好,为该类群真菌的系统演化分析建立了一套快速简易的方法。通过对得到的ITS序列比对分析后发现,形态学鉴定与ITS序列分析两种方法的结果基本一致。
The fungi in the genus of Colletotrichum can cause important plant diseases in varieties of hosts,and cause great lost in agricultural production.Tar spot is a common plant disease which caused by Phyllachora.It has a common occurrence in plants of Phyllostachys and forage grasses.To edit the Chinese flora of Colletotrichum and Phyllachora,the specimens from Northwest China were collected and identified.Phylogenetics was carded out based on our materials.
About 200 Colleotrichum specimens were examined.Five species(C.acutatum,C. subineolum,C.gloeosporioides,C.dematium and C.coccodes) were identified.ITS sequence analysis and phylogenetic analysis were carried out on 27 isolates.The results show that morphological identification and ITS sequence analysis have good consistency.PCR amplification with specific primer CaInt2/ITS4 was carried out on isolates from grapes.The result shows that there are C.gloeosporioides and C.acutatum can cause anthracnose on grapes,and C.acutatum can cause anthracnose on wolfberries.C.acutatum is a new pathogen of anthracnose of grapes and wolfberries.
About 100 Phyllachora specimens were examined.Three species were identified(P. graminis,P.chloris-virgatae,P.graminis var.ellipticta).Clone was carded out on 7 specimens.The sequences of LSU were obtained.According to these sequences,a specific primer Phyl(5'-CRGCGCAAGTCTGGTCAMAGG-3') was designed.PCR amplification with specific primer ITS1/Phyl was carried out on 24 specimens.There is obvious effect on Gramineae.The result shows that morphological identification and ITS sequence analysis have good consistency.
引文
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