白三叶草一种叶斑病病原菌的分离鉴定  

Isolation and Identification of the Pathogen Causing Leaf Spot on Trifolium repens

作　　者：商娜 任爱芝[1] 刘冰 赵培宝[1] SHANG Na;REN Aizhi;LIU Bing;ZHAO Peibao(College of Agronomy,Liaocheng University,Liaocheng,Shandong 252000;Liaocheng Academy of Agricultural Sciences,Liaocheng,Shandong 252000)

机构地区：[1]聊城大学农学院,山东聊城252000 [2]聊城市农业科学院,山东聊城252000

出　　处：《中国农学通报》2022年第4期81-85,共5页Chinese Agricultural Science Bulletin

基　　金：山东省自然科学基金委“长枝木霉菌与植物互作效应子(effector)的分离鉴定”(ZR2020MC125);山东省科技厅“聊红椿果色遗传机制、组培快繁及高效栽培技术集成”(2019GNC106018);山东省科技发展计划“拟康氏木霉及其产生的多肽抗菌素peptaibols在植物病害防治上的应用研究”(2014GNC110020)。

摘　　要：明确在山东省聊城市白三叶草上发现的一种叶斑病的病原菌,以期为该病害的防控提供参考。2019年采集山东省聊城市东昌府区具有叶斑病的白三叶叶片,利用组织分离法分离病原菌,并依据柯赫式法则验证致病性。在形态学观察的基础上,对病原菌进行分子生物学鉴定,构建系统发育树,分析其遗传多样性。从发病叶片中分离得到病原菌(标记为SY1),具有致病性,且与田间发病症状一致。利用真菌通用引物ITS4/ITS5进行PCR扩增和测序,序列分析表明SY1菌株与乔木链格孢(Alternaria arborescens)(序列号KC464334.1)的相似性为100%,且在系统发育树上聚在一起。根据形态特征和ITS基因序列鉴定结果,将白三叶草叶斑病病原菌鉴定为乔木链格孢,为国内首次报道乔木链格孢引起白三叶草叶斑病。An investigation in Liaocheng of Shandong Province shows that a pathogen causing leaf spot was found on Trifolium repens.Therefore,it is of great significance to find out the pathogen species and provide reference for the prevention and control of the disease.The leaves of T.repens with spot disease collected from Liaocheng were used as materials.The pathogenic bacteria were separated by tissue separation method,and the pathogenicity was verified by Koch's rule.Based on morphological observation,molecular biological method was used to identify pathogenic bacteria.The phylogenetic tree was constructed to analyze the fungus genetic diversity.The pathogenic bacteria(labelled SY1)isolated from diseased leaves were pathogenic and consistent with the field symptoms.To further classify the taxonomic status of the pathogen,the generic fungal primers ITS4/ITS5 were used for PCR amplification and sequencing.Sequence analysis showed that the SY1 was 100%similar to Alternaria arborescens(serial number:KC464334.1),and gathered together on the phylogenetic tree.According to the morphological characteristics and gene sequence identification results,the pathogen of T.repens leaf spot was identified as A.arborescens,which was the first report of A.arborescens causing T.repens leaf spot in China.

关 键 词：白三叶草 乔木链格孢 分子生物学鉴定 形态学鉴定 叶斑病 病原菌 

分 类 号：S436.8[农业科学—农业昆虫与害虫防治]