Journal of Agricultural Science and Technology ›› 2017, Vol. 19 ›› Issue (7): 44-49.DOI: 10.13304/j.nykjdb.2016.677

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Identification and ITS Sequence Analysis of Pathogen of Cherry Brown Leaf Spot in Shandong

SUN Yang1, HAN Wei1*, SUN Yugang2, WEI Guoqin2, FU Quanjuan2, ANG Xinghua2, GONG Qingtao2   

  1. 1.Vegetable Research Institute,  Shandong Academy of Agricultural Sciences, Jinan 250100;
    2.Shandong Institute of Pomology, Shangdong Taian 271000, China
  • Received:2016-11-02 Online:2017-07-15 Published:2017-02-20

山东地区樱桃褐斑病病原鉴定及rDNA-ITS序列分析

孙杨1,韩伟1*,孙玉刚2,魏国琴2,付全娟2,杨兴华2,宫庆涛2   

  1. 1.山东省农业科学院蔬菜花卉研究所, 济南 250100; 2.山东省果树研究所, 山东 泰安 271000
  • 通讯作者: 韩伟,副研究员,硕士,主要从事蔬菜品种资源收集与鉴定研究。E-mail:SDSCSHW@163.com
  • 作者简介:孙杨,助理研究员,博士研究生,主要从事植物真菌病害研究。E-mail:sunyang_729@163.com。
  • 基金资助:
    山东省自然科学基金培养基金项目(ZR2014CP017)资助。

Abstract: In order to clarify the pathogenic species of cherry brown leaf spot, this study isolated and purified  16 fungal strains by single spore separation method. It also collected typical morbidity leaf blades from Taian, Weifang and Yantai 3 regions. Identification of cherry brown leaf spot pathogen was carried out by pathogen morphology combined with molecular biology method. The ITS sequence analysis of this pathogen showed that it was aligned with Genbank and showed 97% identity with Passalora arctostaphyli. The results of phylogenetic analysis showed that the pathogen was located at the same branch of phylogenetic tree with Passalora sp. and different from Pseudocercospora sp. According to its morphological characteristics and the sequence of internal transcribed spacer, the pathogen was identified as Passalora circumscissa, and ITS gene sequence was presented to GenBank database(Genbank Accession No.: KT428056).

Key words: cherry, brown leaf spot, ITS sequence analysis, identification

摘要: 为明确引起樱桃褐斑病的病原菌种类,采集泰安、潍坊、烟台3个地区典型发病叶片,用单孢分离法分离、纯化,共获得16个菌株,采用病原菌形态学与分子生物学相结合的方法,鉴定樱桃褐斑病的病原菌。对该病原菌的ITS序列分析显示,其与GenBank中登录的Passalora sp.属的Passalora arctostaphyli相似性达97%;与另一种在甜樱桃叶片中分离的病原菌Pseudocercospora pruni-persicicola相似度仅为85.6%,证明两者并非为同一属真菌。构建系统进化树表明,樱桃褐斑病病原与Passalora sp.属真菌处在同一分支,而与其它Prunus sp.属分离的病原菌处在两个分支,进而通过形态学鉴定和ITS序列分析认为,病原菌为Passalora circumscissa,并将其ITS基因序列提交至GenBank数据库(基因登陆号:KT428056)。

关键词: 樱桃, 褐斑病, ITS序列分析, 鉴定