次郞甜柿炭疽病菌的分离鉴定及其rDNA-ITS序列分析  

Isolation, identifi cation and rDNA-ITS sequence analysis of pathogen of Jiro Persimmon anthracnose

作　　者：余贤美[1] 侯长明[2] 王洁[1] 安淼[1] 王海荣[1] 艾呈祥[1] 

机构地区：[1]山东省果树研究所,山东泰安271000 [2]山东省农业科学院,山东济南250100

出　　处：《经济林研究》2014年第1期45-50,共6页Non-wood Forest Research

基　　金："十二五"农村领域国家科技计划项目(2013BAD14B0504);山东省农业良种工程项目(2010lz010)

摘　　要：为了给生产中柿树次郎甜柿炭疽病的诊断及防治提供理论指导,通过组织分离法和单孢分离法从感病次郞甜柿的果实和嫩梢中获得了2个分离物,菌落及孢子形态特征显示该分离物为炭疽菌。以通用引物ITS6和ITS4,通过PCR扩增,获得了rDNA-ITS基因片段。测序结果显示,从柿的果实和嫩梢分离获得的染病次郎甜柿分离物ITS序列完全一致,片段大小为598 bp,该片段与感病浙江无核柿(AY787483)和新西兰分离物(GQ329690)的同源性分别为100%和99.8%。系统进化树分析结果显示,染病次郎甜柿分离物与浙江无核柿(AY787483,AY791890)和新西兰分离物(GQ329687,GQ329688和GQ329690)处在同一分支,据此,将该病原物鉴定为柿树炭疽菌Colletotrichum horii,并将其ITS基因序列提交到GenBank数据库(基因登录号:JQ957543)。In order to provide a theoretical guidance for diagnosis and control of persimmon anthracnose in production, two isolates were gained from fruits and twigs in infected Jiro Persimmon by using tissue isolation and single spore separation methods were obtained. The morphological characteristics of colonies and spores showed that the isolates belong to Colletotrichum horii., rDNA-ITS gene segments were obtained with the universal primers of ITS6 and ITS4 by PCR amplification method. The sequencing results show that the ITS sequences of the isolates from fruits and twigs are in full accord, and the size of the sequence is 598 bp. The homologies of nucleotide sequence of ITS gene with that of anthracnose pathogens on Zhejiang Wuheshi in （AY787483） and New Zealand isolate （GQ329690） are 100% and 99.8% respectively. The results ofphylogenetic analysis show that the isolates from Jiro Persimmon is located at the same branch of phylogenetic tree with the isolates of Zhejiang Wuheshi （AY787483, AY791890） and New Zealand isolates （GQ329687, GQ329688 and GQ329690）, which indicates that the isolates from Jiro Persimmon can be identified as C. horii., the pathogen of Jiro Persimmon anthracnose disease. The ITS gene sequences were submitted to GenBank Data （Accession No. JQ957543）.

关 键 词：次郎甜柿 柿树炭疽病 病原分离 分子鉴定 RDNA-ITS序列分析 

分 类 号：S665.2[农业科学—果树学]