琯溪蜜柚亚洲柑橘叶点霉与首都叶点霉ITS-RFLP快速鉴定  

A Rapid Identification Method for Phyllosticta citriasiana and Phyllosticta capitalensis in Citrus grandis by ITS-RFLP

作　　者：罗金水[1] 郑域茹[1] 卢松茂[1] 林智明[1] 林艺华[1] LUO Jin-shui;ZHENG Yu-ru;LU Song-mao;LIN Zhi-ming;LIN Yi-hua(Fujian Institute of Tropical Crops,Zhangzhou,Fujian 363001,China)

机构地区：[1]福建省热带作物科学研究所,福建漳州363001

出　　处：《福建农业学报》2018年第2期195-198,共4页Fujian Journal of Agricultural Sciences

基　　金：福建省科技计划项目--省属公益类科研院所基本科研专项(2012GY01);福建省自然科学基金(2010J01113)

摘　　要：亚洲柑橘叶点霉Phyllosticta citriasiana是引起琯溪蜜柚黑斑病的病原菌。首都叶点霉Phyllosticta capitalensis是分离自琯溪蜜柚的一种腐生真菌。在本研究建立2种叶点霉属真菌的ITS-RFLP快速鉴定方法。首先采集平和县琯溪蜜柚样品,分离纯化获得目的菌P.citriasiana和P.capitalensis,提取这2种病菌的菌丝体总DNA,使用ITS1和ITS4引物对扩增rDNA-ITS基因片段,再用限制性内切酶Acc I酶切扩增产物。结果表明,P.citriasiana和P.capitalensis这2种菌的rDNA-ITS基因酶切分别产生了2个和3个特异条带。通过条带的数量和大小使这两种真菌得以区分,该方法快速、简便。Phyllosticta citriasiana is a new pathogen of Citrus grandis which causes black spot on the fruits,while Phyllosticta capitalensis is another fungus which is saprophytic and seprated from Citrus grandis.For distinguishing from each other,a rapid identification method named ITS-RFLP was established in this research.First of all,the target pathogens were seperated from the desease samples which were collected from the Citrus grandis Osbeck orchard at Pinghe city in Fujian.Then the mycelium total DNA were extracted for amplifying the rDNA-ITS gene fragments by using ITS1 and ITS4 primers.And then the amplification products were digested by the restriction enzyme of Acc I.The results showed that the rDNA-ITS of P.citriasiana was digested into two fragments,and that of P.capitalensis into three fragments.Thus,the two kinds of fungi can be distinguished from each other by the fragment number and size.This technology is quite rapid and simple.

关 键 词：琯溪蜜柚 黑斑病 限制性内切酶片段长度多态性分析技术 

分 类 号：S436[农业科学—农业昆虫与害虫防治]