[1]李俊,秦文韬,曹子健,等.卵孢小奥德蘑蛛网病菌PCR检测方法的建立与应用[J].北方园艺,2022,(22):124-131.[doi:10.11937/bfyy.20221350]
　LI Jun,QIN Wentao,CAO Zijian,et al.Establishment and Application of PCR Detection Methods for Cladobotryum varium[J].Northern Horticulture,2022,(22):124-131.[doi:10.11937/bfyy.20221350]
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卵孢小奥德蘑蛛网病菌PCR检测方法的建立与应用

《北方园艺》[ISSN:1001-0009/CN:23-1247/S] 卷: 期数: 2022年22 页码: 124-131 栏目: 出版日期: 2022-11-30
Title:
Establishment and Application of PCR Detection Methods for Cladobotryum varium
作者:
李俊1; 2; 秦文韬1; 曹子健1; 3; 刘宇1; 赵娟1; 王守现1
（1.北京市农林科学院 植物保护研究所，北京市食用菌工程技术研究中心，北京 100097；2.北京农学院 生物与资源环境学院，北京 102206；3.河北工程大学 园林与生态工程学院，河北 邯郸 056038）
Author(s):
LI Jun1; 2; QIN Wentao1; CAO Zijian1; 2; LIU Yu1; ZHAO Juan1; WANG Shouxian1
(1.Institute of Plant Protection，Beijing Academy of Agriculture and Forestry Sciences/Beijing Engineering Research Enter for Edible Mushroom，Beijing 100097;2.College of Bioscience and Resources Environment，Beijing University of Agriculture，Beijing 102206;3.School of Landscape and Ecological Engineering，Hebei University of Engineering，Handan，Hebei 056038)
关键词:
卵孢小奥德蘑; 蛛网病; 枝葡霉; 检测; 特异性
Keywords:
Oudemansiella raphanipes; cobweb disease; Cladobotryum varium; detection; specificity
DOI:
10.11937/bfyy.20221350
文献标志码:
A
摘要:
以卵孢小奥德蘑蛛网病菌（异形枝葡霉）为试材，以异形枝葡霉[STBX]TEF1-α[STBZ]基因为靶标，采用聚合酶链式反应（PCR）方法，设计、合成并筛选出可特异检测异形枝葡霉的PCR引物，基于此建立并应用PCR检测方法，以期为科学防控卵孢小奥德蘑蛛网病提供分子基础。结果表明：利用基于T8F/R引物建立的PCR检测体系对16种枝葡霉属的22个菌株的基因组DNA进行扩增发现，该检测体系特异性强，仅以异形枝葡霉基因组DNA作为模板时，产物呈现出一条467 bp的条带，其它参照菌株及阴性对照均无条带。引物灵敏度验证结果表明，检测体系不受卵孢小奥德蘑组织液的干扰，可检测出最低90 pg·μL-1的异形枝葡霉基因组DNA，土壤中最低检出限为230个孢子·g-1。
Abstract:
Cladobotryum varium，pathogen of cobweb disease on Oudemansiella raphanipes，was used as test material.Targeting [STBX]TEF1-α[STBZ] gene of C.varium，design,synthesized and screened to established the system for detection C.varium using the method of polymerase chain reaction (PCR),in order to provide molecular basis for scientific control of cobweb disease on O.raphanipes.The results showed that the genomic DNA of 22 strains of 16 Cladobotryum taxa were amplified with the PCR detection system established based on T8F/R and found that the system showed strong specificity.Only when genomic DNA of C.varium was used as template，the PCR products showed a specific band of 467 bp.The sensitivity test results displayed that the detection system based on T8F/R was not interfered by the tissue fluid of O.raphanipes，and could detect the lowest of 90 pg·μL-1 C.varium genomic DNA.The minimum detection limit in soil was 230 spores·g-1.

参考文献/References:

［1］HAO Y J，ZHAO Q，WANG S，et al.What is the radicate Oudemansiella cultivated in China?〖KG-*3〗［J］.Phytotaxa，2016，286(1):1-12.［2］江枝和，翁伯琦，林勇.长根菇中蛋白质的营养评价［J］.江西农业大学学报，2003，25(3):439-441.［3］肖淑霞.不同培养料栽培的长根菇氨基酸分析比较［J］.福建农业科技，2003(3):28-29.［4］田龙，庞振凌，杜敏华.富含长根菇营养物质的保健饮料的研制［J］.安徽农业科学，2006，34(15):3793-3794.［5］吴希禹，付永平，李玉.香菇蛛网病病原菌树状枝葡霉生物学特性［J］.菌物学报，2019，38(5):646-657.［6］KIM T S，LEE H W，SONG G W，et al.King oyster mushroom (Pleurotus eryngii) white mold disease caused by Cladobotryum varium［J］.KSM News Lett (Korean Soc Mycol)，1998(11):46.［7］兰玉菲，丛倩倩，崔晓.金针菇蛛网病病原菌鉴定［J］.中国食用菌，2019，38(1):76-79.［8］WANG G Z，GUO M P，BIAN Y B.First report of Cladobotryum protrusum causing cobweb disease on the edible mushroom Coprinus comatus［J］.Plant Disease，2015，99(2):287.［9］BACK C G，LEE C Y，SEO G S，et al.First occurrence of cobweb disease on Hypsizigus marmoreus caused by Cladobotryum varium in Korea［J］.Journal-Faculty of Agriculture Kyushu University，2012，57(2):373-377.［10］曹满堂，李宾，李宏，等.食用菌蛛网病研究进展［J］.食用菌学报，2020，27(3):127-136.［11］TAMM H，PLDMAA K.Diversity，host associations，and phylogeography of temperate aurofusarin-producing Hypomyces/Cladobotryum，including causal agents of cobweb disease of cultivated mushrooms［J］.Fungal Biology，2013，117(5):348-367.［12］CARRASCO J，NAVARRO M J，SANTOS M，et al.Incidence and pathogenicity of Cladobotryum mycophilum，causal agent of cobweb disease on Agaricus bisporus mushroom crops in Spain［J］.Annals of Applied Biology，2016，168(2):214-224.［13］黄建春，宋晓霞，王倩，等.双孢蘑菇蛛网病及其防治技术［J］.食用菌，2017，39(3):61-62.［14］FLETCHER J T，WHITE P F，GAZE R H.Mushroom pest and disease control［J］.Mycologia，2008，83(5):683-684.［15］CARRASCO J，NAVARRO M J，GEA F J.Cobweb，a serious pathology in mushroom crops:A review［J］.Spanish Journal of Agricultural Research，2017，15(2):e10R01.［16］LAN Y，WANG Q，YU C，et al.First report of Cladobotryum mycophilum causing cobweb disease on Agaricus bisporus in China［J］.Plant Disease，2016，100(11):2334.［17］秦文韬，黄晓庆，孔繁芳，等.葡萄霜霉病菌PCR检测方法的建立与应用［J］.植物保护，2014，40(2):99-102.［18］秦文韬，谷彤彤，刘宇，等.杏鲍菇枯萎病菌PCR检测方法的建立与初步应用［J］.北方园艺，2021(7):123-128.［19］WHITE T，BRUNS T，LEE S H，et al.Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics.PCR protocols:A guide to methods and applications［M］.London:Academic Press，1990.［20］REHNER S A，BUCKLEY E.A Beauveria phylogeny inferred from nuclear ITS and EF1-α sequences:Evidence for cryptic diversification and links to Cordyceps teleomorphs［J］.Mycologia，2005，97(1):84-98.［21］郭凤柳，张海颖，于秀梅，等.中国马铃薯疮痂病菌快速PCR检测技术［J］.中国农业科学，2013，46(23):4926-4932.［22］赵志强，张卢慧，郭庆元.甜菜黄萎病病原菌(Gibellulopsis nigrescens)的快速分子检测［J］.中国植保导刊，2021，41(8):20-24.［23］伍文宪，刘勇，黄小琴，等.尖孢镰刀菌分子检测技术的建立与应用［J］.草业学报，2016，25(5):109-115.［24］李承达，张亚鑫，刘权德，等.长枝木霉引发灵芝绿霉病的快速检测体系的建立［J］.天津农业科学，2021，27(7):61-65.

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备注/Memo

第一作者简介：李俊（1998-），女，硕士研究生，研究方向为食用菌病害。E-mail：13439752126@163.com.责任作者：秦文韬（1988-），女，博士，助理研究员，现主要从事真菌病害诊断与综合防控等研究工作。E-mail：qinwentao@ipepbaafs.cn.基金项目：国家自然科学基金青年资助项目（32002106）；北京市农林科学院创新能力建设专项资助项目（KJCX20220415，KJCX20200801）。收稿日期：2022-04-07

更新日期/Last Update: 2023-01-19