草莓褐色叶斑病病原菌Pilidium lythri鉴定及室内防治药剂筛选.pdf
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1、RESEARCHREPORTSdoi:10.16801/j.issn.1008-7303.2023.0063Identification of Pilidium lythri and screening of fungicides in vitroYANYitong1,SUNJunyuan1,LIFuxin1,ZHANGGuijun1,YANZhe1,HUANGZhongqiao2,GAOHuige2,BIYang*,1(1.Key Laboratory for Northern Urban Agriculture of Ministry of Agriculture and Rural Af
2、fairs,Beijing University of Agriculture,Beijing102206,China;2.Department of Plant Pathology,China Agricultural University,Beijing 100193,China)Abstract:Inrecentyears,strawberrytan-brownleafspotdiseasecausedbyPilidium lythriisaseriousnewdiseasethathasoccurredinChina.Between2015and2017,atotalof90sympt
3、omaticsampleswerecollectedfromChangpingDistrict,BeijingCity;ZhuchengCounty,ShandongProvince;andLoudiCity,HunanProvinceinChina,with30samplesfromeachdistrict.Twenty-sixfungalisolateswereidentifiedasP.lythribasedonmorphology,molecularbiology,andphylogeneticanalyses.ThisisthefirstreportofP.lythricauseds
4、trawberrytan-brownspotinShandongandHunanProvincesinChina.Atpresent,therearenoregisteredfungicidesinChinaforcontrollingtan-brownspotdiseaseonstrawberrycausedbyP.lythri.Inthisstudy,thesusceptibilityof26isolatestoeightcommonlyusedfungicidesandnovelfungicideSYP-14288weredeterminedin vitro.Theresultshows
5、thatSYP-14288hasthestrongestinhibitoryactivityonthemycelialgrowthof26isolatesofP.lythri,withanaverageEC50valueof(0.330.06)g/mL,canbeusedasaneffectivefungicideforthecontrolofstrawberrytan-brownleafspotdisease.TheaverageEC50valueswerefrom3.92g/mLto72.58g/mLforepoxiconazole,difenoconazole,prochloraz,te
6、buconazole,chlorothalonil,andmyclobutanil.Amongthem,azoxystrobinhadlowertoxicity,andmancozebhadthelowestinhibitoryactivity.Theresultsofthisstudycanprovideanimportantreferencefortherationaluseoffungicidestocontrolstrawberrytan-brownspotdisease.Keywords:Pilidium lythri;strawberrytan-brownleafspotdisea
7、se;mycelialgrowth;SYP-14288草莓褐色叶斑病病原菌Pilidium lythri鉴定及室内防治药剂筛选闫奕彤1,孙浚源1,李福鑫1,张桂军1,闫哲1,黄中乔2,高慧鸽2,毕扬*,1(1.北京农学院农业农村部北方城市重点实验室,北京102206;2.中国农业大学植物病理学系,北京100193)摘 要:近年来,由病原菌 Pilidium lythri 引起的草莓褐色叶斑病,是一种在中国发生严重的新病害。作者在 2015 年至 2017 年间,从北京市昌平区、山东省诸城市和湖南省娄底市采集有症状的草莓样品共 90 株,每地 30 株,根据形态学特征、分子生物学及系统发育分析,
8、其中 26 株分离物被鉴定为 P.lythri,并且山东和湖南系首次发现由 P.lythri 引起的草莓褐色叶斑病。鉴于目前中国还没有登记用于防治草莓褐色叶斑病的杀菌剂。本研究通过室内毒力测定法测定了26 株 P.lythri 菌株对 8 种常用杀菌剂和 1 种新型杀菌剂双苯菌胺(SYP-14288)的敏感性。结果表明:SYP-14288 对 P.lythri 菌丝生长的抑制活性最强,平均 EC50值为(0.330.06)g/mL,可将其作为防治草莓褐色叶斑病的首选药剂;氟环唑、苯醚甲环唑、咪鲜胺、戊唑醇、百菌清Received:December6,2022;Accepted:June28,2
9、023;Published online:July11,2023.URL:https:/doi.org/10.16801/j.issn.1008-7303.2023.0063http:/ Journal of Pesticide ScienceE-mail:和腈菌唑对 P.lythri 的平均 EC50值在 3.9272.58g/mL 之间,其中菌株对嘧菌酯和代森锰锌的敏感性较低。该研究结果可为合理使用杀菌剂防治由 P.lythri 引起的草莓褐色叶斑病提供参考。关键词:Pilidium lythri;草莓褐色叶斑病;菌丝生长;双苯菌胺中图分类号:S482.2文献标志码:A 0 Introdu
10、ctionStrawberry(Fragaria ananassaDuch.)isanimportantcommercialfruitcropthatiswidelygrownintemperateregionsworldwide.Overthepasttwentyyears,theplantingareaandyieldofstrawberryhavegrownrapidly.AccordingtothelateststatisticsfromFAOSTAT(https:/www.fao.org/faostat/zh/#home)in2020,strawberryplantingareash
11、avereachedmorethan100000hectaresinChina.Meanwhile,strawberryproductionwasupto3000000tonsperyearinChina,rankingtopoftheworld1.Duringthegrowthperiod,strawberriesareaffectedbyavarietyoffungalpathogens,forexample,Botrytis cinereaPers.,Colletotrichumspp.,andVerticillium dahliaeKleb.2.Pilidium lythri,prev
12、iouslynamedPilidium concavumandHainesialythri,hasbeenreportedtocausestrawberrytan-brownrotdiseaseworldwide.Strawberrytan-brownrotdiseasewasfirstreportedinJapan,andsincethenreportedinseveralothercountriessuchasPoland,Brazil,andBelgium3-7.InChina,strawberrytan-brownrotdiseasecausedbyP.lythrihasbeenrep
13、ortedonlyinBeijingsofar8.Inadditiontostrawberry,P.lythricanalsoinfectmorethantenhosts,includingOlea europaea,Fallopia japonica,Hieracium caespitosum,Bergeniacrassifolia,Paeonia suffruticosa,Eucalyptussp.,Rosa rugosa,Prunus domestica,Paeonia lactiflora,Greyia radlkoferi,Aesculus hippocastanum,Vaccini
14、um corymbosumandCornus mas5-6,8-9.P.lythriinfectsstrawberryleaves,stems,fruits,androots.Aftertheinitialacuteinfection,acircular,water-stainedbrownspotemergesinthecenterormarginofstrawberryleaves6-7.Theapplicationofchemicalfungicidesisaneffectivemethodformanagingstrawberrydiseases.However,thedevelopm
15、entoffungicideresistancehascompromisedthiscontrolstrategy.Besides,consi-deringthesafetyoffreshstrawberry,therearealsosomelimitationstotheuseofchemicalfungicidesinthefield.Currently,themanagementofP.lythriheavilyreliesontheapplicationofeffectivefungicidesandhostresistance.However,thereislittleresearc
16、honthechemicalcontrolofstrawberrytan-brownrotdiseasecausedbyP.lythri.ThepurposeofthisstudyistoidentifyandcharacterizeP.lythriassociatedwithtan-brownleafspotdiseaseofstrawberryinChinabasedonmorphological,molecularandpathogeniccharac-teristicsandtoevaluatetheinhibitoryactivityofazoxystrobin,chlorothal
17、onil,epoxiconazole,difeno-conazole,mancozeb,myclobutanil,prochloraz,tebu-conazoleandSYP-14288againstthisfungusin vitro.1 Materials and methods 1.1 MediumPDA:200gpotato,20gdextrose,and15gagarin1000mLdistilledwater.1.2 FungicidesTechnical-gradeazoxystrobin(95%a.i.;SyngentaBiotechnologyCo.,Ltd.,Shangha
18、i,China),chlorothalonil(98%a.i.;HenanChunguangAgrochemicalsCo.,Ltd.,Henan,China),difenoconazole(98%a.i.;HangzhouYulongAgrochemicalsCo.,Ltd.,Zhejiang,China),epoxiconazole(97.8%a.i.;HenanZhongzhouSeedTechnologyDevelopmentCo.,Ltd.,Henan,China),mancozeb(98.5%a.i.;HebeiHesenChemicalCo.,Ltd.,Hebei,China),
19、myclobutanil(98%a.i.;HenanZhongyuanGermplasmFactory,Henan,China),prochloraz(97%a.i.;HangzhouQingfengAgrochemicalsCo.,Ltd.,Zhejiang,China),SYP-14288(97%a.i.;ShenyangResearchInstituteofChemicalIndustry,Shenyang,China),andtebuconazole(98%a.i.;JiangsuFengdengPesticideCo.,Ltd.,Jiangsu,China)weredissolved
20、respectivelyindimethylsulfoxide(DMSO)tomakestocksolutionsandwerestoredat4inthedark.1.3 Diseased sample collection and pathogenisolationAtotalof90diseasedsampleswerecollectedrandomlyfromstrawberryfieldsexhibitingtypicalsymptomsoftan-brownrotdiseaseinChangpingNo.5YANYitongetal:IdentificationofPilidium
21、 lythriandscreeningoffungicidesin vitro1077District,Beijing;ZhuchengCounty,ShandongProvince;andLoudiCity,HunanProvince,Chinain2015-2017,with30samplesfromeachdistrict.Among26isolatesofP.lythri,oneisolate(BJ-4)wasfromBeijingCity,nineisolates(HNLD-1toHNLD-7,HNLD-9,HNLD-12)werefromHunanProvince,and16iso
22、lates(SD17-2toSD17-12,SD14toSD18)werefromShandongProvince(Table1).TheinfectedTable 1 Information of ITS and LSU gene sequence of P.lythri strains used for phylogenetic analysisSpeciesorstrainGenBankNo.ITSGenBankNo.LSUHostCountryReferenceCommonnameLatinnameP.lythriAY487094.1AY487095RoseRosasp.USA11P.
23、lythriAY487097.1AY487098PeonyPaeonia suffruticosaJapan11P.pseudoconcavumKF777184.1KF777236.1AshtreeGreyia radlkoferiSouthAfrica12P.acerinumAY487088AY487089HorsechestnutAesculus hippocastanumNetherlands11P.acerinumAY487091.1AY487092gardensoilHortum terramNetherlands11P.eucalyptorumKT950854.1KT950868.
24、1EucalyptusgrandisEucalyptus robustaFrance13Chaetomella raphigeraAY487076.1AY487077BlueberryVaccinium corymbosumUSA11BJ-4MH322003MH322002StrawberryFragaria ananassaChinaThisstudyHNLD-1MH322005MH322004StrawberryFragaria ananassaChinaThisstudySD17-3MH322016MH322014StrawberryFragaria ananassaChinaThiss
25、tudyHNLD-2MH322006_StrawberryFragaria ananassaChinaThisstudyHNLD-3MH322007_StrawberryFragaria ananassaChinaThisstudyHNLD-4MH322008_StrawberryFragaria ananassaChinaThisstudyHNLD-5MH322009_StrawberryFragaria ananassaChinaThisstudyHNLD-6MH322010_StrawberryFragaria ananassaChinaThisstudyHNLD-7MH322011_S
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