甘藍黑腳病菌之生理特性及其病害防治

本菌在馬鈴薯葡萄糖瓊脂培養基(PDA)上生長緩慢，菌絲初期為白色，隨後逐漸呈橄欖綠或深綠色，隨後產生分生孢子器(pycnidia)及器孢子(pycnidiospore)。

資料載入處理中... 跳到主要內容 臺灣博碩士論文加值系統 ::: 網站導覽| 首頁| 關於本站| 聯絡我們| 國圖首頁| 常見問題| 操作說明 English |FB專頁 |Mobile 免費會員 登入| 註冊 功能切換導覽列 (178.128.221.219)您好！臺灣時間：2022/04/1312:08 字體大小：       ::: 詳目顯示 recordfocus 第1筆/ 共1筆  /1頁 論文基本資料 摘要 外文摘要 目次 參考文獻 紙本論文 論文連結 QRCode 本論文永久網址: 複製永久網址Twitter研究生:曾淑菁研究生(外文):Shu-ChingTseng論文名稱:甘藍黑腳病菌之生理特性及其病害防治論文名稱(外文):StudiesonthephysiologicalcharacteristicsofLeptosphaeriamaculansanditsdiseasecontrol指導教授:郭章信指導教授(外文):Chang-HsinKuo學位類別:碩士校院名稱:國立嘉義大學系所名稱:生物資源學系研究所學門:生命科學學門學類:生物科技學類論文種類:學術論文畢業學年度:100語文別:中文論文頁數:72中文關鍵詞:甘藍菜、小球腔菌、黑腳病外文關鍵詞:BrassicaoleraceaL.、Leptosphaeriamaculans、Blacklegdisease、Phomalingam相關次數: 被引用:0點閱:892評分:下載:0書目收藏:0 甘藍黑腳病(blacklegdisease)分布在溫帶地區，而本省僅發現於高冷蔬菜地區，近年來，本病害嚴重影響甘藍菜之產量，平地則尚未有發生之記錄。

由罹病株經組織分離所得之菌株，經外部形態及ITSrDNA鑑定為Leptosphaeriamaculans，本菌製作孢子懸浮液以種子浸漬法接種確定病原性。

經接種之甘藍種子在幼苗期均可表現病徵，在葉片上產生明顯的灰色病斑，病斑逐漸擴大，終至幼苗倒伏枯死。

本菌在馬鈴薯葡萄糖瓊脂培養基(PDA)上生長緩慢，菌絲初期為白色，隨後逐漸呈橄欖綠或深綠色，隨後產生分生孢子器(pycnidia)及器孢子(pycnidiospore)。

以不同接種種源密度接種甘藍種子之結果顯示，孢子濃度在105-106spores/ml時，具有較高致病力，發病率可達80％以上，濃度在103以下不發病。

測試溫度對病原菌之影響，溫度對菌絲生長以20℃為最佳，5℃-35℃均可生長，40℃以上不生長；在產孢方面，分生孢子器數產生以20℃最佳，10℃-30℃均可形成分生孢子器；孢子發芽最適溫度以25℃為最佳，10℃-30℃孢子均可發芽。

測試不同相對濕度對孢子發芽之影響試驗結果顯示，孢子發芽率隨相對濕度增加而遞減，相對濕度85％時發芽率達82.8％。

以不同培養基培養病原菌結果顯示，病原菌在10％V-8瓊脂、20％V-8瓊脂及PDA上菌絲生長最快；而在產孢上則以PDA為最佳。

測試不同碳及氮素源對病原菌菌絲生長及產孢之影響，供試十一種碳素源中以半乳糖、果糖促進菌絲生長的效果最佳；在供試十三種氮素源中，以蛋白腖、精氨酸促進菌絲生長效果最佳。

測定不同酸鹼值對菌絲生長之影響，結果顯示培養基酸鹼值在4-5之間為最佳。

初步篩選四十三種中草藥粗萃液防治本病害，取50μl粗萃液分別與等量的孢子懸浮液混合，滴於3孔載玻片上，每處理六重複，測試結果顯示以厚朴、香椿、黃岑、肉桂、丁香、麻黃、八角、黃蓮、五倍子、何首烏、百里香、雞屎藤、開脾草及赤山葡萄之粗萃液可顯著抑制病原菌孢子發芽，與對照組相比，孢子發芽率由88％降至1％。

測試拮抗菌對病原菌之抑制效果，結果顯示篩選所得之拮抗細菌BV-1、PWP-1、PWP-2及拮抗酵母菌163、251、333、G14菌株經對峙培養對Leptosphaeriamaculans皆具有抑制效果。

以篩選化學藥劑防治本病害，結果以腐絕、待克利、普克利、依滅列抑制菌絲生長效果最佳，進一步在溫室中以幼苗接種試驗測試，結果以待克利、普克利、依滅列有防治效果，但會有抑制幼苗頂芽生長之現象。

BlacklegdiseasedistributedintheTemperateZones,wereonlyfoundinthealpineareainTaiwan.Inrecentyears,blacklegdiseaseseverelyaffectproductionofcabbage.Butintheflatgrounddidnothaveanyrecordaboutblacklegdisease.ThecausalagentwasidentifiedasLeptosphaeriamaculansthroughthemorphologyandITSrDNAanalysis.Andmakesporesuspensiontocarryoutthepathogenicitytestbyseedsoaking.Allcabbageseedlingsshowedthesymptomsafterseedsinoculation,andproducedthegrayspotinthecotyledon.Thenthegrayspotgraduallymagnify,finallytheseedlingwaswiltanddead.ThemyceliaofpathogengrownonPDAwaswhiteandgraduallyturnedintogreenorolivegreen,thenproducedpycnidiaandpycnidiospore.Thehighestdiseaseincidenceofblacklegdiseaseofcabbagewereover80％whencabbageseedswereincolatedwith105-106spores/ml,butnoincidencewasfoundin103spores/ml.Thephysiologicalcharacteristicstudiesofthepathogenindicatedthattheoptimumtemperaturewasforthemycelialgrowthat20℃,withtherangesfrom5to35℃.Theoptimumtemperatureforpycnidiasporegerminationwasat25℃.Onsporulation,theoptimumtemperaturewasat20℃,andthematurityofpycnidiawerenotsignificantlywhenculturedat10℃-30℃.Theeffectofdifferentrelativehumidityonsporegerminationtoshowthehumiditytoincreaseandsporesgerminationtolow,Thehighergrowthwasover82.8％inthe85％relativehumidity.Theeffectofculturemediaonthemyceliagrowthandpycnidiamaturityofthepathogenwereconducted.Thedatashowedthatthemycelialgrowthon10％V8,20％V8,PDAandsporulationonPDAhadthehighestgrowthrateamongtestedmedia.Elevencarbonsourcesandthirteennitrogensourceswerechosenasnutrientsforinvestigatingtheireffectsonmyceliagrowthandsporulation.Acomparisonofdifferentcarbonsourcesshowedthatmycelialgrowthratewashighestwhengrownongalactoseandfructose.Acomparisonofdifferentnitrogensourcesshowedthatmyceliagrowthratewashighestwhengrowthonbacto.peptoneandarginine.ThebestofpHvalueformyceliagrowthwasatpH4-5.Preliminaryscreening43kindsofcrudeofchineseherbmedicineoninhibitionofsporegerminationofthepathogenwereconducted.Thedatashowedthat14kindsofcrudeofChinesemedicineherbhadthemostsingnificantinhibitiononsporegerminationwithlowerthan1％comparingtothecontrolwith88％.UsingthepairedculturetoscreeningantagonistsforthecontrolofLeptosphaeriamaculansonPDA,datashowedthatantagonistsofbacteria,BV-1,PWP-1,PWP-2andyeast163,251,333,G14weresignificantlyinhibitedmycelialgrowth.Screeningelevenfungicidesforthecontrolofthepathogen,thedatashowedthatfourfungicides,Thiabendazole,Difenoconazole,PropiconazoleandImazalilhadthemostsingnificantinhibiononmycelialgrowth,however,thedatashowedthatonlyDifenoconazole,PropiconazoleandImazalilwassingnificantefficacyonthecontrolofLeptosphaeriamaculansoncabbageseedlingsbutrestrainedterminalbudsgrowth. 中文摘要....................................................Ι英文摘要....................................................Ⅲ目錄........................................................Ⅴ圖表目次....................................................Ⅷ壹、前言....................................................1貳、材料與方法..............................................7一、甘藍黑腳病菌之分離、培養與病原性測定...................7二、不同孢子濃度接種對病原性之影響........................10三、甘藍黑腳病菌對不同品種甘藍之致病力之測定................11四、甘藍黑腳病菌基本生理測定..............................111.溫度對菌絲生長、產孢量、分生孢子器成熟度及孢子發芽之影響....................................................112.不同培養基對菌絲生長、產孢量及分生孢子器成熟度之影響...123.碳素源對菌絲生長、產孢量及分生孢子器成熟度之影響.......134.氮素源對菌絲生長、產孢量及分生孢子器成熟度之影響.......145.不同酸鹼值對菌絲生長之影響…………………………………….146.相對濕度對孢子發芽率之影響…………………………………….15五、甘藍黑腳病防治試驗……………………………………………..151.篩選中草藥粗萃液對甘藍黑腳病之防治效果………………….152.篩選結抗菌對甘藍黑腳病之防治效果………………………….163.篩選化學藥劑對甘藍黑腳病之防治效果……………………….17參、結果…………………………………………………………………….19一、甘藍黑腳病菌之分離、培養與病原性測定………………………….19二、不同孢子濃度接種對病原性之影響………………………………….19三、甘藍黑腳病菌對不同品種甘藍之致病力之測定……………………20四、甘藍黑腳病菌基本生理測定…………………………………………201.溫度對菌絲生長、產孢量、分生孢子器成熟度及孢子發芽之影響……………………………………………………………………202.不同培養基對菌絲生長、產孢量及分生孢子器成熟度之影響……203.碳素源對菌絲生長、產孢量及分生孢子器成熟度之影響…………214.氮素源對菌絲生長、產孢量及分生孢子器成熟度之影響…………215.不同酸鹼值對菌絲生長之影響…………………………………….216.相對濕度對孢子發芽率之影響…………………………………….21五、甘藍黑腳病防治試驗………………………………………………221.篩選中草藥粗萃液對甘藍黑腳病之防治效果………………….222.篩選結抗菌對甘藍黑腳病之防治效果………………………….223.篩選化學藥劑對甘藍黑腳病之防治效果……………………….22肆、討論…………………………………………………………………….24伍、參考文獻……………………………………………………………….28陸、附錄……………………………………………………………………57 1黃振文。

1989。

十字花科。

台灣農家全書。

行政院農業委員會編。

行政院農業委員會出版。

豐年社發行。

台北。

2費雯綺、王玉美編。

2000。

植物保護手冊。

行政院農業委員會農業藥物毒物試驗所。

3鄭安秀、林益昇。

1995。

台灣農家要覽農作篇。

行政院農業委員會台灣農家要覽增修訂再版策劃委員會。

豐年社出版。

台北。

4Badawy,H.M.A.,andHoppe,H.H.,1989.NonspecificphytotoxiceffectsofsirodesminsonhostandnonhostplantsofLeptosphaeriamaculans.J.Phytopathol.127:137–145.5Balesdent,M.-H.,Gall,C.,Robin,P.,andRouxel,T.,1992.Intra-specificvariationinsolublemycelialproteinandesterasepatternsofLeptosphaeriamaculansFrenchisolates.MycologicalResearch96:677-684.6Boudart,G.,1989.AntibacterialactivityofsirodesminPLphytotoxin:Applicationtotheselectionofphytotoxin-deficientmutants.Appl.Environ.Microbiol.55:1555–1559.7Bokor,A.,Barbetti,M.J.,Brown,A.G.P.,MacNish,G.C.,andWood,P.,1975.Blacklegofrapeseed.JournalofAgricultureofWesternAustralia16:7-177.8Barbetti,M.J.,andKhangura,R.K.,1997.DevelopmentsforbettermanagementofblacklegdiseaseinWesternAustralia.Proceedingsofthe11thAustralianResearchAssemblyonBrassicas,AgricultureWesternAustralia,11-15.9Barbetti,M.J.,andKhangura,R.K.,1999.Managingblackleginthedisease-proneenvironmentofWesternAustralia.Proceedingsofthe10thInternationalRapeseedCongress,Canberra,Australia.10Cunningham,G.H.,1927.Dryrotofswedesandturnips:itscauseandcontrol.NewZealandDepartmentofAgriculture.Wellington,NewZealand:NewZealandDepartmentofAgriculture.Bulletin133.11Curtis,P.J.,Greatbanks,D.,andHesp,B.,1977.SirodesminA,B,CandG.Antiviralepipolythiopiperazine-2,5-dionesoffungalorigin:X-rayanalysisofsirodesminAdiacetate.J.Chem.Soc.PerkinI:180–189.12Eckert,M.R.,Rossall,S.,Selley,A.,andFitt,B.D.L.,2009.EffectsoffungicidesoninvitrosporegerminationandmyceliagrowthofthephytopathogensLeptosphaeriamaculansandL.biglobosa(phomastemcankerofoilseedrape).SocietyofChemicalIndustry.66：396-405.13E.P.,P.H.,1972,CMIDescriptionsofPathogenicFungiandBacteriaNo.331.14Evans,I.R.,Kharbanda,P.D.,andHarrison,L.,1995.BlacklegofcanolasurveyinAlberta1994.CanadianPlantDiseaseSurvey75:136.15Gugel,R.K.,andPetrie,G.A.,1992.History,occurrence,impactandcontrolofblacklegofrapeseed.Can.J.PlantPathology14:36–45.16Hammond,K.E.,andLewis,B.G.,1987.TheestablishmentofsystemicinfectioninleavesofoilseedrapebyLeptosphaeriamaculans.PlantPathology36:135–157.17Harman,G.E.,andNash,G.,1978.Soakingbrassicaseedsinfungicidesolutionstoeradicateseedbornefungi:Acomparisonofaqueousandorganicsolventinfusiontechniques.PlantDiseaseReporter.62:5.18Hammond,K.E.,Lewis,B.G.,andMusa,T.M.,1985.AsystemicpathwayintheinfectionofoilseedrapeplantsbyLeptosphaeriamaculans.PlantPathology34:557–565.19Harrison,L.M.,Kharbanda,P.D.,Stevens,R.R.,andCalpas,J.,1995.SpreadanddistributionofvirulentblacklegofcanolainthePeaceRiverRegionofAlbertain1994.CanadianPlantDiseaseSurvey75:135.20Johnson,R.D.,andLewis,B.G.,1994.Variationinhostrange,systemicinfectionandepidemiologyofLeptosphaeriamaculans.PlantPathology43:269-277.21Kharbanda,P.D.,1992.Performanceoffungicidestocontrolblacklegofcanola.CanadianJournaiofPlantPathology.14：169-176.22Kharbanda,P.D.,Yang,J.,Beatty,P.,Jensen,S.,andTewari,J.P.,1999.BiocontrolofLeptosphaeriamaculansandotherpathogensofcanolawithPaenibacilluspolymyxaPKB1.Proceedingsofthe10thInternationalRapeseedCongress,Canberra.23Kowaslska,J.,andRemlein-Starosta,D.,2011.ResearchofnonchemicalmethodsofwinteroilseedrapeprotectioninPoland.JournalofResearhandApplicationsinAgriculturalEngineering.56(3):220-223.24Marcroft,S.J.,Sprague,S.J.,Salisbury,P.A.,andHowlett,B.J.,2004.PotentialforusingresistancetoreduceproductionofpsedotheciaandascosporesofLeptosphaeriamaculans,theblacklegpathogenofBrassicanapus.PlantPathology.53:468-474.25Pedras,M.S.C.,Seguin-Swartz,G.,andAbrams,S.R.,1990.MinorphytotoxinsfromtheblacklegfungusPhomalingam.Phytochemistry29:777–782.26Pedras,M.S.C.,Taylor,J.L.,andNakashima,T.T.,1993.Anovechemicalsignalfromtheblacklegfungus:Beyondphytotoxinsandphytoalexins.J.Org.Chem.58:4778–4780.27Pedras,M.S.C.,1998a.TowardanunderstandingandcontrolofplantfungaldiseasesinBrassicaceae.Rec.Res.Dev.Agric.FoodChem.2:513–532.28Pedras,M.S.C.,1998b.Phytoalexinsfromcrucifers:Mimickingorovercomingplantchemicaldefenses.Rec.Res.Dev.Phytochem.2:259–267.29Pedras,M.S.C.,andBiesenthal,C.J.,1998a.ProductionofthehostselectivetoxinphomalidebyisolatesofLeptosphaeriamaculansanditscorrelationwithsirodesminPLproduction.Can.J.Microbiol.44:547–553.30Pedras,M.S.C.,Smith,K.C.,andTaylor,J.L.,1998b.Productionof2,5-dioxopiperazinebyanewisolatetypeoftheblacklegfungusPhomalingam.Phytochemistry49:1575–1577.31Pedras,M.S.C.,andOkanga,F.I.,1999.Strategiesofcruciferouspathogenicfungi:Detoxificationofthephytoalexincyclobrassininbymimicry.J.Agric.FoodChem.47:1196–1202.32Pedras,M.S.C.,Erosa-Lopez,C.C.,Quail,J.W.,andTaylor,J.L.,1999.Phomalairdenone:Anewhost-selectivephytotoxinfromavirulenttypeoftheblacklegfungusPhomalingam.Bioorg.Med.Chem.Lett.9:3291–3294.33Pedras,M.S.C.,2001.Phytotoxinsfromfungicausingblacklegdiseaseoncrucifers:Isolation,structuredetermination,detection,andphytotoxicactivity.Rec.Res.Dev.Phytochem.,inpress.34Pedras,M.S.C.,andBiesenthal,C.J.,2000a.HPLCanalysesofculturesofPhomaspp.:Differentiationamonggroupsandspeciesthroughsecondarymetaboliteprofiles.Can.J.Microbiol.46:685–691.35Pedras,M.S.C.,andBiesenthal,C.J.,2000b.Vitalstainingofplantcellsuspensioncultures:EvaluationofthephytotoxicactivitiesofthephytotoxinsphomalideanddestruxinB.PlantCellRep.19:1135–1138.36Petrie,G.A.,1995.EffectsofchemicalsonascosporeproductionbyLeptosphaeriamaculansonblackleg-infectedcanolastubbleinSaskatchewan.CanadianPlantDiseaseSurvey.75：1.37Rawlinson,C.J.,Muthyalu,G.,andCayleyG.R.,1984.Fungicideeffectsonlightleafspot,canker,cropgrowthandyiekdofwinteroil-seedrape.GreatBritain.103,613-628.38Rempel,C.B.,andHall,R.,1995.Effectoftimeandrateofapplicationoftriazolefungicidesonincidenceandseverityofblacklegandgrowthandyieldofcanola.DepartmentofEnvironmentalBiology.N1G2W1:5.39Rouxel,T.,Sarniguet,A.,Kollmann,A.,andBosquet,J.F.,1989.AccumulationofaphytoalexininBrassicaspp.inrelationtoahypersensitiveresponsetoLeptosphaeriamaculans.Physiol.Mol.PlantPathol.34:507–517.40Rouxel,T.,Kollmann,A.,Boulidard,L.,andMithen,R.,1991.AbioticelicitationofindolephytoalexinsandresistancetoLeptosphaeriamaculanswithinBrassiceae.Planta184:271–278.41Salisbury,P.A.,Ballinger,D.J.,Wratten,N.,Plummer,K.M.,andHowlett,B.J.,1995.BlacklegdiseaseonoilseedBrassicainAustralia:Areview.Aust.J.Exp.Agric.35:665–672.42Sosnowski,M.,Ramsey,M.,andScott,E.S.,1999.HostpathogeninteractionsbetweenLeptosphaeriamaculansandcanola.Proceedingsofthe10thInternationalRapeseedCongress,Canberra,Australia.43Tewari,J.P.,Shinners,T.C.,andBriggs,K.G.,1997.ProductionofcalciumoxalatecrystalsbytwospeciesofCyathusincultureandininfestedplantdebris.VerlagderZeitschriftfuÈrNaturforschung52c:421-425.44Williams,R.H.,andFitt,B.D.L.,1999.DifferentiatingAandBgroupsofLeptosphaeriamaculans,causalagentofstemcanker(blackleg)ofoilseedrape.PlantPathology48:161–175.45West,J.S.,Biddulph,J.E.,Fitt,B.D.L.,andGladders,P.,1999.EpidemiologyofLeptosphaeriamaculansinrelationtoforecastingstemcankerseverityonwinteroilseedrapeintheUK.AnnalsofAppliedBiology135:535-546.46West,J.S.,Fitt,P.K.,Leech,P.K.,Biddulph,J.E.,Huang,Y.J.,andBalesdent,M.H.,2002.EffectsoftimingofLeptosphaeriamaculansascosporereleaseandfungicideregimeonPhomaleafspotandPhomastemcankerdevelopmentonwinteroilseedrape(Brassicanapus)insouthernEngland.PlantPathology51:454-463.  國圖紙本論文 連結至畢業學校之論文網頁點我開啟連結註:此連結為研究生畢業學校所提供，不一定有電子全文可供下載，若連結有誤，請點選上方之〝勘誤回報〞功能，我們會盡快修正，謝謝！ 推文 網路書籤 推薦 評分 引用網址 轉寄                                                                                                                                                                                                                    top 相關論文 相關期刊 熱門點閱論文 無相關論文   無相關期刊   1. 臺灣牛皮葉衣屬地衣之分類 2. 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