DOC

Streptomyces

By Kim Jenkins,2014-07-19 00:33
17 views 0
StreptomycesStrept

    Streptomyces

JournaloyEnvironmentalSciencesVo1.17.No.1.PP.123125,2005[SSNl00lo742

    ;CNI1262q,X

    ;ArtideID:10010742(2005)01012303CLCnumber:X17Documentcode:A ;Streptomycesavermitilisfrommarine

    ;XIONGLixia,LIJianzhong,WANGHuili

    ;fResearchCenterforEcoEnvironmentalSciences,ChineseAcademyofSciences,Beijing100085,C

    hina.E-mail:1ook99@263.net)

    ;Abstract:Theinsecticidalstrain173wasisolatedfrommarinesourceanditsactivitywasexploredbytheb

    ioassayofbdneshrimpand

    ;Helicoverpaarmigera.Basedonmorphological,physiologicalandmolecularproperties,theinsecticid

    alstrain173wasidentifiedas

    ;Streptomycetesavermitilis,whichisthebestinsecticidalmicroorganismfoundintheterrestrialenviron

    ment.Thetaxonomyofthestrain

    ;173,insecticidalspectrumandpropertiesofthecorrespondinginsecticidalantibioticsarereported.

    ;Keywords:insecticidalmicroorganism;marine;taxonomy;Streptomycesavermitilis

    ;Introduction

    ;Theintensiveuseofchemicalpesticidesisoneofthe ;factorscontributingtohighcropyieldsandlowercommodity ;prices.Itisestimatedthat2.5milliontonsofpesticidesare ;appliedtoagriculturalcropsworldwideannually.However, ;inmostcases,theamountofpesticidecomingindirect ;contactwithtargetpestsisanextremelysmallpercentageof ;theamountapplied,usuallylessthan0.3%,andthus ;99.7%pesticideswent”somewhereelse”intheenvironment

    ;(Pimentel,1995).Contaminationbychemicalpesticideshas ;affectedthequalityofsoil,waterandairand,consequently, ;mayposearisktohuman,floraandfauna(Laura,2004). ;Fortunately,biologicalpesticidesaredistinguishedfrom ;conventionalchemicalpesticidesbytheirnaturaloccurrence, ;nontoxicmodeofactiontothetargetpests,safetyforhuman ;andothernontargetorganisms,reductionofpesticide

    ;residuesinfoodandenvironment,preservationofother ;naturalenemies,andincreasedbiodiversityinmanaged ;ecosystems(Mike,2000;Lacey,2001).

    ;Thepasttwodecadeshaveseensignificantresearch ;interestinattemptstodiscoveranddevelopenvironmental ;friendlypesticidesfromnatureorigin(Prem,2000).Sofar, ;mostoftheinsecticidalmicroorganismshavebeenisolated ;fromentomopathogensandterrestrialenvironment(Leonard,

    ;2000).Oceanisahugedrugresource,nearly6000kindsof ;novelchemicalcompoundshavebeenidentifiedfrommarine ;livingorganisms,amongwhichabout10000compoundsexert ;biologicallyactivities,suchasantitumor,antimicrobeand

    ;anti-virus,etc.(Fenical,1993;Davidson,l995). ;However,themarineenvironmentisstillrarelyexploredfor ;thedevelopmentofbiologicalpesticidesandlittleinformation ;aboutmarinemicroorganismspossessinginsecticidalactivities ;hasbeenreported(Takahashi,1989;Hu,2000).

    ;Inthecourseofscreeningfornewinsecticidal

    ;metabolitesfrommarinemicroorganisms,thestrain173was ;foundtohavestronginsecticidalactivityagainstbothbrine ;shrimpandHelicoverpaarmigra,aswepreviouslyreported ;(Xiong,2004).Inthispaper,thetaxonomyofthestrain ;173,insecticidalspectrumandpropertiesofthe ;correspondinginsecticidalantibioticsarereportedinthis ;paper.

    ;1Materialsandmethods

    ;1.1Microorganism

    ;Thestrain173wasisolatedandmaintainedaspreviously ;described(Xiong,2004).

    ;1.2Taxonomy

    ;Theinsecticidalstrain173wascharacterizedbyits ;morphologicalandphysiologicalproperties(Yan,1975),as ;wellas16SrDNAsequencing.ExtractionofgenomicDNA, ;PCRamplificationof16SrDNAandpurificationofthePCR ;productswasperformedfollowingtheprocedureofChun& ;Goodfellow(Chun,1995),Kim(Kim,1996)andLu(Lu, ;2001)respectively.The16SrDNAofthestrainwasaligned ;manuallywiththeavailableStreptomycesnucleotidesequence ;thatretrievedfromEMBL/GenebankandRDP(Ribosomal ;Databaseeroject;Maidak,1997)usingClustal*1.8 ;program(Thomposon,1997).

    ;1.3Laboratorybioassayofsomeimportantinsectpests ;Helicoverpaarm@era,Plutella”lostllaL.,aphidsand

    ;Spodopteraex@Ha,whichareseriouspestsofmanyimportant ;cropsandclaimmajorshareincroplossesevery)’ear,are

    ;usedasthetestpests.

    ;Testpests:3-instarlarvaeofHelicoverpaarmigera, ;PlutellaxylostllaL.,aphidsand5podopteraex@ua; ;Positivecontrol:1ppmand10ppmavermectinB1.

    ;;Test

    ;solution:thesupernatantoffermentationbrothofthestrain ;173;Thebioassaymethodwasthesameaspreviously

;described(Xiong,2004).

    ;1.4Propertiesoftheinsecticidalsubstancesofthe ;strain

    ;

    ;l24X10NGLi.xiaetalVo1.17

    ;ThepHofelevenportionsofthefermentationbrothwere ;adiustedto2,3,4,5,6,7,8,9,10,11and12

    ;respectivelyforthetestofpHsensitivity.Thefermentation ;brothwereheatedat40oC,60oC,80oC,100oC,121cIfor

    ;30rninrespectivelyforthetestofthermo-stability.The ;insecticidalactivitiesforthesamplesobtainedfromtheabove ;treatmentswereexaminedbybrineshrimpbioassay(Xiong, ;2004).

    ;1.5HPLCanalysis

    ;Thesupernatantandmyceliaoffermentationbrothwere ;extractedrespectivelywithethylacetateforthreetimes.The ;ethylacetateextractsofthesupernatantandthemyceliawere ;concentratedunderdiminishedpressuretodryness.,rIletwo ;residuesweredissolvedinmethano1.TheavermectinBl. ;(92%)wasusedasauthenticsample.HPLCanalysiswas ;carriedoutonShimadzuLC-3Aaccordingtothemethodof ;Thomas(Thomas,1979).

    ;2Results

    ;2.1Taxonomicstudiesofthestrain173

    ;Theculturalandphysiologicalcharacteristicsofstrain ;173grownonvariousmediaat2oCfor14daleshownin ;Table1andTable2respectively.Theorganismisaerobic, ;mesophilicandgram-positive.Thesporechainswere ;rectiflexibilesandsporesweresubsperiealtoovalinshape ;andsmooth.

    ;Table1Culturoldracteristiesofstrainl73

    ;Table2Physiologicalpropertiesofstrain173The16SrDNAsequenceofthestrain173showed100%

    ;similaritytothatofthetypestrainStreptomycesavermitilis ;andthestIain173wasidentifiedasStreptomycesavermitilis? ;2.2Bioassayofsomeimportantinsectpests

    ;ThebioassayresultsareshowninTable3.Thestrain ;173sho,vedstrongactivityagainstHeliothisarmigeralarvae, ;PlutellaxylostllaL.,Spodopteraexiguaandaphids.The ;inseeticidalactivityoffermentationbrothofthestrain173was ;similartothatof10ppmavermeetinBlinviewofthedeath ;timeandmortality,indicatingthatthestrain173isavery ;promisingmarineinsecticidalStreptomycesavermitilis. ;Table3Labbioas~yof3-instarlarvae

    ;ltellcoverpa

;arrmgera

    ;Plutella

    ;ylostllaL

    ;Spodoptera

    ;extgua

    ;Aphids

    ;1ppmavermectin ;10ppmavermectin ;Strain173

    ;1ppmavermectin ;10ppmavermectin ;Strain173

    ;1ppmavermectin ;10ppmavermectin ;Strain173

    ;1ppmavermectin ;10ppmavermectin ;Strain173

    ;18.O5

    ;76.39

    ;84.72

    ;15.27

    ;76.38

    ;79.16

    ;9.73

    ;40.27

    ;66.66

    ;44.44

    ;93.O5

    ;70.82

    ;20.83

    ;79.17

    ;87.50

    ;18.O5

    ;79.16

    ;81.94

    ;l2.5l

    ;43.O5

    ;69.44

    ;47.22

    ;95.83

    ;73.60

    ;40.27

    ;81.94

    ;90.27 ;44.44 ;91.66 ;84.7l ;22.22 ;51.39 ;79.16 ;90.27 ;94.44 ;90.27 ;45.83 ;87.5O ;95.83 ;50.0o ;97.22 ;90.27 ;27.78 ;56.95 ;84.72 ;95.83 ;l0o

    ;95.83 ;79.17 ;91.67 ;91.67 ;81.95 ;91.67 ;90.28 ;37.5 ;69.48 ;83.34 ;91.67 ;91.67 ;91.67 ;87.5O ;1O0

    ;1O0

    ;90.28 ;1o0

    ;9861 ;45.83 ;77.81 ;91.67 ;l0o

;J00

    ;l0o

    ;

    ;No.1Streptomycesavermitilisfrommarine125

    ;2.3Propertiesoftheir~ecticidalantibioticsofthe ;strain

    ;Theinsecticidalsubstancesshowedagoodstabilityina ;broadpHrangefrom3to12,andgoodthermo-stabilityfrom ;40~Cto80~(2,sotheyalegoodcandidatesforindustrial ;applications.

    ;2.4HPLCresults

    ;HPLCanalysisindicatedthatboththesupernatantand ;myceliaoffermentationbrothcontainavermectinBl.Thisis ;differentfromtherelatedreport(Richard,1979)thatclaimed ;avermectinBlmainlyexistsinmycelia.Itiswellknownthat ;avermectinBl,themostpowerfulavermectin,isbroadly ;effectiveagainstmostimportantagriculturalpests(Mrozik, ;1989).

    ;Bycomparisonwiththetaxonomicresultsof

    ;Streptomycesavermitilis(Richard,1979),itwasfoundthat ;thereweremanyobviousdifferencesbetweenthemarine ;insecticidalstrain173andStreptomycesavermitilisfrom ;terrestrialenvironment,suchasmorphologyofsporophores, ;solublepigment,utilizationofraffinoseandSUCroSe,andH2S ;productionetc.Itcanbededucedthatmanydissimilarities ;canbemadebytheStreptomycesavermitilisfrommarineand ;theStreptomycesavermitilisfromterrestrialenvironment. ;Avermectins,isolatedoriginallyfromtheculturebroth ;ofStreptomycesavermitilisfromsoil,areaseriesofunique ;16-memberedmacrolides(Egfrton,1979).Owingtohigh ;efficiency,broadspectrum(Mrozik,1989),lowresidue ;(Forbes,1993),rapiddegradationintheenvironment ;(Bruce,1993)andsafefornon-targetorganisms(Payne, ;1993),theavermectinnaturalproductsarethebest ;biologicalpesticideeverfoundandtheycanreplacethe ;highlytoxicchemicals.Thus,theproducingstrain173from ;marinesourceisaverypromisingandlesspollutedbio- ;pesticidewithsignificantimpo~anceforenvironmental ;protection.

    ;References:

    ;BruceAH,WilliamJA.VandenHeta1..1993.EnvironmentaleffeCIsofthe ;usageofavermectinsinlivestockJj.VetParasitol,48:l09125

    ;ChunJ,GoodfellowM.1995.AphylogeneticanalysisofthegenusNoeardiawith

    ;16SrRNAgenesequence[Jj.IntJSystBacteriol,49:l3951402.

    ;DavidsonBS,1995.Newdimensionsinnaturalproductsresea~h:cultured ;marinemicroorganismslJJ.CurtOpinBiotech,(6):284291.

    ;EgfrtonJR,OstlindDA,BlairLSeta1.,1979.Avermectins,newfamilyof ;potentanth-elmintie8genL~:efficacyoftheB1.component[J].InIJSyst ;Baeteriol,15(3):372378.

    ;FeniealW,1993.ChemicalstudiesofInarinebacteria:developinganewresource ;[J].ChemRev,93:1673--1683.

    ;ForbesB,1993.Areviewofregionalandtemporaluseofavermeetinsincattle ;andhorsesworldwide[J].VetParasitol,48:l928.

    ;HuZY,LiouSZ,HuangYJeta1.,2000.Arapidscreeningassayfor ;insecticidalantibioticsproducedbymarineactinomycetes[J].MarsciBuu, ;19(4):3641.

    ;KimD,ChunJ,SahinNeta1.,1996.Aanlysisofthermophilieeladeswithinthe ;genusStreptomyeesby16SribosomalDNAsequencecomparisons[J].IntJ ;SystBaeleriol,46:581--587.

    ;LaeeyLA,FrutosR,KayaHKeta1.,2001.Insectpathogensasbiological ;controlagents:dotheyhaveafuture?J].BioloControl,21:23(}’--248.

    ;LaumP,MarcoT,EttoreC,2004.Acalculationproceduretoas$~spotential ;envirenmentalriskofpesticidesatthefarmlevelJ].EcolIndicat,4:

    ;Ill123.

    ;LeonardGC,JuliusJM,2O00.Reviewbio.pesticides:areviewoftheiraction, ;applicationsandefficacyJj.PestManageSci,56:65l676.

    ;LuzH,LiuzH,WangLMeta1.,2001.S~charopolysporamsp.nov.

    ;andSaccharopolysporathermophilasp.nov.,novelaetinomycetesfromsoil ;[Jj.IntJ0fSystEnvolBaeteriol,5l:3l9325.

    ;MaidakBL,OlsenGL,Lal~nNeta1.,1997.Theribosomealdatabasepmject ;(RDP)[J].NueleicAcidsRes,25:l09Il1.

    ;MikeN,2000.neregulationofnaturalproductsascroppmteetiortagentS[J]. ;PestManageSci,56:677680.

    ;MrozikH,EskolaP,LinnB0eta1.,1989.Discoveryofnovelavermectilmwith ;unprecedentedinsecticidalactivity[J].Experientia,45:3l53l6.

    ;PayneGT,SoderlundDM,1993.Actionsofavermectinanalogson7- ;aminobutyrieacid(GABA)sensitiveandGABAinsensitivechloridechannels

    ;inmousebrainJJ.PestleBiochemPhys,47:l78184.

    ;PimentelD,1995.Amountsofpesticidesreachingtargetpests:environmental ;impactsandethicsJ].JAsrieEnvironEthics,8:l729.

    ;PremW,2000.Livingsystemsasnaturalcropprotectionagents[J].PestManage ;Sci.56:68l687

    ;RichardWB,BrintonMM,EdwardEB,1979.Avermectins,newfamilyof ;potentanthelmintieagents:producingorganismandfermentation[J]. ;AntimicrobAgentsChemother,15:36l367

    ;TakahashiA,KurasawaS,IkedaD,1989.Ahemicidin,Anewacarieidaland ;antitumorsustaneeI.Taxonomy,fermentation.isolationandphysical

    ;chemicalandbiologicalpropeaies[J].JAntibiotics,42:1556--1561.

    ;ThomasWM,LousC,DouglasJCeta1.,1979.Avermectins.newfamilyof ;potentanthelminticagents:isolationandchromatographicprope~ies[J]. ;AntimicrobAgentsChemother,15:36837l

    ;ThompsonJD,GibsonTJ,P1ewniakFeta1.,1997TheClustalXwindows ;interface:flexiblestrategiesformultiplesequencealignmentaidedbyquality ;analysistools[J].NucleicAcidRes,25:4876--4882.

    ;XiongLX,LiJZ,KongFZ,2004.Streptom)sp.173,aninsecticidal ;microorganismfrommarineJJ.LettApplMicrobiol,38:3237.

    ;YanXC,1975.IdentificationhandbookofStreptomyces[M].Beijing:Science ;Press.

    ;(ReceivedforreviewMarch31,2004.AcceptedMay14,2OO4)

    ;

    ;

Report this document

For any questions or suggestions please email
cust-service@docsford.com