生防细菌BRF21和BRF22鉴定及生物学特征

©1994-2010ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.:(2004AA3CN154);(KZCX32SW2NA204):(1966-),,,,E2mail:[e-mail]21BRF2213,1,1,2,1,2,1(11/,150081;21):BRF21BRF22,7BRF21,;BRF22,,Biolog16SrDNA,BRF21BRF22,BRF21BRF22:;;Biolog;16SrDNA;:Q939111019;S476:A:100529261(2007)0120049206IdentificationandBiologicalCharacteristicsofBiocontrolBacterialStrainsBRF21andBRF22WANGGuang2hua,JINJian,XUMei2na,CHENXue2li,LIUXiao2bing(LabofBlackSoilEcology,NortheastInstituteofGeographyandAgriculturalEcology,AcademiaSinica,Harbin150040,China)Abstract:BRF21andBRF22weregrampositivebiocontrolbacterialstrainsisolatedfromsoybeanrhizo2sphere.Dual2culturetestsshowedthatthetwostrainswerestronglyagainstsevenplantfungalpathogens.Bothstrainsformedendospores.TheendosporeofBRF21andwasBRF22growninthemiddleandattheendofcell,respectively.BRF21hasnoflagellum,andBRF22hasflagellagrowingaroundcell.ResearchresultsshowedthatthebiologicalcharacteristicsofBRF21andBRF22weredifferent.Identificationbyus2ingBiologand16SrDNAsequenceanalysisshowedthatBRF21andBRF22werePaenibacilluspolymyxaandBacillussubtilis,respectively.Keywords:Paenibacilluspolymyxa;Bacillussubtilis;biologidentification;16SrDNAsequence;bio2logicalcharacteristicBRF21BRF22,,9423(1)49-54ChineseJournalofBiologicalControl20072©1994-2010ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.[1,2],Biolog16SrDNA,,1111BRF21BRF22(Fusariumoxysporum)(Fusariumsolani)(Fusariumgraminearum)(Rhizoctoniasolani)(Sclerotiniasclerotiorum)(Fusariumoxgsporumf.sp.lycopersici)(Fusariumoxysporumf.sp.cucumerinum),112BRF21BRF22PDA5mm,215cm,3,2,285d,113[3]114BiologBUG+M+T12h(BRF22)24h(BRF21)(Biolog),,28%,8BiologGP150l,96,30,618h,,,11516SrDNAPCRDNA,27f(52AGAGTTTGATCCTGGCTCAG23)1492r(52GGCTACCTTGTTACGACTT23)PCR[4]PCR(50l):10PCR5l,dNTP5l,27f1492r(25pmol)015l,DNA115l,Ex2Taq(5U/l)0125l,37125lPCR953min;941min,551min,72115min,30;728min,PCR,,27f1492rBlastGenBank(),,PhylogeneticTreeNeighbor2Joining2211BRF21PDA,,0523©1994-2010ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.()PDB,,,118212m,016018m,,(12A),BRF22PDA,,,,,,(013019m)(019212m),,(12B),1BRF21(A)BRF22(B)212,BRF21,,,,,,,NaCl5%PDBBRF22,,,,,,,NaCl7%PDB,NaCl10%PDB213(1),BRF21BRF22,,BRF2131008188mm,BRF22917515100mm1BRF21BRF22(,mm)BRF2181881103A81750165A81670158A81500182A61000182B31000182C31290195CBRF22121000141B91880185C111000191BC151000100A101250196BC121131103B91751126C:(P0101)21421411BiologBiolog2,BRF216h(Paenibacilliuspolymyxa),(SIM)0193,(DIS)1101;18h,015BRF21BRF226h,015,(Bacillus)2141227f1492r16SrDNAPCR,151:BRF21BRF22©1994-2010ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.(Paenibacilliuspolymyxa)019300148611017160(Globicatellasanguinis)010010102931228158(Streptococcussuis)00412710100BRF22C(BacillussubtilisC)014920124061117127B(BacillusamyloliquefaciensB)010980121461647131A(BacillusamyloliquefaciensA)010070108171537162115kbPCRPCRBRF21BRF221433bp1420bpGenBank,BRF21BRF22DQ298737DQ298738BLAST16SrDNAGenBank,PhylogeneticTree,(2):BRF21(AY359614AJ223989AJ320493),98%,BRF21,Biolog;BRF22,(AY881640AY651023AY894690AY553097),99%,BRF222BRF21BRF223BRF21BRF22,,,BRF21BRF22,,Bi2ologMicroStationBiolog,952523©1994-2010ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.[5,6][6]Biolog11,100%,,,,Biolog,BRF21,16SrDNA;BRF22Biolog,(46hSIM0175;1624hSIM015),16SrDNA,16SrDNA98%,,93%95%,[7],BRF2216SrDNA,AY881640AY651023AY894690AY55309799%,BRF22[810],,(iturin)[11],[12]BRF22,,[2],1994,Ash[13],,[1416],BeattyJensen[16],,883897948961D[17]WY110(Magnaporthegrisea)P2,26kD,BRF21,,[1,2],:,[1],,,.BRF21[J].,2003,19(2):73771[2],,,.[J].,2005,21():96-1011[3],.[M].:,20011[4]PolzMF,HarbisonC,CavanaughCM.DiversityandheterogeneityofepibioticbacterialcommunitiesonthemarinenematodeEu2bostrichusdianase[J].AppliedEnvironmentalMicrobiology,1999,65:231-2401[5],.BIOLOG[J].,1996,23(5):264-2671[6],,,.Biolog[J].,2000,20(6):36-381[7]FryNK,WarwickS,SaundersNA,etal.Theuseof16SribosomalRNAanalysestoinvestigatethephylogenyofthefamilyLe2gionellaceae[J].JournalofGeneralMicrobiology,1991,137:215-12221[8],,,.Bs2208[J].,2003,19(4):171-1741351:BRF21BRF22©1994-2010ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.(,110161):B8,;,100%B87811%,,,16SrDNA,:;;;16SrDNA:S435172;S476:A:100529261(2007)0120054206:2006201218:(2001208001);([2002]187);(2004011):(1972-),,,,E2mail:zhaoxx0772@1631com;3,E2mail:[e-mail][9],,PhamH,.B2[J].,2003,19(4):175-1791[10],,.G3[J].,2003,19(4):206-2071[11],.[J].,2005,3:213-2191[12]MartenP,SmallaK,BergG.GenotypicandphenotypicdifferentiationofantifungalbiocontrolstrainbelongingtoBacillussubtilis[J].JournalofAppliedMicrobiology,2000,89:463-4711[13]AshC,PriestFC,CollinsMD.MolecularidentificationofrRNAgroup3bacilli(Ash,Farrow,WallbanksandCollins)usingaPCRprobetest[J].AntonieVanLeeuwenhoek,1994,64:253-2601[14],,,.[J].,