海南生态区植物根际解磷细菌的筛选及分子鉴定

ResearchPaperActaMicrobiologicaSinica49(1):64-71;4January2009ISSN0001-6209;CN11-1995PQhttp:PPjournals.im.ac.cnPactamicrocn:(Rky0502)3Tel:[phone];Fax:[phone];E2mail:[e-mail]:(1969-),,,,,Tel:[phone];E2mail:[e-mail]:2008207204;:2008209213,,3(,571737):Ca3(PO4)2FePO4;,21,22,,,;Ca3(PO4)25d,;Ca3(PO4)232200rPmin6d,200mgPL;415d4,Ca3(PO4)26d,200mgPL(PSBHS),FePO4PSBHS6d,;PSBHS16SrDNA1460bp,,Blast,363,1264514;14PSBHSCa3(PO4)26d,2011062313mgPL,pH(31824134)(r=-018155)14PSBHSFePO46d,1163412mgPL,pH(21875167)(r=-016836)16SrDNA,6PSBHSAcinetobacter,3Pseudomonas,3Serratia,2Enterobacter:;;;:Q939:A:000126209(2009)0120064208,95%Ca2+Fe2+Fe3+Al3+,,5%10%,,25%[1],,,pH,,28,Ca3(PO4)2FePO4,,111111111:21,(Heveabrasiliensis)(Eucalyptussp.)(Acacoasp.)(Phyllostachyssp.)(Musasp.)(Festucalsp.)(Zoysiatenuifolia),020cmpH416615(Bruguieragymnorrhiza)(Xylocarpusgranatum)(Aegicerascormiculatum)(Rhizophoraapiculata)(Excoecariaagallocha)(Ceriopstagal)(Bruguierasexangula)(B.s.vae.rhynchopetala)(Acanthusilicifolius)(Hibiscustiliscus)(Lumnitzerareacemosa)(Sonneratiaalba)(Avicenniamarina)(Sonneratiaovata),,020cmpH219616[2]5,4,19111112::(NA)[3]2(NAC)[4]2(YEM)[5](20%)(50mgPL),pH710NA;NACNA(110mgPL),G-;YEM:(NAM)[5][6]:10g,5g,MgCl26H2O5g,MgSO47H2O0125g,KCl012g,(NH4)2SO4015g,110L,pH710Ca3(PO4)2(19197%)FePO44H2O(13189%)FePO44H2O11211211:,(),2mm,,24h5min,35();,,20g,,50mL,,,(),200rPmin015h,5min,210mL,,10-1,10,10-210-310-410-5,2030cm,,3mm(),,24h11212:10-510-410-3012mL3(NANACYEM),2,323d,NAM,,NAMNA23d,4,(20%)-20113,Ca2P,3,3;325d,,(phosphate2solubilizingbacterialstrain,PSB)PSB:100mL20mLCa2P,PSB,3;32200rPmin6d,8000g10min,,[7],pHpHPSB200mgPL,415d,;45Ca2P,,(Phosphate2solubilizingbacterialstrainswithhighandstableactivity,PSBHS)114PSBHSFe2P56:.P(2009)49(1),,pH115Excel2000,SAS6112Duncanps116[8]PSBHSDNA,(fD:5’2CCGAATTCGTCGACAACAGAGTTTGATCMTGGCTYAG23’,rP:5’2CCCGGGATCCAAGCTTACGGHTACCTTGTTACGACTT23’,M=AC;Y=CT;H=ATC)16SrDNA1460bp,,Blast,PSBHS2211NANACYEM328,363560,446,47,14210212363Ca2P325d,126(12A3),135(12BDR350DR363),Ca2P126PSB32200rPmin6d,4520010mgPL(1812mgPL)PSB45PSB4,Ca2P6d,31PSB,4,14PSB20010mgPL(1),DR379,62613mgPL14PSBHS6d,Ca2PpH31824134(pH6183)pH(X)(Y)r=-018155,1Ca2PFig.1Phosphate2solubilizingcapabilitytestsforbacterialrepresentativestrainswereconductedonsolidmediumwithtricalciumphosphateasthesolePsource.Halozonessurroundingthecoloniesindicatedcorrespondingstrainswithphosphate2solubilizingactivities.16dCa3(PO4)2pHa)Table1AmountsofphosphorusdissolvedfromCa3(PO4)2byPSBHSafterliquidcultivationfor6daysandtheresultingpHvaluesofmediaa)Strainnumberofphosphate2solubilizingbacteriaAmountofdissolvedphosphorusP(mgPL)pHvalueofculturemediumDR379626138.4a31820.08fDR2445321844.0b41030.06eBR0984281232.6c41050.02eBR1153741119.0c41010.03eBR1213571273.1cd41160.02cdAR0453561143.3de41030.06eDR1723421118.5de41020.04eBR1193141056.9de41100.02deDR3043071032.7de41020.05eAR0662921118.6ef41250.07abcCR1502911111.1fg41220.04bcBR12628812103.2de41340.05aDR2062381821.7fg41260.04abCR1422011021.8g41290.02abNon2inoculatedmedium18123.4b)61830.04a)DataareshowedasmeanStd.,andthelettersappendedshowthemultiplecomparisonresultsfordifferentmeans.Meanswiththesameletterarenotsignificantlydifferent(Alpha=0105).b)Amountofdissolvedphosphorusbynon2inoculatedmedium.66YuekunWangetal.PActaMicrobiologicaSinica(2009)49(1)P0101,YXY=135510-24717X213FePO414PSBHSFe2P,32200rPmin6d,Fe2P,1910mgPL4(BR098BR115BR119DR379),DR379,3412mgPL,1612mgPL(2)26dFePO4pHa)Table2AmountsofphosphorusdissolvedfromFePO4byPSBHSafterliquidcultivationfor6daysandtheresultingpHvaluesofmediaa)Strainnumberofphosphate2solubilizingbacteriaAmountofdissolvedphosphorusP(mgPL)pHvalueofmediumDR37934125.4a21870.06iBR09832111.3ab31050.09ghBR11929111.0b31020.08hBR11519163.9c21810.08iBR1216161.1d31130.08ghDR2446121.0d31150.05gDR1724160.8de31550.05fCR1503100.7ef41830.12cDR3042160.7ef41440.04dAR0662160.3ef41000.06eCR1421190.9f31940.05eBR1261170.6f51670.04aAR0451160.5f51640.04aDR2061160.7f51500.09bNon2inoculatedmedium20123.1b)51820.04a)andb).,seeTable1forannotatons.14PSBHS6d,Fe2PpH21815167(pH5182)pH(X)(Y)r=-016836,P0101,YXY=4211640-719658X419103412mgPL,pH21813105,21416SrDNA1540bp,,,16SrDNA,5016SrDNA,,16SrDNA(1450bp)4,16SrDNA1450bp416SrDNA1460bp(),,Blast,3,()(1450bp)4,34,146Acinetobacter,3Pseudomonas,3Serratia,2Enterobacter[9,10]PseudomonasSerratiaEnterobacter,,[11]Acinetobacter,Acinetobacter3200,:,,;3,,,60%80%,;,80%,,[10]10-2911310-22,,10-7molPL10-11molPL,,,(),,1L5g5g,6d,pH61863154,1318mgPL76:.P(2009)49(1)614mgPL;2molPL217mL,2molPL210mL,6d,pH51465196,7016mgPL1412mgPL,pH,,316SrDNABlastna)Table3Blastnsearchresultsforthe16SrDNAsofdifferentphosphate2solubilizingbacteriaa)InformationonsubmittedsequencesSequencesproducingsignificantalignmentsInformationonalignmentsStrainNo.SequencelengthPbpAccessionNo.MicroorganismspeciesSequencelengthPbpAlignmentdomainPbpAccessionNo.ScoreIndentitiesGapsDR3791465gb|EU603505|Serratiamarcescens1464121464dbj|AB24445311|28641461P1465(99%)1P1465(0%)Serratiamarcescens1464121464dbj|AB24443311|2864146