环境科学乳酸菌复合系SFC2处理水稻秸秆的效果高丽娟

©1994-2010ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.(,100094):,,SFC22,.,,,30d,pH318,411.,L22,10116%,312%.(DGGE),SFC22,Lactobacillusplantarum,LactobacillusfermentumLactobacillusparacasei,Enterobactersakazakii,Pantoeaagglomerans,Enterobacterendosymbiont,Pantoeaananatis,.:;;:X712:A:025023301(2007)0621392205:2006208230;:2006211203:(30571088):(1978),,,,E2mail:[e-mail]3,E2mail:[e-mail]cteriaCommunitySFC22TreatedonRiceStrawGAOLi2juan,WANGXiao2fen,YANGHong2yan,GAOXiu2zhi,LBYu2cai,CUIZong2jun(CollegeofAgronomyandBiotechnology,ChinaAgriculturalUniversity,Beijing100094,China)Abstract:Aimedtoutilizericestrawandlessenthepressureofenvironment,thericestrawwasusedasthefermentationmaterial,andalacticacidbacteriacommunitySFC22frommylaboratorywasinoculatedintothericestrawtoinvestigatetheinoculationeffects.After30daysfermentation,theinoculatedfermentedstrawsmeltacid2fragrant,andthepHvaluewas318,whichwaslowerthanthecontrolof411.Furthermore,lacticacidconcentrationwasmorethanthatinthecontrol.EspeciallyL2lacticacidconcentrationwastwotimesmorethaninthecontrol,andthecrudeproteincontentwas10116%higherthanthatinthecontrol,andthecrudefibercontentwas312%lowerthanthatinthecontrol.Fromthepatternsofdenaturinggradientgelelectrophoresis(DGGE),Lactobacillusplantarum,LactobacillusfermentumandLactobacillusparacaseirapidlybecametheadvantageousspeciesintheinoculatedstraws.However,Enterobactersakazakii,Pantoeaagglomerans,Enterobacterendosymbiont,Pantoeaananatis,whichwerepredominateinthecontrols,werenotdetectedintheinoculatedstraws,andthefermentedqualitywasimprovedsignificantly.Keywords:ricestrawfermentation;community;DGGE,.2,[1,2].,,.,,,[2].,6151087108t[3].,5%,30%,[4].,,..,,.,10a:201055%[3].,,,,..[5],,SFC22[6].SFC22,.28620076ENVIRONMENTALSCIENCEVol.28,No.6Jun.,2007©1994-2010ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.[6],(Lactobacillusplantarum)(L1fermentum)(L1paracasei).MRS2S(1L10g,10g,5g,K2HPO42g,2g,5g,20g,801mL,MgSO47H2O0158g,MnSO44H2O0125g,pH612614,11520min)24h.:2004.11212cm,70%(),MRS2S24hSFC221106CFUPg(),,100mL,30.,15,27101530d3.113pH,.015g415mL,20min,012mLHORIBApH(modelB2212,HORIBA,Japan).1146048h,40,(water2solublecarbohydrates,WSC)[7],,,,[8].1151g,2mL,0122m,(GC2MS,modelQP15050,Shimadzu,Japan).:CP2Chirasil2DexCB(25m0125mm);:502min5Pmin100,15Pmin190,2min,18min;:190;:230;:115kV;:(64kPa);:30mLPmin;,1P22,1L.116MRS;SFC22MRS2S(),(310g,1010g,NaCl510g,15g,1L,pH714716,12115min);PDA(2010g,200300g,1510g,1L,pH515517,11520min).,,2,30.117DNADNA2[9].118PCR2DGGEDGGEPCR:357F2GC(GCclamp252CCTACGGGAGGCAGCAG23,GC2clamp52CGCCCGCCGCGCGCGGCGGGCGGGGCGGGGGCACGGGGGG23)517R(52GTGCCAGC(APC)GCCGCGG23)[10]16SrDNAV3,DGGE.PCR(50L):DNA10ng,10PCRGoldBuffer5L,25mmolPLMgCl24L,2mmolPLdNTP5L,45molPL357F2GC517R015L,5UPLAmpliTaqGold(PerkinElmerJapan)012L.PCR:9510min,931min,481min,721min10s,30,723min50s.PCR2%.DGGE:DCodeTM(Bio2Rad,Laboratories,Hercules,CA,USA).6%12%(),015TAE(20mmolPLTris2HClpH813,10mmolPLaceticacid,015mmolPLEDTA),20%60%[100%7molPL,40%()],200V615h,SYBRGreenI(MolecularProbes,Eugene,Ore)30min,[10].DGGEDNA,,QIAquickPCRpurificationkit(QIAGEN),[11].ABIPRISMBigDyeTerminatorCycleSequencingReadyReactionkit(AppliedBiosystems),ABI3130xl(AppliedBiosystemsJapan),AppliedBiosystems.119DGGE(16SrDNAV339316:SFC22©1994-2010ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.)GenBankPEMBLPDDBJ,:DQ5158742DQ515889.2211pHSFC222d,30d,,;,,,.2dpH613410,415,15d318,411,.pH,pH.212;;;;2,32;L2;D2;130dGC2MSFig.1GC2MSspectrometrymapsofvolatileproductsofthericestrawbetweenthecontrolandtheinoculatedwiththeSFC22after30dfermentation30d1.1,6137%6135%,5113%5166%,10116%,33144%32136%,312%.GC2MS30d,2,32L2D2,1..L2D2,30d,,,15d,,().30d1.30dL27114gkg-1,18153gkg-1,159152%,D2113120%,139149%.,8108gkg-1,7182gkg-1,3122%,147146%.pH,,.,,[12].130d()Table1Chemicalanalysesofthefermentedstrawsafter30daysoffermentationandcontrol(DMbasis)P%pH41113180-7150Pgkg-11216630132+139149L2Pgkg-1711418153+159152D2Pgkg-1515311179+113120Pgkg-181087182-3122P11573188+147146P%61376135-0130P%51135166+10116P%3314432136-3120213.2.2,30d,2d,61967113CFUPg,7d,81228182CFUPg,,30d61167156CFUPg.230d,.2d,81787106CFUPg,30d611210CFUPg.,30d,41053161CFUPg.21430dPCR2DGGE,32,230d,16,BLAST,Enterobactersakazakii,Pantoeaagglomerans,EnterobacterendosymbiontPantoeaananatis.493128©1994-2010ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.(L.fermentum)14(L.plantarum),15d5(L.paracasei)7,30d57,15(Unculturedbacterium).30d,L.plantarumL.fermentumL.paracasei,3SFC22.SFC22,,..M2Marker:(20%60%),(6%12%)3PCR2DGGEFig.3DGGEprofilesofthericestrawbetweenthecontrolandtheinoculated3SFC22,,.pH,2dpH613410,415.,pH5,[13],,,[14].23Table2IdentitiesofPCR2DGGEbandsinFig13P%1UnculturedbacteriumAF49935696192UnculturedbacteriumAB254251100103EnterobacteriaceaebacteriumAY53869499154EnterobactersakazakiiAY752