假单胞菌Nwu1_mu对陕北石油污染土壤的生物修复作用研究

2010,29(5):910-917JournalofAgro-EnvironmentScienceNwu1-mu。、、。Nwu1-mu60d86.5%C24~C28C28。Nwu1-mu。X53A1672-2043(2010)05-0910-08Nwu1-mu710069BioremediationofPetroleum-ContaminatedSoilinNorthwestofChinabyPseudomonassp.Nwu1-muLUXin,CHENLi,LIJuan,CHENWu-lingCollegeofLifeScience,NorthwestUniversity,Xi′an710069,ChinaAbstractTostudycharacteristicsandmechanismofpetroleumhydrocarbonsdegradationduringthebioremediationcourse,Pseudomonassp.Nwu1-muwasusedonbioremediationtothepetroleum-contaminatedsoilPCSfromthenorthernofShaanxiProvince,China.ThedegradationeffectofpetroleumhydrocarbonsinPCSwasinvestigatedthroughdetectingquantityofstrains,dehydrogenaseactivity,surfacetension,degradationrateoftotalpetroleumhydrocarbonsTPHanditsconstituentsinbioremediationcourse.DegradativeefficiencyofNwu1-mutoTPHfinallyreached86.5%within60dayswithoutadditionalnutrients,whilesignificantdegradationwasmadetothepetroleumhydrocarbonsconstituentsofC24~C28andC28.DehydrogenaseandbiosurfactantproducedbyNwu1-muplayedimportantrolesinpetroleumhydrocarbondegradation,bothofthemshowedeffectiveaccelerationduringtheremediationcourse.ThePseudomonassp.Nwu1-mushowsremarkablypetroleumhydrocarbonsdegradationcharacteristicswhileusingPCSastheonlynutrientsource.Agreatpotentialforbioremedia－tiononpetroleum-contaminatedsoilofloessplateaucanbeexpectedbyNwu1-mu.KeywordsPseudomonas;petroleumhydrocarbondegradation;dehydrogenase;surfactant;bioremediation2009－09－172009ZKC04-161984—。[e-mail]@263.com[1]。[2]。MarinJA[3]。[4-5]。Bento1241mN·m-1[6]。HarveyP.aeruginosaSB3029540℃1min1%3[7]。。、。Nwu1-mu。11.1。、、16sRNA。0.5～1.0μm×1.5～5.0μm。LB、0.3～0.5μm×0.4～1.0μm1。16sRNA《》[8]Pseudomonassp.Nwu1-mu。20cm3~12mm1.936g·cm-312.69%28.3g·kg-119.26g·kg-11.43g·kg-10.68g·kg-115.63g·kg-1pH7.6。5g2g1g1000mLpH=7.5。30℃、165r·min-1。C10C12C14C16C24C28C32C36500μL·mL-1inHexaneAccustandard。1.212A、B、C、D、E、A′、B′、C′、D′、E′、F、G。1。2.5kg121000mLA、B、C、D、E、G1×105Pa30min。1.1～1.3×108CFU·mL-1A、B、C、D、E、A′、B′、C′、D′、E′。121Figure1Pseudomonasmigulacolonyandmicroscopicmorphology1Table1ExperimentdesignNwu1-muA1%+B1.5%+C2%+D2.5%+E3%+A′1%－B′1.5%－C′2%－D′2.5%－E′3%－F0－G0+9112010525～30℃20%60d3~6dTPHTotalpetroleumhydrocarbonTPH、、、。FG。1.31.3.10.2%、30℃14d[9]。1.3.2、12500r·min-130min6mol·L-1pH2.02Na2SO440℃。0.05mol·L-1NaHCO3pH2.04℃24h12500r·min-115min[10]。TLC//V∶V∶V=65∶15∶201-;2;3[11]。1.3.35gV∶V=2∶120mL1h3。JYW-200A[12]。1.3.41gTris-HCl、0.1mol·L-1、0.5%TTC2mL37±1℃10～20min。25mL4000r·min-15min[13]。1h1mgTFmgTF·g-1·h-1。1.3.5TPHEPA3546USEPA3546MicrowaveExtrac－tion[S]。2gCEM25mLV∶V=2∶1。CEM1200W5min120℃30min。40℃。TPH[14]。35%。1.3.6。ShimazuGC-MS-QP2010-ShimazuGC-2010。DB2130m0.25μm0.32mmFID100℃→8℃·min-1→295℃-16min。-100℃→8℃·min-1→295℃-16minScanm/z33—650。22.1TPHTPH2、3。TPHTPH。。10%TPH[15]。2、3TPH3%TPH2%。Nwu1-mu912295。2%。2%C′C′。2.2TPH60d。TPHFGTPH4。Nwu1-mu5dNwu1-mu。TPH15d20d。。20dTPH50dTPH86.5％。50d。60dTPH2.83％0.382％86.5%。TPH。F35dTPH28.1%35d25%。GTPH20d18.2%TPH12%。TPHNwu1-muTPH。Nwu1-mu。TPHC+FC′。Nwu1-mu、TPH。。2.3TPHTPHTPH91320105TPH。[16]。。Nwu1-mu1.98g·L-1。-。、[17]。、、、、[18][19]。5。Nwu1-muTPH10dTPH。20d4。Nwu1-mu25dTPHTPH。40d31mN·m-1TPH。1Nwu1-mu23Nwu1-muTPH。2.4TPHTPH[20]。[21]。TPH6。5dNwu1-mu。5dNwu1-mu9142957GC/MSFigure7TheGC/MSchromatogramofpetroleumhydrocarbonconstituentsintheundisposedpollutedsoil2.752.502.252.001.751.501.251.00t/minTIC×1065.010.015.020.025.030.035.040.045.0。20~40d5TPH。45d2.71mgTF·g-1·h-150dTPH86.5％。50d。。60dTPH。2.5Nwu1-muNwu1-mu。7。781722263638475557596372C12C14C16C17C21C22C23C24C3424．82％5910111213614151618253033343537414364666768697071120532356272849525462、C12C14C16C17C21C22C23C24C3445．46％267192940445160、、、11．52％34654846215045423932312458、、、18．20％。。7C10C36C10C12C14C16C24C28C32C36、15d45d。8。7、82Nwu1-muC16~C24、C24~C28C28。10dC16~C24、C24~C28C28C14[22]。10～30dC16~C24、C24~C28C28C144、5、6TPH、30～40d40～50d。915201051.251.000.750.500.250.00-0.25t/min×10610.015.020.025.030.035.040.045.050.055.060.02Table2Thequantitativeanalyticresultofgaschromatogramofpetroleumhydrocarbonconstituentscontentinthecontaminatedsoil0d15d30d45d60d/mg·kg-1/mg·kg-1/%/mg·kg-1/%/mg·kg-1/%/mg·kg-1/%C14348406-16.5371-6.413461.726324.6C14~C1695574721.852045.624574.419579.6C16~C244435313329.4266839.8103976.671983.8C24~C283574206242.3141060.661282.921194.1C28134568249.341968.89892.86994.98Figure8ThegaschromatogramofPetroleumHydrocarbonconstituentscontentwithtimea.b.c.15dd.45de.。31Nwu1-mu60d86.5%。2。。3。4。[1]DrachukSV,KoksharovaNV,FirsovNN.Microfloraofsoilspollutedwithpetroleumproducts[J].RussianJournalofEcology,2002,332135-137.[2],,,.[J].,2009,293193-197.ZHANGZi-jian,LIUYong-di,MENGQing-mei,etal.Researchpro－gressesonbiodegradationofpetroleumhydrocarbonpollutants[J].Envi－ronmentalProtectionofChemicalIndustry,2009,293193-197.[3]MarinJA,HernandezT,GarciaC.BioremediationofoilrefinerysludgebylandfarminginsemiaridconditionsInfluenceonsoilmicrobialactiv－ity[J].EnvironmentalResearch,2005,98185-195.[4],,.[J].,2002,29375-78.Nwu1-mu916295QIANXin-ping,YANGYong-rong,MENGQin.Theeffectsofbiosurfactantonmicrobes′growthandmetabolism[J].Microbiology,2002,29375-78.[5],,,.[J].,2003,2