一株菊酯类农药降解菌的分离鉴定及其降解酶基因的克隆Isolation,identificationanddegradinggenecloningofapyrethroids-degradingbacterium摘要：[li的]筛选分离髙效降解菊酯类农纯的光合细菌，研究其降解特性，并对该菌株屮降解酶垄因进行克隆与初步分析.[方法]根据分离菌株的细胞形态结构、活细胞光吸收特征、生理生化特征及其16srDNA序列系统发育分析鉴定降解菌,气和色谱法测定该菌株降解菊酯类农药的能力,PCR方法克隆降解晦基因.[结果]菌株PSB07-21属红假单胞菌属(Rhodopseudomonassp.),其降解最佳条件为3000lx、35°C、pH7,在此条件下培养15d对600mg/L甲氤菊酯、氯氤菊酯、联苯菊酯降解率分别为66.63,43.25,50.18%.经序列测定与分析表明，克隆到1个329bp的ORF,该0RF预测的编码蛋白与20G-Fe(II)oxygenase的同源性为37.0%.添加低浓度Fe(II)对该菌株降解菊酯类农药有一定促进作用.[结论1PSB07-21能冇效的降解多种菊酯类农药，该研究为菊酯类农药生物修复提供理论依据.Abstract：[Objective]Thisstudyaimedtoisolate,identifyandclonedegradinggeneofasyntheticpyrethroidsdegradingbacterium.[Methods]AphotosyntheticbacterialstrainPSB07-21capableofdegradingseveralsyntheticpyrethroidsefficientlywasisolatedbyanenrichmentculture・PSB07-21wasidentifiedbasedonitsmorphology,physiologyandphylogeneticJinalysisof16SrDNAsequenee.Thedegradationabilityofthisstrainwasevaluatedwithgaschromatography・ThedegradinggenewasclonedwithPCR・[Results]PSB07-21wascloselyrelatedtoRhodopseudomonassp・Theoptimumconditionofdegradingsyntheticcypermethrinandbipthenthrinby66.63%,43.25%and50.18%inaconcentrationof600mg/Latday15,respectively・Weclonedaputativegenewhichwas326bplongwith37.0%identicalto20G-Fe(11)oxygenasegene・WhencompensatinglowconcentrationFe(II)inPSBmediumwithsyntheticpyrethroids,thedegradationefficiencyofPSB07-21wasenhanced・[Conclusion]Thestrainhasthepotentialapplicationtosyntheticpyrethroidsbioremediation.作者::张松柏⑴：张德咏⑴2刘勇⑴：罗香文[2]?成飞雪⑵7罗源华⑵Author：SongbaiZhang[1DeyongZhang[1YongLiu[1XiangwcnLuo?FeixucCheng12'YuanhuaLuo[2]作者单位:屮南大学研究生院隆平分院，长沙,410125;湖南省植物保护研究所，长沙,410125湖南省植物保护研究所,长沙,410125刊名：微生物学报1STICPKUJournal：ACTAMICROBIOLOGICASINICA年,卷（期）:2009,49(11)分类号:Q814关键词：红假单胞菌PSB07-21旨菊酯类农药型降解酶基因为克隆旨Keywords：Rhodopseudomonassp.PSB07-21syntheticpyrethroidsdegreiding-enzymegeneclone机标分类号:X17S88在线出版日期:2010年1月8日基金项目:国家高技术研究发展计划（863计划）,国家科技支撑计划参考文献（25条）1.GeorgeN,KaiyanasundaramM.Chemistryofsyntheticpyrethroidinsectlcides-somerecenteidvances[J].JournalofScientific&IndustrialResearch,1994.933-945.2.SahaS,KavirajA.EffectofambienttemperatureanddaylightonthesurvivaloffreshwatercatfishHeteropneustesfoss订is(Bloch,1794)exposedtocypermethrin[J].EnvironmentalEngineeringScience,2009,(02):459-462.doi:10.1089/ees.2008.0047.3.MeekerJD,BarrDB,HauserR.Pyrethroidinsecticidemetabolitesarcassociatedwithserumhonnonelevelsinadultmen[J]・ReproductiveToxicology,2009,(02):155-160.doi:10.1016/j.reprotox.2008.12.012.4.PineMD,HineyJK,LeeB.ThePyrethroidpesticideesfenvaleratesuppressestheafternoonriseofluteinizinghormoneanddelayspubertyinfemalerats[J].EnvironmentalHealthPerspectives,2008,(09):1243-1247.5.RodriguezH,TamayoC,InestrozaJ.Cypermethrineffectsontheadultmiceseminalglands[J].EcotoxicologyandEnvironmentalSafety,2009,(02):658-662.6.UeyamaJ,KimataA,KeimijimeiM.Urinaryexcretionof3-phenoxybenzoicacidinmiddle-agedandelderlygeneralpopulationofJapan[J]•EnvironmentalResearch,2009,(02):175-180.doi:10.1016/j.envres.2008.09.006.7.AlmakkawyHK,MadboulyMD.PersistenceandaccumulationofsomeorganicinsectcidesinNilewalerandfish[J]•ResourcesConservationandRecycling,1999,(1/2):105-115.doi:10.1016/S0921-3449(98)00090-1.8.AntwiFB,PetersonRK.Toxicityof§-phenothrinandresmethrintonon-targetinsects[」]•PestManagementScience,2009,(03):300-305.doi:10.1002/ps.1688.9.GrantRJ,BettsWB.Biodegrachtionofthesyntheticpyrethroidcypermethrininusedsheepdip[J].LettersinAppliedMicrobiology,2003,(3):173-176.doi:10.1046/j.1472-765X.2003.01288.x.10.TallurPN,McgadiVB,NinnekerHZ.BiodegradationofcypcnncthrinbyMicrococcussp.strainCPNl[J].Biodegrndation,2008.77-82.doi:10.1007/s10532-007-9116-8.
