一株好氧反硝化细菌的分离鉴定及反硝化能力

第28卷第7期2008年7月环境科ActaScientiae学学报CircumstantiaeVo1．28．No．7Ju1．，2008杨基先，高珊珊，马放，等．2008．一株好氧反硝化细菌的分离鉴定及反硝化能力[J]．环境科学学报，28(7)：1302—1307YangJX，GaoSS，MaF，eta1．2008．Identificationandphylogeneticanalysisofanisolatedaerobicdenitrifier[J]．ActaScientiaeCircumstantiae，28(7)：1302—1307一株好氧反硝化细菌的分离鉴定及反硝化能力杨基先，高珊珊，马放，苏俊峰，王强哈尔滨工业大学城市水资源与水环境国家重点实验室，哈尔滨150090收稿日期：2007—05—31录用日期：2008—04—24摘要：利用驯化活性污泥的方法筛选出一株好氧反硝化细菌c3，该菌在碳源充足且溶解氧为(4．0±1．0)mg·L条件下，24h内对NO2-一N积累量为l2．03mg·L～，对NO3一N的去除率为91．09％．扫描电镜观察其形态特征为(0．4—0．6)p．m×(1．2—2．0)p．m的短杆菌．16SrDNA序列分析表明．该菌与Pseudomonasstutzeri的相似性为100％．利用MEGA软件对该菌株与已报道的相关微生物进行系统发育分析，并克隆出与好氧反硝化相关的周质硝酸盐还原酶的亚基napA基因，初步判定筛选菌株具有周质硝酸盐还原酶．关键词：好氧反硝化细菌；系统发育分析；周质硝酸盐还原酶文章编号：0253—2468(2008)07—1302—06中图分类号：X172文献标识码：AIdentificationandphylogeneticanalysisofanisolatedaerobicdenitrifierYANGJixian，GAOShanshan，MAFang，SUJunfeng，WANGQiangStateKeyLaboratoryofUrbanWaterResourceandEnvironment，HarbinInstituteofTechnology，Harbin150090Received31May2007；accepted24April2008Abstract：BydomesticatingactivatedsludgeinaSBRreactorwithalternationofanaerobic~aerobicprocesses，thebacteriumPseudomonasstutzeristrainG3wasisolated，whichiscapableofaerobicdenitrificationwithsodiumsucciniteassolecarbonsource．Whentheconcentrationofdissolvedoxygenwas(4．0±1．0)mg·L～，thenitriteaccumulationandtheremovalrateofnitratebystrainG3were12．03mg·L～and91．09％in24hours，respectively；whenC／N：3theremovalrateofnitratebythebacteriumculturedat30％for28hwas72．91％．Themorphologyofthebacteriumwasabrevibacilluswithaveragesizeof(0．4～0．6)p．m×(1．2～2．0)p．masmeasuredbyscanningelectronmicroscopy．Phylogeneticanalysisbasedonpartial16SrDNAandperformedbyMEGAshowedthatG3has100％sequencesimilaritywithPseudomonasstutzeri．Thephylogenetictreeofthestrainanditsneighboringdenitrifiersisgiven．ItspartialnapAgenewascloned，andshowntobeasubunitofperiplasmicnitratereductase(NAR)，whichisconsideredtoberelatedtoaerobicdenitrification．Keywords：aerobicdenitrifyingbacteria；phylogeneticanalysis；periplasmienitratereduetase1引言(Introduction)近年来，国内外的不少研究和报道已能充分证明好氧反硝化菌的存在