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1、瓜实蝇致病菌粘质沙雷氏菌的分离与鉴定海南大学毕业论文(设计)题目:瓜实蝇致病菌粘质沙雷氏菌的分离与鉴定学号:姓名:年级:2010级学院:环境与植物保护学院系别:植物保护专业:植物保护(农药方向)指导教师:完成日期:2014年5月1日III瓜实蝇致病菌粘质沙雷氏菌的分离与鉴定摘要从瓜实蝇虫尸上分离出的一株产紫红色色素的细菌BC-hn212,染病死亡后的瓜实蝇成虫腹部明显发红。采用形态学、生理生化方法对其进行初步鉴定;并提取基因组DNA,16SrDNA基因序列分析。该菌株菌落圆形隆起,光滑粘稠,有轻微异味;24h后菌落颜色加深、气味变为恶臭,显微镜下观察为球形或杆
2、状菌;革兰氏染色阴性,甲基红为阳性,V.P阳性,淀粉水解实验阳性,果聚糖形成阴性,H2S产生阴性,明胶水解阳性,过氧化氢酶阳性,氧化酶阴性。根据形态学于生理生化方法的鉴定结果初步判定为沙雷氏菌属。根据16SrDNA基因序列分析结果,可以确定此株菌株为粘质沙雷氏菌。本实验初步证明粘质沙雷氏菌对于瓜实蝇具有较强致病能力,为进一步研究粘质沙雷氏菌对于昆虫存在致病性提供依据。关键词:瓜实蝇;粘质沙雷氏菌;分离与鉴定III瓜实蝇致病菌粘质沙雷氏菌的分离与鉴定AbstractAstrainisolatedfromthepurplepigmentbacteria(2-1-1
3、),afterexpandingtrainingforhealthmelonfly3to5daysafterdyingbacteria,melonflyabdominalbecameredandmelonflydied.Thentoisolatethebacteria,themorphological,physiologicalandbiochemicalmethodcarriesonthepreliminaryappraisal;ExtractionofgenomicDNA,using16srRNAuniversalprimers,PCRamplificat
4、ionof16srDNAgenefragments,throughmultiplesequencealignmentandhomologyanalysis,constructingphylogenetictrees,serratiabacteriahasthehighesthomology.Thecircularupliftmutantsweresmoothviscousandslightodor;After24hthecolonycolordeepened,anditsmelllikeamess.UsingMicroscopetoobserve,itlook
5、edsphericalorrod-shapedbacteria.Gramwasnegative,andmethylredwaspositive,andValerieplamewaspositive,andstarchhydrolysisexperimentsispositive,andfructanformationwasnegative,andH2Sproducednegative,andgelatinhydrolysatewaspositive,andcatalasewaspositive,andoxidasewasnegative.Accordingto
6、theresultofmorphologyinidentificationofphysiologicalandbiochemicalmethods,wedetermineinitialserratia.Thenweuse16SrDNAgenesequenceanalysis,.Accordingtothecomparedresults,wecertainthatthisstrainofbacteriaisserratiamarcescens.Theexperimentpreliminaryprovesthatserratiamarcescenshasstron
7、gabilityofpathogenicformelonfly,forthefurtherstudyofserratiamarcescens,itprovidesevidencetoprovethatserratiamarcescensarepathogenic.Keywords:Serratiamarcescens;Bactrocera,cucuribitae;IsolationandidentificationIII瓜实蝇致病菌粘质沙雷氏菌的分离与鉴定目录1前言................................................
8、.............11.1瓜实