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1、实用海南大学毕业论文(设计)题目:瓜实蝇致病菌粘质沙雷氏菌的分离与鉴定学号:20100304310059姓名:路冰斌年级: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:杨叶教授文档实用完成日期:2014年5月1日文档实用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
2、微镜下观察为球形或杆状菌;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;分离与鉴定文档实用AbstractAstrainisolatedfromthepurplepigmentbac
3、teria(2-1-1),afterexpandingtrainingforhealthmelonfly3to5daysafterdyingbacteria,melonflyabdominalbecameredandmelonflydied.Thentoisolatethebacteria,themorphological,physiologicalandbiochemicalmethodcarriesonthepreliminaryappraisal;ExtractionofgenomicDNA,using16srRNAu
4、niversalprimers,PCRamplificationof16srDNAgenefragments,throughmultiplesequencealignmentandhomologyanalysis,constructingphylogenetictrees,serratiabacteriahasthehighesthomology.Thecircularupliftmutantsweresmoothviscousandslightodor;After24hthecolonycolordeepened,andi
5、tsmelllikeamess.UsingMicroscopetoobserve,itlookedsphericalorrod-shapedbacteria.Gramwasnegative,andmethylredwaspositive,andValerieplamewaspositive,andstarchhydrolysisexperimentsispositive,andfructanformationwasnegative,andH2Sproduced文档实用negative,andgelatinhydrolysat
6、ewaspositive,andcatalasewaspositive,andoxidasewasnegative.Accordingtotheresultofmorphologyinidentificationofphysiologicalandbiochemicalmethods,wedetermineinitialserratia.Thenweuse16SrDNAgenesequenceanalysis,.Accordingtothecomparedresults,wecertainthatthisstrainofba
7、cteriaisserratiamarcescens.Theexperimentpreliminaryprovesthatserratiamarcescenshasstrongabilityofpathogenicformelonfly,forthefurtherstudyofserratiamarcescens,itprovidesevidencetoprovethatserratiamarcescensarepathogenic.Keywords:Serratiamarcescens;Bactrocera,cucurib
8、itae;Isolationandidentification文档实用目录1前言.............................................................11.1瓜实蝇为害的基本情况..............................