海南花梨木dna条形码序列测定-毕业论文

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1、毕业论文（设计）题冃：海南花梨木DNA条形码序列测定学号：姓名：年级：学院：系别：专业：指导教师：完成日期：摘要DNA条形码作为生命体所固有的遗传物质，客观存在于细胞内的物质，不受人为因素干扰，能准确的反映出物种最基本的信息。为了验证国际生命条形码联盟推荐的MatK,tmH・psbA,ITS2三个候选条形码，在珍贵木材降香黄檀(DalbergiaodoriferaT.Chen)又名花梨木DNA条形码测定屮的冇效性。从基地采摘的降香黄檀叶片中捉取DNA,用特异性引物进行PCR扩增，得到三条候选条形码的目的片段，扩增成功的DNA片段可用于进一步测序分析。以PC

2、R扩增成功率和测序成功率初步评价候选DNA条形码片段，评选出最优的条形码片段，为今后利用DNA条形码鉴定木材和构建木材分子条形码数据库提供理论依据。实验结果：只有ITS2的条形码片段测序成功，得到对应的序列。通过NCBI查询黄檀屈英他物种的ITS2序列，得到：降香黄檀的ITS2分子序列与黄檀屈植物均不同，其中与越南黄檀羌异最小，其次是多裂黄檀。多重比对后构建该序列的进化树结果说明降香黄檀与越南黄檀、多裂黃檀有着较近的亲缘关系，但乂不属于同一植物类群，且三者有着稳定的遗传差异性。建议把ITS2序列作为鉴定降香黄檀的候选条形码序列。关键词：降香黄檀；DNA条形

3、码；ITS2；分子鉴定AbstractDNAbarcodeasthegeneticmatcrialinlifeinherentthatobjectivelyexistedinthecell,withoutartificialinterference.Itcouldaccuratelyreflectthemostessentialinformationforspecies.InordertoverifythreecandidatebarcodesoftheMatK,tmH-psbAandITS2recommendedbyCBOLandtheeffective

4、nessinDalbergiaodorifera'sDNAbarcode.ExtractedDNAofitsleavesfromthebase,usingspecificprimersforPCRamplication.Thepurposeofthethreecandidatesfrombarcodefragmentswereobtainedandthetargetedsegmentwasamplifiedsuccessfullytobeusedforsequencinganalysis.PCRamplificationandsequencingsucce

5、ssratecouldbeusedforthepreliminaryevaluationthebarcodefragmentsofcandidateDNA,Selectedthebestbarcodefragment,whichprovidedtheoreticalbasisforusingDNAidentificationofwoodandwoodconstructionmolecule.Theresultswereasfollows:onlyITS2codedfragmentwassuccessfullysequenced・andthecorrespo

6、ndingsequencewasgained・BysearchedITS2ofotherspecieswhichbelongtoDalbergiafromNCBI.Wegottheconsionconclusionthat:ITS2ofDalbergiaodoriferaweredifferent,withDalbergiatonkinensisminimum,followedbyDalbergiarimosaMultiplecomparisonafterbuildingthesequenceoftheevolutionarytreeresultshowe

7、dthatDalbergiaodoriferaDalbergiatonkinensisandDalbergiarimosahadthecloserelationship.Thoughtheyarenotbelongtothesamegroupofplants,andtheyhaveastablegeneticdiversity.SoITS2sequencecanbeucsdtoidentifiedcandidatebarcodeofDalbergia.Keyvrords:DalbergiaodoriferaT.Chen;DNAbarcoding;ITS2

8、;molecularidentificationIT目录1前言11