保护遗传学方法在生物多样性监测和评价领域的应用

本文是一篇生物多样性论文范文,关于生物多样性本科毕业论文,关于保护遗传学方法在生物多样性监测和评价领域的应用相关毕业论文参考文献格式范文。适合生物多样性及大学学报及自然科学方面的的大学硕士和本科毕业论文以及生物多样性相关开题报告范文和职称论文写作参考文献资料下载。

【摘 要】根据环境保护部文件《中国生物多样性保护战略和行动指南（2010-2030）》,遗传资源的持续丢失已经成为威胁中国生物多样性的三大问题之一,因此遗传多样性监测计划的重要性需要在将来的环境监测工作中得到体现.在两条环境保护部标准《区域生物多样性评价标准（HJ623-2011）》和《生物遗传资源采集技术规范（HJ628-2011）》开始贯彻实施后,遗传资源的鉴别和收集工作成为生物多样性评价项目中不可缺少的内容.本文综述了细胞学标记技术和DNA分子标记技术在植物品种鉴别研究中的前沿性应用,并在此基础上讨论了DNA标记技术在生物多样性监测与评价工作中大规模应用的可行性.

【关 键 词】生物多样性；细胞学标记；DNA分子标记

【Abstract】AccordingtotheChineseBiodiversityConservationStrategyandActionPlanning（2010-2030）,thecontinuouslossofgeicresourcesbeesoneofthreethornyissuesthreateningbiodiversityconservationinChina,whichhighlightsthesignificanceofgeicdiversitymonitoringplaninthefuture.AfterbothStandardfortheAssesentofRegionalBiodiversity（HJ623-2011）andRegulationfortheCollectionofGeicResources（HJ628-2011）eintoforce,identificationandcollectionofgeicresourcesbeesessentialinbiodiversityassesentprojects.ThisreviewsummarizesthefrontapplicationofbothcytologicalmarkerandDNAmolecularmarkertechniquestodistinguishplantvarieties,andconsequentlythefeasibilityoflarge-scaleapplicationofDNAmarkertechniqueonfuturebiodiversitymonitoringandassesentprojectsisdiscussed.

【Keywords】Biodiversity；Cytologicalmarker；DNAmolecularmarker

0Introduction

Asoneofthreelayersofbiodiversity,whichincludesecosystem,speciesandgeics,geicdiversityisthediversityofgeicfactorsthatdeterminethetraitsofanisandtheirbinations,sothatbeesthebasisofspeciesandecosystemdiversity[1].Itisinevitableforaspeciesofpoorgeicdiversitytomovetowardstheextinctioninnaturalselectionprocess[2].

AfteraseriesofenvironmentalpolicyhasbeenworkedoutbycentregovernmentofChina,suchasChineseBiodiversityConservationStrategyandActionPlanning（2010-2030）,StandardfortheAssesentofRegionalBiodiversity（HJ623-2011）andRegulationfortheCollectionofGeicResources（HJ628-2011）,itisessentialforenvironmentalengineerstoincludegeicdiversityinbiodiversitymonitoringandassesentprojects,andcollectionandidentificationofgeicresourcesinthenaturedefinitelybeesthefirststepofthiswork.Inpresent,identificationofplantvarietieainlyreliesonthebiologicaltraitsofplants[3],whicharesusceptibletoenvironmentalconditionsandtime-consumingwhenthosebiologicaltraitsareartificiallycultivatedandobservedinexperimentland[4].However,thedevelopmentofDNAmarkertechnologyprovidesaquickerandmoreaccuratesolutionforenvironmentalengineerstodistinguishdifferentsub-populationsofaplantspeciesinthenature,particularlywhenidentificationofeconomictraitsisnotessentialinbiodiversityassesentwork.ThisreviewsummarizesbothcytologicalmarkerandDNAmolecularmarkerforthedifferentiationofplantcultivarsinrecentyears.1CytologicalMarker

Duetoitshighstabilityandreproducibility,karyotypebeesoneoftheuniquechromosomeinformationtodistinguishdifferentspecies,populationsofthesamespeciesandtoidentifythehybrids.Karyotypeparameters,mainlyincludingtheabsolutelengthandrelativelengthofchromosome,armratio,centromereindex,chromosomeploidyandasymmetryindex,arefrequentlyanalyzedbybotaniststostudythevariationinchromosomenumberandstructurebetweenspecies,theoriginofspeciesandthegeicevolution[4].

1.1Traditionalsquashtechnique

Zhangetc[5]analyzedkaryotypeofthreeFritillarithunbergiicultivarsbasedontraditionalsquashtechnique.ThekaryotypeformulaofF.thunbergii（Xiaye,Kuanye,Duozi）variedamongthreevarieties,indicatingthefeasibilityofgeicidentificationofFritillarithunbergiicultivars.Thekaryotypeofallthevarietieswereclassifiedinto3Btype,andheterozygosityofhomologouschromosomewerefoundinbothF.thunbergii（Xiaye）andF.thunbergii（Duozi）.

ThekaryotypeofthreediploidoatspecieswasstudiedbyLiuetc[6]withapplicationoftraditionalsquashtechnique.BothkaryotypeformulaandasymmetryindexofAvenastrigosa,Avenahispanica,Avenabreviswerecalculatedforparison,revealingmoreadvancedevolutioninkaryotypeforA.strigosa,followedbyA.abrevisandA.hispanica.Threediploidoatspecieswereeffectivelydistinguishedbyabinationofbothkaryotypeformulaandasymmetryindex.

Thetraditionalslice-makingmethodwithmicrographtechnologywasadoptedbyDaietc[7]tostudythecytologybasiorcultivaridentificationofSecalecerealesubsp.segetale.ThreepopulationsofSecalecerealesubsp.segetale（89R4,89R14,89R60）andonevarietySecalecerealeL.（H36）wereselectedtoconductkaryotypeanalysis.Karyorypeformulae,asymmetryindexandasymmetricalkaryotypecoefficientwereprovidedandparedamongthesevarietiesinthisresearch,whichshowedrichdiversityinchromosomemorphology.

TraditionalsquashingmethodwasadoptedbyLiuetc[8]toanalyzethekaryotypeof7R.hybridacultivarsand5R.rugosacultivars.Accordingtotheresults,alltheR.hybridacultivarsweretetraloid（2n等于4x等于28）,exceptthatR.hybrida‘Elmshorn’wastriploid（2n等于3x等于21）,whileallthe5R.rugosacultivarswerediploid（2n等于2x等于14）.Anumberofkaryotypeparameters,includingkaryotypeformula,chromosomerelativelength,ratioofthelongestchromosometotheshortestoneinlength,armratio,asymmetryindexandcentromereindex,wereinterpretedasbiomarkeroridentificationofvarietiesandcorrespondinglythegeicdistancewasanalyzed,revealingthatdistinctdifferencesinbothkaryotypeandploidylevelsexistedbetweenR.hybridaandR.rugosacultivarsandR.rugosacultivarsappearedtobemoreadvancedinkaryotypeevolution.21cultivars’karyotypeofornamentalGinkgowasstudiedbyGaoetc[9]withearmethod.Thekaryotypeofallcultivarswasreportedtobeidentical,andtherelativelengthofchromosomevariedfrom4.31%to15.34%forthefemalecultivars,aswellas4.37%to17.12%forthemale.Forapproximately83.33%ofallthevarietiesinthisresearch,thearmratioofchromosomewasabove2：1,whichbelongedtoasymmetric3Btype.Clusteranalysiswasconductedonthebasisofkaryotypecalculation,showingthatthemeanarmratioorlengthratioofornamentalGinkgocultivarswassignificantlydifferentfromoriginalGinkgoBiloba,andconsequentlytheoriginality,evolutionandclassificationofthesecultivarswerediscussed.

Intotal6varietiesofHippophaeRhamnoidesL.wereselectedbyLietc[10]toanalyzekaryotypecharacteristicsofchromosomes,including4strainromRussiaand2strainromChina.Karyotypeformula,asymmetryindex,centromereindexandratioofthelongestchromosometotheshortestoneinlengthwereparedandcontrastedbetweenthesevarieties,providingthebasiortheidentificationandevolutionaryanalysisofHippophaeRhamnoidesL.varieties.Accordingtotheasymmetryindex,sixofthesecultivarswereclassifiedintomiddlecentromereorsub-middlecentromere,withkaryotypetypesas2Aor2B.

40typicalandstablevarietiesofChineselarge-floweredchrysanthemumwerechosentocarryoutcytologicalkaryotypeanalysiorinvestigationofgeicdifferences[11].1-4satellitechromosome（s）werereportedinapproximately35%ofthecultivars,withincreasingpossibilityofsatellitechromosomewhenchromosomenumberincreased.Th

本文是一篇生物多样性论文范文,关于生物多样性本科毕业论文,关于保护遗传学方法在生物多样性监测和评价领域的应用相关毕业论文参考文献格式范文。适合生物多样性及大学学报及自然科学方面的的大学硕士和本科毕业论文以及生物多样性相关开题报告范文和职称论文写作参考文献资料下载。

WildRosaspecies,whicharebroadlyfoundintheXinjiangUygurautonomousregionofChina,possesanyimportantunknowneconomictraits.Yuetc[12]collectedkaryologicaldatafrom13samplesofsevenwildRosataxa（R.berberifolia,twobotanicalvarietiesofR.spinosissima,R.platyacantha,R.beggeriana,R.acicularis,andR.laxa）,whichwereeasilydistinguishedbykaryotypeparametersofchromosomeploidy,asymmetryindex,centromereindex,anddistributionofrelativelengths.Thekaryologicaldataprovidedprehensivecytogeicresourcetoanalyzethetaxonomy,evolutionandspeciationinthegenusRosaaswellastoidentifysuitablecultivarorbreedingprograms.1.2Fluorescenceinsituhybridization（FISH）technique

Fluorescencebindingtechnologywithfluorescentdyes,whicharecapableofrevealingATorGCDNAsequencesonchromosomes,candistinguishdifferenttypesofheterochromatinonthechromosomes.Forexample,DAPI（4',6-diamino-2-pheny-lindoledihydrochloride）resultsintheappearanceofATrichregiononchromosomes,whereasCMA（ChromomycinA3）canrevealtheGCrichregion[13].Fluorescenceinsituhybridization（FISH）techniqueprovidestheaccuratemappinginformationofrDNAprobesonthechromosome,whichbeesthemoreeffectivemarkerstodistinguishchromosomesofplants[14].Sheetc[15]analyzedthemitoticmetaphasechromosomesofArachishypogaeaL.speciesbyusingabinationofDAPI+bandingtechnologyanddoublefluorescenceinsituhybridization（FISH）techniquewithboth5Sand45SrDNAprobes.Onthebasisofthechromosomemeasurements,DAPI+bandsandrDNAFISHsignals,thechromosomesofArachishypogaeaL.wereaccuratelypairedandarranged,leadingtoamolecularcytogeickaryotypeindetail.

However,DAPIbandingpatternsvariesbetweendifferentplantspecies.Xuetc[16]paredDAPIfluorescentbandingpatternsamongdifferentplantspecies,indicatingthatfluorescentbandswereobviouslyobservedinmaizeandpeanutspecies,followedbysesameandloofahwhoseDAPIbandswererelativelyweaker.However,noclearDAPIbandscouldbeidentifiedinsoybeanchromosomes.

2DNAMolecularMarker

DNAmolecularmarkertechnologieorplantvarietyidentificationmainlyincludeRFLP,RAPD,ISSR,AFLP,SNPandSSR.However,therankingofthesemolecularmarkertechniquesbasedonprehensiveeffectivenessisAFLP>SSR>RAPD>RFLP,whichhasbeeninternationallyrecognizedinthe92thASHSconference[17].ThisreviewsummarizestherecentdevelopmentofbothSSRandAFLPmarkertechnologyforvarietydifferentiation.

2.1SSRmarker

EST-SSRmolecularmarkertechniquewasconductedbyZhaoetc[18]toidentify12Chinesecabbagecultivars.Basedonexpressedsequencetags（ESTs）ofChinesecabbageinGenBank,30pairsofscreenedSSRprimersweredesignedandsynthesized,resultingin21pairsofEST-SSRprimerswhichwereeffectivelyamplified,butonly10pairsofEST-SSRprimerswerehighlypolymorphic.Accordingtotheidentificationresultsandthemappingdifference,10pairsofprimerswithhighpolymorphiweredesignedas2setsofmultiplexEST-SSRmarkerstodistinguishthese12Chinesecabbagevarieties,withsatiactorypolymorphicrateof88.9%and97.0%respectively,aswellashighpolymorphiinformationcontentof0.910%.Laietc[19]selected26inbredlinesand54testvarietieortheexaminationofdistinctness,uniformityandstability（DUS）ofthesevarietiesbyadoptingSSRmarkers.49pairsofSSRprimerswerescreenedfrom952pairsintotal,basedonthecriteriaofrichnessofpolymorphiinformationcontent（PIC）,theclearnessofPCRbandsandconvenienceofdifferentalleleidentification.49pairsofSSRprimersledto57lociwith311allelesidentifiedintotal.Theeragenumberofallelesperlocuswas5.5,rangingfrom2to13,withameanPICof0.53.Clusteranalysisshowedthatalltestvarietieswereclearlydistinguishedby49markerswhenthegeicsimilaritycoefficientwassetas0.93.

Inordertoproviderobustreferencefortheidentificationofbarleyvarietiesandoidcounterfeitandinferiorvarieties,Wangetc[20]selected29barleystandardvarietiesandgeicdiversitywasanalyzedbyDUStesting.28pairsofhighlypolymorphicSSRprimerswerechosen,leadingto125alleleeasuredintotal.Eachpairofpolymorphicprimersdetectedanerageof4.46alleles,withpolymorphiinformationcontent（PIC）varyingfrom0.81to0.25andaneragePICof0.62among28pairs.

Thespecificityandstabilityof123representativericevarietieswereanalyzedbyTianect[21]basedonSSRfingerprintingprofiles,andthevalueofSSRcoremarkerschoseninthisstudywasexamined.24pairsofprimersdetected138allelesintotal,with12locidetectedinsinglecultivarand21locisuccesullydistinguishingjaponicaandindicaricevarieties.Onthebasisofgeicsimilaritycoefficientsetas0.96fortheclassification,alltestedvarietiesshowedtheiruniquespecificitybyclusteranalysis,whichindicatedthat24pairsofSSRcoreprimerswasabletoeffectivelyidentify123varietiesofrice.

2.2AFLPmarker

SixpairsofAFLPprimerswithrichpolymorphiwerescreenedbyLietc[22]toconductfingerprintinganalysisontwoChinesecabbagesamples（label587and586）aswellasastandardsample.Euclideandistancescoefficientofeachsamplewasestimated,indicatingthatdistinctdifferencewaoundbetweenthesample587andstandardsample,withthepolymorphibandrateof31.7%.Consequentlyvariety587wasidentifiedasadifferentvarietyfromthestandardsample.Inparison,variety586showedconsistentPCRbandswiththestandardsample,whichwasconsequentlyidentifiedasthesamevarietyasthestandardsample.ThisresearchdemonstratedthatAFLPwascapableofprovidingreliabledifferentiationtechnologyforplantcultivars.Intotal14samplesofeightvarietiesandsixwildpopulationsofToxicodendronvernicifluumfromShaanxiwerechosenbyWeietc[23]forthedevelopmentofvarietyidentificationtechnique.BothmorphologicalandAFLPmolecularmarkerswereexaminedwith26morphologicalcharacterindexesand8AFLPprimers（EcoRⅠ+3/MseⅠ+3）.Multivariatestatisticanalysiswasconductedonmorphologicalmarkers,resultinginrincipleponentindex（PCI）.ThefistPCIincludedtheratioofpetalandanther,lengthtowidthofthefifthlobular,thelengthanddiameteroffilament；thesecondPCIcoveredthelengthofpoundleafandpetioleofpoundleaf,thenumbersofleaflet,thefifthlobular,andthetoplobular；andthethirdPCIw

本文是一篇生物多样性论文范文,关于生物多样性本科毕业论文,关于保护遗传学方法在生物多样性监测和评价领域的应用相关毕业论文参考文献格式范文。适合生物多样性及大学学报及自然科学方面的的大学硕士和本科毕业论文以及生物多样性相关开题报告范文和职称论文写作参考文献资料下载。

Wenetc[24]triedtodistinguish26jujubecultivarsand1sourjujubebyadoptingfluorescent-labeledAFLPmarkers.8AFLPprimerpairswerechosen,leadingto886AFLPmarkersidentifiedintotal.AmongtheseAFLPmarkers,112markerswereidentifiedasuniquebandorspecificvarieties,whereas60markersweredeletionbandorspecificvarieties,leadingtoeffectiveidentificationofjujubecultivars.

Songetc[25]chosen90cultivarsofChinesecabbagerom7differentproductionareas,anddevelopedfingerprintingtechniquebasedonAFLPmarkerortheidentification.Intotal20pairsofAFLPprimersweredesignedtoexaminethegeicpolymorphiofthesecultivars,andAFLPprimersvariedbroadlyintermsofdifferentiationcapacityofChinesecabbagevarieties.ThenumberofpolymorphicbandsthatweredetectedbyAFLPprimersdifferedfrom9to32.Abinationofprimers（E-ACA/M-CTG）resultedin71amplifiedbands,including32polymorphicbands,whicheffectivelydistinguishedallofthe90varieties.Inparison,thegeicpolymorphibetweenindividualsofthesamevarietywasalsoexaminedbyAFLPmarkertechnique.Twohybridcultivars（Beijingxin2andJingxiawang）ofChinesecabbagewereselectedand10individualswerechosenfromeachcultivar.TheAFLPbandsshowedconsistencebetweenindividualsofthesamevariety,exceptthatoneofBeijingxin2differedfromtheothers.2.3Capillaryelectrophoresiswithfluorescencedetection

Comparedwithpolyacrylamidegelelectrophoresisandsilverstainingtechnique,capillaryelectrophoresiswithfluorescencedetectionmethodioreautomatedandprogrammed.Thesystemsoftwareofcapillaryelectrophoresiswithfluorescencedetectionisabletocalibratethedifferencesbetweencapillaryelectrophoresis,andreducetheartificialandsystematicerrors,whichconsequentlyimprovesthestabilityandrepeatabilityofvarietyidentificationtests[26].Fengetc[3]screened58SSRprimerstoidentify14Poplarvarietiesbyapplicationofcapillaryelectrophoresiswithfluorescencedetection,whichincluded4varietiesofPopulusdeltoids,5varietiesofPopulusnigra（including3transgenicvarieties）and4hybridvarieties.Theresultsshowedthatthe4varietiesofP.deltoids,5varietiesofP.nigra,and4hybridvarietieswereeffectivelyidentifiedby4primers,5primers,and4primersrespectively,withsignificantdifferenceobservedattheSSRlocibetweenP.deltoidesandP.nigra.DifferentSSRgenotypeswerealsoidentifiedbetweenthetransgenicandnon-transgenicvarieties.

3ConclusionandImplicationforBiodiversityMonitoringandAssesent

InparisontotheDNAmolecularmarker,cytologicalmarkertechniquesresultinlesspolymorphiforthesub-populations’differentiationofaplantspecies,butobviouslyreducethecostofthiswork,oncebiodiversitymonitoringandassesentprojectsareimplementedatlargescale.Consequently,cytologicalmarkerwouldbemoresuitableasthemainsolutionforenvironmentalengineerstoconductgeicresourcecollectionwork,basedonwhichDNAmolecularmarkerwouldbeeaplementarysolution.Capillaryelectrophoresiswithfluorescencedetectionmethodcertainlyleadstohigheraccuracyandstabilityforidentificationtests.Nevertheless,therelativelycheaperfacilitiesrequiredbypolyacrylamidegelelectrophoresisandsilverstainingtechniquewouldbemoreacceptableinpractice,whichhasbeenadoptedbyrecentNationalStandardsincludingProtocolofPurityIdentificationforSoybeanVarietyusing-SSRMolecularMarkers（NY/T1788-2009）,aswellasGenuinenessandPurityVerificationofPotatoSeedTuber-SSRMolecularMarker（GB/T28660-2012）.

CollectionandstorageofsamplinglocationinformationaswellasphotosofplantmorphologicalcharactersareusuallynecessaryforthegeicresourcecollectionworkasindicatedbyRegulationfortheCollectionofGeicResources（HJ628-2011）,andGIStechnologyprovidesasupportivetoolforthecollectionandstorageofbothlocationinformationandfieldsamplingphotos[27]inthisprocess.【参考文献】

[1]李昂,葛颂.植物保护遗传学研究进展[J].生物多样性,2002（1）：61-71.

[2]C,A.J.,H.J.L.ConservationGeics,CaseHistorieromNature[M].Chapman&Hall,NewYork,1996.

[3]冯锦霞,等.利用荧光SSR标记鉴别杨树品种[J].林业科学,2011（6）：167-174.

[4]周延清,张改娜,杨清香.生物遗传标记与应用[M].化学工业出版社,2008.

[5]张彦南,等.浙贝母主要栽培品种类型花粉形态及染色体核型研究[J].中国中药杂志,2013（19）：3265-3270.

[6]刘伟,张宗文,吴斌.加拿大引进的二倍体燕麦种质的核型鉴定[J].植物遗传资源学报,2013（1）：141-145.

[7]代明,等.新疆杂草黑麦染色体核型分析[J].麦类作物学报,2013（3）：440-444.

[8]刘佳,等.7个月季和5个玫瑰品种的核型分析[J].西北农林科技大学学报：自然科学版,2013（5）：165-172.

[9]高进红,等.银杏观赏品种染色体核型分析[J].山东农业大学学报：自然科学版,2005（1）：19-24.

[10]李洪梅,等.核型分析技术在沙棘品种进化研究中的应用[J].济南大学学报：自然科学版,2013（1）：97-101.

[11]ZHANG,Y.,M.ZHU,S.DAI.Analysisofkaryotypediversityof40Chinesechrysanthemumcultivars[J].JournalofSystematicsandEvolution,2013,51（3）：335-352.

[12]Yu,C.,etal..KaryotypeAnalysisofWildRosaSpeciesinXinjiang,NorthwesternChina[J].JournaloftheAmericanSocietyforHorticulturalScience,2014.139（1）：39-47.

[13]T.,S.A..Chromosomebanding[M].London：UnwinHyman,1990.

[14]佘朝文,宋运淳.植物荧光原位杂交技术的发展及其在植物基因组分析中的应用[J].武汉植物学研究,2006（4）：365-376.

[15]佘朝文,张礼华,蒋向辉.花生的荧光显带和rDNA荧光原位杂交核型分析[J].作物学报,2012（4）：754-759.

[16]徐延浩,高伟,张文英.不同作物染色体DAPI荧光显带的研究[J].吉林农业科学,2013（2）：27-28+51.

[17]郑成木.植物分子标记原理与方法[M].湖南科学技术出版社,2003.

[18]赵新,等.复合EST-SSR标记在大白菜品种鉴定中的应用[J].生物技术通报,2013（1）：107-110.

[19]赖运平,等.利用SSR标记筛选DUS测试中甘蓝型油菜近似品种[J].分子植物育种,2013（2）：174-184.

[20]王艳平,等.大麦DUS测试标准品种的遗传多样性分析及指纹图谱的构建[J].麦类作物学报,2013（2）：273-278.

[21]田大刚,等.123份水稻重要品种的SSR核心标记指纹分析[J].分子植物育种,2013（1）：20-29.

[22]李丽,郑晓鹰.AFLP分子标记应用于白菜品种鉴定[J].分子植物育种,2006（5）：685-689.

[23]魏朔南,等.应用植物形态学和AFLP分子标记鉴别陕西漆树品种[J].西北植物学报,2010（4）：665-671.

[24]文亚峰,何钢,张江.枣优良品种分子鉴别系统的开发[J].中南林业科技大学学报,2007（6）：119-121.

[25]宋顺华,郑晓鹰.AFLP分子标记鉴别大白菜品种[J].分子植物育种,2005（3）：381-387.

[26]易红梅,等.玉米品种SSR标记毛细管电泳荧光检测法与变性PAGE银染检测法的比较研究[J].华北农学报,2006（5）.

[27]H,L.Y.,GISAPPLICATIONINRESEARCHOFWILDLIFEHABITATCHANGE――AcasestudyoftheGiantPandainWolongNatureReserve：51-60[Z].

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