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分子性别鉴定引物的优化及在雀形目鸟类中的应用(英文).docx

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分子性别鉴定引物的优化及在雀形目鸟类中的应用(英文) OptimizationofMolecularSexingPrimersandtheirApplicationinPasserineBirds Introduction: MolecularsexingbasedonPCRamplificationofsex-specificDNAregionshasbecomeacommontoolinavianresearch,enablingnon-invasivesexingofbirds,whichisparticularlyimportantforspecieswithcrypticsexualdimorphism.Inthisstudy,weaimedtooptimizemolecularsexingprimersforpasserinebirdsandapplythemtoinvestigatesexratiosandsexualdimorphisminasubsetofPasseriformes. Methods: Weamplifiedandsequencedafewsex-linkedgenesandidentifiedpotentialsex-specificregionsinZandWchromosomesinafewpasserinespecies.Basedonthesesequences,wedesigned2setsofPCRprimerstargetingZ-specificandW-specificregions,respectively.WetestedthespecificityandefficiencyoftheseprimersonasmallsubsetofgenomicDNAsamplesextractedfromknown-sexbirdsofvariouspasserinefamilies. WethenappliedtheseprimerstoanalyzeDNAsamplesfrom141passerinebirdscollectedfromdifferenthabitatsandrepresenting51speciesin5families(Paridae,Sittidae,Regulidae,Certhiidae,Fringillidae).WeassignedthesexofeachindividualbasedonPCRproductswiththeexpectedsizeofZ-orW-specificregions.Wecomparedthesexratiosandquantifiedsexualsizedimorphism(SSD)ofdifferentspeciesandfamiliesbasedonbodymassandtarsuslengthmeasurements. Results: OuroptimizedPCRprimersshowedhighspecificityandsensitivityforsexingpasserinebirds,withclearseparationofZ-andW-specificPCRproductsonagarosegels.Weobtainedrobustresultsforsexingall141birds,withamale-biasedsexratioof67.4%(95malesand46females)acrossallspeciesandfamilies.However,wefoundsignificantvariationsinsexratiosamongdifferentspeciesandfamilies,rangingfrom100%male(1species)to92.3%female(1species).TheSSDpatternsalsovariedgreatlyamongfamiliesandspecies.Insomespecies,femaleswerelargerthanmales,whileinotherspecies,maleswerelargerortherewasnosignificantdifferenceinbodysizebetweenthesexes. Discussion: Ourresultsdemonstratetheeffectivenessofouroptimizedsexingprimersinenablingaccurateandnon-invasivesexingofpasserinebirds.Themale-biasedsexratioinoursam