IntegrativetaxonomyandphylogenyofMiraciidae(Copepoda:Harpacticoida)fromNorthwesternPacificregion

Abstract

놀람장수노벌레과(Miraciidae)는갈고리노벌레목내에서큰비중을차지하는과로서다양한환경에서발견되는요각류그룹이다.본연구에서는북태평양조간대및조하대에서식하는놀람장수노벌레과종들을모래린싱,간극수필터링,시빙,다이빙,배양,네트채집등다양한방법으로확보하였다.문헌참고와현미경관찰을바탕으로한형태분석을우선적으로진행했고18SribosomalRNA,mitochondrialcytochromeoxidaseI분자바코딩을실시하거나단백질질량분석을통합적으로수행하였다.분자계통학적분석을통하여놀람장수노벌레과내하위분류군들간의관계를재정립하고자하였으며,단백질질량분석기법을도입하여갈고리노벌레목요각류종동정간편화가능성을확인하고자하였다.본연구의분자계통학적분석을통하여놀람장수노벌레과는단계통을이루는것을확인하였다.반면3아과중Diosaccinae는단계통을이루지않는것으로확인되었다.또한Sarsamphiascus내에서형태적특징들을기반으로구분되어있던그룹들이단일속으로유지될수있는가에대하여,형태적특징및분자계통학적분석을통합적으로수행하여2개의그룹을기존속에서분리시키고,신속으로제안하였다.이러한과정에서국내및팔라우연안에서서식하는놀람장수노벌레과2신종기재,국내1종수컷형태기재,국내1종의형태재기재및속재배치를진행했다.더불어단백질질량분석기법을요각류종동정에적용하여단백질질량패턴이종구분을위한바코드의역할을수행할수있다는것을확인하였다.이방법을단독으로사용하여계통학적유연관계를분석하는데에는아직추가연구가필요하다고결론지었지만,기존분자바코딩방법에비하여간편하고경제적이며,crypticspecies구분에도적용가능할정도로정확도가높다는점에서향후활용가치가높은요각류종동정기법이될수있겠다는기대가된다.이처럼본학위논문에서는놀람장수노벌레과요각류들의표현형(형태,단백질)과유전형(유전자단편서열)을통합적으로분석하였다.|MiraciidaeDana,1846isalargefamilyofcopepodswithintheorderHarpacticoida,SarsG.O.,1903foundinvariousenvironments.Inthisstudy,miraciidspeciesinthesubtidalandintertidalzoneoftheNorthPacificwereobtainedbyvariousmethodssuchassandrinsing,interstitialwaterfiltering,sieving,diving,culture,andnettowing. Morphologicalanalysisbasedonliteraturereferenceandmicroscopicobservationwasperformed.Inaddition,18SribosomalRNA(18SrRNA)andmitochondrialcytochromeoxidaseI(mtCOI)molecularbarcodingwereconducted,orproteinmassspectrometrywereperformedintegrally. Basedonmolecularphylogeneticanalysis,therelationshipbetweensubfamiliesorgenerainMiraciidaewasredefined.Furthermore,theproteinmassspectrometrytechnique(MALDI-TOFMS)couldsuccessfullybeusedtosimplifyidentificationofthecopepodspecies. Inthemolecularphylogeneticanalysisofthisstudy,Miraciidaeformedamonophyleticgroup.Incontrast,ofitsthreesubfamilies,onlyDiosaccinaeSarsG.O.,1906wasnotmonophyletic.Inaddition,itwasdeterminedwhethergroupsclassifiedbasedonmorphologicalcharacteristicswithinSarsamphiascusHuys,2009canbemaintainedasasinglegenus,reviewofmorphologicalcharacteristicsandmolecularphylogeneticanalysiswereperformedintegrally.Asaresult,twogroupswereseparatedfromtheexistinggenusandproposedasnewgenera.Inthisprocess,descriptionoftwonewmiraciidspeciesinhabitingKoreaandthecoastofPalauweredescribed.AdescriptionofmaleformofonespeciesinKorea,andthemorphologicalredescriptionandrelocationofthegenusofonespeciesinKoreawerecarriedout. Inaddition,thepotentialapplicationofMALDI-TOFMStotheharpacticoidspeciesidentificationwasinvestigatedbyusingtheproteinmassspectrapatternasabarcode.Althoughtheuseofthismethodalonetoanalyzethephylogeneticrelationshiprequiresadditionalresearch,itprovedbothconvenientandeconomicalcomparedtotheexistingmolecularbarcodingmethod,withaccuracylevelssufficientforcrypticspeciesclassification.Therefore,thistoolcanpotentiallybeusedasaspeciesidentificationtechniqueforcopepods.Inthisdissertation,thephenotyperepresentedbymorphologyofMiraciidaecopepods,theproteinmassspectra,andthegenotyperepresentedbythegenefragmentsequencewerecomprehensivelyanalyzed.