Phylogeny of Austropotamobius pallipes (Lereboullet, 1858) based on the analysis of 16S and COI mitochondrial DNA markers

Abstract

Filogenetski odnosi unutar populacija slatkovodnih rakova vrste Austropotamobius pallipes (Lereboullet, 1858) još nisu u potpunosti razjašnjeni. Novija istraživanja ukazuju na postojanje vrste-kompleksa, odnosno postojanje dva ili više taksona u okviru sadašnje vrste A. pallipes koji bi se u budućnosti mogli zasebno izdvojiti na razinu vrste. Cilj ovog istraživanja je dokazati ovu hipotezu i odrediti filogenetski status jedinki uzorkovanih na teritoriju Hrvatske. U ovom diplomskom radu istražuju se filogenetski odnosi 76 jedinki vrste Austropotamobius pallipes uzorkovanih na 22 lokacije u Hrvatskoj, Bosni i Hercegovini i Austriji. Istraživanje je provedeno na temelju varijabilnosti u DNA sekvencama mitohondrijskih gena za 16S rRNA i COI. Genske sekvence ekstrahirane iz uzorkovanih jedinki uspoređene su sa homolognim sekvencama korištenim u ranijim istraživanjima na ovim životinjama širom Europe. Za analizu rezultata primijenjeni su statistički filogenetski programi koji se baziraju na stanjima karaktera i matrici genske udaljenosti. Dobiveni rezultati potvrdili su pretpostavku da se unutar kompleksa vrste A. pallipes nalaze dva taksona koji odgovaraju razini vrste te nekoliko taksona koji odgovaraju razini podvrste; omogućili su bolje razumijevanje filogenetskih odnosa između populacija rakova u Hrvatskoj i populacija u ostatku Europe te su omogućili određivanje vremena odvajanja pojedinih populacija na istraženom području jugoistočnog Balkanskog poluotoka.Phylogenetic relations within populations of Austropotamobius pallipes (Lereboullet, 1858) are yet not fully understood. Recent research reveals a possible existence of speciescomplex, that is, a multiple taxa existence within a single species that is today known as Austropotamobius pallipes. The aim of this study is to verify that hypothesis and if necessary suggest a differentiation of the species-complex in to two separate species. The second intention is to put the Croatian crayfish in to the right phylogenetic context. Phylogenetic relations between 76 A. pallipes individuals, sampled from 22 localities in Croatia, Bosnia and Herzegovina and Austria were studied in this graduation thesis. The analysis of A. pallipes populations is based on DNA sequence variations of mitochondrial genes for 16S rRNA and COI. The extracted gene sequences were compared with homologues sequences that were used in other studies of this species all over Europe. Phylogenetic studies were carried out using statistical phylogenetic analysis by means of different character state and distance matrix methods. The results of this research showed that the species-complex Austropotamobius pallipes consists of two distinct species and a few subspecies. Also they are a contribution to a better understanding of relations between Croatian and European populations and they specify the time of divergence within population in the Southeast Balkan Peninsula

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