Isolation and characterization of mitochondrial dna from the endangered white-clawed crayfish austropotamobius pallipes pallipes, lereboullet, 1858

Abstract

Mitochondrial DNA (mtDNA) variation in the white-clawed crayfish, Austropotamobius pallipes pallipes, was examined by restriction endonuclease analysis of samples obtained from three geographical locations representing two very different habitats : the Crochatière and the Magot for brooks and the Pinail for ponds. This study used a new type of molecular marker in crayfish. The method of mtDNA extraction was not based on the clear lysate method or on ultracentrifugation and used no end-labelling detection. It is discussed according to the literature about marine crustaceans. MtDNA was digested with 6 endonucleases. The molecule size of the mtDNA of the white-clawed crayfish was approximately 17750 ±580 base pairs. Two restriction enzymes produced polymorphic digestion patterns, defining a total of three haplotypes (1-3). There was a shared haplotype 2 between individuals among the two brooks. The haplotype 3 was only found in individuals obtained from ponds of Pinail.