Genetic and Morphological Systematic Studies on the Crayfish Austropotamobius pallipes (Decapoda: Astacidae)

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Abstract
Morphological criteria and mitochondrial DNA (mtDNA) were analyzed to examine the taxonomic and systematic relationships of the endangered crayfish species Austropotamobius pallipes. Morphological data confirmed the difficulty of differentiating the 3 subspecies: A. p. pallipes, A. p. italicus, and A. p. lusitanicus. MtDNA analysis by RFLP using 6 endonucleases revealed 5 haplotypes and allowed unequivocal identification of the subspecies. The nucleotide sequence divergence among haplotypes characteristic of each subspecies ranged from 10.93-13.6% and may be explained by an ancient geographical separation of each subspecies. This distribution of genetic variation agrees with the presumption of 3 refugial regions during the glaciation periods, possibly the Iberian Peninsula, southern France, and the Balkans. UPGMA analysis from nucleotide distances placed A. p. italicus and A. p. lusitanicus in a cluster separate from A. p. pallipes, suggesting that A. p. italicus and A. p. lusitanicus are more closely related.

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