Mitochondrial DNA length variation in DicentrurchusZubrux is the outcome of the simultaneous variation in the copy number of two tandem repeat arrays located in the D-loop region. The repeated sequences, named R17 and R48, are 17 and 48 bp long, respectively. On the basis of 209 individuals collected in eight Mediterranean localities, length variation and its partitioning between the two repeat arrays have been characterized and quantified by means of frequency distributions of repeat copy number and genetic diversity estimates. The median copy numbers are 4 and 11 for the R17 and R48 repeat arrays, respectively. For both repeat arrays, continuity in step variation and the occurrence of both frequent and rare arrangements was observed. More than 50% of the individuals are heteroplasmic for up to four variants. Within populations, heteroplasmy ranges between 33.3% and 70.0% of the individuals. A large proportion of the total gene diversity (29.1%) occurs within individuals and the greatest proportion (66.6%) is found between individuals within populations, while only 4.3% is due to variation between populations. Although the interpopulation component of the diversity is rather low, the significant differentiation of the two Eastern populations from all the other Western samples suggests the occurrence of a phylogeographic component of the genetic divergence between the Mediterranean populations, which is only recognizable on a large geographic scale.

Three specific satellite DNA families can be detected in the genome of the cave cricket Dolichopoda schiavazzii. The pDoP102 and the pDsPv400 families are species specific for D. schiavazzii; the pDoP500 family is probably present in all Dolichopoda species. The three satellite DNA families were characterized from individuals of three isolated populations of D. schiavazzii with respect to nucleotide sequence, sequence complexity, sequence variability, and copy number. This unique data set on satellite DNAs of D. schiavazzii seems to allow one to test the significance of theoretical approaches to the mode of evolution of noncoding, tandemly arranged satellite DNA. At least for satellite DNAs of D. schiavazzii two clear trends were observed: (1) sequence variability increases with copy number and (2) the repeat length decreases with copy number. The first trend is in good agreement with the theory but the second is not. Thus, a revision of the models is proposed.

BackgroundNon-coding satellite DNA (satDNA) usually has a high turn-over rate frequently leading to species specific patterns. However, some satDNA families evolve more slowly and can be found in several related species. Here, we analyzed the mode of evolution of the pDo500 satDNA family of Dolichopoda cave crickets. In addition, we discuss the potential of slowly evolving satDNAs as phylogenetic markers.ResultsWe sequenced 199 genomic or PCR amplified satDNA repeats of the pDo500 family from 12 Dolichopoda species. For the 38 populations under study, 39 pDo500 consensus sequences were deduced. Phylogenetic analyses using Bayesian, Maximum Parsimony, and Maximum Likelihood approaches yielded largely congruent tree topologies. The vast majority of pDo500 sequences grouped according to species designation. Scatter plots and statistical tests revealed a significant correlation between genetic distances for satDNA and mitochondrial DNA. Sliding window analyses showed species specific patterns of variable and conserved regions. The evolutionary rate of the pDo500 satDNA was estimated to be 1.63-1.78% per lineage per million years.ConclusionsThe pDo500 satDNA evolves gradually at a rate that is only slightly faster than previously published rates of insect mitochondrial COI sequences. The pDo500 phylogeny was basically congruent with the previously published mtDNA phylogenies. Accordingly, the slowly evolving pDo500 satDNA family is indeed informative as a phylogenetic marker.

The satellite DNA family pDoP102 is species specific for the cave cricket Dolichopoda schiavazzii, an endemic species of mainland and insular Tuscany. It consists of numerous tandemly arranged repeats, 102 bp in length, and evolved most probably after cladogenesis of D. schiavazzii from the D. baccettii-aegilion group within the last 2.3 +/- 0.8 million years. A sequence comparison of 31 clones (53 repetition units) from three isolated populations reveals a very high degree of sequence homogeneity within the species with no evidence for any specific population features. This appears to be in contrast to the results of allozyme analyses which account for a relatively old evolutionary divergence of the Elba island population from the mainland ones. Since the assumption of actual gene flow and recent colonization is rejected, the observed sequence homogeneity is hypothesized to be maintained by recombination processes preventing fixation of newly introduced mutations on pDoP102 sequence clusters.

Dolichopoda cave crickets are found in caves in the northern Mediterranean region from the Pyrenees in the west to the Caucasus in the east. In this study we analysed the phylogeny within the genus Dolichopoda using parts of the mitochondrial cytochrome oxidase I and 16S ribosomal genes, and explored biogeographic patterns through a dispersal-vicariance analysis (DIVA). Phylogenetic analyses grouped the 15 species into the four geographically restricted main lineages corresponding to the Caucasus, Greece, the Pyrenees and Italy, respectively. The species occur largely in allopatry. The Caucasian and Greek species were basal in the phylogeny, as was the clade including the nine Italian species, which grouped into two major lineages, one mainly including species from western Italian coastal regions and islands, and the other including species with a predominantly inland distribution. Thus it seems likely that there have been two main immigrations into Italy followed by multiple consecutive speciation events. The DIVA analysis supported the assumption of an east-west migration route, and indicated that there have been four major dispersal events. Since the statistical support for the basal node connecting D. remyi and D. hussoni with the west Mediterranean species is low, alternative interpretations for the colonization of the Mediterranean, namely parallel colonization of the main areas of the current Dolichopoda distribution, i.e. the Caucasus, Greece, Italy, and the Pyrenees is also possible. Particular emphasis was put on the D. geniculata-laetitiae species complex. D. geniculata included several recently diverged lineages and constitutes a paraphyletic species complex, also embracing the closely related D. laetitiae.