Teresa Capriglione

Evolution of sex-chromosomes in lacertid lizards

The occurrence and form of sex chromosomes were investigated with the aid of C-banding and 4′-6-diamidino-2-phenylindole (DAPI) staining in 13 species of lacertid lizards. The results obtained show the presence in five species of a female heterogamety in which the two sex chromosomes have the same shape and size, but the W differs from the Z in being almost entirely heterochromatic. This condition is clearly similar to that found in some snakes and considered to be an early stage of differentiation of sex chromosomes by Singh et al. (1976, 1980). A more evolved condition may be that found in three other species in which the W is distinctly smaller than the Z. A third situation is that found in all Podarcis species which, even though they are considered to be among the more evolved species in the family, possess two sex chromosomes that are indistinguishable. In general, the situation in lacertids may be compatible with the hypothesis of sex chromosome evolution put forward by Singh et al. (1976, 1980). However a differentiation mechanism of this kind does not seem to be well established in lacertids, and is probably not the only mechanism that is in operation in this family.

Mitochondrial DNA variation in the caramote prawn Penaeus (Melicertus) kerathurus across a transition zone in the Mediterranean Sea

In this study we analysed mitochondrial DNA variation in Penaeus kerathurus prawns collected from seven locations along a transect across the Siculo–Tunisian region in order to verify if any population structuring exists over a limited geographical scale and to delineate the putative transition zone with sufficient accuracy. Partial DNA sequences of COI and 16S genes were analysed. In contrast to the highly conservative 16S gene, the COI sequences exhibited sufficient diversity for population analysis. The COI gene revealed low levels of haplotype and nucleotide diversities. The size of the annual landings of this commercial species suggests large population sizes. Hence, the low genetic diversity detected in this study could indicate a possible reduction in effective population sizes in the past. We detected significant genetic differentiation between eastern and western populations likely due to restricted gene flow across the Siculo–Tunisian boundary. We discuss the different evolutionary forces that may have shaped the genetic variation and suggest that the genetic divide is probably maintained by present-day dispersal limitation.

Characterization of sex determination and sex differentiation genes in Latimeria.

Genes involved in sex determination and differentiation have been identified in mice, humans, chickens, reptiles, amphibians and teleost fishes. However, little is known of their functional conservation, and it is unclear whether there is a common set of genes shared by all vertebrates. Coelacanths, basal Sarcopterygians and unique “living fossils”, could help establish an inventory of the ancestral genes involved in these important developmental processes and provide insights into their components. In this study 33 genes from the genome of Latimeria chalumnae and from the liver and testis transcriptomes of Latimeria menadoensis, implicated in sex determination and differentiation, were identified and characterized and their expression levels measured. Interesting findings were obtained for GSDF, previously identified only in teleosts and now characterized for the first time in the sarcopterygian lineage; FGF9, which is not found in teleosts; and DMRT1, whose expression in adult gonads has recently been related to maintenance of sexual identity. The gene repertoire and testis-specific gene expression documented in coelacanths demonstrate a greater similarity to modern fishes and point to unexpected changes in the gene regulatory network governing sexual development.

Ancient tetraploidy and slow molecular evolution in Scaphiophryne: ecological correlates of speciation mode in Malagasy relict amphibians

Karyotypes of three microhylid frog species of the Malagasy relict genus Scaphiophryne were studied: Scaphiophryne gottlebei, S. madagascariensis and S. spinosa. The latter two showed a plesiomorphic ranoid karyotype of 2n = 26. In contrast, tetraploidy was demonstrated in S. gottlebei, which constitutes an exceptional state among Malagasy amphibians. A combination of different banding techniques and of rDNA-FISH provided evidence for allopolyploidy in the species and for a completed subsequent functional and structural diploidization. Phylogenetic analysis of mitochondrial 16S rDNA sequences revealed a significant deceleration of nucleotide substitution rates in Scaphiophryne. The tetraploidy of S. gottlebei probably occurred early in their radiation. Ecological and behavioural patterns of Scaphiophryne probably favoured intraspecific gene flow and hybridization events, thereby leading to slow molecular substitution rates and to allopolyploid chromosome speciation in S. gottlebei.

Different genomic evolutionary rates in the various reptile lineages

Although Reptiles occupy a strategic position among terrestrial vertebrates, studies of the composition and evolution of their genome are scarce. The cytogenetic analysis of nearly 1400 species evidenced different karyotypical evolutionary rates and different G-banding structures in turtles and crocodiles on the one hand and squamates on the other. A similar dichotomy was also identified through the study of the quantitative and compositional characteristics of the genome. The different evolutionary rates of chromosome morphology and genome size and composition and the diversification of coding and non-coding sequences bear an interesting relationship to the number of extant species and the extinction rates of the reptilian orders and suborders studied, suggesting a large role for such different evolutionary rates in the phylogenesis of this class. The different molecular and structural organisation of chromosomes could be an important, though by no means the sole, factor affecting the genomes evolutionary rate.

Repetitive DNA and polyploidy in selachians

1. The DNA reassociation kinetics have been studied in 6 selachian species: Raja asterias, Raja montagui, Dasyatis violacea, Torpedo marmorata, Torpedo ocellata and Oxynotus centrina. 2. The results obtained show that the genomes of the two Torpedo, Dasyatis and Oxynotus are polyploid if compared with those of the two rays, though this finds no correspondence in the diploid chromosome number. 3. The phenomena of polyploidization would often be followed by wide chromosome rearrangements and by a progressive divergence of the various repetitive DNA sequences. 4. The existence of polyploidy in almost all the main superorders of living Selachians suggests that this mechanism of genomic evolution may have played an important role in the phylogeny of this class.

Chromosomal and molecular analysis of some repeated families in Discoglossus Otth, 1837 (Anura, Discoglossidae): Taxonomic and phylogenetic implications

Abstract We conducted a cytogenetic and molecular analysis in six out of the eight Discoglossus taxa CD. pictus pictus, D. p. auritus, D. sardus, D. montalentii, D. galganoi galganoi and D. g. jean‐neae) All the taxa possessed 2n = 28 biarmed chromosomes, except D. pictus pictus where the 13th pair was acrocentric, and the NORs were located on the short arm of the 7th pair in all the taxa but D. galganoi, where they were on the short arm of the 13th pair. Heterochromatin was detected by banding methods (C‐band‐ing, base‐specific fluorochromes, replication patterns and digestions with restriction enzymes). The techniques used were able to discriminate among all the Discoglussus taxa studied, identifying several heterochromatin families with marked differences in chromosome location and/or in the behaviour of the banding techniques used. The differentiation of these heterochromatin families can have either a phylogenetic (as is the case with A+T rich cen‐tromeric heterochromatin) or an adaptive (G+C rich in...

Genome composition and tandemly repetitive sequence at some centromeres in the lizard Podarcis s. sicula Raf.

A detailed study on the genome of the lizard P. sicula has been carried out using restriction enzyme analysis followed by identification and cloning of a repetitive DNA fraction. The results show that P. sicula generally possesses a quite homogeneous genome composition, with a single tandemly repetitive sequence family that is easily visualized after digestion of genomic DNA with Taq I. The cloned repeating unit of this satellite (260 bp) has been designed pLCSl. In-situ hybridisation shows that this satellite is localized in the centromeric region. Dot blot experiments show that sequences similar to pLCSl are present in other species of the same family of lizards.

Karyology of the Antarctic scallop Adamussium colbecki, with some comments on the karyological evolution of pectinids.

Karyotype, location of the nucleolar organiser region (NOR) and heterochromatin presence and composition were studied in the Antarctic scallop Adamussium colbecki Smith, 1902. The karyotype exhibits 2n = 38 chromosomes with 11 pairs of metacentrics, 5 of submetacentrics, one subtelocentric and two telocentrics. Ag–NOR, CMA3, DA/MM and NOR–FISH evidenced paracentromeric NORs on the short arm of 2nd pair chromosomes. Digestion with three restriction endonucleases followed by sequential staining with Giemsa, CMA3 and DAPI evidenced on all chromosomes centromeric heterochromatin positive for both DAPI and CMA3. In situ hybridisation analysis showed the presence of an AT-rich satellite DNA in the centromeric heterochromatin of several chromosomes. A mosaicism was detected in the germinal cell lines of one specimen, as in six of the 20 plates examined the set had 37 chromosomes with a missing pair of telocentrics and an unpaired metacentric. Comparison of the chromosome sets of all the pectinids studied to date and comparison with a phyletic tree obtained from molecular mitochondrial genes studies yielded good agreement between karyotype morphology and taxonomic classification.

Karyological and genetic variation in Middle Eastern lacertid lizards, Lacerta laevis and the Lacerta kulzeri complex: a case of chromosomal allopatric speciation.

AbstractKaryological (standard and C, Ag-NOR and Alu-I banding methods) and mtDNA analyses (cytochrome b and 12S rRNA) were conducted on specimens from eight allopatric populations of the Lacerta kulzeri complex. Parallel analyses were performed for comparison on Lacerta laevis specimens. Karyological and molecular studies support the morphological and ethological evidence indicating the specific separation between Lacerta laevis and Lacerta kulzeri In the Lacerta kulzeri complex, chromosomal analysis substantiated an interpopulation differentiation roughly along a north–south trend, mainly regarding the sex chromosome morphology and heterochromatin.The cytochrome b and 12S rRNA gene analyses showed minor genetic differences that were considerably smaller than those commonly found in genetically isolated populations. The L. kulzeri populations from Barouk, Druze and Hermon show a mean genetic distance that, in other saurians, characterises subspecies.The conditions found in L. laevis and L. kulzeri are reminiscent of Kings model of chromosomal primary allopatry and support the hypothesis that in these lacertid lizards chromosome variations can become fixed before the accumulation of the genetic mutations.