A phylogeny for the Old World newts, genus Triturus: biochemical and behavioural data

A reconstruction is presented of the phylogenetic relationships withing Triturus . Two different data sets are used. Protein electrophoretic data were scored for 19 presumptive gene loci in nine species (Rafinski & Arntzen, 1987) and a set of 16 behaviour characters from the courtship of Triturus was scored for the same nine species. When these are analysed in concernt, a single, fully resolved and robust solution is found. Consequently, two subgenera are recognized: Triturus and Palaeotriton , the former including all te medium- and large-bodied species: T. alpestris, T. vittatus, T. marmoratus, T. cristatus and its three former subspecies ( T. carnifex, T. dobrogicus and T. Karelini ); the latter comprising the small-bodied species: T. bascai, T. italicus, T. helveticus, T. vulgaris and T. montandoni . Within the subgenus Triturus, T. alpestris and T. vittatus are sister taxa to the remaining members of the subgenus. Similarly, T. boscai and T. italicus are a monophyletic group, sister to T. vulgaris, T. helveticus and T. montandoni . Alternative solutions may be found on the basis of a quantitative analysis of the behavioural characters employed in this study. An alternative phylogenetic tree is included as an example.     

Distribution of satellite DNA TkS1 in genomes of salamanders (Salamandridae)

Genomes of 22 species and subspecies of salamandrids (Triturus, Cynops, Neurergus, Notophthalmus, Pachytriton, and Pleurodeles ) were studied. Satellite TkS1 (32-33 bp) was found in all examined species, both in direct and reversed orientations. The use of single-primer PCR with oligonucleotides, which are homologous to the most conservative part of satellite TkS1, allowed us to reveal DNA fragments, flanked by single copies (or small tandem repeats) of satellites TkS1 having various (5' and 3') orientation. Such fragments were observed in Pleurodeles waltl, Triturus a. alpestris, T. vulgaris lantzi, T. v. vulgaris, Neurergus crocatus and T. vulgaris graecus. The length of amplified DNA fragments in three subspecies of T. vulgaris differed. This might be connected with unequal amounts and different distribution of 5' and 3' copies of satellite TkS1 in their genomes. Patterns of DNA amplification in Triturus a. alpestris and Neurergus crocatus were quite similar. Only two species (Triturus a. alpestris and T. v. vulgaris) had approximately equal amounts and similar distribution of 5' and 3' copies of satellite TkS1 in their genomes.     

Phylogeography of two European newt species--discordance between mtDNA and morphology

The newts Triturus vulgaris and Triturus montandoni are sister species that exhibit contrasting levels of intraspecific morphological variation. Triturus vulgaris has a broad Eurasiatic distribution encompassing both formerly glaciated and unglaciated areas and shows substantial morphological differentiation in the southern part of its range, while T. montandoni, confined to the Carpathians, is morphologically uniform. We analysed sequence variation of two mtDNA fragments of the total length of c. 1850 bp in 285 individuals of both species collected from 103 localities. Phylogenetic analysis of 200 unique haplotypes defined 12 major clades, their age estimated at c. 4.5-1.0 million years (Myr). Most of the older clades were found in the southern part of the range, and also in central Europe, mainly in Romania. The distribution of mtDNA clades points to the existence of several glacial refugia, located in the Caucasus region, Anatolia, the Balkan Peninsula, Italy, and more to the north in central Europe. The concordance between mtDNA based phylogeny and the distribution of T. vulgaris subspecies was weak. Triturus montandoni haplotypes did not form a monophyletic group. Instead they were found in six clades, in five of them mixed with T. vulgaris haplotypes, most likely as a result of past or ongoing hybridization and multiple introgression of mtDNA from T. vulgaris to T. montandoni. Patterns of sequence variation within clades suggested long-term demographic stability in the southern groups, moderate and relatively old demographic growth in the populations inhabiting central Europe, and high growth in some of the groups that colonized northern parts of Europe after the last glacial maximum.     

Microhabitat selection and feeding relations of smooth and warty newts, Triturus vulgaris and T. cristatus, at an upland pond in mid-Wales

The utilization of microhabitat and food resources by Triturus vulgaris and T. cristatus was studied in an upland pond in mid-Wales. From April to June T. vulgaris was more evenly distributed across the pond than T. cristatus , which showed a preference for the middle of the pond and the bottom of the water column. From July to September however, when most T. vulgaris had left the pond, T cristatus was evenly distributed within the water column and a much higher proportion was captured around the shorelines. Both species appear to be generalist predators with diets limited by prey size. Zooplankton were the most important prey for T. vulgaris , while leeches formed the bulk of the diet of T. cristatus . However, T. cristatus did not select larger-sized zooplankton than T. vulgaris . Large body size therefore enables T. cristatus to consume a wider range of prey sizes than T. vulgaris . Microhabitat niche overlap was greater than feeding niche overlap between the species.     

The effect of pH on embryonic and larval development in smooth and palmate newts, Triturus vulgaris and T. helveticus

The distribution of smooth newts, Triturus vulgaris , and palmate newts, Triturus helveticus , in north-west Europe is related to geology and water quality. This study compared the development of the eggs and larvae of the two species under sublethal acidic and neutral conditions. Newt embryos raised under low pH hatched at an earlier stage of development, at a smaller size, and before those raised under neutral conditions. T. vulgaris hatched at a smaller size than T. helveticus , but pH did not affect the species differentially. Larvae of both species grew to a larger size under neutral than under acid conditions. Larvae raised in heterospecific pairs grew at least as well as those raised in conspecific pairs. Feeding was depressed under acid conditions, and reduced growth may therefore be associated with changes in the behaviour of newt larvae and their prey.     

Sexual size and shape evolution in European newts (Amphibia: Caudata: Salamandridae) on the Balkan Peninsula

We used a phylogenetic perspective in an examination of the direction and extent of sexual dimorphism in body size and body shape in European newts from the Balkan Peninsula (alpine newts, Mesotriton alpestris; crested newts, Triturus cristatus superspecies; smooth newts, Lissotriton vulgaris). We found a strong, female‐biased sexual size dimorphism (SSD) in the analysed clades of alpine newt, whereas within crested newts we found a less stringent female‐biased SSD in Triturus carnifex, Triturus macedonicus and Triturus karelinii, and no significant SSD in T. cristatus or Triturus dobrogicus. Among the smooth newts, we found male‐biased SSD in Lissotriton vulgaris vularis and Lissotriton vulgaris greacus and no SSD in Lissotriton vulgaris meridionalis. Most of these newts also exhibit a significant sexual dimorphism in body shape, which varied more randomly than body size, regardless of SSD level. Female and male body size as well as the degree of SSD displayed statistically significant phylogenetic signal, while sexual dimorphism in body shape was phylogenetically independent. The relationship between independent contrast data for female size and male size indicated that SSD in European newts could be driven by a disproportionate increase in female size as increase in female size was not accompanied by a proportional increase in male size.     

The effect of pH on feeding behaviour in newt larvae (Triturus: Amphibia)

The feeding responses of three species of newt larvae were compared under circumneutral and sublethal acid conditions. Under acid conditions (pH 4.5) feeding behaviour was suppressed in palmate newts, Triturus helveticus, and smooth newts. T. vulgaris , but not in crested newts, T. cristatus. At low pH, approach and orientation towards food occurred in T. helveticus and T. vulgaris , but snapping was inhibited; T. cristatus snapped and consumed food immediately it was offered under the same conditions. These differences are not consistent with the apparent greater tolerance of T helveticus for acidified ponds. The observations suggest that the chemosensory system of T. helveticus and T. vulgaris may be impaired at low pH.     

Predation and competition within an assemblage of larval newts (Triturus)

The impact of crested newts (Triturus cristatus) on the smaller-bodied palmate and smooth newts (T helveticus and T vulgaris) was studied during the larval stages using a combination of field and laboratory experiments In pond enclosures T cristatus larvae had no effect on the two smaller species over the first four weeks of development By eight weeks, however. T cristatus had achieved a size advantage which enabled it to eliminate T helveticus and severely reduce T vulgaris by predation In laboratory trials under food-limited conditions, T helveticus and T vulgaris were slightly smaller when raised with T cristatus, suggesting that this predatory effect was complemented by interspecific competition during early development Predation of the smaller species started when T cristatus reached a threshold size of c 27 mm No reciprocal effects on T cristatus growth or survival were observed Although T cristatus may be a significant predator of congeneric species in natural ponds, other factors, such as differences in microhabitat selection, higher-order predator-prey interactions and the occasional desiccation of pond habitats may facilitate coexistence between the species     

Influence of introduced fish Percottus glenii (Odontobutidae, Pisces)

Monitoring of 28 waterbodies has been carried out since 1994 in the region of reserve "Lake Glubokoe" (Moscow region, Russia). It was revealed that species diversity as well as abundance of larval amphibians correlate negatively with presence of introduced fish, rotan, Perccottus glenii (Odontobutidae). Newts (Triturus cristatus, T. vulgaris) and frogs (Rana temporaria, R. arvalis, R. lessonae) as a rule are not capable to breed in waterbodies colonised by rotan. In contrast, toads (Bufo bufo) breed successfully in such sites. Larvae of toads are comparatively less edible for rotan and pass their metamorphosis. Persistence of amphibians to predation of rotan decreases in the row: B. bufo (R. temporaria, R. arvalis and R. lessonae), T. vulgaris, T. cristatus. The Crested newt (T. cristatus) is the most endangered species and could extinct in next years.     

Rare hybridization and introgression in smooth and palmate newts (Salamandridae: Triturus vulgaris and T. helveticus)

This paper describes the occurrence of hybridization and introgression in two species of amphibians (the newts Triturus helveticus and Triturus vulgaris ) in mid-Wales, northern France and western France. A single aberrant adult male with intermediate phenotype was found. The multivariate analysis of 14 morphometric and two meristic characters supported its hybrid status. Electrophoretic analysis of 42 protein loci showed a genetic distance of 0.57 ± 0.14 Nei units between the species and revealed 15 diagnostic loci. The aberrant specimen was heterozygous at 11 of those and most likely to be a F₁ hybrid. Four marker loci appeared homozygous, suggesting the presence of enzymatically non-active ('null') alleles. The analysis of (maternally inherited) mitochondrial DNA showed the hybrid to be the offspring of a T. helveticus mother (and a T. vulgaris father). This observation does not conform to expectations based on the species composition in the pond from which the hybrid was collected. No F₁ hybrids were observed in a large sample (n > 5000) of larvae, recently metamorphosed newts and adults using two diagnostic protein loci. Occasionally alleles characteristic for one species were observed in the gene pool of the other species, suggesting the presence of bidirectional introgression. However, the frequency of alien genes was low (maximally 0.07%) which renders it difficult to rule out alternative explanations conclusively. The increase in total genetic variation in T. helveticus and T. vulgaris due to gene flow between them is negligible.     

Molecular cytogenetics of the ribosomal (18S + 28S and 5S) DNA loci in primitive and advanced urodele amphibians.

In the present work we performed a cytogenetic analysis of the ribosomal (18S + 28S and 5S) loci in amphibian species belonging to the advanced family Salamandridae (genera Triturus, Salamandra, and Salamandrina) and in the primitive hynobiid Salamandrella keyserlingii (family Hynobiidae). In each analyzed karyotype the 5S rDNA sites appear to be stable, and definite in number, while an intraspecific variability both in number and chromosomal location of the 18S + 28S rDNA loci has been found in some Triturus species. In particular, an evolutionary trend toward a large intraspecific variability of the 18S + 28S rDNA loci has been found in the T. vulgaris species group. A structural analysis of the ribosomal repetition units demonstrates the occurrence of a length polymorphism within the 18S + 28S rDNA repeats in the examined species of the family Salamandridae; however, this polymorphism is rather limited, even in those Triturus species characterized by high intragenomic variability of the ribosomal sites. We show that in T. vulgaris meridionalis the variant repetition units actually segregate with individual chromosomes. This implies that they are not intermingled in the ribosomal clusters.     

Alternative life-history pathways: paedomorphosis and adult fitness in European newts (Triturus vulgaris and T. alpestris)

Paedomorphs and metamorphs of the smooth newt ( Triturus vulgaris ) and alpine newt ( Triturus alpestris ) were compared with respect to body size, age structure, age at sexual maturity, survivorship, and female and male fecundity. Paedomorphs were significantly smaller than metamorphs, except for the alpine newt males. Non-significant differences between morphs in both species in terms of the life span, age of sexual maturity, survival rates and male fecundity were found. The relationships concerning female-fecundity parameters were not so straightforward. The total number of oocytes was significantly higher in smooth-newt paedomorphs, while in the alpine newt the difference was insignificant. When ovary mass was compared, significant differences appeared only in the alpine newt, in favour of metamorphic females. Oviductal egg size was similar in both morphs of T. vulgaris . The maintenance of both life-history strategies in the species studied is discussed in the light of these findings.     

Differential effects of pH and temperature on embryonic development in the British newts (Triturus)

Changes in the water quality and temperature relationships of ponds may affect the structure of amphibian assemblages. The survival, time to hatching, hatching size and hatching stage of newt embryos were studied in the three British species ( Triturus cristatus, T. helveticus and T. vulgaris ), at two temperatures and two pHs. All T. cristatus embryos failed to hatch at pH 4.5, whereas over 80% of T. helveticus and T. vulgaris embryos hatched successfully at the same pH. At pH 7.5, T. cristatus survival was the same as the other two species, after the 50% mortality due to the homomorphism of chromosome 1 was taken into account. Temperature had no effect on survival of embryos. Time to hatching was two to four times longer at 12°C than at 17°C. Low pH shortened development time in T. vulgaris but not in T. helveticus . Low pH, but not temperature, affected size at hatching, with T. helveticus and T. vulgaris embryos emerging at a smaller size and earlier stage of development under acidic conditions. This reduction of size at low pH affected T. vulgaris more than T. helveticus . We predict that T. cristatus embryos will be the most vulnerable of the three species to acidification in nature. Warm ponds will result in rapid embryonic development, but T. helveticus and T. vulgaris larvae hatching in acid ponds will do so at a smaller size and earlier stage of development. The pattern of vulnerability to acidification within amphibian assemblages may change during ontogeny.     

On the biological significance of certain morphological characters in males of the Smooth newt Triturus vulgaris and of the Palmate newt Triturus helveticus (Urodela: Salamandridae)

The males of Triturus vulgaris and Triturus helveticus can be differentiated by a number of morphological characters whose biological function is unknown. It is suggested that these characters enhance the effectiveness of male displays performed during sexual behaviour. This hypothesis is discussed in relation to the taxonomic and genetic relationships between the two species.     

Population ecology of Trituras cristatus and T. vulgaris (Urodela) in SW Sweden

The population ecology of newts was investigated between 1969 and 1974 in two localities near Gothenburg, SW Sweden: the Gunnebo park pond (about 300 m²) where Triturus cristatus Laurenti and T. vulgaris L. are sympatric, and a rock-pool (6 m²) in Billdal with only T. vulgaris. At Gunnebo, a population of about 350 breeding T. cristatus was found with about as many males as females but males being more aquatic than females. An annual survival of 0.7–0.8 was estimated for the adults. The T. vulgaris population at the same locality was estimated to about 250–300 breeding adults with a sex ratio possibly in favour of males. The survival of adults was found to be extremely low and a very small percent of the adults seems to breed more than once. The Billdal T. vulgaris population was estimated to contain nearly 70 breeding adults with a sex ratio very near to 1:1. The annual survival of adults was found to be 0.5. The different survival of the two T. vulgaris populations are discussed with particular reference to predation. The age at which newts breed for the first time seems to vary; for T. vulgaris five years may well be the normal age. Factors that may limit the sizes of the populations, possible reasons for differences in frequency of occurrence in water, and possible predators are discussed.     

Sperm morphology of salamandrids (Amphibia, Urodela): implications for phylogeny and fertilization biology

Mature spermatozoa belonging to four salamander species, Salamandrina terdigitata, Triturus alpestris, Triturus carnifex and Triturus vulgaris, have been investigated by electron microscopy. The sperm ultrastructure of these species was compared with that of previously examined urodeles (36 species and 20 genera) and with that of anurans and caecilians. Many phylogenetic considerations may be inferred as a consequence of comparative spermatology. Urodela appears to be a monophyletic order characterized by three sperm synapomorphies: the acrosomal barb, nuclear ridge and marginal filament. Cryptobranchoidea are confirmed to form a monophyletic suborder having two synapomorphic characters: absence of mitochondria in the tail, and cylindrical shape of the tail axial rod. Within the family Salamandridae, sperm morphology confirms the phylogenetic distance between Salamandrina and Triturus, as already pointed out on the basis of molecular and morphological characters. The very complex ultrastructure of spermatozoa confirms a previous opinion that internal fertilization is the ancestral condition of the Amphibia.     

Limitation of similarity and feeding habits in three syntopic species of newts (Triturus, Amphibia)

Differences in body size between both species and sex were observed to be close to Hutchinsonian ratios in an assemblage of three species of newts (Triturus italicus, T alpestris and T carnifex) in southern Italy According to feeding, differences in body size were not related with a strict food niche segregation The diet of larger newt included indeed all the prey eaten by smaller newts but showed new prey types Differences between species were more marked than between sexes of the same species, despite a strong sexual dimorphism in body size Larger newts foraged in a wider range of microhabitats and exhibited a greater diversity of foraging tactics Triturus italicus fed essentially on plankton and chironomids in shallow water at the banks of the pond Triturus alpestris foraged also in deeper water, particularly on lsopods in vegetation Prey fallen on the water surface constituted the main part of T carnifex diet According to recent data on newt evolution, speciation consisted in the reduction of body size Our data show that this process has been related with a specialization of the diet and of habitat use     

A molecular assessment of phylogenetic relationships and lineage accumulation rates within the family Salamandridae (Amphibia, Caudata)

We examine phylogenetic relationships among salamanders of the family Salamandridae using approximately 2700 bases of new mtDNA sequence data (the tRNALeu, ND1, tRNAIle, tRNAGln, tRNAMet, ND2, tRNATrp, tRNAAla, tRNAAsn, tRNACys, tRNATyr, and COI genes and the origin for light-strand replication) collected from 96 individuals representing 61 of the 66 recognized salamandrid species and outgroups. Phylogenetic analyses using maximum parsimony and Bayesian analysis are performed on the new data alone and combined with previously reported sequences from other parts of the mitochondrial genome. The basal phylogenetic split is a polytomy of lineages ancestral to (1) the Italian newt Salamandrina terdigitata, (2) a strongly supported clade comprising the "true" salamanders (genera Chioglossa, Mertensiella, Lyciasalamandra, and Salamandra), and (3) a strongly supported clade comprising all newts except S. terdigitata. Strongly supported clades within the true salamanders include monophyly of each genus and grouping Chioglossa and Mertensiella as the sister taxon to a clade comprising Lyciasalamandra and Salamandra. Among newts, genera Echinotriton, Pleurodeles, and Tylototriton form a strongly supported clade whose sister taxon comprises the genera Calotriton, Cynops, Euproctus, Neurergus, Notophthalmus, Pachytriton, Paramesotriton, Taricha, and Triturus. Our results strongly support monophyly of all polytypic newt genera except Paramesotriton and Triturus, which appear paraphyletic, and Calotriton, for which only one of the two species is sampled. Other well-supported clades within newts include (1) Asian genera Cynops, Pachytriton, and Paramesotriton, (2) North American genera Notophthalmus and Taricha, (3) the Triturus vulgaris species group, and (4) the Triturus cristatus species group; some additional groupings appear strong in Bayesian but not parsimony analyses. Rates of lineage accumulation through time are evaluated using this nearly comprehensive sampling of salamandrid species-level lineages. Rate of lineage accumulation appears constant throughout salamandrid evolutionary history with no obvious fluctuations associated with origins of morphological or ecological novelties.     

The evolution of the adult body form of the crested newt (Triturus cristatus superspecies,Caudata, Salamandridae)

We characterized the adult body form of the crested newt (Triturus cristatus superspecies) and explored its evolution. From seven morphometric traits, we determined that body size, interlimb distance and head width define the body form. None of the morphometric traits showed a phylogenetic signal. Three body‐shape morphotypes (Triturus dobrogicus + T. cristatus, Triturus carnifex + Triturus macedonicus and Triturus karelinii + Triturus arntzeni) and three body‐size morphotypes (T. dobrogicus, T. cristatus and all other crested newts) could be recognized. The ancestral phenotype (a large body with a short trunk and a wide head) characterized T. karelinii and T. arntzeni. Triturus carnifex and T. macedonicus had a somewhat different phenotype (large body and wide head, accompanied by mild body elongation). The most derived phenotype included body size reduction and more pronounced body elongation in T. cristatus and, especially, in T. dobrogicus. Body elongation occurred by trunk lengthening but not head and tail lengthening. Additionally, contrary to other tetrapods, evolutionary axis elongation in crested newts was followed by a decrease in body size. We advocate the hypothesis that ecology drives the evolution of body form in crested newts.