Testing the evolutionary constraints of metamorphosis: The ontogeny of head shape in Triturus newts

In vertebrates with complex, biphasic, life cycles, larvae have a distinct morphology and ecological preferences compared to metamorphosed juveniles and adults. In amphibians, abrupt and rapid metamorphic changes transform aquatic larvae to terrestrial juveniles. The main aim of this study is to test whether, relative to larval stages, metamorphosis (1) resets the pattern of variation between ontogenetic stages and species, (2) constrains intraspecific morphological variability, and (3) similar to the “hour‐glass” model reduces morphological disparity. We explore postembryonic ontogenetic trajectories of head shape (from hatching to completed metamorphosis) of two well‐defined, morphologically distinct Triturus newts species and their F1 hybrids. Variation in head shape is quantified and compared on two levels: dynamic (across ontogenetic stages) and static (at a particular stage). Our results show that the ontogenetic trajectories diverge early during development and continue to diverge throughout larval stages and metamorphosis. The high within‐group variance and the largest disparity level (between‐group variance) characterize the metamorphosed stage. Hence, our results indicate that metamorphosis does not canalize head shape variation generated during larval development and that metamorphosed phenotype is not more constrained relative to larval ones. Therefore, metamorphosis cannot be regarded as a developmental constraint, at least not for salamander head shape.     

Chromosome 1 in crested and marbled newts (Triturus)

The heteromorphic chromosomes 1 of Triturus cristatus carnifex and T. marmoratus were studied in mitotic metaphase after staining with the Giemsa C-banding technique and with the fluorochromes, DAPI (AT-specific) and mithramycin (GC-specific). They were also examined in the lampbrush form under phase-contrast before fixation and after fixation and staining with Giemsa. Chromosomes 1 of T.c. carnifex are asynaptic and achiasmatic throughout most of their long arms. They are also heteromorphic in most of their long arms for the patterns of Giemsa and fluorochrome staining and the distribution of distinctive lampbrush loops. The heteromorphic regions correspond to the regions that are asynaptic and achiasmatic. They stain more strongly with mithramycin and more weakly with DAPI than the remainder of the chromosomes, signifying that their DNA is relatively rich in GC. The patterns of staining with Giemsa and fluorochromes and the distributions of distinctive lateral loops vary from one animal to another in the same species and even in the same population. The asynaptic and achiasmatic regions of chromosomes 1 in T. marmoratus extend throughout the whole of the long arms and well beyond the heterochromatic region. Chiasmata form only in the short arm and occasionally in the short euchromatic segment at the tip of the long arms. The staining patterns of chromosomes 1 in T. marmoratus differ from those in T.c. carnifex although, like carnifex, their DNA is relatively GC-rich. The chromosomes 1 of T. marmoratus are more submetacentric than those of T.c. carnifex. In T. marmoratus chromosome 1B is about 12% shorter than 1A. There is a short paracentric inversion heterozygosity in the long arm of chromosome 1B in T. marmoratus which probably accounts for the lack of chiasmata in the euchromatin that separates the centromere from the start of the heterochromatin. In both carnifex and marmoratus, embryos that are homomorphic for chromosome 1 arrest and die at the late tailbud stage of development. The same applies to F₁ hybrid embryos T.c. carnifex x T. marmoratus, and this has permitted identification of chromosomes 1A and 1B in both species. There is no correspondence between patterns of Giemsa or fluorochrome staining of the heteromorphic regions of chromosome 1 and any feature of the lampbrush chromosomes. However, the short euchromatic ends of the long arms of chromosomes 1 in both species are distinguished in the lampbrush form by a series of uniformly small loops of fine texture associated with very small chromomeres. The Giemsa C-staining patterns of both chromosomes 1A and 1B are different in each of the four subspecies of T. cristatus. T.c. karelinii stands out by having unusually large masses of Giemsa C-staining centromeric heterochromatin on all but 1 of its 12 chromosomes. A scheme is proposed for the evolution of chromosome 1 in T. cristatus and T. marmoratus, based on all available cytological and molecular data.     

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     

Major cranial changes during Triceratops ontogeny

This is the first cranial ontogenetic assessment of Triceratops, the well-known Late Cretaceous dinosaur distinguished by three horns and a massive parietal-squamosal frill. Our analysis is based on a growth series of 10 skulls, ranging from a 38 cm long baby skull to about 2 m long adult skulls. Four growth stages correspond to a suite of ontogenetic characters expressed in the postorbital horns, frill, nasal, epinasal horn and epoccipitals. Postorbital horns are straight stubs in early ontogeny, curve posteriorly in juveniles, straighten in subadults and recurve anteriorly in adults. The posterior margin of the baby frill is deeply scalloped. In early juveniles, the frill margin becomes ornamented by 17-19 delta-shaped epoccipitals. Epoccipitals are dorsoventrally compressed in subadults, strongly compressed and elongated in adults and ultimately merge onto the posterior frill margin in older adults. Ontogenetic trends within and between growth stages include: posterior frill margin transitions from scalloped to wavy and smooth; progressive exclusion of the supraoccipital from the foramen magnum; internal hollowing at the base of the postorbital horns; closure of the midline nasal suture; fusion of the epinasal onto the nasals; and epinasal expansion into a morphologically variable nasal horn. We hypothesize that the changes in horn orientation and epoccipital shape function to allow visual identity of juveniles, and signal their attainment of sexual maturity.     

Use of a magnetic compass for Y-axis orientation in premetamorphic newts (Triturus boscai)

Experiments were carried out to investigate whether premetamorphic larvae of Boscas newt (Triturus boscai) are capable of using the geomagnetic field for Y-axis orientation (i.e., orientation toward and away from shore). Larvae were trained outdoor in two different training configurations, using one training tank aligned along the magnetic north–south axis, with shore facing north, and another training tank positioned with its length along the east–west axis, with shore located west. After training, premetamorphic newts were tested in an outdoor circular arena surrounded by a pair of orthogonally aligned cube-surface coils used to alter the alignment of the Earths magnetic field. Each newt was tested only once, in one of four magnetic field alignments: ambient magnetic field (i.e., magnetic north at North), and three altered fields (magnetic north rotated to East, West, South). Distributions of magnetic bearings from tested larvae indicated that they oriented bimodally along the magnetic direction of the trained Y-axis. These findings demonstrate that T. boscai larvae are sensitive to the geomagnetic field and can use it for orienting along a learned Y-axis. This study is the first to provide evidence of Y-axis orientation, accomplished by a magnetic compass, in larval urodeles.     

Female alpine newts (Triturus alpestris) mate initially with males signalling fertility benefits

Gametic asymmetry implies that females invest more per gamete than males do and thus sperm is considered to be a relatively cheap resource. However, contrary to this classic view, sperm has been shown to be frequently in short supply; hence, selection favouring females that mate for fertility benefits should occur. For this reason, we determined whether males signalling fertility are preferred by female newts of the species Triturus alpestris . We performed paired female–male trials using unmated and previously inseminated females to determine potential criteria for female interest in a courting male, to establish what factors lead to successful mating and to assess the importance of female choice for direct and indirect benefits. We found that female interest in any potential mate and mating success decreased once mating had occurred. Furthermore, we detected an increase in spermatophore deposition rate and rapid spermatophore transfer in encounters that resulted in a successful mating. The results obtained indicate that female alpine newts are attracted to males showing signs of relatively high fertility and that females exhibit a decreased propensity to mate once initial sperm reserves have been acquired. Our results support the theory of initial female choice for fertility benefits.  © 2007 The Linnean Society of London, Biological Journal of the Linnean Society , 2007, 91 , 483–491.     

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.     

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.     

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.     

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 comparative analysis of predator-induced plasticity in larval Triturus newts

Species that occupy similar habitats are expected to show convergent phenotypes. If habitats are defined by the presence of predators, then traits that modify vulnerability to predation, including predator-induced phenotypic plasticity, should be similar within habitats. We tested this idea using larvae of six syntopic newt species belonging to the two Triturus clades. Behavioural plasticity induced by odonate predators was strongly dissimilar between the two main clades but similar within them. Morphological plasticity was variable among species, even between one pair of closely related species. A predation experiment tested whether differences between clades could be caused by differences in body size. Size-specific vulnerability differed between newts in the small-bodied and large-bodied clades, indicating that similar predators may affect the two clades differently. The results showed both similarity and dissimilarity in predator-induced phenotypic plasticity in syntopic larval newts although theory suggests that divergence is unlikely in such ecologically similar species.     

High aquatic niche overlap in the newts Triturus cristatus and T. marmoratus (Amphibia,Urodela)

We studied spatial niche metrics of large-bodied newts (Triturus cristatus and T. marmoratus) in three breeding ponds in western France. Adults and larvae were sampled with underwater funnel traps. Larvae were identified to the species with diagnostic microsatellite DNA markers. The distribution of adult T. cristatus and T. marmoratus across pond regions differed in one out of six cases, no differences were observed between larvae (two ponds studied). Niche overlap and niche breadth indices across resource states defined as pond regions or individual traps were high (Schoener's C: pond regions 0.60–0.98, traps 0.35–0.71; Levins' B: pond regions 0.71–0.98, traps 0.35–0.76). Adults of large-bodied newts significantly differed in resource use from small-bodied newts (T. helveticus). The results are discussed in view of the occurrence of interspecific breeding attempts, and the unpredictable ecological characteristics of newt breeding ponds.     

Molecular and morphological evidence of hybridization between newts Triturus vulgaris and T. montandoni (Caudata: Salamandridae) in Slovakia

Natural hybridization and introgression is recorded between the newt species Triturus vulgaris and T. montandoni in Slovakia. To confirm a hybrid status of two putative hybrids, morphological and Randomly Amplified Polymorphic DNA (RAPD) markers specific for T. vulgaris and T. montandoni were used. The individuals found in Zbojská (Veporské vrchy Mts, Slovakia; analyzed morphologically and genetically) and Zubrohlava (Oravská kotlina basin, Slovakia; analyzed only morphologically) possessed markers of both species. Segregation of RAPD markers together with a model-based Bayesian analysis revealed that the specimen from Zbojská belonged to later generation hybrids.     

Cytology, embryology, and evolution of the developmental arrest syndrome in newts of the genus Triturus (Caudata: Salamandridae)

We have examined embryonic development in three species (T. carnifex, T. cristatus, and T. marmoratus) of European newts of the genus Triturus (subgenus Neotriton) in which developmental arrest occurs in embryos that are homomorphic for a chromosomal heteromorphism involving chromosome 1 (Horner and Macgregor: J. Herpetol., 19:261-270, 1985). Embryonic arrest occurred during tailbud stages in all three species, but at a slightly earlier stage in T. marmoratus. Two phenotypes were identified among the arrested embryos. One of these is indistinguishable in embryonic morphology from normal embryos at all stages up to the time of arrest, but the other is characterized by a protruding yolk plug, which persists from the late gastrula/early neurula stage to the tailbud arrest stage and apparently interferes with normal morphogenesis. Evidence is presented that the two arrested phenotypes, which occur in approximately equal numbers, represent embryos that carry the two alternative homomorphic chromosome pairs of chromosome 1 heteromorphism. We conclude that developmental arrest reflects a balanced lethal heterozygosity probably resulting from an unequal exchange of genic material between the homologues of chromosome 1 which occurred in a common ancestor of the Neotriton species.     

The ontogeny of feeding behavior and cranial morphology in the leopard shark Triakis semifasciata (Girard 1854): A longitudinal perspective

While much is known about the ecological and functional morphology of feeding in lower vertebrates, studies of elasmobranch feeding over ontogeny are broadly lacking. In this study, the ontogeny of feeding behavior and morphology were investigated in neonatal and young-of-the-year leopard sharks Triakis semifasciata using morphometric measurements of growth and high-speed videography in a longitudinal study. Five food types were used during filming sessions to facilitate differentiation of modulation and variation over ontogeny. Functional aspects of muscle and buccal volume scaling were investigated through dissection. Growth over ontogeny was shown to influence numerous kinematic variables, while intra- and inter-individual variability was the dominant factor affecting feeding behavior. Modulation of feeding behavior based on food size and elusiveness was present for timing variables and predator motion during the strike, but not for food motion or the relative extent of buccal expansion. Allometric growth occurred in all aspects of external cranial morphology measured, resulting in a shallower head profile, anterior displacement of the mouth, and relatively larger jaw musculature over ontogeny. While the degree to which morphology constrains or enhances behavior cannot be directly quantified, variability in behavior greatly exceeds variability in morphology over early ontogeny. Maintenance of a behaviorally and morphologically versatile feeding apparatus throughout ontogeny is proposed to enhance the exploitation of resources and facilitate a diverse diet in T. semifasciata under variable environmental conditions.