Streambed microstructure predicts evolution of development and life history mode in the plethodontid salamander Eurycea tynerensis

Background

Habitat variation strongly influences the evolution of developmentally flexible traits, and may drive speciation and diversification. The plethodontid salamander Eurycea tynerensis is endemic to the geologically diverse Ozark Plateau of south-central North America, and comprises both strictly aquatic paedomorphic populations (achieving reproductive maturity while remaining in the larval form) and more terrestrial metamorphic populations. The switch between developmental modes has occurred many times, but populations typically exhibit a single life history mode. This unique system offers an opportunity to study the specific ecological circumstances under which alternate developmental and life history modes evolve. We use phylogenetic independent contrasts to test for relationships between a key microhabitat feature (streambed sediment) and this major life history polymorphism.

Results

We find streambed microstructure (sediment particle size, type and degree of sorting) to be highly correlated with life-history mode. Eurycea tynerensis is paedomorphic in streams containing large chert gravel, but metamorphoses in nearby streams containing poorly sorted, clastic material such as sandstone or siltstone.

Conclusion

Deposits of large chert gravel create loosely associated streambeds, which provide access to subsurface water during dry summer months. Conversely, streambeds composed of more densely packed sandstone and siltstone sediments leave no subterranean refuge when surface water dries, presumably necessitating metamorphosis and use of terrestrial habitats. This represents a clear example of the relationship between microhabitat structure and evolution of a major developmental and life history trait, and has broad implications for the role of localized ecological conditions on larger-scale evolutionary processes.     

Cytogenetic evolution in the plethodontid salamander genus Aneides

Chromosomal variation in the plethodontid salamander genus Aneides was analyzed using C-banding, to visualize heterochromatin distribution, as well as silver and fluorochrome banding, to reveal nucleolus organizer regions. Evidence is presented that Aneides has undergone pronounced karyological divergence from a Plethodon-like ancestor. It is concluded that this divergence reflects accelerated rates of chromosomal rearrangements in Aneides relative to Plethodon. These rearrangements have involved mainly pericentric inversions that have occurred in one or more of the three smallest pairs of chromosomes in the karyotype. Karyological variation within and between species of Aneides was used to analyze phylogenetic relationships within the genus. This analysis revealed at least two karyologically distinct species that had not been previously recognized by morphological criteria.     

Chromosome variation in the plethodontid salamander,Aneides ferreus

Karyotype variation in the plethodontid salamander, Aneides ferreus, has been analysed. 358 individuals from 14 populations, representing the major portion of the range of this salamander, have been karyologically examined. In A. ferreus, n=14. When the chromosomes are arranged in a decreasing relative length series, the karyotype is heteromorphic with respect to chromosome number 13, which may be either telocentric (T) or subtelocentric (ST). Variation in the heteromorphism over the range of the species is sex related, and probably also reflects relative population sizes. The heteromorphism in the isolated populations of A. ferreus on Vancouver Island, British Columbia, Canada, resembles a WZfemale/ZZmale sex chromosome dimorphism, suggesting the possibility that chromosome number 13 may be involved in sex determination in this population. The possibility that chromosome number 13 is involved in sex determination in all populations of A. ferreus is discussed. Our data suggest that the ancestral A. ferreus karyotype was homomorphic for T (T/T), and that the ST was derived from the T by a pericentric inversion. In peripheral populations, only the W homologue has been affected, whereas in central populations both the W and the Z chromosomes have been rearranged. Comparisons are made with other species of Aneides for which karyological information is available, and it is concluded that chromosome rearrangements have played an important role in the evolution of the genus. In C-banded chromosomes of A. ferreus, staining is most intense at the centromere regions of the larger chromosomes and is absent only in some of the smaller chromosomes. Implications of this C-banding pattern are discussed.     

Sperm storage and degradation in the spermathecae of the salamander Eurycea Cirrigera

Spermathecae are exocrine glands in the roof of the female cloaca that store sperm. Cytological and histochemical data indicate that the one type of secretion into the lumen is a glycoprotein. After a period of stasis in the summer, production of the secretion is initiated in the fall, coincident with an increase in ovarian follicular size. By the time of maximal follicular development and most intense mating activity in March and April, the spermathecal epithelium is filled with secretory granules. The secretory material is released into the lumen, enveloping the sperm. Many sperm remain in the spermathecae after oviposition, and most of these sperm are degraded in the spermathecal epithelium or pass through interruptions in the spermathecal walls caused by desquamation. Sperm in contact with the stromal environment are phagocytized by leukocytes. Some sperm, however, may survive in the lumen until at least the following fall. These sperm retain normal cytology, but whether or not they remain fertile and intact until a subsequent ovipository cycle is unknown.     

Epidermal keratinization in the salamander and a comparison with other amphibia

In a variety of amphibians examined the stratum corneum was one cell in depth, although in Xenopus it was up to three cells deep. The flattened horny cells were closely fused together along their lateral membranes to form a continuous sheet. Disulphide bonds of keratin were most concentrated in the peripheral cytoplasm, but the interiors of the cornified cells were sufficiently well keratinized to prevent more than slight enzymatic cytolysis of the normal cell components. Characteristically large, weakly stainable, non-shrunken nuclear remnants were found in the salamander and frog horny layers, but the clawed toad had small pyknotic (parakeratotic) nuclei. The mature amphibian keratinocytes contained free fats, bound phospholipids, calcium and sulphydryl groups, together with acid phosphatase and non-specific esterase. Cornification appears to begin by a process of separate individual cell keratinization and lateral membranes of neighbouring cells only later become fused together. This differs from the process in higher vertebrates in which the cells undergoing keratinization form a uniform transitional layer in the epidermis. In the amphibian epidermis neighbouring cells occur in different stages of keratinization.     

Organization of the vomeronasal organ in a plethodontid salamander

Salamanders in the family Plethodontidae show a unique behavior (nose-tapping) and have unique structures (nasolabial grooves) that may be used specifically to convey chemicals to the vomeronasal organ. The nasal structure of Plethodon cinereus was studied to determine if there is enhanced development of the vomeronasal organ compared with other salamander families that would correlate with use of these unique features. The vomeronasal organ in salamanders is found in a ventrolateral diverticulum of each main olfactory organ. P. cinereus has a more anteriorly placed vomeronasal organ within the diverticulum, and the posterior limit of each nasolabial groove is adjacent to the anterior limit of the vomeronasal organs. This suggests that the grooves deliver chemicals preferentially to the vomeronasal organs instead of to the main olfactory organs. In addition, the vomeronasal sensory epithelium is thickest anteriorly and is at its thinnest at about the level corresponding to the location of the vomeronasal organ in other salamander families. These adaptations suggest a specific mechanism of odorant delivery to the vomeronasal organ in plethodontid salamanders not found in other salamander families.     

Modular evolution of the Cetacean vertebral column

Modular theory predicts that hierarchical developmental processes generate hierarchical phenotypic units that are capable of independent modification. The vertebral column is an overtly modular structure, and its rapid phenotypic transformation in cetacean evolution provides a case study for modularity. Terrestrial mammals have five morphologically discrete vertebral series that are now known to be coincident with Hox gene expression patterns. Here, I present the hypothesis that in living Carnivora and Artiodactyla, and by inference in the terrestrial ancestors of whales, the series are themselves components of larger precaudal and caudal modular units. Column morphology in a series of fossil and living whales is used to predict the type and sequence of developmental changes responsible for modification of that ancestral pattern. Developmental innovations inferred include independent meristic additions to the precaudal column in basal archaeocetes and basilosaurids, stepwise homeotic reduction of the sacral series in protocetids, and dissociation of the caudal series into anterior tail and fluke subunits in basilosaurids. The most dramatic change was the novel association of lumbar and anterior caudal vertebrae in a module that crosses the precaudal/caudal boundary. This large unit is defined by shared patterns of vertebral morphology, count, and size in all living whales (Neoceti).     

Interdigital webbing and skin morphology in the neotropical salamander genus Bolitoglossa (amphibia; plethodontidae)

Arboreal salamanders of the Neotropical genus Bolitoglossa are characterized by palmate, fully webbed feet. The feet act as adhesive structures enabling the salamanders to cling to smooth surfaces. Scanning electron microscopy of skin epithelium and light microscopy of serial sections show exceptionally smooth cell surfaces and increased numbers of mucous glands on the soles of the feet. These features enhance the abilities of the feet to adhere by means of surface tension and suction. They are part of a set of morphological characteristics that may have been produced as a result of paedomorphosis.     

Trout affect the density, activity and feeding of a larval plethodontid salamander

1. Ecologists have struggled to describe general patterns in the impacts of predators on stream prey, particularly at large, realistic spatial and temporal scales. Among the confounding variables in many systems is the presence of multiple predators whose interactions can be complex and unpredictable. 2. We studied the interactions between brook trout (Salvelinus fontinalis) and larval two‐lined salamanders (Eurycea bislineata), two dominant vertebrate predators in New England stream systems, by examining patterns of two‐lined salamander abundance in stream reaches above and below waterfalls that are barriers to fish dispersal, by measuring the effects of trout on salamander density and activity using a large‐scale manipulation of brook trout presence, and by conducting a small‐scale laboratory experiment to study how brook trout and larval two‐lined salamanders affect each other's prey consumption. 3. We captured more salamanders above waterfalls, in the absence of trout, than below waterfalls where trout were present. Salamander density and daytime activity decreased following trout addition to streams, and salamander activity shifted from aperiodic to more nocturnal with fish. Analysis of stomach contents from our laboratory experiment revealed that salamanders eat fewer prey with trout, but trout eat more prey in the presence of salamanders. 4. We suggest that as predators in streams, salamanders can influence invertebrate prey communities both directly and through density‐ and trait‐mediated interactions with other predators.     

On the environmental control of oocyte maturation in a plethodontid salamander

Summary The relationship between condition and oocyte maturation was experimentally studied in Plethodon cinereus. Gravid females with immature oocytes were collected in September and maintained on different feeding regimes for 180 days. An index of salamander condition was derived. There was no significant relationship between the number of maturing oocytes and condition of the salamander at the end of the experiment, but oocyte size was signifiantly related to condition. A significant regression relationship was found for oocyte number and initial body weight, suggesting that condition may affect oocyte number early in the reproductive cycle. The implications of environmental induction of oocyte number for affecting clutch size and fitness are discussed.     

Effect of temperature,photoperiod, and feeding on the rate of tail regeneration in a semiaquatic plethodontid salamander

Salamander tail autotomy improves survival, but loss of the tail can subsequently be costly. For example, burst swimming speed is significantly slower after autotomy in desmognathan salamanders, which may increase predation risk in aquatic habitats. However, any long-term cost of tail loss is contingent on the rate of tail regeneration. To examine variation among seasons and environments in the cost of tail autotomy, we tested the effect of temperature, photoperiod, and feeding on tail-length re-growth in the semiaquatic plethodontid salamander Desmognathus conanti. Eight experimental groups (n=15 each, equivalent in body size) were tested. After acclimation for four weeks at one of two temperatures (either 10 °C or 20 °C) and one of two photoperiods (either L:D 9.5:14.5 h or 14.5:9.5 h), 60% of the tail length was autotomized for each individual. After autotomy, each experimental group was maintained under unique conditions of temperature (either 10 °C or 20 °C), photoperiod (either L:D 9.5:14.5 h or 14.5:9.5 h), and feeding (either fasting or weekly feeding). The length of the regenerated tail portion for each individual was measured each week until the group with the fastest re-growth had regenerated 50% of the lost tail length. Low temperature had a large, negative effect, fasting had a small, negative effect, but photoperiod had no significant effect on tail re-growth. The large thermal effect resulted from a combination of delayed initiation of tail-length re-growth and reduced regeneration rate thereafter at low temperature. We conclude that the cost of salamander tail autotomy differs among seasons and environments based on variation in temperature and food availability.     

Influence of intraspecific density and cover on home range of a plethodontid salamander

Summary Hypotheses about the influence of intraspecific density and available cover on home range size (=mean activity radius) of a plethodontid salamander (Desmognathus monticola) were tested by field experimentation from June 1980 to October 1981. A second order mountain stream in southwestern North Carolina was marked into five sections (two control and three experimental) and three replicated treatments were performed: Treatment I, low and high density additions of D. monticola; Treatment II, addition of cover objects (rocks); Treatment III, addition of cover and D. monticola.In Treatment I, home ranges of D. monticola on high density addition plots increased significantly relative to those on control and low density addition plots. Mean stomach content biomass and number of prey items of salamanders from high density plots were less than controls in all comparisons throughout the experiment but only one comparison revealed a statistically significant difference. Mean stomach contents of salamanders from low density plots were less than controls in 60% of all comparisons and only one statistically significant difference was found. Home range size was not affected significantly by Treatments II and III.Results of these experiments suggest that home range size of D. monticola may vary as a function of the number of conspecifics and cover objects; i.e., when the number of D. monticola increases where cover is limited, home range size increases.     

Gene lineages and eastern North American palaeodrainage basins: phylogeography and speciation in salamanders of the Eurycea bislineata species complex

Contemporary North American drainage basins are composites of formerly isolated drainages, suggesting that fragmentation and fusion of palaeodrainage systems may have been an important factor generating current patterns of genetic and species diversity in stream-associated organisms. Here, we combine traditional molecular-phylogenetic, multiple-regression, nested clade, and molecular-demographic analyses to investigate the relationship between phylogeographic variation and the hydrogeological history of eastern North American drainage basins in semiaquatic plethodontid salamanders of the Eurycea bislineata species complex. Four hundred forty-two sequences representing 1108 aligned bases from the mitochondrial genome are reported for the five formally recognized species of the E. bislineata complex and three outgroup taxa. Within the in-group, 270 haplotypes are recovered from 144 sampling locations. Geographic patterns of mtDNA-haplotype coalescence identify 13 putatively independent population-level lineages, suggesting that the current taxonomy of the group underestimates species-level diversity. Spatial and temporal patterns of phylogeographic divergence are strongly associated with historical rather than modern drainage connections, indicating that shifts in major drainage patterns played a pivotal role in the allopatric fragmentation of populations and build-up of lineage diversity in these stream-associated salamanders. More generally, our molecular genetic results corroborate geological and faunistic evidence suggesting that palaeodrainage connections altered by glacial advances and headwater erosion occurring between the mid-Miocene and Pleistocene epochs explain regional patterns of biodiversity in eastern North American streams.     

Complex secretory products in the oviduct of the plethodontid salamander Bolitoglossa dofleini (Amphibia, Urodela)

The ultrastructure of the secretory granules in the cells of the subdivisions of the oviduct in the neotropical plethodontid salamander Bolitoglossa dofleini was studied by transmission electron microscopy. In addition, we applied the cationic dye Cuprolinic Blue (CB) at different electrolyte concentrations to demonstrate proteoglycans, and the pyrogallol red-copper (PR-C) method to stain proteins at the ultrastructural level. The entire oviduct is lined by a simple epithelium that contains ciliated and microvillous cells in the first subdivision, the aglandular pars recta; microvillous cells show a moderate secretory activity. The following pars convoluta is differentiated into five glandular subdivisions and the aglandular “uterine portion”. Especially in the glandular parts, the epithelium is arranged in longitudinal folds. At their crests ciliated and microvillous cells similar to those in the pars recta occur. Gland cells are crowded with secretory granules that differ in their structural complexity (with and without electron-dense spheres or masses; elaborated, homogeneous or granular matrix; spherical; distorted) along the various subdivisions. Further, as suggested by the CB-technique, the cranial subdivisions contain large amounts of sulphated proteoglycans that decrease in the caudal direction. Carboxylated proteglycans appear to be present in all subdivisions examined. Electron-dense spheres of secretory granules are largely free of CB-precipitates, but stain more or less intensely with PR-C. The ultrastructure of the pars recta, and especially the “uterine portion” indicates transporting capability. The epithelial cells of the “uterus” have coated pits and a considerable amount of lysosome-like bodies.     

Eurycea bislineata (Green), the Two-Lined Salamander, a New Host of Myxidium serotinum Kudo & Sprague, 1940 (Myxosporida, Myxidiidae)

SYNOPSIS. Myxidium serotinum was found in 51 of 58 (87.9%) specimens of the two-lined salamander, Eurycea bislineata , from West Virginia. This constitutes a new host record. No specimens of the order Salientia, or other species of the order Caudata were infected. Eurycea bislineata appears to be the normal host of M. serotinum , which is specific for the gallbladder in a variety of amphibian hosts. Apparently, larval E. bislineata becomes infected with M. serotinum , and the parasite does not become a mature trophozoite until after the tadpoles metamorphose to the adult salamander. Certain host hormonal factors may be important to the maturation of M. serotinum.     

Territorial behavior of the plethodontid salamander Plethodon kentucki: influence of habitat structure and population density

To investigate the possible influence of variation in ecological and demographic factors on the spatial organization of the terrestrial plethodontid salamander Plethodon kentucki, I conducted a 3-year capture-recapture study and determined home-range characteristics and spatial relationships of individuals at two field sites that differed in predominant cover type and population density. Home ranges of adults were fixed and the home ranges of same-sex adult neighbors were mostly exclusive. The spatial arrangement of adult home ranges exhibited overall regularity or regularity within aggregations, whereas the distribution of juvenile home ranges was usually random. Analysis of nearest-neighbor sex indicated a positive intersexual association of adult home ranges. Removal studies provided evidence for defense of adult home ranges only at the high-density site. The distribution of home ranges was influenced by the presence of cover objects, but there was no significant relationship between adult body size and percent of home-range area with cover. Males overlapped the home ranges of gravid females significantly more often than those of non-gravid females, indicating that the distribution of gravid females had a strong influence on the distribution of male home ranges. In laboratory tests, increased male-male aggression during the breeding season suggests that males may compete for access to mates. At the high-density site, larger males may have benefited by having greater reproductive success than smaller males because they were more dominant and their home ranges overlapped a greater number of gravid-female home ranges. My results indicate that habitat structure and population density may influence the spatial organization and mating system of P. kentucki. Received: 26 May 1997 / Accepted: 9 October 1997     

Developmental morphology of the shoot in Weddellina squamulosa and implications for shoot evolution in the Podostemaceae

BACKGROUND AND AIMS: In angiosperms, the shoot apical meristem produces a shoot system composed of stems, leaves and axillary buds. Podostemoideae, one of three subfamilies of the river-weed family Podostemaceae, have a unique 'shoot' that lacks a shoot apical meristem and is composed only of leaves. Tristichoideae have been interpreted to have a shoot apical meristem, although its branching pattern is uncertain. The shoot developmental pattern in Weddellinoideae has not been investigated with a focus on the meristem. Weddellinoideae are in a phylogenetically key position to reveal the process of shoot evolution in Podostemaceae. METHODS: The shoot development of Weddellina squamulosa, the sole species of Weddellinoideae, was investigated using scanning electron microscopy and semi-thin serial sections. KEY RESULTS: The shoot of W. squamulosa has a tunica-corpus-organized apical meristem. It is determinate and successively initiates a new branch extra-axillarily at the base of an immediately older branch, resulting in a sympodial, approximately plane branching pattern. Large scaly leaves initiate acropetally on the flanks of the apical meristem, as is usual in angiosperms, whereas small scaly leaves scattered on the stem initiate basipetally in association with the elongation of internodes. CONCLUSIONS: Weddellinoideae, like Tristichoideae, have a shoot apical meristem, leading to the hypothesis that the meristem was lost in Podostemoideae. The patterns of leaf formation in Podostemoideae and shoot branching in Weddellinoideae are similar in that these organs arise at the bases of older organs. This similarity leads to another hypothesis that the 'branch' in Weddellinoideae (and possibly Tristichoideae) and the 'leaf' in Podostemoideae are comparable, and that the shoot apical meristem disappeared in the early evolution of Podostemaceae.