Reproductive interference between Rana dalmatina and Rana temporaria affects reproductive success in natural populations

Experimental evidence suggests that reproductive interference between heterospecifics can seriously affect individual fitness; support from field studies for such an effect has, however, remained scarce. We studied reproductive interference in 25 natural breeding ponds in an area where two ranid frogs, Rana dalmatina and Rana temporaria, co-occur. The breeding seasons of the two species usually overlap and males of both species are often found in amplexus with heterospecific females, even though matings between heterospecifics produce no viable offspring. We estimated species abundance ratios based on the number of clutches laid and evaluated fertilization success. In ponds with low spatial complexity and a species abundance ratio biased towards R. temporaria, the average fertilization success of R. dalmatina eggs decreased, while this relationship was not detectable in spatially more complex ponds. Fertilization success of R. temporaria did not decrease with increasing relative numbers of heterospecifics. This asymmetry in fitness effects of reproductive interference may be attributed to R. temporaria males being more competitive in scramble competition for females than R. dalmatina males. Our study is among the first to demonstrate that in natural breeding populations of vertebrates interference among heterospecifics has the potential to substantially lower reproductive success at the population level, which may in turn affect population dynamics.     

Laich-Fressen durch Kaulquappen als mögliche Ursache spezifischer Biotoppräferenzen und kurzer Laichzeiten bei europäischen Froschlurchen (Amphibia,Anura)

Zusammenfassung In einem Kunstweiher aufgezogene Kaulquappen von Rana temporaria fressen in der Versuchsanordnung von Abb. I den Laich folgender sympatrischer Anuren-Arten: Rana temporaria (Kannibalismus), Rana esculenta, Rana ridibunda, Bufo bufo, Bufo calamita, Bombina variegata und Hyla arborea (Laichräubern). — Daß die Kaulquappen der früh im Jahr laichenden Rana temporaria später gelegten arteigenen und artfremden Laich fressen, wirkt als Selektionsdruck in folgende Richtungen: Rana temporaria züchtet sich selbst eine kurze Laichzeit. Spät in seichtem Wasser laichende Arten werden im Biotop von Rana temporaria unterdrückt (Bombina variegata, Bufo calamita, Hyla arborea). Rana esculenta, Rana ridibunda und Bufo bufo sind dem Laichräubern durch Kaulquappen wenig ausgesetzt, weil sie in größeren Wassertiefen laichen. Zudem ist Bufo bufo ebenfalls ein Frühlaicher. Alytes obstetricans steht außer Konkurrenz, weil er die Eier nicht ins Wasser setzt. — Untersuchungen über die Biotoppräferenzen und Laichzeiten der betroffenen Arten bestätigen diese Interpretation.

---

Summary Tadpoles of Rana temporaria which have been raised in an artificial pond prey in the experimental situation shown in Fig. 1 upon the spawn of the following sympatric European Anura species: Rana temporaria (cannibalism), Rana esculenta, Rana ridibunda, Bufo bufo, Bufo calamita, Bombina variegata and Hyla arborea (predation). — Rana temporaria is an early breeder. Thus selection pressure acts in the following ways: Rana temporaria establishes for itself an explosive breeding pattern. Species which breed later in the year in shallow water are suppressed in the biotope of Rana temporaria and develop other biotope preferences (Bombina variegata, Bufo calamita, Hyla arborea). Rana esculenta, Rana ridibunda and Bufo bufo spawn in deeper water. Therefore they are scarcely exposed to the tadpoles of Rana temporaria. Moreover Bufo bufo, too, is an early breeder. Alytes obstetricans laying no spawn in the water is not endangered. — An analysis of biotope preferences and breeding times of the species involved confirms these interpretations.

---

---

Mit Unterstützung des Schweizerischen Nationalfonds zur Förderung der wissenschaftlichen Forschung.     

Freeze tolerance evolution among anurans: Frequency and timing of appearance

Despite numerous mechanistic studies on physiological responses supporting freeze tolerance in anurans, few have addressed the evolutionary significance of this trait. We thus investigated the phylogenetic relationships among anuran species whose freeze tolerance has been assessed and in combination with new data on freezing tolerance of two closely related species of the European brown frogs (Rana temporaria and Rana dalmatina). The species we studied exhibited short survival times in frozen state (around 8 h for both species). Phylogenetic analysis suggests that freeze tolerance evolved at least two times among Ranidae and one or two times among Hylidae and never in Bufonidae. Furthermore, in order to assess the timing of divergence of this character we used a relaxed molecular clock created, and found that the most recent separation between a freeze tolerant species and a freeze intolerant species dates from 15.9 ± 7.6 Myr (Rana arvalis and R. temporaria). The comparison between these two species thus represents the best current model to understand freeze tolerance evolution. Addressing the evolution of this trait with such large-scale approaches will not only improve our understanding of cold hardiness strategies, but might also create a framework guiding future comparative studies.     

Polymorphic microsatellite DNA loci identified in the common frog (Rana temporaria,Amphibia, Ranidae)

We developed seven microsatellite loci in the common frog, Rana temporaria. There were between 2 and 27 alleles per locus and the expected heterozygosities ranged from 0.28 to 0.96 in a sample of frogs collected in the French Alps. Adding these seven markers to the 15 previously available microsatellite loci for this species should facilitate studies of genetic structure of Rana temporaria populations at a fine geographical scale.     

Molecular phenotyping of maternally mediated parallel adaptive divergence within Rana arvalis and Rana temporaria

When similar selection acts on the same traits in multiple species or populations, parallel evolution can result in similar phenotypic changes, yet the underlying molecular architecture of parallel phenotypic divergence can be variable. Maternal effects can influence evolution at ecological timescales and facilitate local adaptation, but their contribution to parallel adaptive divergence is unclear. In this study, we (i) tested for variation in embryonic acid tolerance in a common garden experiment and (ii) used molecular phenotyping of egg coats to investigate the molecular basis of maternally mediated parallel adaptive divergence in two amphibian species (Rana arvalis and Rana temporaria). Our results on three R. arvalis and two R. temporaria populations show that adaptive divergence in embryonic acid tolerance is mediated via maternally derived egg coats in both species. We find extensive polymorphism in egg jelly coat glycoproteins within both species and that acid‐tolerant clutches have more negatively charged egg jelly – indicating that the glycosylation status of the jelly coat proteins is under divergent selection in acidified environments, likely due to its impact on jelly water balance. Overall, these data provide evidence for parallel mechanisms of adaptive divergence in two species. Our study highlights the importance of studying intraspecific molecular variation in egg coats and, specifically, their glycoproteins, to increase understanding of underlying forces maintaining variation in jelly coats.     

Karyotype analysis in Stylidium crossocephalum (Angiospermae:Stylidiaceae)

Qualitative studies of karyotypes were carried out on 79 plants of Stylidium crossocephalum from 28 collection sites. The sample included 7 distinct karyotypic classes which could be resolved into 5 clearly distinguishable haploid karyotypes or genomes. Three genomes, J, K and L, were localised in an area covering a southern portion of the species range while the other two genomes, A and B, were found in a region 90km to the north. Two different genome heterozygotes were found, one characterised by a J/K karyotype, the other by an A/B karyotype. —Karyotypic studies of a synthetic hybrid and its two parents showed that the chromosomes of the constituent genomes of the hybrid could be readily and confidently discerned in the karyotypes of the parents. These studies demonstrated that the identification of genomes by the qualitative attributes of their constituent chromosomes constitutes a reliable analytical technique. — Statistical analyses using arm ratio and percentage length measurements of chromosomes from plants allocated to the two groups of genomes confirmed the differences between marker chromosomes and demonstrated additional statistically significant differences among the non marker chromosomes. Some of these statistical differences may be attributable to technical problems inherent in the karyotyping procedures. In two instances, however, they could be convincingly argued as real, demonstrating a further component of chromosome variation in S. crossocephalum.     

A comparative study of physiological and structural changes at the myoneural junction in two species of frog after transection of the motor nerve

Summary Structural and functional behaviour of motor end-plates after transection of the motor nerve has been studied in two species of frog: Rana esculenta and Rana temporaria. The physiological results show that in both species there is a transient cessation of spontaneous activity followed by a resumption of miniature end-plate potentials (min. e.p.p.s.) after denervation. The characteristics of these potentials (frequency, distribution of amplitudes, time-course) are similar in the two species. However, some differences have been observed: Firstly, the period of silence lasts for 2–4 days in the case of Rana temporaria whereas it is prolonged to about 15 days in Rana esculenta. Secondly, the resumption of min. e.p.p.s. is gradual and after the 10th day of denervation remains constant in Rana temporaria. It is inconstant independent of the period of denervation in Rana esculenta. The morphological results show that the Schwann cell is constantly in contact with the post-synaptic membrane after about 6 days of denervation in both species. It is suggested that either the Schwann cell is capable of functioning for a limited period of time in Rana esculenta or is activated to produce min. e.p.p.s. only in certain cases.     

Zur Feinstruktur des Organon vasculosum hypothalami des Frosches (Rana temporaria)

Zusammenfassung Das Organon vasculosum hypothalami von Rana temporaria wurde licht-, fluoreszenz- und elektronenmikroskopisch untersucht. Es besteht aus einem kleinen Kerngebiet catecholaminhaltiger Nervenzellen, deren apikale Zellausläufer das Ependym durchbrechen und im Ventrikel ein Geflecht bilden.

---

The fine structure of the organon vasculosum hypothalami of the frog (Rana temporaria)

Summary The organon vasculosum hypothalami of Rana temporaria was investigated with the light-, fluorescence-, and electron microscope. It consists of a small nucleus of catecholamine containing nerve cells, the apical processes of which perforate the ependymal lining and form an intraventricular plexus.

---

---

Mit dankenswerter Unterstützung durch die Deutsche Forschungsgemeinschaft.     

Thermal influences on the habitat preference and the diurnal activity in three European Rana species

The diurnal behaviour of frogs was recorded quantitatively in a habitatlike experimental environment by a new method of automatic registration via thermocouples. Three species with different habitat preferences during their summer activity period were chosen: the terrestrial common frog Rana temporaria LINNÈ and the semiaquatic water frog Rana lessonae CAMERANO and Rana ridibunda PALLAS. The activity, the location and the skin temperatures of these frogs were recorded continuously in four different temperature ranges (8.1° C–31.0° C) within the temperature span of the summer activity period. The thermal requirements and behavioural adaptations to the habitat of each species were analysed. The experimental results coincided with field observations and showed some details not yet known. The northern species R. temporaria exhibited a distinctive behavioural thermoregulation including cooling behaviour at high temperatures and warming behaviour at low temperatures during the day. In all temperatures tested these frogs remained on land changing their preferred location from the open area to the hiding places. The main activity period shifted from night to day with decreasing temperatures. The more southern species R. lessonae preferred higher temperatures than the other species showing basking behaviour during the day independent of the ambient temperature. With decreasing temperatures the preferred location changed from the shore to the water and the activity during the night almost disappeared. The closely related R. ridibunda tolerated high temperatures too, but only rarely basked. This species, however, reacted contrarily to decreasing temperatures: it changed its preference from the water to the land. This behavioural differenciation between the water frog species may be of importance for the actual habitat preference of these central european water frogs.     

Dimorphic nucleolar organizer regions in the frog Rana blairi.

The nucleolar organizer-specific staining procedure, ammoniacal silver (Ag-AS), has been used to study the distribution and size of the nucleolar organizer regions (NORs) in chromosomes of the frog Rana blairi (Mecham, Littlejohn, Oldham, Brown and Brown). The somatic metaphase karyotype of this frog is similar to that of other frogs of the Rana pipiens species complex, numerically (2n=26) and morphologically. Secondary constrictions are detectable in untreated Giemsa-stained metaphase preparations as achromatic gaps in the long arms of a pair of submetacentric chromosomes (no. 10). These constrictions are the only regions which are deeply stained with the Ag-AS method and are thus identified as the nucleolar organizer regions (Ag-NORs). In each of the three individuals, the Ag-NORs as visualized on the homologues are of unequal length.     

The Chromosomes of some octodontids with special reference to Octodontomys (Rodentia; Hystricomorpha)

The chromosomes of three species (Octodontomys gliroides, Octodon degus and Ctenomys talarum) of octodontid hystricomorph rodents are compared. — The diploid numbers are 38, 58 and 48 respectively. No polymorphic chromosomes were noted in the specimens available. The sex chromosomes are heteromorphic and pair end to end in meiosis. — The karyotypes are compared on the basis of the extreme specialisation of the habitat of the three species. The asymmetric karyotype of Octodontomys can be compared more readily with that of a Ctenomys ancestor than with that of its present-day relative, Octodon.     

Chromosome evolution in Australian rodents

The chromosome complements of 188 specimens of 29 species of Australian murid rodents belonging to the subfamilies Pseudomyinae and Hydromyinae and the Uromys/Melomys group have been compared. At least one specimen of 18 different species was successfully C-banded. — The autosomal complements of many (9) diverse Pseudomyinae, one species of Melomys and one Hydromyinae proved to be identical, comprising 48 elements in the diploid set, the two smallest autosomal pairs of which are metacentric. No other karyotype is common to more than one species. From this we conclude that these three groups have been derived from a common ancestor which also possessed such a karyotype. The genus Zyzomys is exceptional since it possesses only 44 elements and lacks the two smallest metacentrics. — Karyotypic evolution within this apparently single phyletic line has been remarkably conservative, only three rearrangements being required to derive the most divergent karyotype. Moreover most of the observed rearrangements involve pericentric inversions and only one example of a fusion was found. Considerable differences in heterochromatin content, as determined by C-banding, occur between species however. Two species proved exceptional in this respect, namely Notomys cervinus and Uromys caudimaculatus. N. cervinus possesses numerous heterochromatic short arms. In U. caudimaculatus, there is a striking difference between northern and southern populations; in the former heterochromatin is present principally in the telomeric areas of the conventional A-chromosomes whereas in the latter it is found as separate supernumerary chromosomes. — In contrast to the autosomes, the X and Y chromosomes show high inter- and intra-specific variability in both size and morphology. All of this variability can be explained in terms of variation in heterochromatin content. Moreover the amount of heterochromatin in the X and Y chromosomes is highly correlated both within and between species. The Y chromosome of Uromys caudimaculatus is, however, distinctive in that it lacks C-banding.     

Comparative enzymological study of catalytic properties of liver monoamine oxidases in frogs

Comparative enzymological study of catalytical properties of monoamine oxidase (MAO) of liver of the marsh frog Rana ridibunda and common frog Rana temporaria has revealed certain features of similarity and differences between these enzymes. The MAOs from both studied biological sources show catalytic properties resembling those of the classical MAO of terrestrial vertebrates: they deaminate tyramine, tryptamine, serotonin, and benzylamine and do not deaminate histamine, have sensitivity to clorgyline, the specific inhibitor of the MAO A form, and deprenyl, the specific inhibitor of the MAO B form, and are not inhibited by 10⁻² M semicarbazide. Based on data of substrate-inhibitor analysis, a suggestion is put forward about the existence of two molecular forms of the enzyme in liver of the studied frog species. Quantitative interspecies differences have been revealed between liver MAO of Rana ridibunda and Rana temporaria in values of kinetic parameters of reactions of deamination of several substrates and in sensitivity to the inhibitors, deprenyl and clorgyline. In the species Rana temporaria the MAO activity in reaction of deamination of serotonin and benzylamine were virtually identical, whereas in the species Rana ridibunda these parameters for serotonin were almost one order higher than for benzylamine. In the species Rana ridibunda, selectivity of action of deprenyl was expressed many times weaker, while selectivity of the clorgyline—one order of magnitude stronger than in the species Rana temporaria. The catalytic activities towards all studied substrates of liver MAO of both studied amphibian species were several times lower as compared with the enzyme of rat liver.     

Le caryotype de l'Amphibien Urodèle Euproctus asper (Dugès)

Résumé Chez le Salamandridé Euproctus asper, le caryotype analysé à partir de testicules adultes et d'épiderme larvaire montre que le nombre chromosomique diploïde est de 24. Ces chromosomes se rangent en trois groupes principaux à l'intérieur desquels la plupart des éléments peuvent s'identifier individuellement grâce à leur indice centromérique.Le classement des chromosomes en trois groupes reste possible pour les figures haploïdes de 2^e division méïotique spermatocytaire. Dans ce dernier cas on peut encore, à l'intérieur de deux des groupes, identifier individuellement chacun des éléments.Il ne semble pas y avoir d'hétérochromosomes morphologiquement différenciés dans cette espèce.

---

The karyotype of Euproctus asper (Urodela, Salamandridae) was analyzed from squash preparations of adult testes and larval epidermis. The diploid chromosome number is 24. These chromosomes can be classified into three groups. Most of the elements included in these groups can be identified individually by means of their length and arm-ratio. The classification into three groups remains possible for second meiotic division chromosomes. In the latter case, it is still possible to identify individually each chromosome in two of the three groups. There is no evidence of heteromorphic pairs in the present species.

     

Über die Sauerstoffbindung durch Hämoglobin vom Frosch (Rana esculenta und Rana temporaria)

Zusammenfassung Die Sauerstoffdissoziationskurven vom Blute von Rana esculenta und Rana temporaria wurden bei 20° bei verschiedenen Kohlensäurespannungen bestimmt.Für Rana esculenta wurde obendrein die O₂-Bindung bei 12° untersucht.Die für diese Versuche benutzten Apparate konnten mit Hilfe einer Spende der Rockefeller Foundation angeschafft werden.     

Evidence for chromosome number reduction and chromosomal homosequentiality in the 24-chromosome Korean frog Rana dybowskii and related species

The karyotype of the Korean frog Rana dybowskii with its pattern of C-band heterochromatin distribution was numerically analyzed. There are 2n = 24 chromosomes in the karyotype representing a reduction in number from the typical 2n = 26 chromosome karyotype of Rana. The karyotype shows other evidence of reorganization relative to 26-chromosome species. The chromosomes grade smoothly in size from largest to smallest without the two size classes that are characteristic for 26-chromosome species. In contrast to many 26-chromosome species, there are few centromeric C-bands but many interstitial ones. C-bands for each homologous chromosome pair are distinctive. A prominent secondary constriction is located on one of the smallest chromosomes, chromosome 11, in a position similar to that seen in most 26-chromosome species. The karyotype of R. dybowskii is compared to those of other species of Rana known to have 2n = 24 chromosomes; it is most similar to that of R. chensinensis, less so that of R. ornativentris and less still to that of R. arvalis in terms of the positions of centromeres and secondary constrictions. C-bands as well as secondary constrictions in the karyotypes of these frogs show evidence of chromosomal homosequentiality. The process and possible consequences of chromosome number reduction from an ancestral 26-chromosome karyotype is also evident in the karyotypes of these closely allied palearctic frogs. Pericentric inversions followed by fusion of two small elements apparently produced a new chromosome, chromosome 6, occurring originally among northeast Asian populations.     

Apoptosis in larval and frog skin of Rana pipiens,R. catesbeiana,and Ceratophrys ornata

Apoptosis (programmed cell death) occurs during normal development of anurans in organs such as gills, gut, and tail. For example, apoptotic cells have been reported in the luminal epithelium along the length of the digestive tract of both larvae and frogs; however, timing of the peak number of such cells varies in different species. The purpose of the present study was to ascertain whether apoptosis also varies by species during metamorphic restructuring of the skin (as larval epithelium is replaced by adult epidermis). To determine this, cross‐sections of dorsal skin from representative larval stages and frogs of Rana pipiens, R. catesbeiana, and Ceratophrys ornata were incubated with monoclonal antibody against active caspase‐3, one of the main enzymes in the apoptotic cascade. We observed apoptotic cells in the epidermis of the skin of the three species and found that such cells were more numerous in larval stages than in frogs and more abundant in the two ranid species than in C. ornata. These results contribute to our understanding of metamorphic changes in anuran skin. J. Morphol. 275:51–56, 2014. © 2013 Wiley Periodicals, Inc.     

The late replication banding patterns of chromosomes are highly conserved in the genera Rana,Hyla, and Bufo (Amphibia: Anura)

Late replication banding and C-banding analyses were performed on the metaphase chromosomes of six species and one subspecies of Palearctic water frogs, genus Rana. Although C-banding patterns showed interspecific or intersubspecific variation, late replication banding patterns of all 13 chromosome pairs of these species were homologous. Minor differences of banding patterns were observed only in chromosomes 2, 7 and 13. Close comparison of the late replication banding patterns with those of three non-water frog species of Rana, and one each of Hyla and Bufo, provided important information on interspecific and intergeneric variability. In the Rana species, the banding patterns of all 13 pairs were homologous except for those some regions of 8 pairs. In one species each of Hyla and Bufo that was examined, the six large chromosome pairs (Nos. 1-6) showed banding homologies. Furthermore, among the Rana, Hyla and Bufo species the four large chromosome pairs (Nos. 1-3, 5 of Rana and Hyla, and Nos. 1, 3–5 of Bufo) shared banding homologies. These results show that the large chromosomes have been highly conserved in the evolutionary history of the three genera.     

Lipidkugeln im Plexusepithel von Rana temporaria L.

Zusammenfassung Das Plexusepithel von Rana temporaria L. enthält zwei verschiedene Typen von Lipidkugeln, die einen Durchmesser bis zu 8 erreichen können. Diese Einschlüsse haben keine Zytomembran. Die Struktur der Einschlußkörper wird mit Hinweisen auf die Fixierung und den Stoffwechsel diskutiert.

---

Lipid droplets in the epithelium of the choroid plexus of Rana temporaria L.

Summary The epithelium of the choroid plexus of Rana temporaria L. contains two different types of spherical lipid inclusions up to 8 in diameter. The droplets do not have a limiting membrane. The preservation and metabolism of the lipid bodies are discussed.

Mit Unterstützung durch die Deutsche Forschungsgemeinschaft.     

KARYOTYPIC STUDIES IN SOLANUM SECTION BASARTHRUM (SOLANACEAE)

Mitotic chromosomes of 450 individuals, representing 59 accessions of 18 of the 22 species of Solanum sect. Basarthrum were studied. Statistical analyses of chromosome length, genome length, and centromere position yielded estimates of karyotype composition and asymmetry. A generalized karyotype of the section shows that most of the chromosomes are metacentric (44%) or submetacentric (53%). A general trend emerged: the greater the total genome length, the more asymmetric the karyotype. However, karyotype asymmetry does not seem to be based on major chromosomal rearrangements. There is variation in species groups in the karyotype formula, and chromosome length differences were useful in distinguishing series. Patterns of cytological variation reinforce the systematic arrangement of the taxa based on morphological, chemical, and molecular studies. Chromosomal differences characterize most of the species. Five species bear subtelocentric chromosomes, two have telocentrics, and two have satellites. The genomes of the staminate and pistillate plants of the dioecious S. appendiculatum are identical. Of the species proposed as progenitors of the domesticated cultigen S. muricatum, S. caripense is the most similar karyotypically.