Food supply modifies the trade-off between past and future reproduction in a sexual parasite–host system (<Emphasis Type="Italic">Rana esculenta,Rana lessonae</Emphasis>)

Life history theory is concerned with the costs of survival, growth and reproduction under different ecological conditions and the allocation of resources to meet these costs. Typical approaches used to address these topics include manipulation of food resources, followed by measures of subsequent reproductive traits, and measures of the relationship between current and future reproductive investment. Rarely, however, do studies test for the interaction of past investment, present resource availability and future investment simultaneously. Here, we investigate this interaction in females of a sexual parasite–host system consisting of the hybridogenetic frog Rana esculenta (E) and one of its parental species Rana lessonae (L). We kept females from each of two groups (with or without previous reproduction) under two food treatments (low or high) and regularly recorded their growth as well as their body condition and hormone titres as measures of future reproductive condition. After keeping them in hibernation until the following spring, we exposed the females to males, recorded whether they spawned or not and related this response to their condition in the previous autumn. Past reproduction negatively affected growth during summer and condition during autumn which, in turn, reduced the following year’s reproductive output. These costs of previous reproduction were less pronounced under the high than under the low food treatment and lower in R. lessonae than in R. esculenta. Increasing food supply improved reproductive condition more in L than in E females. These species differences in reproductive costs and food requirements provide a mechanistic explanation for why E females skip annual reproduction almost twice as often as L females. Since R. esculenta is a sexual parasite that depends on R. lessonae for successful reproduction, these species-specific life history patterns not only affect individual fitness but also the spatial structure and temporal dynamics of mixed LE populations.     

ASYMMETRIC COMPETITION IN MIXED POPULATIONS OF TADPOLES OF THE HYBRIDOGENETIC: RANA ESCULENTA COMPLEX

Hybridogenetic Rana esculenta tadpoles display tolerance to extreme environmental conditions and fit criteria of the “general-purpose” genotype. A trade-off between generality and competitive ability is assumed to occur in asexual species, but the evidence remains unclear. The purpose of my experiment was to test the competitive ability of hemiclonal hybrid Rana esculenta tadpoles relative to the parental species Rana lessonae. Mixed and single genotype populations of R. esculenta and R. lessonae tadpoles were reared at three densities in artificial ponds. Survival of R. esculenta was higher than for R. lessonae tadpoles, but did not differ among densities. Body size at metamorphosis was the same between genotypes, but decreased with increasing density. Larval period was not affected by density, but R. esculenta tended to metamorphose earlier than R. lessonae. Percentage of individuals metamorphosing was higher for R. esculenta at both medium and high densities, but the same as R. lessonae at the low density. The difference in survival, body size, and larval period between tadpoles reared in single and mixed genotype populations was unaffected by genotype or density. The difference in the percentage of metamorphs, however, was strongly affected. The percentage of hybrids metamorphosing was 9% above the responses of single genotype populations at the highest density. Conversely, the percentage of R. lessonae metamorphosing was 12% below the responses of single genotype populations at the same density. Hybrid success in this experiment further supports the criterion of a “general-purpose” genotype without assumptions of reduced competitive ability.     

ADAPTIVE GENETIC VARIATION IN GROWTH AND DEVELOPMENT OF TADPOLES OF THE HYBRIDOGENETIC RANA ESCULENTA COMPLEX

The distribution and proportion of the sexual species Rana lessonae to the hemiclonal hybrid R. esculenta among natural habitats suggests that these anurans may differ in adaptive abilities. I used a half-sib design to partition phenotypic and quantitative genetic variation in tadpole responses at two food levels into causal variance components. Rana lessonae displays strong phenotypic variation across food levels. Growth rate is strictly determined by environmental factors and includes weak maternal effects. Larval period and body size at metamorphosis both contain moderate levels of additive genetic variance. The sire x food interactions and the lack of environmental correlations indicate that adaptive phenotypic plasticity is present in both of these traits. In contrast, R. esculenta displays less phenotypic variation across food levels, especially for larval period. Variation in body size at metamorphosis is underlain by genetic variation as shown by high levels of additive genetic variance, yet growth rate and larval period are not. Significant environmental correlations between larval period at high food level and growth, larval period, and body size at low food, indicate phenotypic plasticity is absent. A positive phenotypic correlation between body size at metamorphosis and larval period for R. lessonae at both food levels suggests a trade-off between growing large and metamorphosing quickly to escape predation or pond drying. The lack of a similar correlation for R. esculenta at the high food level suggests it may be less constrained. Different levels of adaptive genetic variation among larval traits suggest that the sexual species and the hybridogenetic hemiclone differ in their abilities to cope with temporally and spatially heterogeneous environments.     

Ecology of the Hybridogenetic Rana esculenta Complex: Differential Oxygen Requirements of Tadpoles

Because of intrinsic demographic load induced by hybridogenesis (infertility of homotypic hybrid matings), the maintenance of hybrid lineages supposes that they present better performances (heterosis) than their host species which allows them to coexist on a long-term basis. However, this necessity of high fitness can be relaxed if a relative niche partitioning occurs between the taxa, each of them differing in their ecological optima. In the waterfrog hybridogenetic complex (Rana esculenta complex), recent studies have revealed that hybrids show intermediate distribution between parental species across a gradient of river influence (that is related to a gradient of oxygen levels), and intermediate performances of their tadpoles with regard to oxygen availability (hypoxia). In investigating oxygen consumption rates, survival time in anoxia, and metabolite contents in the three forms of the complex, the present study confirms intermediate characteristics of hybrid tadpoles (R. esculenta) when compared to both parental lineages (R. lessonae and R. ridibunda). Whereas R. ridibunda tadpoles were the most sensitive to anoxia, R. lessonae tadpoles were the most tolerant. Because oxygen requirements of the hybrid proved to be intermediate, no heterosis was detected. These results confirm the hypothesis of the intermediate niche hypothesis to explain the coexistence of R. lessonae and R. esculenta and the success of the hybridogens.     

Morphometric and DNA investigations into European water frogs (Rana kl. esculenta Synklepton (Anura, Ranidae)) from different population systems

The population specific variability of diploid and triploid R. kl. esculenta individuals was investigated by means of morphometric methods (canonical discriminant analysis, UPGMA cluster analysis) and DNA fingerprinting. As a result of the morphometric investigations, as well as of the DNA investigations, a clear separation of single populations was possible. However, no correlations between the morphometry and different population systems could be recognized. Clear morphometric differences could be seen between diploid ♀♀ and ♂♂ and triploid ♀♀ on the one hand, and triploid ♂♂ on the other. While the diploid ♀♀ and ♂♂ and the triploid ♀♀ were located in the intermediate area between the parental species R. lessonae and R. ridibunda according to their morphometric parameters, the triploid ♂♂ showed a great overlap with R. lessonae. Up to now, this phenomenon has not been explained. The first results of the DNA investigations provided further hints at the high inter-individual and population-specific variability of R. kl. esculenta. R. kl. esculenta individuals of the R. lessonae/esculenta population Toter See could be distinguished from conspecific individuals of the R. ridibunda/esculenta-♀♀ population Alte Oder according to their fingerprint patterns. Moreover, in the R. lessonae / esculenta population, the fingerprints or the diploid R. kl. esculenta-♀♀ and the investigated R. lessonae-♀ were very similar. Furthermore, in this population, many R. kl. esculenta genotypes resemble R. lessonae in their morphometric features. This finding suggests the occurrence of recombination in R. kl. esculenta. In general, every population seems to have its own genetic background. A classification of water-frog populations according to population systems is only possible under certain conditions.     

A genetic mechanism of species replacement in European waterfrogs?

Introduced Rana ridibunda currentlyreplace the native waterfrogs R. lessonaeand R. esculenta in several areas ofcentral Europe. The unusual reproductive systemin waterfrogs of the Rana esculentacomplex suggests that this replacement may bedriven by a genetic mechanism: Ranaesculenta, a hybrid between R. ridibundaand R. lessonae, eliminates the lessonae genome from the germline and clonallytransmits the ridibunda genome(hybridogenesis). Hybrids form mixedpopulations with R. lessonae (L-E-system)in which they persist by backcrossing with theparental species. Matings between hybrids areunsuccessful, because their ridibundagenomes contain fixed recessive deleteriousmutations. When introduced into a L-E-system,R. ridibunda can mate with both nativetaxa, producing R. ridibunda offspringwith R. esculenta, and R. esculentaoffspring with R. lessonae (primaryhybridizations). If primary hybrids arehybridogenetic, they produce viable R.ridibunda offspring in matings with otherhybrids, because their clonal genomes areunlikely to share the deleterious allelespresent in the ancient clones. Thus, R.ridibunda will increase in the population atthe expense of both native taxa, eventuallyleaving a pure R. ridibunda population.We provide three lines of evidence for thisprocess from a currently invaded population inSwitzerland: (1) Primary hybridizations takeplace, as roughly 10% of hybrids in thepopulation possess ridibunda genomesderived from the introduced frogs. (2)Hybridogenesis occurs in primary hybrids,although at a low frequency. (3) Many hybrid ×hybrid matings in the population indeed produceviable offspring. Hence, the proposed geneticmechanism appears to contribute to the speciesreplacement, although its importance may belimited.     

Patterns of protein synthesis in oocytes and early embryos of Rana esculenta complex

Summary We have used isotopic labelling and both one-and two-dimensional electrophoretic procedures to analyse the protien synthesis patterns in oocytes and early embryos of three phenotypes of the European green frogs. The results demonstrated that protein patterns of Rana ridibunda and R. esculenta are identical, but that they differ from those of R. lessonae. Progeny of the lethal cross R. esculenta × R. esculenta showed a distinct delay in the appearance of stage-specific proteins during early embryogenesis. The heat-shock response of R. ridibunda and R. esculenta oocytes was found to be identical, but different from that of Xenopus laevis. The implications of these findings, with respect to hybridogenesis in R. esculenta complex and variations in the regulations of heat shock genes in different amphibian species, are discussed.     

Premeiotic chromosome doubling after genome elimination during spermatogenesis of the species hybrid Rana esculenta

Summary Gamete production in the hybridogenetic species hybrid Rana esculenta (Rana ridibunda X Rana lessonae) is preceded by a premeiotic elimination of the R. lessonae genome and subsequent duplication of the remaining R. ridibunda genome, so that only ridibunda chromosomes enter a quasi normal meiosis, and only ridibunda gametes are formed. This is demonstrated by differences in genome specific centromere fluorescence and electrophoretic patterns between somatic and gonadal tissue.     

Hybrid zones between two genetically differentiated forms of the pond frog Rana lessonae in southern Italy

Two distinct population groups of the pond frog Rana lessonae were detected in peninsular Italy and Sicily by multilocus electrophoresis: one group inhabits the peninsula down to northern Calabria, the second occurs in southern Calabria and on Sicily. Fixed alternative alleles distinguish the two groups at 5 of the 25 loci examined; marked allele frequency differences were observed at two additional loci. On average, the two groups differ by a Nei's standard genetic distance of 0.4. A wide hybrid zone (about 120 km) occurs between the two groups, with high genotypic diversity and absence of pure parental genotypes in central Calabria. Patterns of allozyme variation suggest that at least two distinct contact and hybridization events occurred, one in the Catanzaro, the other in the Crati-Sibari plains, about 70 km to the north. Geological evidence indicates that these areas correspond to two main marine-flooded grabens that would have repeatedly interrupted or reduced genetic exchange during Plio-Pleistocene times. The finding of a fixed difference at the Mdhp-1 locus between Sicilian and Calabrian R. lessonae witnesses their continuing differentiation following their last separation by definitive opening of the Strait of Messina, about 50 000 years ago. The wide hybrid zone, the diversity of genotypes and the agreement with Hardy-Weinberg expectations suggest complete hybrid fertility. Different patterns of introgression were observed at the various loci. The pattern of allelic variation at loci in R. lessonae is paralleled by the pattern of variation in lessonae genomes of the sympatric hemiclonal hybrid Rana esculenta, into which lessonae genomes are introduced each generation as a result of hybridogenesis.     

Seasonal variation in use of winter roosts by five bat species in south-east Finland

We studied seasonal variation in the use of winter roosts by five bat species (Eptesicus nilssonii, Myotis brandtii/mystacinus, Myotis daubentonii and Plecotus auritus) in south-east Finland during the winters of 2003/2004 and 2004/2005. At the beginning of the bat hibernation season all species used higher temperatures and humidity than by the season’s end. Hibernacula were at their coldest in mid-hibernation season and became warmer towards the end of the season. However, no species hibernated in warmer locations at the end of the season than in mid-season. Results suggest that bats tend to use different strategies throughout the hibernation season, minimizing the cost of hibernation early in the season by hibernating in warmer locations and minimizing energy expenditure later in the season by hibernating in colder locations. M. brandtii/mystacinus were found in locations with stable temperature and humidity, moving to increasingly stable conditions (chambers, crevices, clusters, ceiling) towards spring. All other species hibernated in more variable microclimates throughout the hibernation season.     

Genetic compatibility between sexual and clonal genomes in local populations of the hybridogeneticRana esculenta complex

Summary Hybridogenetic species possess a hybrid genome: half is clonally inherited (hemiclonal reproduction) while the other half is obtained each generation by sexual reproduction with a parental species. We addressed the question of whether different hemiclones of the hybridogenetic water frogRana esculenta are locally adapted for genetic compatibility with their sexual parental hostRana lessonae. We artificially crossedR. esculenta females of three hemiclones (GUT1, GUT2 and GUT3) from a pond near Gütighausen, Switzerland and one hemiclone (HEL1) from near Hellberg, Switzerland each toR. lessonae males from both populations. We also created primary hybrids by crossing the sameR. lessonae males from both populations toR. ridibunda females from Pozna, Poland (POZ). Tadpoles were then reared in the laboratory at two food levels to assess their performance related to early larval growth rate, body size at metamorphosis and length of the larval period. Tadpoles from hemiclones GUT1, GUT3 and POZ had higher growth rates than those from hemiclones GUT2 and HEL1 at the low food level, but at the high food level all growth rates were higher and diverged significantly between hemiclones GUT2 and HEL1. Tadpoles from the intrapopulational crosses GUT2 × GUT and HEL1 × HEL were larger at metamorphosis than those from the interpopulational crosses GUT2 × HEL and HEL1 × GUT. A high food level increased the size at metamorphosis in all tadpoles. A high food level also decreased the days to metamorphosis and tadpoles from GUT1, GUT3 and POZ had the shortest larval period whereas those from GUT2 and HEL1 had the longest. These results indicate that the differential compatibility of clonal genomes may play an important role in hybridogenetic species successfully using locally adapted sexual genomes of parental species and that interclonal selection is likely important in determining the distribution of hemiclones among local populations.     

Distribution and habitat use of water frog hybrid complexes in France

Hybrid zones are either distributed along clines or in a mosaic of patches. This distribution may depend upon variation in taxon habitat use. Habitat use and distribution of diverse taxa of water frogs (Rana ridibunda, R. lessonae, R. perezi, R. kl. grafi and R. kl. esculenta) in France are analysed to determine whether water frog complexes conform to the mosaic or clinal model. Biogeographical scenarios may be invoked in order to explain the distribution of water frogs. However, the distribution of R. perezi and R. kl. grafi, being restricted to regions characterized by Mediterranean or Oceanic climatic conditions, suggests that these frogs do not endure cold winters. R. ridibunda is widespread in Southern France and its distribution suggests multiple introductions. It is concluded that water frogs conform to the mosaic zone model rather than to the tension zone model because: (i) taxa exhibited differences in habitat use, (ii) pure parental species were documented and (iii) hybrids are not unfit relative to parental species.     

Karyological and serological studies in Rana lessonae,R. ridibunda and in their hybrid R. ‘esculenta’ (Amphibia,Anura)

The karyotypes of the three water frog forms: Rana lessonae, R. ridibunda and R. esculenta were analysed from bone marrow cell preparations of animals captured in several localities of the GDR. In the three forms chromosome morphology was similar (5 large and 8 small pairs), although differences in the relative length of most elements were found; R. esculenta chromosomes were always intermediate.One of the small pairs (Chr. 12) was found to be metacentric in R. lessonae and submetacentric in R. ridibunda. Most R. esculenta individuals examined had one meta-and one submetacentric 12th element, indicating the hybrid nature of this form. However 16.6% esculenta proved to be homozygous for either the metacentric or the submetacentric chromosome 12, while 13% lessonae individuals and 7.7% ridibunda were heterozygous for this element.By starch gel electrophoresis an analysis was undertaken of serum proteins from water frogs coming from regions in which the forms occur together (sympatric populations) and from zones in which only one of them lives (allopatric populations).In Rana lessonae, where only one allele had been previously described, two different alleles were found in animals coming from the GDR.If genetic polymorphism is excluded, between 6.1% and 9.1% individuals from sympatric lessonae and ridibunda populations showed introgression of an albumin allele. No gene introgression was found in allopatric lessonae populations from the Leningrad region or in ridibunda from Alma Ata and southern Bulgaria.     

Variation in Sex Ratio and Evolutionary Rate of Hemiclonal Rana esculenta Populations

In many plant and animal taxa mutation rates are higher in males than in females. As a result, the evolutionary speed of genes depends on how much time they spend in either sex. Usually, this time differs between genes located on sex chromosomes but not between those on autosomes. Here we present an unusual system with a partially sex-linked inheritance of autosomes: the hemiclonal frog Rana esculenta (E) which is originally a hybrid between the sexual species R. lessonae (L) and R. ridibunda (R). Rana esculenta excludes the L genome prior to meiosis, produces eggs or sperm containing an unrecombined R genome and restores hybridity by mating with R. lessonae (‘hybridogenesis’). Matings between L males and E females result in offspring with an even sex ratio, whereas the reverse combination produces only daughters. The extent of the resulting female bias and the proportion that R alleles have spent in either sex depend on the relative survival (b) and the relative reproductive contribution (a) of E males vs. E females. In this paper, we analyze mathematically how different combinations of a and b influence the sex ratio in R. esculenta populations and, combined with the male/female mutation rate ratio (α), the evolutionary rate of the clonally transmitted R genome. We find that this rate is higher than in an asexual population and lower than in a sexual one. Hence, clonal diversity through new mutations is more easily achievable than in purely asexual species. In contrast, the occurrence and accumulation of deleterious mutations is lower than in a comparable sexual species. We conclude that these intermediate mutation rates improve the ecological and evolutionary potential of hemiclonal organisms, and we draw attention to the implications for the use of microsatellites. Co-ordinating editor: L. Hurst     

Variation of plastic responses to oxygen availability within the hybridogenetic Rana esculenta complex

The hypothesis of superiority of hybrid phenotype to explain the maintenance of hybridogenesis (exclusion of one parental genome in the germline before meiosis) was investigated in the Rana esculenta complex. Survival and growth of progenies from each parental and hybrid lineages were compared across contrasted conditions of oxygen availability (this factor is suspected to influence habitat utilization of each form of the complex). Whereas growth and development were strongly affected by constant hypoxic conditions in the R. ridibunda lineage, they were not altered in the R. lessonae lineage. Because the performance of the hybrid (R. esculenta) lineage proved to be intermediate between the parental species, no heterosis effect was detected. These results supported the intermediate niche hypothesis as an alternative to the heterotic hybrid superiority hypothesis. The ecological and evolutionary implications are then discussed.     

Heat resistance of the skeletal muscle in Western Palearctic green frogs (<Emphasis Type="Italic">Rana esculenta</Emphasis> complex)

Heat resistance of the gastrocnemius muscle was studied in five species of the Rana esculenta complex. It was similar in R. bedriagae, R. lessonae, and in the European form of R. ridibunda; while North African R. saharica demonstrated a lower heat resistance. No heterosis was expressed in R. esculenta, a clonal hybrid of R. lessonae and R. ridibunda, for the heat resistance of the muscle. Moreover, this species demonstrated low heat resistance at the highest test temperature (42°C). Comparison of diploid and triploid R. esculenta syntopically occurring in the same water bodies demonstrated no differences between them, thus, suggesting that polyploidy has no effect on this parameter at least in this case.     

Mating calls of water frogs (Ranidae) from Lake Skutari,Yugoslavia, and the relationship to water frogs of other regions1

According to the results of a bioacoustic analysis of the mating calls recorded at the northern end of Lake Skutari, Yugoslavia, this region is inhabited by three water frog phenotypes: Rana ridibunda, R. lessonae and hybrids of these two species. The mating call of R. ridibunda consists of an average of 3.55 pulse groups at all water temperatures. Other parameters of the call, however, are greatly influenced by temperature. The conclusion that the second phenotype is the species R. lessonae was based on the characteristics of the mating call. At a water temperature of 20°C, a call consists of 29 pulse groups repeated at a rate of 24 Hz. Analysis revealed almost complete agreement between this mating call and those of Central and Eastern European R. lessonae. The systematic implications of this finding are discussed. The mating calls of the hybrids of R. ridibunda and R. lessonae are intermediate between those of the parents.     

Vocal response of male European water frogs (Rana Esculenta complex) to mating and territorial calls

The responses of male European water frogs (the two species Rana lessonae and Rana ridibunda and their hybrid, Rana esculenta) to playback of their mating and territorial calls were studied during the mating season.In order to select biologically relevant intensities for the presentation of the recorded calls, the sound pressure of the calls produced by the frogs themselves was established prior to the experiment. At a distance of 1 m the most intense calls were those of R. ridibunda, with a sound pressure of 110 dB (peak SPL). The smaller males of R. esculenta gave calls about 5 dB lower in intensity. The calls of R. lessonae, the smallest phenotype, were still less intense, 10 dB lower than those of R. ridibunda.The territorial calls of all three phenotypes elicited territorial calls in all of the males tested, as a rule accompanied by approach to the sound source. The sound pressure required to elicit a vocal response was nearly the same for each of the three different territorial calls. Sometimes encounter calls and warning calls were given in addition to territorial calls.When the mating calls were presented at low intensity, in some cases the males responded with their own mating calls. Mating calls at higher intensity elicited the same behavior that appeared following presentation of territorial calls, but significantly higher sound pressures were required to elicit such a response to mating calls than to territorial calls. The males of R. lessonae and R. esculenta did not respond to the mating calls of R. ridibunda, and each of them had significantly lower thresholds to the mating call of its own phenotype than to that of the other. The males of R. ridibunda responded only to conspecific mating calls.The vocal-response thresholds are compared with those of the electrodermal response reacting to the same stimuli. The significance of the different calls of the European water frogs is discussed.     

Behavioural response and winter survival of mated and unmated diapausing females of the Tetranychus atlanticus-urticae complex (Acari, Tetranychidae)

Behavioural responses were studied in mated and unmated females of a Dutch strain of the spider mite Tetranychus urticae which were destined to enter diapause. Observations were confined to the period between acquirement of the winter coloration by the females (a few days after the last moult) and their entering of artificial hibernation sites offered to the mites on the host leaves. Unmated females were found to stay longer on the leaf surface than mated females; they entered the hibernation sites significantly later than mated females. The delay in the search for hibernation sites shown by unmated females may be seen as a behavioural adaptation to enhance the chance of being fertilized before hibernation. Winter survival of mated and unmated diapausing females of the same strain of mites was studied both in the laboratory at a constant temperature of 2±1°C and outdoors under natural climatic conditions in Amsterdam during the winter of 1990–1991. Survival was high under both conditions for mated as well as unmated females; no significant differences in survival were found between both types of female. Observations on post-diapause females of Tetranychus atlanticus (a mite belonging to the T. urticae complex) sampled from strawberry fields near Moscow in spring, showed that at most 10% of the females of this natural spider mite population were unmated. Both mated and unmated females had survived winter temperatures of -28 to -30°C.     

Winter diet analysis in Rhinolophus euryale (Chiroptera)

We investigated the winter food of Mediterranean horseshoe bats (Rhinolophus euryale) in four winter cave roosts in southern Slovakia and northern Hungary and investigated the relationship between food and ambient temperature. The bats were active during the whole winter period and they produced excrement throughout the entire hibernation period, even when outside temperatures dropped below zero. The guano was in two forms, containing (1) prey items and (2) non-prey items. The identifiable items belonged to lepidopteran species, but only one was identified, on the basis of the genital fragments, the moth Colotois pennaria, which was the main prey species in autumn and early winter. Our results shed light on the extraordinarily high level of activity in this bat species during winter hibernation, which in temperate regions is a strategy that enables bats to survive when prey is reduced or absent. In R. euryale, the torpor in the course of hibernation is not continuous and our results help to explain how energy losses caused by bat movements are covered.