COMPETITION AMONG TADPOLES OF COEXISTING HEMICLONES OF HYBRIDOGENETIC RANA ESCULENTA: SUPPORT FOR THE FROZEN NICHE VARIATION MODEL

Vertebrate animals reproducing without genetic recombination typically are hybrids, which have large ranges, are locally abundant, and live in disturbed or harsh habitats. This holds for the hemiclonal hybridogenetic frog Rana esculenta: it is widespread in Europe and commonly is found in disturbed habitats such as gravel pits. We hypothesize that its widespread occurrence may either be the result of natural selection for a single hemiclone acting as a broadly adapted “general-purpose” genotype, or of interclonal selection, which maintains multiple hemiclones that each are relatively narrowly adapted and perform differently across environments, that is, the Frozen Niche Variation model. We tested these competing hypotheses using 1000-L outdoor artificial ponds to rear tadpoles of the parental species (Rana lessonae [LL] and Rana ridibunda [RR]) alone, and each of three hemiclones of Rana esculenta (GUT1, GUT2, GUT3) alone, and in mixed hemiclonal populations from hatching to metamorphosis. Tadpoles of three coexisting hemiclones from a single natural population (near Gütighausen, Switzerland) were reared in both two- and three-way mixtures in equal total numbers at high and low density. For each species and hemiclone, the proportion of tadpoles metamorphosing decreased as the density of tadpoles increased, with the three hemiclones spanning the range of values exhibited by the two parental species. LL and GUT1 tadpoles produced the highest proportion of metamorphs, whereas tadpoles of RR produced the fewest metamorphs at both densities. GUT1 tadpoles also produced the largest metamorphs at low density, GUT2 and GUT3 tadpoles produced smaller metamorphs than did GUT1 tadpoles at the low density, but the three hemiclones did not differ from each other at high density. The parental species (LL and RR) were intermediate in metamorphic size to the hemiclones at low density, but all genotypes converged on a similar size at high density. Length of the larval period also was affected by density, but its effect was dependent on genotype. GUT1 tadpoles had the shortest larval period at the low density, but larval period was longer and not different between GUT1, GUT3, and LL at high density. RR tadpoles had the longest larval period at both densities. The most dramatic results were that three genotypes (GUT1, GUT2, and RR) maintained rank order and increased days to metamorphosis from low to high density, whereas two genotypes (GUT3 and LL) changed rank order and decreased days to metamorphosis from low to high density. Mixtures of hemiclones in two- and three-way combinations facilitated the proportion of tadpoles metamorphosing for GUT1 and GUT2 at both densities, but only at the low density for GUT3 tadpoles. Results from this experiment are incompatible with the General-Purpose Genotype model as a global explanation of hybrid abundance in these frogs. Alternatively, the Frozen Niche Variation prediction of general performance superiority of clonal mixtures relative to single clone populations is strongly supported. The data confirm that fitness advantages of hemiclones change, depending on the environment, such that in temporally and spatially heterogeneous habitats like ponds, frequency-dependent selection among hemiclones may promote coexistence in hemiclonal assemblages. Yet, differential dispersal or colonization ability and historical factors affecting hemiclone distribution may also be important in shaping patterns of clonal coexistence.     

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.     

FIXATION OF DELETERIOUS MUTATIONS IN CLONAL LINEAGES: EVIDENCE FROM HYBRIDOGENETIC FROGS

Abstract The hemiclonal waterfrog Rana esculenta (RL genotype), a bisexual hybrid between R. ridibunda (RR) and R. lessonae (LL), eliminates the L genome from its germline and clonally transmits the R genome (hybridogenesis). Matings between hybrids produce R. ridibunda offspring, but they generally die at an early larval stage. Mortality may be due to fixed recessive deleterious mutations in the clonally inherited R genomes that were either acquired through the advance of Muller's ratchet or else frozen in these genomes at hemiclone formation. From this hypothesis results a straightforward prediction: Matings between different hemiclones, that is, between R. esculenta possessing different R genomes of independent origin, should produce viable R. ridibunda offspring because it is unlikely that different clonal lineages have become fixed for the same mutations. I tested this prediction by comparing survival and larval performance of tadpoles from within‐ and between‐population crossings using R. esculenta from Seseglio (Se) in southern, Alpnach (Al) in central, and Elliker Auen (El) in northern Switzerland, respectively. Se is isolated from the other populations by the Alps. Enzyme electrophoresis revealed that parents from Se belonged to a single hemiclone that was different from all hemiclones found north of the Alps. Parents from Al also belonged to one hemiclone, but parents from El belonged to three hemiclones, one of which was indistinguishable from the one in Al. Rana esculenta from Se produced inviable tadpoles when crossed with other hybrids of their own population, but when crossed with R. esculenta from Al and El, tadpoles successfully completed metamorphosis, supporting the hypothesis I tested. Within‐population crosses from Al were also inviable, but some within‐population crosses from El, where three hemiclones were present, produced viable offspring. Only part of the crosses between Al and El were viable, but there was no consistent relationship between hemiclone combination and tadpole survival. When backcrossed with the parental species R. ridibunda, hybrids from all source populations produced viable offspring. Performance of these tadpoles with a sexual and a clonal genome was comparable to that of normal, sexually produced R. ridibunda tadpoles. Thus, in the heterozygous state, the deleterious mutations on the clonal R genomes did not appear to reduce tadpole fitness.     

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.     

Feeding behaviour, food consumption, and growth efficiency of hemiclonal and parental tadpoles of the Rana esculenta complex

1. Clonally reproducing species are often assumed to lack sufficient genetic variability to evolve specific local adaptations to cope with environmental perturbation and competition from sexual species. Yet, many asexuals are extremely successful judged by abundance and wide range, suggesting high competitive abilities in resource exploitation.
2. In this study, food use and its effects on larval growth in a water frog system consisting of the two parental sexual species, Rana lessonae (Camerano 1882) and Rana ridibunda (Pallas 1771), and three different coexisting hemiclones of their hybrid, Rana esculenta (Linnaeus 1758) were investigated.
3. R. esculenta tadpoles spent 18·6% more time feeding than did tadpoles of either parental species, but feeding time was not affected by interspecific mixture.
4. R. esculenta tadpoles consumed 50·8% more food over the whole test period than did tadpoles of the two parental species.
5. R. esculenta tadpoles exhibited higher growth rates than did tadpoles of either parental species.
6. R. lessonae tadpoles had the highest and R . ridibunda tadpoles the lowest growth efficiencies with the R. esculenta tadpoles ranging between the two parentals.
7. The results obtained indicate that hemiclonal hybridogenetic R . esculenta tadpoles display significant phenotypic variation among coexisting hemiclones as well as out-perform tadpoles of the parental sexual species R. lessonae and R . ridibunda. The primary mechanism for success of the hybrid tadpoles is probably behavioural, through increased feeding time and food consumption, and not physiological via growth efficiency.     

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.     

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.     

PERFORMANCE OF TADPOLES FROM THE HYBRIDOGENETIC RANA ESCULENTA COMPLEX: INTERACTIONS WITH POND DRYING AND INTERSPECIFIC COMPETITION

The performance of three genotypes (LL, LR, RR) of tadpoles resulting from the hybrid mating system of Rana lessonae (phenotype L, genotype LL) and Rana esculenta (phenotype E, genotype LR) was determined in artificial ponds. The effects of interspecific competition and pond drying on growth, development, and survival of tadpoles were used to measure the performance of genotypes and the relative fitness of offspring. Among the three genotypes, tadpoles from the homogametic mating RR had the lowest survival, growth, and development under all environmental conditions. Body size of the LL and LR genotype tadpoles at metamorphosis was reduced by competition and pond drying. Days to metamorphosis were also higher for the LL and LR genotype tadpoles in competition ponds. The proportion of individuals metamorphosing of each genotype was differentially lowered by competition and pond drying. The LL genotype produced more metamorphs than the LR genotype in the constant water level ponds, but the LR genotype produced more in drying ponds. In competition ponds, the LR genotype produced more metamorphs than the LL genotype, but the LL genotype produced more metamorphs in ponds without competition. The RR genotype produced no metamorphs in any of the experimental environments. Increased performance of LR offspring from the heterogametic mating, in harsh conditions, and reduced performance of RR offspring from the homogametic mating, even under favorable conditions, relative to the parental genotype (LL) suggests that the population dynamics of this hybridogenetic system is strongly dependent on mate choice in mixed populations and the subsequent pond environment females select for oviposition and larval development.     

Heterozygous fitness effects of clonally transmitted genomes in waterfrogs

The European waterfrog Rana esculenta (RL‐genotype) is a natural hybrid between R. ridibunda (RR) and R. lessonae (LL) and reproduces by hybridogenesis, i.e. it eliminates the L‐genome from the germline and produces gametes only containing the clonally transmitted R‐genome. Because of the lack of recombination, R‐genomes are prone to accumulate spontaneous deleterious mutations. The homozygous effects of such mutations become evident in matings between hybrids: their offspring possess two clonal R‐genomes and are generally inviable. However, the evolutionary fate of R. esculenta mainly depends on the heterozygous effects of mutations on the R‐genome. These effects may be hidden in the hybrid R. esculenta because it has been shown to benefit from spontaneous heterosis. To uncouple clonal inheritance from hybridity, I crossed R. esculenta with R. ridibunda to produce nonhybrid offspring with one clonal and one sexual R‐genome, and compared their survival and larval performance with normal, sexually produced R. ridibunda tadpoles. Because environmental stress can enhance the negative effects of mutation accumulation, I measured the performance at high and low food levels. There was no indication that tadpoles with a clonal genome performed worse at either food level, suggesting that at least in the larval stage, R. esculenta benefits from heterosis without incurring any costs because of heterozygous effects of deleterious mutations on the clonally transmitted R‐genome.     

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.     

Interacting effects of predation risk and food availability on larval anuran behaviour and development

Age and size at metamorphosis are two important fitness components in species with complex life cycles. In anurans, metamorphic traits show remarkable phenotypic plasticity, especially in response to changes in growth conditions. It is also possible that the perception of risk directly determines changes in larval period and the size of metamorphs. This study examines how the perception of predation risk affects the timing of and size at metamorphosis in common frogs (Rana temporaria). I raised tadpoles at two risk levels (fish-conditioned water or unconditioned water) crossed with the availability or lack of food at night (all tadpoles had food available in the day). Tadpoles reacted to chemical cues from predatory fish by decreasing activity. A novel behavioural result was a predation×food interaction effect on refuge use, which also accounted for most of the predator main effect: predation risk only caused increased refuge use in the night-starved treatment. Despite these behavioural modifications, the perception of predation risk did not affect growth rate and mass at metamorphosis in a simple way: the effects of food regime on growth and size at metamorphosis were dependent on the level of predation risk as revealed by significant predation×food interaction effects. Tadpoles who had food withheld at night metamorphosed at the smallest size, suggesting a negative relationship between size at metamorphosis and refuge use. Tadpoles raised in fish-conditioned water had longer larval periods than those in unconditioned water, but these differences were significant only if food was available at night. These results conflict with the hypotheses that tadpoles should reduce their larval period or growth rates (and hence metamorphose at a smaller size) as the risk of predation increases. In contrast to predation risk, food availability strongly affected the length of the larval period: night-starved tadpoles metamorphosed relatively early with or without fish stimulus. Thus, early metamorphosis resulted from periods of low food availability, but not from a heightened ”perceived risk” of predation. This example counters the hypothesis of acceleration of the developmental rate (which shortens the time to metamorphosis) as a mechanism to escape a risky environment. Received: 18 August 1999 / Accepted: 10 January 2000     

Geographical and ecological distributions of frog hemiclones suggest occurrence of both 'General-Purpose Genotype' and 'Frozen Niche Variation' clones

Asexuals often occupy broad geographical and ecological ranges. Two models have been proposed to explain the ubiquity of asexuals: the General‐Purpose Genotype (GPG) and the Frozen Niche Variation (FNV) model. According to these models, asexuals differ in their ecological niche width and may occupy narrow specialist niches or ubiquitous niches. A thousand water frogs from 37 different populations located in France in different habitats were studied, and two (hemi)clonal hybrid types were identified genetically, Rana esculenta and R. grafi. Altogether, 13 hemiclones were identified both in R. grafi and R. esculenta. Three of these were geographically and ecologically widely distributed, and usually very common in populations. In contrast, the remaining 10 hemiclones had small geographical ranges and were restricted to special habitat types, suggesting ecological niche specialization. The results suggest that in hybridogenetic water frogs GPG and FNV hemiclones coexist.     

Mode of hybridogenesis and habitat preferences influence population composition of water frogs (Pelophylax esculentus complex,Anura: Ranidae) in a region of sympatric occurrence (western Slovakia)

Coexistence of sperm‐dependent asexual hybrids with their sexual progenitors depends on genetic and ecological interactions between sexual and asexual forms. In this study, we investigate genotypic composition, modes of hybridogenetic gametogenesis and habitat preferences of European water frogs (Pelophylax esculentus complex) in a region of sympatric occurrence. Pelophylax esculentus complex comprises parental species P. ridibundus and P. lessonae, whose primary hybridization leads to hybridogenetic lineages of P. esculentus. Hybrids clonally transmit one parental genome and mate with the other parental species, forming a new generation of hybrids. In the region of western Slovakia, we found syntopic occurrence of diploid and triploid hybrids with P. lessonae, syntopic occurrence of all three taxa as well as the existence of pure P. ridibundus populations. All triploid hybrids were exclusively male possessing one ridibundus and two different lessonae genomes (RLL). Sex ratio in diploid hybrids was substantially female‐biased. Irrespective of the population composition, diploid hybrids excluded the lessonae genome from their germ line and produced ridibundus gametes. Contrarily, RLL males unequivocally eliminated the ridibundus genome and produced diploid lessonae sperms. Perpetuation of RLL males in studied populations is most likely achieved by their mating with diploid hybrid females. The composition of water frog populations is also shaped by taxon‐specific habitat preferences. While P. ridibundus preferred larger water bodies (gravelpits, fishery ponds, dead river arms), P. lessonae was most frequently found in marshes and smaller sandpits. Pelophylax esculentus occupied predominately similar habitats as its sexual host P. lessonae.     

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.     

Factors influencing the composition of mixed populations of a hemiclonal hybrid and its sexual host

Hemiclonal/hybridogenetic hybrids combine demographic superiority of asexuals and genetic diversity of sexuals, but their need for backcrossing with a parental species tightly couples them with this sexual host. How can systems like this persist in ecological and evolutionary time? Two discrete‐time mathematical models describing the complex life cycle and mating system of hybridogenetic waterfrogs (Rana esculenta) identified four factors and their interactions as important. Although female mating preferences, in combination with differences in fecundity, determine species coexistence, differences in larval competitiveness seem to be more important for the hybrid‘s actual frequency. However, coexistence is possible even when host and hybrid are equally fecund and competitive. Dispersal and competition interact in their influence on species composition, but ecological and reproductive dispersal has opposing effects. In ecological terms our results explain the remarkable stability of observed species ratios over time within natural hybridogenetic populations, and indicate why the species composition can vary so widely between localities. In evolutionary terms they explain the old age of these and other hybridogenetic systems. They also suggest interesting consequences for other tightly coupled systems.     

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.     

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.     

Energy systems concepts and self-organization: a rebuttal

Tadpoles in small, ephemeral pools whose duration and food content are unpredictable can potentially encounter substantial variation in diet composition and availability. We compared the effects of 10 days of food deprivation occurring early, midway and late in ontogeny on the metamorphic size and bioenergetic properties of Hyla chrysoscelis tadpoles. Tadpoles fed throughout ontogeny were controls. Metamorphs from tadpoles starved early and midway in ontogeny had the same snout-vent length and dry mass as controls, but the time to metamorphosis was extended by 8 and 19% respectively. Metamorphs of tadpoles starved late in development attained 85% of the length and 55% of the mass of controls, metamorphosed at the same time as controls, and suffered mortality 15 times greater than other treatments, perhaps because they were near the absolute minimum necessary level of energy reserves. There were no significant differences in percent organic matter, percent tissue water, condition index, and protein or glycogen concentrations between any experimental and control treatments. If food deprivation occurred early in development, the tadpoles caught up to the size of controls, but an extended developmental time would increase the risk of predation or habitat loss. If food reductions occur late in development, perhaps magnified by pond desiccation, tadpoles are stimulated to metamorphose at the same time as controls but at a smaller size. The bioenergetic composition of tadpoles at metamorphosis is unaffected by time of food deprivation.