Sex reversal and primary sex ratios in the common frog (Rana temporaria)

Sex reversal has been suggested to have profound implications for the evolution of sex chromosomes and population dynamics in ectotherms. Occasional sex reversal of genetic males has been hypothesized to prevent the evolutionary decay of nonrecombining Y chromosomes caused by the accumulation of deleterious mutations. At the same time, sex reversals can have a negative effect on population growth rate. Here, we studied phenotypic and genotypic sex in the common frog (Rana temporaria) in a subarctic environment, where strongly female‐biased sex ratios have raised the possibility of frequent sex reversals. We developed two novel sex‐linked microsatellite markers for the species and used them with a third, existing marker and a Bayesian modelling approach to study the occurrence of sex reversal and to determine primary sex ratios in egg clutches. Our results show that a significant proportion (0.09, 95% credible interval: 0.04–0.18) of adults that were genetically female expressed the male phenotype, but there was no evidence of sex reversal of genetic males that is required for counteracting the degeneration of Y chromosome. The primary sex ratios were mostly equal, but three clutches consisted only of genetic females and three others had a significant female bias. Reproduction of the sex‐reversed genetic females appears to create all‐female clutches potentially skewing the population level adult sex‐ratio consistent with field observations. However, based on a simulation model, such a bias is expected to be small and transient and thus does not fully explain the observed female‐bias in the field.     

Isolation and characterization of 22 polymorphic microsatellite loci for the Barrow's goldeneye (Bucephala islandica)

We describe primers and polymerase chain reaction conditions to amplify 22 microsatellite loci from the Barrow's goldeneye (Bucephala islandica). The primers were tested on 27 individuals from a population breeding in British Columbia, Canada. The developed primer pairs yielded an average of 6.11 alleles per locus (range 2-12), an average observed heterozygosity of 0.70 (range 0.07-0.96) and a polymorphic information content of 0.07-0.88.     

Eight new tetranucleotide microsatellite loci for the agile frog (Rana dalmatina)

We describe eight new polymorphic tetranucleotide microsatellite loci isolated from the agile frog (Rana dalmatina). In 25 individuals from the Nature Reserve Lüneburger Heide (Lower Saxony, Germany), the number of alleles per locus ranged from four to nine and average observed heterozygosities from 69.1% to 80.7%. No evidence for linkage disequilibrium was found and none of the loci showed significant deviation from Hardy-Weinberg expectations. These microsatellite DNA markers are suitable tools for addressing population genetics issues in this endangered species.     

Molecular characterization of major histocompatibility complex class II alleles in the common frog, Rana temporaria

Major histocompatibility complex (MHC) class II genes, which play a major role in the immune system response, are some of the most polymorphic genes in vertebrates. We developed polymerase chain reaction primers for part of the second exon of an expressed MHC class II gene in the common frog, Rana temporaria. We genotyped this locus in five frog populations in southeast England and detected eight alleles in 215 individuals. Five or six alleles were detected in each population with a maximum of two alleles per individual, indicating that only a single locus was amplified. We also inferred the possible existence of a null allele. There were 23 variable nucleotide sites (out of 136) and 13 variable amino acid sites (out of 44), many of which corresponded to amino acids involved in antigen recognition. We detected a significant excess of nonsynonymous substitutions at antigen binding sites, indicating that this gene is under positive selection. The level of variation we found was similar to that in other amphibian MHC class II loci, such as those in Bombina bombina, Xenopus laevis and Ambystoma tigrinum.     

A test of the effect of supplemental UV-B radiation on the common frog, Rana temporaria L., during embryonic development

The release of certain man-made chemicals has led to recurrent, seasonal destruction of ozone in the upper atmosphere, allowing more solar radiation in the UV-B waveband to reach the Earth. Consequently, many amphibians may suffer increased exposure to UV-B at various stages in their lives. Embryonic stages of species which spawn in the spring, in shallow, open water, are at high risk of increased exposure. We exposed newly fertilized eggs of one such species, Rana temporaria L., to sunlight with and without supplemental UV-B. We used outdoor arrays of lamps to simulate the increase in UV-B which might result from previously documented ozone depletion. From immediately after fertilization to when hatchlings began feeding, ambient solar UV-B, weighted for DNA-damaging potential, was supplemented by ≈ 81% in 1995 and 113% in 1996. These levels of supplementation approximated the increase in solar UV-B expected to result from losses of 21% and 25%, respectively, of the total amount of ozone in the atmospheric column, relative to pre-ozone-depletion values. We found no evidence that these additions of UV-B radiation increased the incidence of mortality or overt developmental abnormality among embryos. We stress the need for appropriate dosimetry in studies of effects of UV-B on organisms.     

Isolation and characterization of 100 polymorphic microsatellite loci for the Siberian jay (Perisoreus infaustus)

We describe primers and polymerase chain reaction conditions to amplify 100 microsatellite loci from the Siberian jay (Perisoreus infaustus). The primers were tested on two geographically separated Finnish populations. The developed primer pairs yielded an average of 4.72 alleles per locus (range one to 17) and an average observed heterozygosity of 0.55 (range 0.04 to 1).     

Isolation and characterization of 17 polymorphic microsatellite loci for the three-toed woodpecker (Picoides tridactylus)

We describe primers and polymerase chain reaction conditions to amplify 17 di‐, tri‐ and tetranucleotide microsatellite loci from the three‐toed woodpecker (Picoides tridactylus). The primers were tested on 26 to 30 individuals from a single population breeding in southern Finland. The developed primer pairs yielded an average of 7.6 alleles per locus (range two to 15), an average observed heterozygosity of 0.69 (range 0.07 to 0.97), and an average polymorphic information content of 0.68 (range 0.06 to 0.90).     

First-generation linkage map for the common frog Rana temporaria reveals sex-linkage group

The common frog (Rana temporaria) has become a model species in the fields of ecology and evolutionary biology. However, lack of genomic resources has been limiting utility of this species for detailed evolutionary genetic studies. Using a set of 107 informative microsatellite markers genotyped in a large full-sib family (800 F1 offspring), we created the first linkage map for this species. This partial map-distributed over 15 linkage groups-has a total length of 1698.8 cM. In line with the fact that males are the heterogametic sex in this species and a reduction of recombination is expected, we observed a lower recombination rate in the males (map length: 1371.5 cM) as compared with females (2089.8 cM). Furthermore, three loci previously documented to be sex-linked (that is, carrying male-specific alleles) in adults from the wild mapped to the same linkage group. The linkage map described in this study is one of the densest ones available for amphibians. The discovery of a sex linkage group in Rana temporaria, as well as other regions with strongly reduced male recombination rates, should help to uncover the genetic underpinnings of the sex-determination system in this species. As the number of linkage groups found (n=15) is quite close to the actual number of chromosomes (n=13), the map should provide a useful resource for further evolutionary, ecological and conservation genetic work in this and other closely related species.     

Isolation and characterization of 15 polymorphic microsatellite DNA loci from Argyrosomus japonicus (mulloway), a new aquaculture species in Australia

Fifteen microsatellite loci were isolated for population genetic studies of mulloway (Argyrosomus japonicus), a commercial/recreational fish species in Southern Australia. A genomic library was screened randomly for di- tri- and tetranucleotide repeats. Fifteen microsatellite marker loci were developed that were highly polymorphic (allele number ranged from four to 18). Observed and expected heterozygosities ranged from 0.17 to 1 and from 0.27 to 0.91, respectively. These markers have proven useful for estimating genetic variation and for evaluating population structure across the species' natural distribution. They also provide powerful tools for optimizing hatchery practices to conserve genetic diversity.     

Isolation and characterization of microsatellite loci for the European tree frog (Hyla arborea)

We developed 11 new microsatellite markers for the European tree frog (Hyla arborea), and tested patterns of polymorphism in 54 adults (27 males and 27 females) from two ponds close to Lausanne (Western Switzerland). One marker was sex linked and two pairs displayed linkage disequilibrium. Comparisons of allele numbers with heterozygosity values support a stepwise-mutation model at neutral equilibrium, with mutation rates spanning nearly two orders of magnitude. These markers will prove useful for population genetic studies and fine-scale investigations requiring genetic assignment techniques.     

Isolation of microsatellite loci from the coqui frog, Eleutherodactylus coqui

Thirteen microsatellite loci were isolated from the coqui frog (Eleutherodactylus coqui) and optimized for future research. The loci were screened across 37 individuals from two Puerto Rican populations. Loci were variable with the number of alleles per locus ranging from three to 38. Polymorphic information content ranged from 0.453 to 0.963 and observed heterozygosity for each population ranged from 0.320 to 0.920.     

Isolation of ten tetranucleotide microsatellite loci in the Northern Wheatear (Oenanthe oenanthe)

We isolated 10 polymorphic microsatellite DNA loci from the Northern Wheatear (Oenanthe oenanthe) and optimized these for future studies in population genetics and behavioural ecology. The loci were screened for polymorphism using 107 individuals from one population in Germany. The primers amplified loci with high numbers of alleles ranging from two to 20 alleles per locus.     

Isolation and characterization of polymorphic microsatellite loci for the northern flicker (Colaptes auratus; Aves)

To investigate the mating system of northern flickers (Colaptes auratus), we developed primers for 14 microsatellite loci and screened them in 68 unrelated adults and their offspring. All markers were highly polymorphic with 9 to +36 alleles per locus. One marker was Z‐chromosome linked; one marker exceeded the size standard range and could not be analysed further. We checked the other 12 markers for Mendelian inheritance in 36 broods for which the social parents were known. Seven markers showed evidence for the presence of null alleles, and three of those showed significant deviation from Hardy–Weinberg equilibrium. Markers were generally unlinked.     

Isolation and characterization of polymorphic microsatellite loci in the green leafhopper Empoasca vitis Goethe (Homoptera)

Eight dinucleotide microsatellite loci were isolated and characterized within the green leafhopper Empoasca vitis (Goethe) using an enrichment cloning procedure. Primers were tested on 171 individuals collected in the southwest of France from the vine plants. The identified loci were polymorphic, with allelic diversity ranging from two to 18 alleles per locus. Observed heterozygosities were from 0.021 to 0.760. These microsatellite markers should prove to be a useful tool for estimating the population genetic structure, host-plant specialization and migration capacity of this insect.     

Isolation and characterization of eight polymorphic microsatellite loci in the rainforest canopy tree,Syzygium sayeri (Myrtaceae)

Eight polymorphic microsatellite loci were isolated from the rainforest canopy tree, Syzygium sayeri, in order to study parentage and subsequently pollen dispersal among individuals in wild populations. Screening of one natural population (n = 64) mapped in a 500 × 500 m area at Cape Tribulation, north Queensland, Australia, yielded two to 11 alleles per locus with observed levels of heterozygosity ranging from 0.07 to 0.70. One locus was significantly out of Hardy–Weinberg equilibrium after Bonferroni correction for multiple tests. These loci should provide a useful tool in further understanding the dispersal patterns of this species.