Characterization of microsatellite markers for Couch's spadefoot toad (Scaphiopus couchii) and cross‐amplification in other species of the family Scaphiopodidae

I developed 12 di‐ and tetranucleotide microsatellite loci for Couch's spadefoot toad (Scaphiopus couchii). These loci have 3–37 alleles per locus and observed heterozygosities ranging from 0.157 to 0.941 among 85 individuals from four populations. Global and within‐population exact tests do not reveal departure from Hardy–Weinberg expectations and all loci pairs are in linkage equilibrium. These independent markers will be useful for studies of population structure and kinship in this commonly studied amphibian. Additionally, several of these loci may be applicable for studies of other North American toads of the family Scaphiopodidae.     

Five polymorphic microsatellite markers for the Great Plains toad,Bufo cognatus

We characterize five polymorphic microsatellite loci for the Great Plains toad, Bufo cognatus, which are being used as markers in a study of the genetics and structure of toad populations inhabiting playa lakes on the Llano Estacado plateau of Texas. Observed heterozygosity (H_O) estimates for the toads ranged from 0.56 to 0.90, and expected heterozygosity (H_E) estimates ranged from 0.85 to 0.95.     

Characterization of microsatellite markers for Thoropa taophora (Anura, Cycloramphidae), a frog endemic to the Brazilian Atlantic Rain Forest

Here we characterize nine microsatellite loci for Thoropa taophora collected from a coastal and an island population in southeastern Brazil. The number of alleles per locus ranged from three to 11 for the coastal population and from two to eight for the island population. Observed heterozygosities ranged from 0.308 to 0.778 for the coastal population and from zero to 0.737 for the island population. One locus did not conform to Hardy-Weinberg expectations on the coast, while two were in disequilibrium on the island. These markers are currently being used to examine genetic differentiation and patterns of dispersal in T. taophora.     

Novel tetranucleotide microsatellite DNA markers for the wood frog,Rana sylvatica

Fifteen tetranucleotide microsatellite loci were identified and characterized for wood frogs (Rana sylvatica) collected from three vernal pools in the southeastern US. These markers revealed a high degree of genetic diversity (nine to 34 alleles per locus), heterozygosity (30.6–92.3%) and allelic heterogeneity (69% of comparisons were statistically significant). Considerable differentiation among populations was observed as genetic distances (chord) ranged between 0.40 and 0.55 and all F_ST values (0.02–0.05) were statistically significant. Genotypic assignment tests correctly classified 103 of 113 individuals to their respective collection. These markers should prove useful for investigating fine‐scale population structure and metapopulation dynamics.     

Interspecific microsatellite markers for the study of pinniped populations

Microsatellites have rapidly become the marker of choice for a wide variety of population genetic studies. Here we describe 20 pinniped microsatellite markers which have been tested across 18 pinniped species. The majority of these markers have broad utility in all pinnipeds and provide a strong base for detailed population genetic studies in the Pinnipedia.     

Isolation and characterization of 19 polymorphic microsatellite DNA markers in the Japanese brown frog (Rana japonica)

We report the isolation and characterization of 19 polymorphic microsatellite DNA markers in the Japanese brown frog (Rana japonica). These markers were tested in 24 individuals each collected from three distinct populations in Ichikai-machi, Tochigi Prefecture. The number of observed alleles per locus ranged from 3 to 24 across all populations, and the values of observed and expected heterozygosities ranged from 0.130 to 1 and from 0.125 to 0.941, respectively. These microsatellite loci will be useful for investigating the intraspecific genetic variation and population structure of this species.     

Polymorphic microsatellite markers for the Mojave desert tortoise, Gopherus agassizii

We describe primers and polymerase chain reaction (PCR) conditions to amplify 14 tri- and tetranucleotide microsatellite loci for the Mojave desert tortoise (Gopherus agassizii). Across three populations (87 individuals) located in the Mojave Desert, USA, the markers yielded a range of four to 33 alleles and an average observed heterozygosity of 0.733 (range 0.433 to 0.933). We neither detected linkage disequilibrium between any pair of loci nor did we find a consistent pattern of deviation from Hardy-Weinberg equilibrium. These microsatellites are designed for PCR multiplexing, and provide higher throughput capacity to aid in conservation genetics studies for this threatened species.     

Characterization of microsatellite markers for the endemic sand dune lizard,Sceloporus arenicolus

Here we characterize and report on the genetic variability of eight microsatellite loci for the endemic sand dune lizard, Sceloporus arenicolus, that amplified consistently for individuals throughout the species’ range. The number of alleles per locus was high (mean = 13.25) and observed heterozygosities ranged from 0.354 to 0.808. With the exception of a single locus, the loci did not deviate from Hardy–Weinberg expectations. This set of markers is currently being used to examine population structure and landscape genetics in S. arenicolus, a habitat specialist with a restricted range and patchy distribution.     

Fourteen new microsatellite markers for Atlantic herring Clupea harengus

Fourteen new microsatellite loci were developed and tested on Atlantic herring Clupea harengus with 39 individuals from Iceland and 49 individuals from Norway. The microsatellites, which contain di, tri and tetranucleotide repeats, are polymorphic (7-30 alleles), with observed heterozygosity ranging between 0·69 and 1·00 and expected heterozygosity between 0·55 and 0·97.     

Polymorphic microsatellite markers in the painted dragon lizard,Ctenophorus pictus

We isolated eight polymorphic microsatellite markers from the painted dragon, Ctenophorus pictus, from a genomic library simultaneously enriched for a suite of di‐, tri‐ and tetranucleotide repeat units. Genetic variation was assessed in 40 individuals from populations in Victoria and South Australia. The number of alleles per locus ranged from four to 45 with observed heterozygosity ranging from 0.23 to 0.95. These markers will be useful for studies of natural and sexual selection and the effects of habitat fragmentation in semiarid Australia.     

Polymerase chain reaction primers for microsatellite loci in the Common Toad Bufo bufo

The Common Toad Bufo bufo is a wide‐ranging species, with a distribution encompassing much of Europe and Asia. Few molecular studies have been undertaken on this species, and only one polymorphic microsatellite locus has been identified. Therefore, little is known about the genetic variability within and between B. bufo populations. The value of such information is essential for monitoring the species and its environment. This paper reports the characterization of 15 B. bufo microsatellite loci using individuals from a Spanish and UK sample.     

Sixteen microsatellite loci for the Bufo boreas group

The four species that comprise the Bufo boreas group of toads are critically imperiled in all or portions of their geographical ranges. We present data from 16 microsatellite loci isolated from B. boreas that cross‐amplify in these four species. These markers have proven useful in the analyses of population structure and conservation genetics, and provide a powerful tool for future researchers who seek to understand the conservation genetics of these rare toad species. Polymorphism was assessed for 339 individuals from seven populations representing the four species. All loci were polymorphic (X? = 8, range of four to 19 alleles). Three loci were not in Hardy–Weinberg equilibrium (HWE) in one population, and one of these loci was out of HWE in a second population (P < 0.003 after Bonferroni correction for multiple tests). However, there were no systematic deviations from HWE across all study populations. Small populations in fragmented habitat appear to explain the existing HWE deviations.     

Fifteen polymorphic microsatellite markers for the giant spiny frog, Paa spinosa

We characterized 15 polymorphic microsatellite loci for the giant spiny frog, Paa spinosa. These loci were highly polymorphic when screened in 38 individuals from two different populations, with nine to 23 alleles per locus. The range of observed and expected heterozygosities was 0.231-0.916 and 0.296-0.944, respectively. These polymorphic loci will be useful for assessing genetic diversity, population structure, gene flow, population assignment and determining paternity in the giant spiny frog.     

Polymorphic tetranucleotide microsatellite markers in the Caribbean spiny lobster,Panulirus argus

Ten tetranucleotide microsatellite loci are described for the Caribbean spiny lobster Panulirus argus. Loci were polymorphic (4–15 alleles per locus) and exhibited high levels of expected (0.553–0.921) and observed heterozygosity (0.469–0.906) from samples caught off Belize and Puerto Rico coasts. No significant departure from Hardy–Weinberg equilibrium conditions were observed for any locus. All microsatellite loci should be useful for assessing population discrimination for this valuable marine animal currently subjected to excessive fishing efforts.     

Polymorphic microsatellite markers for endangered spotted suslik,Spermophilus suslicus

Spotted suslik Spermophilus suslicus is one of the most endangered mammal species in Poland. Over the last 50 years, it lost more than 90% of its populations and about 70% of its individuals. In order to establish a conservation and reintroduction program, the knowledge of population structure of the species is crucial. We have developed nine polymorphic microsatellite loci to analyse the population structure. Six of the primer sets also amplify polymorphic markers in the European suslik Spermophilus citellus.     

Ten polymorphic microsatellite DNA markers for the platypus, Ornithorhynchus anatinus

We identified and optimized 10 microsatellite loci for the platypus, Ornithorhynchus anatinus (Monotremata: Ornithorhynchidae), and screened 21 individuals from the southern tablelands area of New South Wales, Australia. Each polymorphic locus possessed between two and 12 alleles with observed heterozygosities between 0.118 and 0.950. The intent of this effort was to provide informative loci for studies on the population genetics of this species.     

Ten microsatellite markers for the freshwater diatom Sellaphora capitata

Ten polymorphic microsatellite markers were developed for the benthic freshwater diatom Sellaphora capitata and tested on 40 isolates from a Belgian pond. Genotyping was very successful (95%). The number of alleles per locus ranged from three to 12 (mean 6.6) and expected heterozygosities from 0.2 to 0.86 (mean 0.67). This is the first time that microsatellite markers have been developed for a freshwater or benthic diatom.     

Development of polymorphic microsatellite markers for the fungal tree pathogen Cryphonectria eucalypti

Polymorphic microsatellite DNA markers were developed from a single spore isolate of Cryphonectria eucalypti collected from a Eucalyptus stem canker in South Africa. Markers were obtained using the enrichment technique known as fast isolation by AFLPs of sequences containing repeats (FIASCO). Ten polymorphic markers were isolated, of which, two were discarded due to their high polymorphism in the flanking region. The mean number of alleles produced by the remaining eight markers from 20 isolates was 7.25, and alleles per locus ranged from four to 12. The markers will be used to study populations of C. eucalypti.