Characterization of 24 microsatellite loci in delta smelt, Hypomesus transpacificus, and their cross-species amplification in two other smelt species of the Osmeridae family

We characterized 24 polymorphic tetranucleotide microsatellite loci for delta smelt (Hypomesus transpacificus) endemic to the San Francisco Bay Estuary, CA, USA. Screening of samples (n = 30) yielded two to 26 alleles per locus with observed levels of heterozygosity ranging from 0.17 to 1.0. Only one locus deviated from Hardy–Weinberg equilibrium, suggesting these individuals originate from a single panmictic population. Linkage disequilibrium was found in two pairs of loci after excluding the locus out of Hardy–Weinberg equilibrium. Twenty‐two primer pairs cross‐amplified in wakasagi smelt (Hypomesus nipponensis), and 15 primer pairs cross‐amplified in longfin smelt (Spirinchus thaleichthys).     

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.     

Characterization of 23 Tri‐ and tetranucleotide Microsatellite Loci in the Brown Tiger Prawn,Penaeus esculentus

Twenty‐three unique tri and tetra‐nucleotide microsatellites were isolated and characterized from the genome of the brown tiger prawn Penaeus esculentus. Nine of the 23 microsatellite loci were found to be polymorphic and were tested across a sample of wild caught Penaeus esculentus. Good fits to Hardy–Weinberg expectations were observed for six of the markers when tested in a population of at least 180 individuals. The total number of alleles for these six markers ranged from eight to 19, with an average total of 13. These values will make them useful for future stock identification and population genetics studies.     

Development of polymorphic microsatellite markers for the Steere's Liocichla (Liocichla steerii)

For the Steere's Liocichla (Liocichla steerii) we developed seven polymorphic microsatellite markers from a partial genomic library enriched for GATA, tetranucleotide repeats. Among the 27–28 individuals screened, allelic diversity ranged from six to 19 alleles per locus and the observed heterozygosity of each locus ranged from 0.250 to 0.889. All loci conformed to Hardy–Weinberg expectations except for two that appeared to be Z‐Iinked; such Z‐Iinked loci can be used in studies of male dispersal.     

Development and validation of microsatellite markers for the tiny dragonfly, <Emphasis Type="Italic">Nannophya pygmaea</Emphasis> (Odonata: Libellulidae), which is endangered in South Korea

The tiny dragonfly, Nannophya pygmaea (Odonata: Libellulidae), is listed as a second-degree endangered wild animal in South Korea. The application of molecular markers to assess genetic diversity and population relationships can provide information necessary to establish an effective conservation strategy. In this study, we developed 12 microsatellite markers specific to N. pygmaea using the NextSeq 500 platform. Forty individuals of N. pygmaea collected from three currently known localities in South Korea were genotyped to validate these markers and to preliminarily assess population genetic characteristics. The observed number of alleles, observed heterozygosity, and expected heterozygosity at a locus ranged from 2 to 9, 0.421–1.0, and 0.508–0.766 in a population with the largest sample size (20 individuals), respectively, thereby validating the suitability of the markers for population analysis. Five of 12 loci showed significant deviation from the Hardy–Weinberg equilibrium in the population. Our preliminary data indicate an absence of inbreeding in all populations and an absence of obvious genetic difference. The microsatellite markers developed in this study will be useful for studying the population genetics of N. pygmaea collected from other regions worldwide, including additional sites in South Korea.     

Isolation and characterization of 150 novel microsatellite markers for Zhikong scallop (Chlamys farreri)

We isolated and characterized 150 novel microsatellite markers of Zhikong scallop (Chlamys farreri) from three simple sequence repeat‐enriched libraries constructed with (GA)₁₅ and (CA)₁₅. The polymorphism was assessed with 48 individuals, and the result showed the number of allele ranged from two to 30, with an average of 8.4 alleles/locus. The values of observed and expected heterozygosities ranged from 0.0791 to 0.9878 and from 0 to 1.0000, respectively. Sixty‐five loci showed significant departure from Hardy–Weinberg equilibrium, and 14 locus pairs displayed linkage disequilibrium. These markers are therefore potentially useful for conservation studies, population structure assessment, ecological analyses and linkage map construction.     

Characterization of microsatellite DNA markers for a mahseer species,Tor tambroides (Cyprinidae) and cross‐amplification in four congeners

This study reports the isolation and characterization of microsatellite DNA markers in a mahseer species, Tor tambroides (Pisces, Cyprinidae). Of a total of 14 loci evaluated, 10 were polymorphic in T. tambroides samples, with an average of 2.86 alleles per locus. Deviations from Hardy–Weinberg equilibrium were observed at one locus and there was no indication of linkage disequilibrium among loci. A high level of cross‐amplification among four congeners was achieved, with 12 loci successfully amplifying and 11 loci showing polymorphism in at least one other species. These markers will be a useful resource for population genetic studies and broodstock management of closely related mahseer species.     

Isolation and characterization of gene-derived single nucleotide polymorphism (SNP) markers in Scylla paramamosain

Single nucleotide polymorphisms (SNPs) are thought to be well suitable for genetic and evolutionary studies. In this study, we reported the first set of SNP markers in a commercially important crab species, Scylla paramamosain. A total of 12,500 base pairs high quality DNA sequences were obtained from 15 genes, and thirty-seven SNPs were identified, representing one SNP every 338 base pairs. Twenty-four SNPs were successfully genotyped in a single population. All loci had two alleles and the minor allele frequency ranged from 0.02 to 0.44. The observed and expected heterozygosity ranged from 0.04 to 0.59 and from 0.04 to 0.50, respectively. No significant departures from Hardy–Weinberg equilibrium at each locus was found. The linkage disequilibrium was detected in six loci pairs, but absent after sequential Bonferroni correction. These SNP markers will provide a useful addition to the genetic tools for genetic and evolutionary studies for S. paramamosain.     

Ten polymorphic tetranucleotide microsatellite markers isolated from the Anolis roquet series of Caribbean lizards

The Anolis roquet series of Caribbean lizards provides natural replicates with which to examine the role of historical contingency and ecological determinism in shaping evolutionary patterns. Here, we describe 10 polymorphic tetranucleotide microsatellites to facilitate studies on population differentiation and gene flow. All loci successfully amplified in several species from this series. Genotyping 96 individuals from two A. roquet populations demonstrated the markers’ suitability as population genetic markers: genetic diversity was high (9–22 alleles per locus); there were no instances of linkage disequilibrium; and, with one exception, all genotypic frequencies conformed to Hardy–Weinberg equilibrium expectations.     

Characterization of microsatellite loci isolated in Mountain Plover (Charadrius montanus)

Primers for 15 microsatellite loci were developed for Mountain Plover, a species whose distribution and abundance have been reduced drastically in the past 30 years. In a screen of 126 individuals collected from four breeding locales across the species’ range, levels of polymorphism ranged from two to 13 alleles per locus. No two loci were found to be linked, although one locus revealed significant departures from Hardy–Weinberg equilibrium. These microsatellite loci can be used in population genetic studies, ultimately aiding in management efforts for Mountain Plover. Additionally, these markers can potentially be used in studies investigating the mating system of Mountain Plover.     

Isolation and characterization of 18 polymorphic microsatellite markers for the “Female Ginseng” Angelica sinensis (Apiaceae) and cross-species amplification

Angelica sinensis (Apiaceae) is an endangered alpine herb that is widely used as a medicinal plant in traditional Chinese medicine (TCM). Wild populations of A. sinensis have become quite rare in China. Thus, population genetics studies of this species are urgently needed for its effective conservation and sustainable use. However, to date, no microsatellite loci have been isolated in A. sinensis. To address this issue, we isolated 18 polymorphic loci and genotyped 120 individuals collected from 6 populations. The number of alleles per locus ranged from 1.2 to 5.5, and the average was 2.4. The observed and expected heterozygosity per locus for a population varied, respectively, from 0.000 to 0.983 (averaged at 0.198) and from 0.066 to 0.661 (averaged at 0.333). Deviation from the Hardy–Weinberg equilibrium (p < 0.01) was observed for 4 to 14 loci in various populations. These microsatellite markers were cross-amplified in 10 species affinis, and 7 loci were successfully amplified in all species. These microsatellite markers are useful for genetic studies, the conservation management of A. sinensis, and identification of A. sinensis.     

Isolation and characterization of microsatellite markers for Viola mirabilis (Violaceae) using a next‐generation sequencing platform

We isolated and characterized microsatellite loci in Viola mirabilis (Violaceae), an endangered species from South Korea. Twenty‐three polymorphic microsatellite loci were developed and tested in Korean, Chinese and Japanese populations. The number of alleles per locus varied from two to eight. The observed and expected heterozygosities within the three populations were 0.000–0.625 and 0.469–0.695, respectively. A total of six loci in the Korean population, one locus in the Chinese population and seven loci in the Japanese population deviated from Hardy–Weinberg equilibrium. We expect that these newly developed microsatellite markers will contribute to understanding the phylogeography and population genetics of V. mirabilis, which will aid in developing conservation strategies for this species.     

Isolation and characterization of fourteen polymorphic microsatellite markers in the viperine snake Natrix maura

Nineteen polymorphic microsatellite loci were identified and developed for Natrix maura. Polymorphism was assessed for 120 individuals sampled across four sampling sites from the French Pyrenees Mountains. The number of alleles per locus ranged from 3 to 15, and expected heterozygosity per locus ranged from 0.227 to 0.863. We tested for deviation from Hardy–Weinberg equilibrium and linkage disequilibrium and assessed the presence of null alleles for all loci, resulting in a selection of 14 high‐quality polymorphic markers. These markers will be extremely useful in identifying fine‐scale genetic structures and providing insight into conservation management plans of this species.     

Isolation and characterization of 18 polymorphic microsatellite markers in Chinese mandarin fish Siniperca chuatsi (Basilewsky)

Eighteen microsatellite markers were isolated and characterized using an enrichment protocol in the Chinese mandarin fish Siniperca chuatsi (Basilewsky), a commercially important piscivorous fish in China. Out of 48 pairs of primers designed, 18 loci exhibited polymorphism with three to six alleles (mean 4.4 alleles/locus) and average observed heterozygosity ranged from 0.633 to 0.833 (mean 0.748) in a test population from Dongting Lake of China. Except for two loci, all other 16 loci were in the Hardy–Weinberg equilibrium. These markers would be useful for such studies as population genetics, ecology and selective breeding of the Chinese mandarin fish in future.     

Microsatellite markers isolated from barn swallows (Hirundo rustica)

Fifteen polymorphic microsatellite DNA loci were isolated from barn swallow (Hirundo rustica) and optimized for future studies of radiation‐induced mutations in populations from Ukraine. The loci were screened for variability among 25 individuals from two populations. The primers amplified loci with relatively high numbers of alleles ranging from five to 32 alleles per locus and polymorphic information content from 0.481 to 0.951. Observed heterozygosity varied from 0.458 to 0.960. None of the loci showed deviations from Hardy–Weinberg equilibrium in either population.     

Characterization of six microsatellite DNA loci for Sorex arizonae

Sorex arizonae is a rare species that occupies a narrow range of habitat types in several mountain ranges of New Mexico, Arizona and Northern Mexico. Here we identify and characterize six microsatellite loci for this species. We screened 63 individuals from four different localities from New Mexico and Arizona to analyse genetic variability. Alleles ranged from three to 16. Heterozygosity ranged from 40% to 78%. Most polymorphic loci were in Hardy–Weinberg equilibrium with the exception of one locus. Primers appear to have reasonable cross‐species applicability as five loci amplified in another shrew species (Sorex monticolus).     

Development and characterization of novel di‐ and tetranucleotide microsatellite markers in Pacific cod (Gadus macrocephalus)

The Pacific cod (Gadus macrocephalus) supports large commercial fisheries in the northern Pacific Ocean and the Bering Sea. Here we characterize 10 polymorphic microsatellite loci isolated from enriched genomic libraries. Loci were screened on a sample of 96 spawning adults. The average number of alleles per locus was 25.3 (range 12–44), with expected heterozygosities (H_E) ranging from 0.54 to 0.97. No significant deviations of genotypic proportions from Hardy–Weinberg equilibrium or linkage equilibrium were detected. These markers will be used in future studies of population structure and mixed stock analysis of this important gadid species.     

Identification and characterization of 21 polymorphic microsatellite loci from the mycophagous fly Drosophila neotestacea

Drosophila neotestacea is a mushroom‐feeding fly that is common in the temperate and boreal forests of North America. Here I describe the isolation and characterization of 21 polymorphic microsatellite loci from the D. neotestacea genome. In a sample of flies from Rochester, New York, the expected and observed heterozygosities ranged from 0.344 to 1.000 and from 0.384 to 0.923, respectively. Of the 21 markers, six were likely X‐linked, seven showed a departure from Hardy–Weinberg equilibrium, and none were found to be in linkage disequilibrium. These loci will facilitate future ecological and population genetic studies of D. neotestacea.     

Short microsatellite DNA markers for the red fox (Vulpes vulpes)

Seven short microsatellite loci (< 165 bp) were characterized for red foxes for the amplification of degraded DNA extracted from historical samples. Following polymerase chain reaction (PCR) using primers developed in the domestic dog, red fox‐specific primers were designed within the flanking regions. The number of detected alleles ranged between six and 15 alleles and the expected heterozygosities ranged between 0.67 and 0.92. No deviations from Hardy–Weinberg equilibrium were detected for any of the markers.     

Polymorphic microsatellite DNA markers for Banksia attenuata (Proteaceae)

We developed 11 polymorphic microsatellite DNA markers for an Australian native shrub Banksia attenuata. The number of alleles per locus in 50 individuals varied from five to 18, observed and expected heterozygosities ranged from 0.300 to 0.740 and from 0.537 to 0.918, respectively. Six loci showed no significant deviation from Hardy–Weinberg equilibrium (P > 0.05), and null alleles appear to exist at locus BA‐B1. All loci showed independent inheritance.