Isolation and characterization of microsatellite loci in Onychostoma sima

Seventeen polymorphic microsatellite loci have been isolated from an (AAAG) _n enriched genomic library of Onychostoma sima. These loci were tested against 32 wild captured individuals for polymorphism. The number of observed alleles per locus ranged from 2 to 10. The expected and observed heterozygosity values ranged from 0.25 to 0.96 and from 0.26 to 0.87, respectively. Seven loci showed significant deviation from Hardy–Weinberg Equilibrium and linkage equilibrium were detected in three loci. These polymorphic loci described for O. sima will be valuable as genetic markers for use in population structure, breeding, and conservation studies.     

Isolation and characterization of ten microsatellite loci for the Chong’an moustache toad Leptobrachium liui (Anura: Megophryidae)

We isolated and characterized ten polymorphic microsatellite loci in the Chong??an moustache toad Leptobrachium liui, nine novel loci designed for this species and one locus from cross-species amplification. Loci were screened in 43 individuals of a single population. The number of alleles per locus ranged from 3 to 17, on average of 10. The average observed and expected heterozygosity per locus ranged from 0.512 to 0.884 and from 0.580 to 0.898, respectively. No significant departures from Hardy?CWeinberg equilibrium were found after sequential Bonferroni corrections. Three loci, CHA8, CHA12, and VIB-D5, were detected to be linked. These polymorphic loci will be useful for conservation genetics and phylogeography studies of this endangered species.     

Isolation and characterization of microsatellite markers for Glytothorax zanaensis

Fifteen polymorphic microsatellite loci were isolated from an enriched genomic library of Glyptothorax zanaensis. The polymorphism of these loci was tested on a population of 78 individuals captured from Nujiang River. The number of observed alleles per locus ranged from 3 to 8. The expected and observed heterozygosity values ranged from 0.578 to 0.820 and 0.623 to 1.000, respectively. Two loci significantly deviated from Hardy–Weinberg equilibrium after Bonferroni correction. No significant linkage disequilibrium was detected. These microsatellite markers will be useful in studies of population genetic structure of G. zanaensis.     

Isolation and characterization of twenty-six microsatellite loci for the tetraploid fish Dabry’s sturgeon (Acipenser dabryanus)

To evaluate the population genetic diversity as a means of devising conservation strategies, we constructed (CA)n and (CT)n enriched genomic libraries for Acipenser dabryanus, a narrowly endemic and endangered species in China. Twenty-six polymorphic microsatellite markers were developed using the fast isolation by AFLP of sequences containing repeats (FIASCO) protocol. Between 3 and 13 alleles per locus were observed. Mean expected heterozygosities (H _E) and Shannon-Wiener Diversity Indices (H′) per locus ranged from 0.303 to 0.806, and from 0.480 to 1.893, respectively. The microsatellite markers described here are valuable tools for the population genetics research of A. dabryanus.     

Isolation and characterization of ten microsatellite loci of endangered Anderson’s crocodile newt, Echinotriton andersoni

Due to an originally small distribution range and recent habitat loss, Anderson??s crocodile newt (Echinotriton andersoni) has been steadily declining in number. For effective conservation of this species, a greater amount of genetic information is needed. Here, we isolated ten microsatellite loci of E. andersoni using three different methods, and polymorphism of these 10 loci were evaluated for 27 individuals collected from three islands. The total number of alleles per locus ranged from 3 to 22, and the expected heterozygosity ranged from 0 to 0.876. Taken together, our findings suggest that these novel loci will be applicable for conservation genetic studies across varying scales.     

Isolation and characterization of microsatellite loci in Darwin’s fox (Lycalopex fulvipes) and cross-amplification in other canid species

Darwin’s fox (Lycalopex fulvipes) is one of the most endangered canid of the world. It occurs only in two small populations in the South of Chile. We obtained molecular markers useful for population level studies of the Darwin’s fox by screening partial genomic DNA libraries enriched for microsatellite repeats. We developed 11 microsatellite loci and genotyped 24 individuals from one of the two known populations. The loci had between 2 and 4 alleles and observed heterozygosities ranged from 0.041 to 0.608. All of these pairs of primers also successfully amplified DNA from two other Lycalopex Chilean foxes and two other canids. These microsatellite markers will be useful to assess the genetic diversity of the species.     

Isolation and characterization of microsatellite loci in the Reevese’s butterfly lizard Leiolepis reevesii (Agamidae)

We characterize 25 polymorphic microsatellite loci isolated from Leiolepis reevesii genomic libraries. Thirty-four individuals were collected from two populations, eighteen from a mainland population in Xuwen, Guangdong, and the remaining 16 from an island population in Haikou, Hainan. These markers revealed a relatively high degree of genetic diversity (2?C18 alleles per locus) and heterozygosity (H _O ranged from 0.000?C0.938, and H _E ranged from 0.160?C0.954). Eight loci exhibited significant deviations from Hardy?CWeinberg equilibrium (HWE) after sequential Bonferroni correction due to heterozygote deficiencies, but only one locus (L311) with low heterozygosity in one population did significantly deviate from HWE in the other. Inbreeding may explain the deviations from HWE. There was no evidence of linkage disequilibrium among pairs of loci in the samples. These microsatellite markers will be useful for future studies focusing on gene flow, population structure and evolutionary history of L. reevesii.     

Isolation and characterization of nine microsatellite loci in the endangered Worthen’s Sparrow (Spizella wortheni)

We report the isolation and characterization of nine microsatellite markers from Worthen’s Sparrow (Spizella wortheni), an endangered bird species endemic to northeastern Mexico. We tested the markers in 32 samples of the species collected near Saltillo Coahuila, Mexico. All markers were polymorphic, with 3–31 alleles per locus, and no difference was found between the observed and expected heterozygosity when applying a sequential Bonferroni correction and a table-wide significance level of 0.05. We found no evidence for linkage disequilibrium between the markers and estimated a null allele frequency of 0.00–0.05. The new markers will allow elucidating the genetic structure and life history of Worthen’s Sparrow and guiding conservation efforts for this endangered species.     

Isolation and characterization of 20 microsatellite loci in Neverita didyma (R?ding 1798)

Neverita didyma is a commercially important and over-exploited species. In order to investigate its genetic diversity and population structure, 20 polymorphic microsatellite loci were isolated and characterized. The number of alleles per locus ranged from 2 to 20, and the observed and expected heterozygosities varied from 0.276 to 1.000 and from 0.331 to 0.939, respectively. These microsatellites will be a useful tool for fisheries management and conservation programme for this species.     

Isolation and characterization of twelve microsatellite loci of endangered Ishikawa’s frog (Odorrana ishikawae)

Due to an originally small distribution range and over-exploitation of primary forest, Ishikawa’s frog (Odorrana ishikawae) has been steadily declining in number. For effective conservation of this species, a greater amount of genetic information of this species is needed. Here, we isolated and characterized 12 microsatellite loci of O. ishikawae using two different methods. Loci were screened from two populations on the Amami (N = 44) and Okinawa (N = 9) Islands. The total number of alleles per locus ranged from 2 to 30, and the observed heterozygosity ranged from 0 to 0.922. Notably, high F _ST values for all examined loci were observed between the two populations. Taken together, our findings suggest that these novel loci will be applicable for conservation genetic studies across varying scales.     

Isolation and characterization of 16 microsatellite markers from a rare and endemic species, Scutellaria austrotaiwanensis (Lamiaceae)

Scutellaria austrotaiwanensis is a new species first described in 1997. It is a rare and endemic species in southern Taiwan. In this study, 16 polymorphic microsatellite loci were isolated from Scutellaria austrotaiwanensis using a magnetic bead enrichment method. Several genetic indices, including observed heterozygosity (Ho), expected heterozygosity (He), number of effective alleles (Ne), and polymorphism information content (PIC), were calculated. Deviation of heterozygosity was detected by the fixation index (Fis) and Hardy–Weinberg equilibrium was tested by possibilities (P _HWE) using a chi-squared test. Allele numbers ranged from 4 to 8, with observed heterozygosity ranging from 0.611 to 1.000. Five microsatellite markers showed deficient heterozygosity by Fis > 0. No loci exhibited a departure from Hardy–Weinberg equilibrium, possibly due to a stable population. No loci pairs revealed significant linkage disequilibrium. The microsatellite markers reported here should be useful to evaluate the population structure of S. austrotaiwanensis for conservation efforts.     

Isolation and characterization of microsatellite loci in two non-native hydromedusae in the San Francisco Estuary: Maeotias marginata and Moerisia sp.

We characterized 10 new microsatellite markers in each of two species of hydromedusae, Maeotias marginata and Moerisia sp. Genetic diversity was estimated using 20–41 individuals collected from Suisun Marsh within the San Francisco Estuary, CA. Allelic richness ranged from 5–9 in M. marginata and 2–10 in Moerisia sp. with average expected heterozygosities of 0.71 and 0.57 respectively. One locus in M. marginata and two in Moerisia sp. deviated from Hardy–Weinberg equilibrium expectations, likely due to null alleles.     

Isolation and characterization of polymorphic microsatellite loci from the endangered Tarim schizothoracin (Schizothorax biddulphi Günther)

Tarim schizothoracin (Schizothorax biddulphi Günther) is an extremely endangered freshwater fish in China. In this paper, 15 polymorphic microsatellite loci were characterized in this species. The number of alleles ranged from 4 to 14, the observed heterozygosities (H _O) were from 0.7609 to 1.0000, and the expected heterozygosities (H _E) ranged from 0.7273 to 0.9194. The polymorphism information content was from 0.6684 to 0.9020. Only two loci deviated significantly from the Hardy?CWeinberg equilibrium, and none of the loci combinations showed significant linkage disequilibrium. These microsatellite loci are useful in investigating the genetic variability in Tarim schizothoracin.     

Isolation and characterization of polymorphic microsatellite loci in Ping’s juniper Juniperus pingii

Juniperus pingii (Ping’s juniper) is a threatened conifer species endemic to China, and a good knowledge of its population genetics would be essential to formulate effective conservation strategies. In this study, ten polymorphic microsatellite loci were isolated, and characterized for their utility using 58 individuals from two geographically distinct populations. In population XGLL, the number of alleles per locus (N _A) ranged from four to 11, with an average of 6.2 per locus; the observed (H _O) and expected heterozygosity (H _E) ranged from 0.205 to 0.854 and from 0.423 to 0.754, with an average of 0.347 and 0.527, respectively. In population BM, N _A, H _O and H _E ranged from four to eight, from 0.198 to 0.454 and from 0.339 to 0.853, with an average of 5.8, 0.294 and 0.556, respectively. Following the sequential Bonferroni correction, only one locus (Jp07) was found to deviate significantly from Hardy–Weinberg equilibrium within population XGLL, and no significant linkage disequilibrium was detected in any of these loci within either population. These polymorphic microsatellite loci would be useful for population genetic studies of this conifer species.     

Development and characterization of seventeen microsatellite loci for the Peter’s tent-roosting bat (Uroderma bilobatum)

In the Neotropics, Peter’s tent-roosting bat (Uroderma bilobatum) is an important keystone species. U. bilobatum promotes plant community diversity and secondary succession, and is becoming more abundant in human-modified habitats where it roosts in non-native plants. Although this change in roosting preferences can have detrimental consequences to their populations, to date, no study has quantified this phenomenon from a population genetic perspective. To investigate effect of human-modified habitats on group cohesion, population structure, and genetic diversity, we developed and characterized 17 microsatellite loci. Primer pairs were screened on 187 individuals from 12 different social groups. Levels of polymorphism ranged from 3 to 14 alleles. Of the 17 loci, 3 revealed significant departures from Hardy–Weinberg equilibrium and evidence of null alleles. Use of these markers will allow researchers to understand effects to population genetic structure and diversity due to changes in habitat use, which is important information to form the best correct management strategy and to ensure perpetuity of an important seed disperser.     

Isolation and characterization of eight polymorphic microsatellite loci for Natterer’s bat, Myotis nattereri (Vespertilionidae, Chiroptera)

Eight new microsatellite loci were isolated and characterized for the Natterer’s bat Myotis nattereri from a microsatellite-enriched genomic library. The usefulness of these markers was assessed by screening a sample comprising 100 specimens collected from throughout the species range in Europe. Both moderately and highly polymorphic loci were identified with 3–17 alleles segregating per locus (mean 8.1 SE ± 0.048). No evidence for departure from HWE or linkage disequilibrium among loci was observed. These markers will provide a valuable addition to the molecular toolbox currently available for studies of population genetic structure, parentage and social organisation of M. nattereri and related species.     

Isolation and characterization of microsatellite markers from the Chinese endemic species,Beal’s-eyed turtle (Sacalia bealei)

Beal’s-eyed turtle (Sacalia bealei) is an endangered species with important medicinal values and effective measures should be taken to protect this species. Development of molecular markers that could be used in population genetic assessment is necessary for the conservation of endangered species. In this investigation, we isolated 14 microsatellite DNA markers from an enriched library. The number of alleles at each locus was between 7 and 17. He and Ho ranged from 0.7958 to 0.9324 and 0.6333 to 0.9697 respectively. The PIC value ranged from 0.757 to 0.913, indicated all the loci are high polymorphism. No significant linkage disequilibrium was detected and only one locus deviated from Hardy–Weinberg equilibrium. These loci could provide useful information for the genetic analyses of S. bealei populations.     

Development and characterization of 24 microsatellite loci for the Illinois chorus frog (Pseudacris illinoensis) and Strecker’s chorus frog (P. streckeri)

The Illinois chorus frog (Pseudacris illinoensis), originally described as a subspecies of Strecker’s chorus frog (P. streckeri), has a disjunct distribution in Illinois, Missouri, and Arkansas. Populations of P. illinoensis are threatened by habitat loss caused by current agricultural practices, but this taxon has not yet been assessed for the IUCN Red List. We developed 24 microsatellite loci to study the genetic structure of P. illinoensis and P. streckeri, investigate the evolutionary history of this species complex, and help inform conservation efforts. The number of alleles ranged from 4 to 13, observed heterozygosities from 0.214 to 0.889, and expected heterozygosities from 0.476 to 0.899, indicating these markers will be useful for studying population genetic variation.     

Isolation and characterization of eight novel microsatellite loci in whisker sheatfish (Micronema bleekeri Günther, 1864)

We describe the isolation and characterization of eight microsatellite loci from whisker sheatfish (Micronema bleekeri Günther, 1864). Genetic variability was assessed using 35 individuals. The number of alleles per locus ranged from two to four. The observed heterozygosity (H _o) ranged from 0.072 to 0.637, whereas the expected heterozygosity (H _e) ranged from 0.174 to 0.725. All loci conformed to the Hardy–Weinberg expectation (HWE). Linkage disequilibrium was not significant for any pair of loci. These loci should prove useful for population genetic studies of whisker sheatfish and other siluriforme species.     

Isolation and characterization of 12 novel microsatellite loci in Hume’s Pheasant, Syrmaticus humiae

Hume’s Pheasant, Syrmaticus humiae is a rare and endangered species of pheasants. As a result of habitat loss, poaching and human disturbance, its wild populations have decreased dramatically, and it was listed as a globally near-threatened species by BirdLife/IUCN. Here, we describe the first set of twelve polymorphic microsatellite markers isolated from S. humiae genomic libraries using an enrichment protocol which is able to enrich AC and AG motifs. All the loci were screened from a group of 16 individuals sampled from Guangxi province, and each locus showed a relatively high polymorphism (ranged from 4 to 13 alleles). The observed and expected heterozygosity values are ranged from 0.2 to 0.938 and from 0.303 to 0.925, respectively. Four loci showed deviation from Hardy–Weinberg equilibrium and no linkage disequilibrium was found between pairs of loci. These informative microsatellite markers will be a useful molecular tool for further studies of the population genetic structure and evolutionary history of S. humiae.