Isolation and characterization of microsatellite loci in the black rat snake (Elaphe obsoleta)

We obtained molecular markers useful for population level studies of the black rat snake (Elaphe obsoleta) by screening genomic DNA libraries enriched for dinucleotide, tetranucleotide, and pentanucleotide microsatellite repeats. Following sequencing of the positive clones, 11 pairs of primers were designed for polymorphic loci and their variability assessed in > 350 individuals from four populations in North America. The loci had between 9 and 40 alleles and observed heterozygosities ranged from 0.071 to 0.87. Some of these pairs of primers also successfully amplified DNA from two other snake species.     

Characterization of polymorphic microsatellite loci for the Tawny Pipit Anthus campestris and their utility in other steppe birds

New microsatellite loci for Tawny Pipit Anthus campestris were isolated from a genomic library. We were able to unambiguously score six loci: two were dinucleotide, one trinucleotide, two tetranucleotide and one pentanucleotide that turned out to be sex-linked. Four out of six loci were polymorphic with 7–23 alleles in our population and an observed heterozygosity ranging between 0.286 and 0.936. Cross-utility of these markers was tested in other 17 steppe-bird species of six families. In addition, 16 microsatellite loci developed for other species were tested for cross-species amplification in A. campestris. Eight microsatellite markers were successfully amplified; seven of them were polymorphic with 2–43 alleles and an observed heterozygosity of 0.040–0.863. Overall, 14 functional locus markers have been characterized for A. campestris that could be useful for future studies of paternity, genetic variability and population structure.     

Microsatellite loci in the Propertius duskywing,Erynnis propertius (Lepidoptera: Hesperiidae), and related species

Fifteen polymorphic dinucleotide microsatellite loci were characterized for Erynnis propertius using an enrichment protocol. The number of alleles varied from nine to 28 for a sample of 24 individuals. Observed heterozygosities ranged from 0.25 to 0.96. Homozygote excess was detected for 10 loci. Twelve markers successfully amplified in related Erynnis species and eight loci were polymorphic in at least one other species.     

Eight highly polymorphic microsatellite loci for the Australian gecko Heteronotia binoei

Eight highly polymorphic dinucleotide microsatellite loci were developed for the Bynoe's gecko, Heteronotia binoei. Across the species as a whole, expected heterozygosities for the loci range from 0.59 to 0.92, with observed numbers of alleles ranging from 13 to 27. All eight loci successfully amplify in each of the three most widespread sexual chromosome races of Heteronotia binoei, and with the exception of one locus in one race all are polymorphic. All eight loci also amplify in hybrid parthenogenetic Bynoe's geckos, in several other sexual chromosome races, and in related Heteronotia species.     

Development and characterization of novel tetra‐ and dinucleotide microsatellite markers for the French Polynesia black‐lipped pearl oyster,Pinctada margaritifera

Ten microsatellite loci were developed for the black‐lipped pearl oyster Pinctada margaritifera with a magnetic bead enrichment protocol. These tetra‐ and dinucleotide markers were polymorphic, with 10 to 43 alleles observed in 97 individuals from two Tuamotu atoll populations. Most loci revealed significant genic differentiation between the two populations and also exhibited some degree of heterozygote deficiencies, probably due to the presence of null alleles. These loci should be very useful to describe genetic structure, genetic variability and reproductive success in the various aquaculture and wild populations of pearl oyster in French Polynesia.     

Isolation and characterization of microsatellites in the sea hibiscus (Hibiscus tiliaceus L., Malvaceae) and related hibiscus species

Microsatellite loci were isolated from the sea hibiscus (Hibiscus tiliaceus L., Malvaceae), a pantropical plant with sea‐drifted seeds. This study describes six dinucleotide microsatellite loci for which the primers produced clear and polymorphic amplification patterns with different levels of variability (between three and nine alleles). Six markers were amplified in four other species of hibiscus, greatly increasing the utility of these markers.     

Isolation and characterization of nine polymorphic microsatellite loci from the desert locust,Schistocerca gregaria

Because of the scarcity of polymorphic genetic markers, only a few genetic studies on the population structure of the desert locust, Schistocerca gregaria, have been carried out. We isolated and characterized nine polymorphic dinucleotide microsatellite loci. These markers were evaluated using individuals from Niger and Senegal. Seven of these microsatellite markers are also applicable to the nongregarious subspecies Schistocerca gregaria flaviventris. Cross‐species applicability was limited to one of the loci in the sister species S. americana and in the locust Locusta migratoria.     

Isolation and characterization of microsatellite DNA loci from Russell's snapper (Lutjanus russellii)

A total of 37 microsatellite loci from the Russell's snapper, Lutjanus russellii, were successfully isolated and characterized. Thirty‐four loci were polymorphic in L. russellii samples. Twenty of the 37 markers did not differ significantly from Hardy–Weinberg expected genotype proportions. Significant linkage disequilibrium was detected in one pairwise comparison. The numbers of alleles and observed heterozygosities in polymorphic loci ranged from two to 16 and from 0.41 to 0.95, respectively. These markers will be useful for studying the population genetic structure of this species.     

Isolation and characterization of 13 microsatellite loci in Rhinolophus pusillus (least horseshoe bat) with cross-amplification in five related species

We used the enriched genomic library method to isolate and characterize dinucleotide microsatellite loci in the least horseshoe bat, Rhinolophus pusillus. Seventeen loci were obtained and tested on 31 individuals sampled from Guangxi Province in southern China. Thirteen of these markers were polymorphic with expected heterozygosity ranging from 0.821 to 0.909. A total of 164 alleles were detected and the number of alleles per locus ranged from 9 to 16 (mean 12.6). These polymorphic markers will be used to assess population structure in R. pusillus. In addition, successful cross-amplification in five congeneric bat species suggests most of these markers will also be useful for studying related species.     

Characterization of microsatellite loci in the stick insects Bacillus rossius rossius,Bacillus rossius redtenbacheri and Bacillus whitei (Insecta: Phasmatodea)

Five microsatellite markers were obtained from a dinucleotide enriched genomic library of the stick insect Bacillus rossius rossius. The markers were tested in three species of Bacillus. All loci were polymorphic when tested across species. The number of alleles at each locus was low (maximum four alleles), but different allelic patters were observed among the species.     

Isolation and characterization of microsatellite markers in Ijima's leaf warbler,Phylloscopus ijimae (Aves: Sylviidae)

Microsatellite markers were isolated from Ijima's leaf warbler, Phylloscopus ijimae. From an enriched genomic library and using the fast isolation by AFLP of sequences containing repeats (FIASCO) method, 10 polymorphic loci were obtained. Four to 12 alleles were identified in an analysis of 23 Ijima's leaf warblers, with the degree of heterozygosity ranging from 0.35 to 0.87. The markers were tested in four species of Phylloscopus and two other non‐Phylloscopus species of Sylviidae. Some loci were successfully amplified and showed polymorphism in Phylloscopus species, whereas amplification in the non‐Phylloscopus species was less successful.     

Polymorphic microsatellite loci in the black‐and‐gold chromis,Neoglyphidodon nigroris (Teleostei: Pomacentridae)

We describe 15 polymorphic dinucleotide microsatellite loci in the black‐and‐gold chromis, Neoglyphidodon nigroris (Cuvier: Teleostei: Pomacentridae). Microsatellites were isolated from a partial genomic library that was enriched for CA repeat motifs. The 15 loci yielded between two and 23 alleles per locus in a sample of 16 fish from two forereef sites off the island of Hoga, Wakatobi National Park, Indonesia. Observed and expected heterozygosities varied from 0.13 to 1.00 and 0.24 to 0.98, respectively. These markers should allow us to discriminate between closely related individuals and also to assess the connectedness of populations of N. nigroris that inhabit different reefs.     

Characterization of microsatellite loci isolated from the blacktip shark and their utility in requiem and hammerhead sharks

We isolated and characterized 16 microsatellite loci from the blacktip shark, Carcharhinus limbatus, and tested cross‐species amplification in 11 Carcharhinus species and five additional shark genera. Thirty‐six (1.6%) and 180 (48%) colonies were positive for dinucleotide repeat motifs from unenriched and enriched libraries, respectively. Heterozygosities of polymorphic loci ranged from 0.04 to 0.96 with two to 22 alleles per locus. Amplification products were observed at nine to 13 loci (five to 11 of which where polymorphic) in 10 Carcharhinus species. Several loci were also polymorphic in each of the additional genera examined.     

Development of polymorphic microsatellite from RAPD bands of black sea bream Acanthopagrus schlegeli

Eight pairs of simple sequence repeat markers were developed from random amplified polymorphic DNA product in black sea bream Acanthopagrus schlegeli. Twenty microsatellites were selected for designing microsatellite primers, of which eight gave working primer pairs. They had between three and seven alleles. Observed and expected heterozygosities varied from 0.65 to 0.90, and from 0.58 to 0.82, respectively. Eight additional fish species assessed for cross‐species amplification revealed between two and five positive amplifications and between zero and three polymorphic loci per species.     

Microsatellite markers isolated from polyploid wood‐sorrel Oxalis alpina (Oxalidaceae)

Twelve polymorphic microsatellite loci were isolated from polyploid alpine wood‐sorrel, Oxalis alpina (Oxalidaceae), and optimized for future studies of its breeding system. The loci were screened for variability among 72 individuals from Pinos Altos, New Mexico. The primers amplified loci with allele number ranging from two to 17 per locus and with estimates of Nei's genetic diversity varying from 0.10 to 0.99. These primers provide an opportunity to use polymorphic DNA markers to study the causes of breeding system variability in this species.     

Isolation and characterization of 10 polymorphic dinucleotide microsatellite markers for llama and guanaco

We isolated and characterized five polymorphic dinucleotide microsatellite markers from llama (Lama glama) and five from guanaco (Lama guanicoe). All loci were assayed on wild llamas and guanacos from Argentina, as all of the primers were able to amplify in both species.     

Characterization of 17 polymorphic microsatellite loci in the Anise swallowtail,Papilio zelicaon (Lepidoptera: Papilionidae), and their amplification in related species

Fifteen polymorphic dinucleotide and two trinucleotide microsatellite loci were identified in the Anise swallowtail, Papilio zelicaon, from DNA genomic libraries enriched for simple sequence repeats. Allele numbers varied from eight to 29, with an excess of homozygotes observed for nine loci. This homozygosity is a feature of other lepidopteran microsatellites and is probably due to null alleles. Sixteen markers were amplified successfully in other representatives of Papilio with 11 loci retaining polymorphism in at least one species. These results suggest that the microsatellites reported here may be appropriate for measuring population genetic structure in a number of Papilio species.     

Isolation and characterization of twelve polymorphic microsatellite markers in bottlenose dolphins (Tursiops truncatus)

We developed polymerase chain reaction primers for 12 dinucleotide microsatellite loci in the bottlenose dolphin, Tursiops truncatus. Seven markers were obtained after hybridization screening, and five following random genome sequencing. Orthologous positions were computed for nine markers on the bovine genome and for seven on the human genome. The markers are distributed across chromosomes and found in different types of DNA regions. All 12 loci are polymorphic for Tursiops. Five loci were also polymorphic in the related species Stenella frontalis and the more distantly related river dolphin, Inia geoffrensis, indicating these markers will be informative across the Delphinidae and other cetacean taxa.     

Characterization of microsatellite loci in Kearney's bluestar (Amsonia kearneyana) and cross‐amplification in other Amsonia species

Kearney's bluestar (Amsonia kearneyana) is a highly endangered herbaceous perennial in the family Apocynaceae. The species is found only in the Baboquivari Mountains of southern Arizona. We report the isolation and development of 12 microsatellite loci for Kearney's bluestar. Numbers of alleles ranged from two to four and observed heterozygosities ranged from 0.20 to 0.80 in the nine loci found to be polymorphic in the test population. All loci were also tested for cross‐amplification in five other Amsonia species representing two subgenera from the southwestern United States. Some loci that were not polymorphic in the Kearney's bluestar were polymorphic in other species.     

Isolation of 12 polymorphic microsatellite loci in the phytopathogenic fungus Alternaria brassicicola

Twelve polymorphic microsatellite markers were isolated from the phytopathogenic fungus Alternaria brassicicola, the causal agent of black spot of crucifers. An enrichment protocol was used to isolate microsatellite loci, which were then analysed in a collection of 46 isolates sampled from seven different countries. The number of alleles detected in 12 loci ranged from two to 10 (mean 3.5). Investigation of cross‐species amplifications showed that the designed primers were specific to A. brassicicola.