Identification of polymorphic microsatellite markers from RAPD product in turbot (Scophthalmus maximus) and a test of cross‐species amplification

Five polymorphic microsatellite loci have been isolated and characterized from random amplified polymorphic DNA product in turbot, Scophthalmus maximus. Twelve microsatellites were selected for designing microsatellite primers, of which five gave working primer pairs. They had between four and nine alleles. Observed and expected heterozygosities varied from 0.76 to 0.90 and from 0.63 to 0.83, respectively. Five additional fish species assessed for cross‐species amplification revealed between one and three positive amplifications and between zero and two polymorphic loci per species.     

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.     

Isolation and characterization of the first microsatellite loci from the order Psocoptera in the economically important pest insect Liposcelis decolor (Pearman) and cross‐species amplification

A total of 11 microsatellite loci from the invasive insect pest Liposcelis decolor were isolated and characterized of which six loci were polymorphic. A population survey involving a total of 30–192 individuals per locus from five populations revealed a range of four to seven alleles per locus and moderate observed heterozygosities (0.183–0.565), highlighting the utility of these loci in further population genetic studies. Cross‐species amplifications were successful for two to 11 loci in five other Liposcelis species also of international economic importance.     

Five new microsatellite loci for Oliver flounder (Paralichthys olivaceus) from an expressed sequence tag (EST) library and cross‐species amplification

Five microsatellite loci have been isolated and characterized from a cDNA library of Oliver flounder, Paralichthys olivaceus. All loci were found to be polymorphic and had between four and 10 alleles. Observed and expected heterozygosities varied from 0.70 to 0.90 and from 0.52 to 0.80, respectively. Five additional fish species assessed for cross‐species amplification revealed between zero and three positive amplifications and between zero and two polymorphic loci per species.     

Microsatellite markers from an expressed sequence tag library of half‐smooth tongue sole (Cynoglossus semilaevis) and their application in other related fish species

Microsatellite markers have been developed from a cDNA library of half‐smooth tongue sole, Cynoglossus semilaevis. Twenty‐five microsatellites were selected for designing microsatellite primers, of which 11 gave working primer pairs. They had between four and 12 alleles. Observed and expected heterozygosities varied from 0.60 to 0.90 and from 0.57 to 0.88, respectively. Five additional fish species assessed for cross‐species amplification revealed between one and four positive amplifications and between 0 and four polymorphic loci per species.     

Isolation and characterization of microsatellite markers in Calomys musculinus (Muridae,Sigmodontinae, Phyllotini), the natural reservoir of Junin virus

Calomys musculinus, one of the most abundant rodent species in central Argentina, is the reservoir of Junin virus, the aetiological agent of Argentine hemorrhagic fever. We isolated six polymorphic microsatellite loci for microgeographical studies of population structure in this species. Amplification of these loci in 36 individuals from three natural populations revealed five to 14 alleles per locus and expected heterozygosities from 0.426 to 0.868. Cross‐species amplifications suggest that primers designed for these loci may be useful in other closely related species of the tribe Phyllotini, but not in species of other more distantly related tribes of the subfamily Sigmodontinae.     

Isolation and characterization of microsatellite loci in the great bustard,Otis tarda

With the aim to study population genetics of the endangered great bustard, Otis tarda, dinucleotide microsatellite loci were isolated using an adapted hybrid‐capture enrichment protocol. This work reports the characterization of a set of six polymorphic microsatellite markers within the great bustard (n = 52). Results from cross‐species amplifications in several other members of the family Otididae demonstrate that five primer pairs also successfully amplified homologous loci outside the species Otis tarda.     

Isolation and characterization of polymorphic microsatellite loci in Wuchang bream (Megalobrama amblycephala)

Wuchang bream (Megalobrama amblycephala) is an economically important fish in China. From a (GT)₁₃‐enriched genomic library, 20 microsatellites were developed. Nine of these 20 loci were polymorphic in a test population with allele numbers ranging from two to four, and the observed and expected heterozygosities ranging from 0.2609 to 0.7826 and from 0.3739 to 0.7546, respectively. In the cross‐species amplifications, six of these nine loci were also polymorphic in white amur bream (Parabramis pekinensis). These polymorphic microsatellite loci are potentially useful for population genetics of Wuchang bream and its closely related species.     

Isolation,characterization and cross‐salmonid amplification of 31 microsatellite loci in the lake whitefish (Coregonus clupeaformis,Mitchill)

Coregonine fish represent the most successful evolutionary lineage of salmonids with Coregonus as the most speciose salmonid genus inhabiting numerous postglacial lakes across the northern hemisphere. We isolated and characterized 31 polymorphic microsatellite loci in Coregonus clupeaformis with an average number of 5.3 alleles per locus (range three to eight) and an overall expected heterozygosity of 0.74 ± 0.11. Two loci revealed significant linkage associations through analyses of mapping families. Six additional salmonid taxa assessed for cross‐species amplification revealed between 18 and 26 positive amplifications and between two and 12 polymorphic loci per species.     

Characterization of five microsatellite loci in the pine shoot beetle Tomicus piniperda (Coleoptera: Scolytidae)

Tomicus piniperda is a bark beetle that causes damage to various pine species across a wide geographical range. We developed five microsatellite polymorphic markers using an enrichment protocol. All loci could be successfully amplified with no evidence of null alleles and will be useful for population genetic studies. Cross‐species amplifications show that at least some of the markers could be useful in four other Tomicus species.     

Isolation and characterization of polymorphic microsatellite loci from an EST‐library of red sea bream (Chrysophrys major) and cross‐species amplification

Microsatellite markers have been developed from a complementary DNA (cDNA) library of red sea bream, Chrysophrys major. Twenty‐eight microsatellites were selected for designing microsatellite primers, of which 11 gave working primer pairs. Observed and expected heterozygosities varied from 0.33 to 1.00 and from 0.38 to 0.83, respectively. Five additional fish species assessed for cross‐species amplification revealed between one and six positive amplifications and between 0 and 6 polymorphic loci per species.     

Characterization of novel microsatellite loci in rare minnow (<Emphasis Type="Italic">Gobiocypris rarus</Emphasis>) and amplification in closely related species in Gobioninae

Rare minnow (Gobiocypris rarus) is an endangered small fish endemic to upper reach of the Yangtze River. From a (GT)n enriched genomic library, 32 microsatellites were isolated and characterized. Nineteen of these loci were polymorphic in a test population with alleles ranging from 2–7, and observed and expected heterozygosities from zero to 0.8438, and 0.2679 to 0.8264, respectively. In the cross-species amplifications, 13 out of 19 polymorphic loci were found to be also polymorphic in at least one of the 7 closely related species of the subfamily Gobioninae. These polymorphic microsatellite loci should provide sufficient level of genetic diversity to evaluate the fine-scale population structure in rare minnow and its closely related species for the conservation purpose.     

Microsatellite DNA markers for a grasshopper: Prionotropis hystrix rhodanica (Orthoptera,Pamphagidae)

Microsatellite loci were isolated from Prionotropis hystrix rhodanica, an endangered grasshopper species, endemic to a steppic area southeast of France. Six polymorphic loci were obtained from an enriched partial genomic libray. A high genetic diversity was observed at all loci, with an observed number of alleles ranging from seven to 29, and an observed heterozygosity between 0.71 and 0.90. Test of cross‐amplifications on two closely related taxa are presented.     

Isolation and identification of 21 microsatellite loci in the Copper redhorse (Moxostoma hubbsi; Catostomidae) and their variability in other catostomids

Catostomidae represent an important family of freshwater fishes mainly distributed in North America, but also found in Eurasia. This paper describes the development of microsatellite DNA markers for a highly threatened member of this family, the Copper redhorse (Moxostoma hubbsi), as well as cross‐catostomids amplifications. 168 tetra‐nucleotide loci were screened to develop 21 polymorphic markers, with an average number of 8.5 alleles per locus and observed heterozygosity ranging between 0.52 and 1.00. Successful amplification was obtained for 12 other members of the family at between seven to 19 loci, with between two to 18 loci being polymorphic per species.     

Isolation and characterization of polymorphic microsatellite loci from RAPD product in half-smooth tongue sole (Cynoglossus semilaevis) and a test of cross-species amplification

Eleven polymorphic microsatellite loci have been isolated and characterized from random amplified polymorphic DNA product in half-smooth tongue sole, Cynoglossus semilaevis. Twenty-one microsatellites were selected for designing microsatellite primers, of which 11 gave working primer pairs. They had between three and 12 alleles. Observed and expected heterozygosities varied from 0.53 to 0.93, and from 0.52 to 0.80, respectively. Five additional fish species assessed for cross-species amplification revealed between one and three positive amplifications and between zero and three polymorphic loci per species.     

Isolation and characterization of polymorphic microsatellite loci from a dinucleotide-enriched genomic library of seven-band grouper (Epinephelus septemfasciatus) and cross-species amplification

Seven-band grouper (Epinephelus septemfasciatus) is a commercially important fishery species. Sixty-six microsatellite loci were isolated from a dinucleotide-enriched genomic library of E. septemfasciatus. Twelve of these loci were polymorphic in a test population with alleles per locus ranging from two to five, and observed and expected heterozygosities per locus from 0.04 to 1.00 and from 0.28 to 0.76, respectively. Three loci significantly deviated from Hardy–Weinberg equilibrium and no significant linkage disequilibrium was found between pairs of loci after Bonferroni correction. Cross-species amplification of these polymorphic microsatellite loci was performed in additional two related species. These polymorphic microsatellite loci would be useful for investigating genetic resource of E. septemfasciatus and other related species. Lili Zhao and Changwei Shao have contributed equally.     

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.     

Microsatellite loci for studies of population differentiation and range expansion in Solidago sempervirens L. (Asteraceae)

For studies of population differentiation and range expansion in the seaside goldenrod, Solidago sempervirens, we identified hypervariable molecular markers by screening genomic libraries enriched for microsatellite motifs. We designed primers that reliably amplified nine polymorphic loci. High polymorphism in a population from Delaware Bay, USA suggests that the loci will be useful in population studies. The success of cross‐amplifications in 11 species of Asteraceae varied among loci and did not appear to reflect phylogenetic relationships within Solidago.     

Characterization of nuclear DNA microsatellite markers in the ant Cataglyphis cursor

The ant Cataglyphis cursor is exceptional in that unmated workers are potentially able to lay both male and female eggs. We characterized eight pairs of primers for microsatellite loci, developed from genomic DNA for this species. Variability was tested with DNA from 19 workers and all eight loci were highly polymorphic, displaying 5–10 alleles and a high level of heterozygosity. Cross‐species amplifications indicate that these microsatellites might be useful in genetic studies of other species belonging to the genus Cataglyphis.     

Microsatellite markers characterized in the barn owl (Tyto alba) and of high utility in other owls (Strigiformes: AVES)

We have identified 15 polymorphic microsatellite loci for the barn owl (Tyto alba), five from testing published owl loci and 10 from testing non‐owl loci, including loci known to be of high utility in passerines and shorebirds. All 15 loci were sequenced in barn owl, and new primer sets were designed for eight loci. The 15 polymorphic loci displayed two to 26 alleles in 56–58 barn owls. When tested in 10 other owl species (n = 1–6 individuals), between four and nine loci were polymorphic per species. These loci are suitable for studies of population structure and parentage in owls.