Isolation and characterization of microsatellite DNA primers in burrowing owl (Athene cunicularia)

We developed polymerase chain reaction (PCR) primers to amplify 22 microsatellite DNA loci in burrowing owls (Strigiformes: Athene cunicularia). The seven most easily amplified and scored polymorphic loci are presented. These show 3?19 alleles per locus and expected heterozygosities of 0.067–0.712. Inheritance at all loci conformed to Mendelian expectations among six owl families, and six loci conform to Hardy–Weinberg expectations.     

Characterization and isolation of DNA microsatellite primers for Arapaima gigas,an economically important but severely over‐exploited fish species of the Amazon basin

Arapaima gigas is a CITES II listed species, but it is also one of the most important food fish of the Amazon basin with a historically very large commercial and subsistence fishery. By the 1980s it became commercially extinct in many areas, and in early 2001 the Brazilian government banned all fishing, although this has not prevented illegal fishing. Therefore we developed 14 variable microsatellite markers for A. gigas to gain a knowledge of its population structure, which is needed to implement a sound management policy, and also to provide us with the forensic tools to manage and monitor this over‐exploited fish.     

Isolation and characterization of polymorphic microsatellite DNA markers in the brown sole,Pleuronectes herzensteini

New microsatellite DNA markers from brown sole were developed and characterized. Fourteen primer sets were designed from 40 microsatellite regions. Eight of 14 loci exhibited variations comprising 8–31 alleles. Observed and expected heterozygosities ranged from 0.611 to 0.833 and from 0.647 to 0.968 among 36 individuals, respectively. Phz3, Phz8 and Phz12 significantly deviated from Hardy–Weinberg equilibrium, and there was a significant linkage disequilibrium between Phz2 and Phz12. Seven of eight loci conformed to the Mendelian manner of inheritance in a full‐sib family. Seven to four loci of three related species were cross‐amplified by primers for brown sole.     

Isolation and characterization of microsatellite loci for storm-petrels

Primers were developed for 10 microsatellite loci for two species of Oceanodroma storm-petrels. Variability was tested in 27 O. castro and 22 O. monteiroi from the Azores, and 24 O. leucorhoa from Norway. At least six loci amplified reliably and were polymorphic in each species. The number of alleles per locus averaged 4.6, and observed heterozygosities averaged 0.41. Most primers also yielded polymerase chain reaction products in O. tethys, O. hornbyi and Pterodroma phaeopygia. These loci are being used to assay population genetic structure in storm-petrels.     

Isolation and characterization of polymorphic DNA microsatellite markers from Schistosoma japonicum

The ability of microsatellite loci to reveal genetic diversity within the trematode Schistosoma japonicum is demonstrated. Eleven novel microsatellite markers were isolated, assessing variability within 80 S. japonicum individuals from three Chinese and one Philippine population. Novel primers were also tested upon S. mansoni and S. haematobium. Eight primers showed polymorphic amplification from all S. japonicum groups, three amplified S. mansoni DNA, but none amplified S. haematobium DNA. Only six of 27 previously isolated S. mansoni‐specific primers amplified S. japonicum DNA. Allelic diversity and observed heterozygosity ranged from 4 to 21 and 0.05 to 0.82, respectively, suggesting high variability.     

Isolation and characterization of polymorphic microsatellite markers in the migratory freshwater fish Prochilodus costatus

We isolated six polymorphic microsatellite loci in the migratory freshwater fish Prochilodus costatus, which is an endemic species and important fisheries resource from São Francisco river basin, Brazil. We have evaluated the applicability of these loci to study genetic variation in wild population of this fish. Thus, based on the genotypes of 32–48 individuals, we detected two to 21 alleles per locus, observed and expected heterozygosities ranging from 0.19 to 0.89 and from 0.17 to 0.92, respectively. These polymorphic markers should provide efficient tools to study population genetic structure of this fish.     

Isolation of polymorphic microsatellite markers for the tetraploid Dipteryx odorata, an intensely exploited Amazonian tree species

Dipteryx odorata is an intensely exploited Amazonian tree legume. Microsatellite markers were developed to study the genetic structure, gene flow and reproductive biology of D. odorata. Eight highly polymorphic microsatellite markers were isolated from enriched repeat libraries screened for microsatellite repeats. An average of 16 alleles and 0.964 phenotype diversity per locus were found in 76 individuals from the Tapajos National Forest, in the state of Pará in the Brazilian Amazon.     

Isolation and characterization of microsatellite DNA markers in the malaria vector Anopheles maculipennis

The Anopheles maculipennis complex includes the most important malaria vectors of the western Palearctic. Anopheles maculipennis s.s., one member of this complex, is a reported vector in the Middle East. Here we describe the isolation of 15 microsatellite polymorphic loci from the An. maculipennis s.s. genome, displaying a high among individual diversity (0.37–0.77) in a sample from France. Three loci displayed a significant departure from Hardy–Weinberg proportions, suggesting a substantial frequency of null alleles. The remaining 12 loci are good candidates for further genetic studies in this species.     

Isolation and characterization of microsatellite DNA markers in the malaria vector Anopheles sacharovi

The mosquito Anopheles sacharovi, a member of the A. maculipennis complex, is an important malaria vector in the Middle East. Here we describe the isolation of 15 microsatellite polymorphic loci from the A. sacharovi genome, displaying a high among‐individual diversity (0.30–0.92) in a sample from Turkey. Seven loci displayed a significant departure from Hardy–Weinberg proportions, suggesting a substantial frequency of null alleles. The remaining eight loci are good candidates for further genetic studies in this species.     

Isolation and characterization of microsatellite DNA markers in the malaria vector Anopheles funestus

Screening of the Anopheles funestus genomic DNA library detected 18 new sequences with dinucleotide tandem repeats. Primers were designed to amplify the loci and 14 out of 18 gave a repeatable and scorable amplification. Deviations from Hardy–Weinberg expectations were tested for each locus in a sample of 30 wild Anopheles funestus females. No heterozygote deficiency was detected for 11 loci of 14, thus revealing the absence of null alleles. The number of alleles per locus ranged from 5 to 15, and observed heterozygosity from 0.13 to 0.85.     

Isolation and characterization of microsatellite DNA markers in the malaria vector Anopheles nili

The mosquito Anopheles nili is widespread across tropical Africa and appears to be the major vector of malaria in some rural forested areas of central Africa. Here we describe the isolation of 11 microsatellite polymorphic loci from the A. nili genome, displaying a high among‐individual diversity (0.58–0.96) in samples from west Africa. Two loci displayed a significant departure from Hardy–Weinberg proportions across all samples, suggesting a substantial frequency of null alleles. The remaining nine loci are good candidates for further genetic studies in this species.     

Isolation and characterization of 13 polymorphic nuclear microsatellite primers for the widespread Indo-Pacific three-spot damselfish, Dascyllus trimaculatus, and closely related D. auripinnis

A set of 13 simple sequence repeat markers was developed from D. trimaculatus genomic DNA, tested for D. auripinnis and characterized using 40 individuals per species. All the loci were polymorphic with a number of alleles ranging from three to 30. Observed heterozygosities varied from 0.23 to 0.89 for D. trimaculatus and from 0.11 to 0.85 for D. auripinnis. Early results show that these will be powerful markers for the study of ecological and evolutionary mechanism in this coral reef fish species complex.     

Isolation and characterization of microsatellite DNA markers in the Pacific abalone,Haliotis discus hannai

We developed 17 new microsatellite markers in Haliotis discus hannai. All loci were found to be polymorphic with an average of 13.1 alleles per locus (range 3–28). The mean observed and expected heterozygosities were 0.77 (range 0.17–1.00) and 0.79 (range 0.42–0.96), respectively. Six loci deviated significantly from Hardy–Weinberg proportions, and thus should be used with caution. The high variabilities revealed in this study suggest that these microsatellite loci should provide useful markers for studies of trait mapping, kinship and population genetics.     

Identification and characterization of 17 microsatellite primers for the tick,Ixodes ricinus,using enriched genomic libraries

Ixodes ricinus is the main vector for important infectious diseases in both humans and in animals. Microsatellite loci were isolated from a dinucleotide‐enriched library made from I. ricinus sampled in Norway. Seventeen polymorphic microsatellites were further characterized among 24 individuals sampled from an island in the Oslofjord region. The number of observed alleles ranged from two to 17 and the observed heterozygosities between 0.10 and 0.83. Analysis of family materials gives evidence of non‐Mendelian inheritance of several of the characterized loci, among which most could be explained by presence of null alleles.     

PERMANENT GENETIC RESOURCES: Isolation and characterization of microsatellite DNA loci from the southern flounder, Paralichthys lethostigma

Paucity of polymorphic molecular markers in southern flounder (Paralichthys lethostigma) has been a major limitation in genetic improvement of this important economic fish. Hence, we constructed a repeat-enriched genomic library from P. lethostigma. A total of 39 new microsatellites were identified, for which 33 primer pairs were designed. After validating and scoring, 10 of these loci were polymorphic in a test population with the range of alleles from two to nine per locus. The observed and expected heterozygosities ranged from 0.2500 to 0.9000 and from 0.4469 to 0.8514, respectively. These polymorphic microsatellites will be useful for genetic diversity analysis and linkage map construction for P. lethostigma.     

Isolation and characterization of microsatellite markers for the seagrass Cymodocea nodosa

In order to study the spatial patterns of genetic diversity of a clonal marine angiosperm, the seagrass Cymodocea nodosa, microsatellite markers were obtained by screening a genomic library enriched for the (CT) dinucleotide motif. Of 38 primer pairs defined, 15 amplified polymorphic microsatellites and are described. These loci identified a number of alleles ranging from two to seven, and showed expected heterozygosity ranging from 0.35 to 0.76, when a group of 40 individuals from Cadiz Bay in Spain was analysed. Additionally, we describe here the multiplexing conditions for 12 of these loci.     

Characterization and isolation of DNA microsatellite primers for the tropical tree,Symphonia globulifera Linn. f.

Five highly polimorphic microsatellite markers for Symphonia globulifera were developed from enriched repeat libraries screened for microsatellite repeats. An average of 5.8 alleles per locus and expected heterozygosity ranging from 0.649 to 0.806 were found in a sample of 12 S. globulifera individuals from the Tapajos National Forest, in the state of Pará in the Brazilian Amazon. These loci are being used for analysis of precise paternity testing, estimation of gene flow and parentage coefficients in a managed forest in Pará, Brazil, as part of the Dendrogene Project.     

Isolation and characterization of microsatellite markers in the Serra Spanish mackerel, Scomberomorus brasiliensis

Thirteen nuclear-encoded microsatellites from a genomic DNA library of Serra Spanish mackerel, Scomberomorus brasiliensis, were isolated and characterized. The microsatellites include 10 perfect repeats (eight tetranucleotide and two dinucleotide) and three imperfect repeats (two tetranucleotide and one dinucleotide). An additional five microsatellites, isolated originally from two congeneric species (S. cavalla and S. niphonius), were characterized in S. brasiliensis. Serra Spanish mackerel support artisanal fisheries along the Caribbean and Atlantic coasts of Central and South America, from Belize to Brazil.     

Isolation and characterization of microsatellite loci in Piaractus mesopotamicus and their applicability in other Serrasalminae fish

The subfamily Serrasalminae (Characidae, Teleostei) is an endemic Neotropical group of fishes distributed along the Amazon, Paraná‐Paraguay and Orinoco Basins. The pacu (Piaractus mesopotamicus) is found in all major rivers that comprise the Pantanal of Mato Grosso. In order to investigate population genetic structure within the group, an enriched microsatellite library was constructed. Eight polymorphic were loci screened, with an average of 6.5 alleles per locus. Five loci exhibited greater than 60% heterozygosity. Additionally, a high level of cross‐species amplification was obtained.