Isolation and characterization of polymorphic microsatellite DNA markers in the Japanese eight‐barbel loach,Lefua echigonia

We isolated and characterized 17 polymorphic microsatellite loci for the Japanese eight‐barbel loach, Lefua echigonia, an endangered freshwater species in streams including agricultural canals in Japan. The number of observed alleles for each locus ranged from two to nine, and the values of observed and expected heterozygosities ranged from 0.125 to 0.844 and from 0.148 to 0.876, respectively. All loci conformed to Hardy–Weinberg equilibrium. Genetic linkage disequilibrium analysis between all pairs of the loci showed no significant departure from the null hypothesis between any of the loci.     

Ten polymorphic autosomal microsatellite loci for the Eider duck Somateria mollissima and their cross‐species applicability among waterfowl species (Anatidae)

We describe the isolation and characterization of the first microsatellite loci specifically developed for the Common Eider duck, Somateria mollissima. Our 10 loci show Mendelian inheritance and no linkage disequilibrium between any pair of loci. In the Eider duck, we observed between two and 16 alleles per locus, an expected heterozygosity between 0.31 and 0.97, and an observed heterozygosity between 0.14 and 1.00. Most primers also yield reproducible results in several other anatid species. These loci comprise a set of autosomal molecular markers for various applications, from moderately polymorphic loci suitable for population studies to highly polymorphic loci for pedigree analysis in waterfowl.     

Isolation and characterization of polymorphic microsatellite DNA markers in the Omono type of ninespine stickleback,genus Pungitius

We isolated and characterized 25 polymorphic microsatellite loci for the Omono type of ninespine stickleback, genus Pungitius, an endangered species in streams including agricultural canals in Japan. The number of observed alleles per locus ranged from two to 15 within 32 individuals each collected from three populations in the Omono River, and the values of observed and expected heterozygosities ranged from 0.031 to 0.906 and from 0.031 to 0.856, respectively. All loci conformed to the Hardy–Weinberg equilibrium. Genetic linkage disequilibrium analysis between all pairs of the loci showed no significant departure from the null hypothesis between any of the loci.     

Development and characterization of microsatellite markers in Populus euphratica (Populaceae)

In this study, we developed 12 microsatellite loci for Populus euphratica and checked their variability in 27 individuals. The number of alleles for 12 loci ranged from five to nine, the observed and expected heterozygosities ranged from 0.32 to 0.48 and from 0.53 to 0.67. Frequencies of null alleles of all loci are not significantly greater than zero. None of the loci showed significant deviation from Hardy-Weinberg equilibrium. No significant linkage disequilibrium was observed between pairs of studied loci. These markers are useful in studies of population genetic structure of P. euphratica.     

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.     

Microsatellite loci for Dawson's burrowing bee (Amegilla dawsoni) and their cross‐utility in other Amegilla species

Microsatellite loci were isolated from Dawson's burrowing bee, Amegilla dawsoni, a species endemic to Western Australia. Twelve polymorphic loci were found with an observed number of alleles ranging from two to 24 and observed heterozygosities between 0.17 and 0.85. These 12 loci were tested for amplification in three additional species of Amegilla. The loci will be used for sex determination and the examination of mating frequency in this species.     

Isolation of 21 polymorphic microsatellite markers for the Virginia opossum (Didelphis virginiana)

Twenty-one polymorphic microsatellite markers were developed for the Virginia opossum (Didelphis virginiana). The number of alleles ranged from two to 13 and observed heterozygosities ranged from 0.464 to 0.964. Significant heterozygote deficiencies were observed at three loci and null alleles were detected at five loci. Evidence for gametic disequilibrium was observed between three sets of paired loci after a sequential Bonferroni correction was applied. These markers will enable us to investigate the mating tactics, movement behaviour and social structure of Virginia opossum populations inhabiting fragmented agricultural landscapes.     

Development and characterization of 13 novel microsatellite loci from the flat-headed bat (Tylonycteris pachypus) with cross-species amplification in closely related taxa

13 novel microsatellite loci was isolated using the enriched library method from genomic DNA of the flat-headed bat (Tylonycteris pachypus). These loci showed high levels of genetic polymorphism testing on 54 individuals sampled from Guangxi province, China. The number of observed alleles per locus ranged from 2 to 15. The expected and observed heterozygosity values ranged from 0.170 to 0.900 and from 0.185 to 0.944, respectively. One loci departed significantly from Hardy-Weinberg equilibrium (HWE) after Bonferroni correction and no Linkage disequilibrium was found between any pairs of loci. In addition, these loci were tested in the sister species, Tylonycteris robustula, seven loci amplified successfully and were also polymorphic.     

Microsatellite loci for the Australian field cricket Teleogryllus oceanicus and their cross‐utility in Teleogryllus commodus

Microsatellite loci were isolated from the Australian field cricket Teleogryllus oceanicus. Seven polymorphic loci were found with an observed number of alleles ranging from eight to 17 and observed heterozygosities between 0.26 and 0.94. One locus was found to be X‐linked. These seven loci were also tested for amplification and polymorphism in the congeneric species Teleogryllus commodus. The loci will be used for paternity studies in these species.     

Extensive ribosomal DNA amplification during Andean common bean (Phaseolus vulgaris L.) evolution

The extent of 5S and 45S ribosomal DNA (rDNA) variation was investigated in wild and domesticated common beans (Phaseolus vulgaris) chosen to represent the known genetic diversity of the species. 5S and 45S rDNA probes were localized on mitotic chromosomes of 37 accessions by fluorescent in situ hybridization (FISH). The two 5S rDNA loci were largely conserved within the species, whereas a high variation in the number of 45S rDNA loci and changes in position of loci and number of repeats per locus were observed. Domesticated accessions from the Mesoamerican gene pool frequently had three 45S rDNA loci per haploid genome, and rarely four. Domesticated accessions from Andean gene pool, particularly from the race Peru, showed six, seven, eight or nine loci, but seven loci were found in all three races of this gene pool. Between three and eight loci were observed in accessions resulting from crosses between Andean and Mesoamerican genotypes. The presence of two to eight 45S rDNA loci in wild common beans from different geographic locations indicates that the 45S rDNA amplification observed in the Andean lineage took place before domestication. Our data suggest that ectopic recombination between terminal chromosomal regions might be the mechanism responsible for this variation.     

Primers for 12 polymorphic microsatellite DNA loci from the guppy (Poecilia reticulata)

Twelve polymorphic microsatellite DNA loci were isolated from genomic guppy DNA. We assessed the level of genetic diversity for these loci using individuals from five native Trinidadian populations. All the loci were polymorphic although there were considerable differences among loci and populations in allele frequencies. Within populations, the number of alleles ranged from 1 to 22 and observed heterozygosities ranged from 0.000 to 0.990. Allele frequencies differed substantially between populations suggesting divergence.     

Characteristics of 11 polymorphic microsatellite markers in the red imported fire ant, Solenopsis invicta Buren

We have characterized 11 polymorphic microsatellite loci in the invasive ant Solenopsis invicta. Primer pairs were evaluated on fire ants collected from monogyne mounds in Lauderdale County, Mississippi. The observed and effective number of alleles ranged from two to six and from 1.31 to 2.64, respectively. The observed and expected heterozygosity values ranged from 0.1613 to 0.7826 and from 0.1491 to 0.6242, respectively. The polymorphism information content of the microsatellites ranged from 0.1482 to 0.6208. Probability tests indicated significant deviations from the Hardy-Weinberg equilibrium at three loci. Pairwise tests did not detect linkage disequilibrium between any pair of loci.     

Fourteen polymorphic microsatellite loci characterized in the house sparrow Passer domesticus (Passeridae,Aves)

We characterized 14 polymorphic microsatellite loci in the house sparrow Passer domesticus. Four loci were isolated from house sparrow genomic libraries and 10 loci were identified by testing 100 loci that had been originally isolated in other passerine species. Loci were characterized in 37–54 unrelated sparrows from British and Norwegian populations. Each locus displayed between two and 31 alleles, with the observed heterozygosity ranging between 0.30 and 0.91.     

Linkage Disequilibrium between Allozymes in Natural Populations of Lodgepole Pine

Pairwise linkage disequilibrium values (D) were estimated for 14 allozyme loci in two natural populations of lodgepole pine (Pinus contorta ssp. latifolia). Maternal multilocus genotypes were inferred from samples of (haploid) megagametophytic seed-endosperms. Coupling/repulsion double heterozygotes were distinguished for closely linked pairs of loci. Assays of seven of the loci in seed embryos allowed estimates of D for these loci in the outcross pollen pool (estimates of outcrossing rates indicate no significant departures from random mating in either population). No disequilibrium was observed between unlinked loci in either maternal genotypes or outcross pollen. However, significant disequilibrium was observed within and between gametes for some allelic combinations of four tightly linked loci; the assumption of random association of gamete types within individuals is thus invalid for some loci in lodgepole pine. Possible causes of the observed D were examined using the noncentrality parameter of the general noncentral chi square distribution. We concluded, from estimates of population size, linkage and measurements of population substructure, that neither drift nor population subdivision was responsible for the significant values of D which were observed and that epistatic selection was the most likely cause of the disequilibrium observed.     

Development of polymorphic EST-derived SSR markers for the shrimp, Fenneropenaeus chinensis

In our study, we evaluated simple sequence repeat (SSR) markers from expressed sequence tags (ESTs) of Fenneropenaeus chinensis. Characteristics of 11 EST-SSR loci were investigated using 34 F. chinensis individuals. The number of alleles per locus ranged from 4 to 9. The observed heterozygosity ranged from 0.118 to 0.826, while the expected heterozygosity ranged from 0.623 to 0.847. Two loci (T2 and T12) conformed to the Hardy–Weinberg equilibrium. There was no LD observed between all pairs of EST-SSRs loci. The PIC values of 11 microsatellite loci were higher than 0.5. These loci and markers will be useful for population genetics and systemic evolution of F. chinensis.     

Isolation and characterization of microsatellite loci in the whale shark (Rhincodon typus)

In preparation for a study on population structure of the whale shark (Rhincodon typus), nine species-specific polymorphic microsatellite DNA markers were developed. An initial screening of 50 individuals from Holbox Island, Mexico found all nine loci to be polymorphic, with two to 17 alleles observed per locus. Observed and expected heterozygosity per locus ranged from 0.200 to 0.826 and from 0.213 to 0.857, respectively. Neither statistically significant deviations from Hardy-Weinberg expectations nor statistically significant linkage disequilibrium between loci were observed. These microsatellite loci appear suitable for examining population structure, kinship assessment and other applications.     

Characterization of 38 microsatellite loci in the European blackbird, Turdus merula (Turdidae, AVES)

We characterized 38 microsatellite loci in the European blackbird, Turdus merula. Thirty-seven loci were identified by testing 242 loci that had been originally isolated in other avian species. One additional locus was isolated from a European blackbird genomic library. All loci were characterized in 20-29 blackbirds from a population in the Czech Republic and displayed between two and 16 alleles, with observed heterozygosity ranging from 0.04 to 1.00. Thirty-seven loci could be assigned a chromosome location in the zebra finch (Taeniopygia guttata) genome based on sequence homology.     

Characterization of 20 microsatellite loci in the long‐tailed tit Aegithalos caudatus (Aegithalidae,AVES)

We characterized 20 microsatellite loci in the long‐tailed tit, Aegithalos caudatus. Polymorphic loci were identified by testing 114 loci that had been originally isolated in other avian species. The loci were characterized in 23–163 unrelated long‐tailed tits from a British population and displayed between two and 42 alleles, with observed heterozygosity ranging between 0.09 and 0.97.     

Characterization of eight polymorphic microsatellite loci from the Bengalese finch (Lonchura striata var. domestica)

Eight polymorphic microsatellite loci were isolated and characterized from the Bengalese finch (Lonchura striata var. domestica). In analyses of 25 individuals, the number of alleles ranged from two to four, and observed heterozygosity ranged between 0.05 and 0.73. At four loci, the observed heterozygosity of the Bengalese finches was significantly different from the expected heterozygosity. Primer sets were also tested in Javan munia (Lonchura leucogastroides), and the same eight loci were successfully amplified. In analyses of 20 unrelated individuals, the number of alleles ranged from one to seven, and the observed heterozygosity ranged from 0 to 0.56. In Javan munia, the observed heterozygosity differed significantly from the expected heterozygosity in only one locus.