New polymorphic microsatellite loci in the house sparrow, Passer domesticus

We developed 13 new polymorphic microsatellite loci in the house sparrow (Passer domesticus), which exhibited from 2 to 15 alleles. Observed and expected heterozygosities ranged from 0.17 to 0.77 and from 0.35 to 0.85, respectively. We detected no linkage disequilibrium between loci. Allele frequencies supported Hardy-Weinberg equilibrium for 8 loci out of 13 after Bonferroni correction. Combined with loci previously isolated in the house sparrow, these new microsatellite markers provide valuable tools to study population genetics of this species.     

Eight new tetranucleotide microsatellite loci for the agile frog (Rana dalmatina)

We describe eight new polymorphic tetranucleotide microsatellite loci isolated from the agile frog (Rana dalmatina). In 25 individuals from the Nature Reserve Lüneburger Heide (Lower Saxony, Germany), the number of alleles per locus ranged from four to nine and average observed heterozygosities from 69.1% to 80.7%. No evidence for linkage disequilibrium was found and none of the loci showed significant deviation from Hardy-Weinberg expectations. These microsatellite DNA markers are suitable tools for addressing population genetics issues in this endangered species.     

Enzyme polymorphisms in the House Sparrow, Passer domesticus

Methods are described for the visualization of 20 enzymes from the house sparrow, Passer domesticus, in starch gels and by iso-electric focussing. Many of the methods are applicable to other avian species. Two samples of sparrows from the east midlands of England have been analysed. The patterns that are manifest in the gels can be ascribed to between 33 and 35 loci, of which the same nine are polymorphic in both samples. The level of heterozygosity in these birds is about 15%, which at first sight is higher than an introduced population in Australia. However, this difference disappears when homologous loci are compared.     

Microsatellite resources for Passeridae species: a predicted microsatellite map of the house sparrow Passer domesticus

We identified microsatellite sequences of potential utility in the house sparrow (Passer domesticus) and assigned their predicted genome locations. These sequences included newly isolated house sparrow loci, which we fully characterized. Many of the newly isolated loci were polymorphic in two other species of Passeridae: Berthelot's pipit Anthus berthelotii and zebra finch Taeniopygia guttata. In total, we identified 179 microsatellite markers that were either isolated directly from, or are of known utility in, the house sparrow. Sixty-seven of these markers were designed from unique sequences that we isolated from a house sparrow genomic library. These new markers were combined with 36 house sparrow markers isolated by other studies and 76 markers isolated from other passerine species but known to be polymorphic in the house sparrow. We utilized sequence homology to assign chromosomal locations for these loci in the assembled zebra finch genome. One hundred and thirty-four loci were assigned to 25 different autosomes and eight loci to the Z chromosome. Examination of the genotypes of known-sex house sparrows for 37 of the new loci revealed a W-linked locus and an additional Z-linked locus. Locus Pdoμ2, previously reported as autosomal, was found to be Z-linked. These loci enable the creation of powerful and cost-effective house sparrow multiplex primer sets for population and parentage studies. They can be used to create a house sparrow linkage map and will aid the identification of quantitative trait loci in passerine species.     

Microsatellite markers for Spergularia media (L.) C. Presl. (Caryophyllaceae) and their cross-species transferability

We developed 11 polymorphic microsatellite markers for Spergularia media (Caryophyllaceae), a perennial halophyte of coastal salt meadows and continental areas of western Eurasia. The number of alleles per locus observed in a single population of 20 individuals from the German North Sea coast ranged from 3 to 20. Observed and expected heterozygosities ranged from 0.200 to 0.850 and from 0.278 to 0.936 respectively. Observed heterozygosities were lower than expected heterozygosities at all loci, presumably as a consequence of inbreeding. All markers cross-amplified in the closely related S. salina. Of these, nine were polymorphic in the heterologous species. Only two primer pairs generated PCR products in S. diandra.     

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.     

Characterization of polymorphic microsatellite loci in the endangered Miho spine loach (Iksookimia choii) and cross-species amplification within the Cobitidae family

The Miho spine loach (Iksookimia choii) is an endangered freshwater fish endemic to Korea. To aid a conservation programme for this species, 20 polymorphic microsatellite loci were isolated and characterized in 67 individuals from three populations. The number of alleles per locus ranged from 8 to 33. Within populations, the observed and expected heterozygosities ranged from 0.59 to 1.00 and from 0.56 to 0.96, respectively, indicating its usefulness in population genetics studies. Cross-species amplification of the loci was tested in six species of the family Cobitidae, and 18 loci were amplified in two or more related species.     

Isolation and characterization of microsatellite loci from Corydalis ambigua (Papaveraceae)

Primers for seven unlinked and highly polymorphic microsatellite loci were developed for Corydalis ambigua to investigate its mating system and population genetics. This species can be used to explore the impact of an alien bumblebee on plant reproduction. Genetic diversity and other population genetic parameters were estimated in two populations with and without the alien bumblebee. The number of alleles per locus ranged from 4 to 20, and observed heterozygosities ranged from 0.355 to 0.969. These markers can be applied to study mating systems and population genetics in C. ambigua.     

Twenty microsatellite DNA markers for the Venus clam (<Emphasis Type="Italic">Cyclina sinensis</Emphasis> Gmelin)

The first set of 20 polymorphic microsatellite loci was isolated and characterized in the Venus clam Cyclina sinensis for population studies. Number of alleles per polymorphic locus varied from 6 to 18. The observed and expected heterozygosities ranged from 0.464 to 1.000 and from 0.739 to 0.938, respectively. These informative markers obtained in this study will offer insights to the assessment of population structure and conservation genetics in the future.     

Isolation and characterization of six polymorphic microsatellite markers in the freshwater turtle Mauremys rivulata (Testudines: Geoemydidae)

Six polymorphic microsatellite loci containing dinucleotide repeats were developed for the freshwater turtle Mauremys rivulata. The number of alleles ranged from five to 18. The observed and expected heterozygosities ranged from 0.19 to 0.79 and 0.46 to 0.90, respectively. These markers may serve as a valuable tool for population genetics analyses and provide information on the evolutionary history of the species.     

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.     

Characterization of 15 tetranucleotide microsatellite markers in the ringtail (Bassariscus astutus)

We characterized 15 polymorphic tetranucleotide microsatellite markers for the ringtail, Bassariscus astutus. We tested these loci in 21 individuals captured in Arizona and Texas and found six to 19 alleles per locus. Observed and expected heterozygosities ranged from 0.381 to 1.000 and from 0.381 to 0.941, respectively. All loci were in Hardy-Weinberg equilibrium, and none were in linkage disequilibrium. These markers may be used to investigate population genetics and mating patterns in this species.     

Characterization of nine microsatellite loci for the tree species Parapiptadenia rigida

Parapiptadenia rigida, locally known as angico, is a tropical tree common in the semideciduous Brazilian forest. Its wood is naturally resistant to insect attack and is useful for construction. Extracts from the tree have medicinal properties. We characterized nine microsatellite loci for P. rigida. Thirty-five alleles were detected in a sample of 45 individuals from 3 different populations, with an average of 3.9 alleles per locus. The average polymorphic information content ranged from 0.099 to 0.640. Observed and expected heterozygosities varied from 0.111 to 0.489 and from 0.106 to 0.707, respectively. One locus exhibited significant deviation from Hardy-Weinberg equilibrium and four pairs of loci showed significant linkage disequilibrium. All nine primers were tested for cross-amplification in species from the Fabaceae-Mimosoidea family, yielding a transferability success rate of 7 loci in Stryphnodendron adstringens to 0 transferred loci in Pithecellobium incuriale and Inga marginata. These microsatellites will be valuable to study population genetics of this and other species where primer transferability was detected.     

Nuclear and chloroplast SSR markers in Paeonia delavayi (Paeoniaceae) and cross-species amplification in P. ludlowii

? Premise of the study: Microsatellite primers were developed for Paeonia delavayi and P. ludlowii (Paeoniaceae) to study their population genetics and phytogeography. ? Methods and Results: Nine polymorphic nuclear microsatellite loci were isolated from an enriched library of P. delavayi and primers were designed. The number of alleles per locus ranged from two to 16; the observed and expected heterozygosities ranged from 0.014 to 0.687 and 0.042 to 0.875, respectively. Six polymorphic chloroplast microsatellite loci were identified in P. delavayi and primers were provided. The number of alleles per locus ranged from two to six and the polymorphic information content ranged from 0.08 to 0.716. Both nuclear and chloroplast primers were successfully applicable to P. ludlowii. ? Conclusions: The markers developed here will facilitate analyses of genetic diversity, population genetic structure, phytogeographical patterns, and conservation for P. delavayi and P. ludlowii.     

Development of microsatellite markers from an ectomycorrhizal fungus, <Emphasis Type="Italic">Tricholoma matsutake</Emphasis>, by an ISSR-suppression-PCR method

An inter-simple sequence repeat (ISSR)-suppression-PCR technique established to develop microsatellite markers of plant species was applied to an ectomycorrhizal fungus, Tricholoma matsutake. Six polymorphic SSR markers were developed. All six polymorphic SSR markers were single-locused and co-dominant. Alleles produced by these six single-locused markers ranged from two to nine per locus and the expected heterozygosities were calculated as values from 0.098 to 0.803. The results indicated that the ISSR-suppression-PCR technique was effective and applicable to the development of microsatellite markers from ectomycorrhizal fungi. Furthermore, the six microsatellite loci did not amplify DNA from any other ectomycorrhizal species investigated, except for Tricholoma nauseosum (Swedish matsutake) and Tricholoma fulvocastaneum, suggesting that population genetics and reproduction of T. matsutake could be investigated by the SSR markers developed in the present study.     

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.     

Development and characterization of microsatellite loci in Mormon crickets (Anabrus simplex,Orthoptera, Tettigoniidae)

Anabrus simplex is an economically significant crop pest in the western United States and is a model organism for studying the influence of sex role reversal on sperm allocation and utilization patterns and population genetics. We isolated seven polymorphic microsatellite loci in Anabrus simplex, and within population allele numbers ranged from 10 to 25. High polymorphism is not unusual for Orthopteran insects, although observed heterozygosities ranged from 0.24 to 0.91 and were lower than expected heterozygosities, suggesting null alleles. These microsatellites will greatly facilitate studies of postcopulatory reproductive isolation in nuptial gift‐giving insects and historical phylogenetics in the Rocky Mountains.     

Isolation and Characterization of Twelve Microsatellite Loci for Rockfish (Sebastes)

We describe the first microsatellites for rockfishes in the diverse genus Sebastes. Clones containing microsatellites were isolated from the genomic library of a quillback rockfish, Sebastes maliger. Twelve microsatellites are characterized; six of these are polymorphic in quillback rockfish, and eight are polymorphic in at least one rockfish species on which they were tested. The number of alleles per variable locus ranged from 4 to 15 and averaged 6.8. The expected heterozygosities ranged from 0.38 to 0.79 and averaged 0.60 in these loci. These loci should prove valuable in studies examining species identification, population genetics, hybridization, paternity, kinship, and microsatellite evolution. Received September 8, 1998; accepted November 23, 1998.     

Development of tetranucleotide microsatellite markers for the cushion star, Asterina gibbosa, and cross-species amplification

We describe nine polymorphic tetranucleotide microsatellite loci from the starfish, Asterina gibbosa. Loci were isolated from a partial genomic library that had been enriched for AAAC repeat sequences. Number of alleles per locus ranged from two to 14 in a sample of 85 individuals from three populations (two from Spain and one from the UK). Observed and expected heterozygosities per population ranged from 0.000 to 0.400 and from 0.040 to 0.784, respectively. All loci presented significant heterozygote deficits in one or more populations. Eight of these loci were amplified and variable in A. pancerii and A. phylactica. These loci will be used to study population structure in A. gibbosa.     

Isolation and characterization of microsatellite DNA loci in sea bass,Lates calcarifer Bloch

Polymerase chain reaction (PCR)‐based isolation of microsatellite arrays (PIMA) technique was used to isolate seven polymorphic microsatellite loci in sea bass, Lates calcarifer Bloch. A total of 62 samples of wild and cultivated sea bass collected from a few populations within Peninsular Malaysia were used in the study. For seven polymorphic loci, the number of alleles ranged from four to nine and locus heterozygosities ranged from 0.710 to 1.000. The loci will be useful for studying population structure, genetic variability of wild and hatchery stocks of L. calcarifer and broodstock management purposes.