Ten new microsatellite loci for the yellowhammer (Emberiza citrinella) and their cross‐species applicability among related taxa

We describe isolation and characterization of the first microsatellite loci specifically developed for a very common European passerine bird, the yellowhammer Emberiza citrinella. Nine out of our 10 loci are polymorphic within the species E. citrinella. Number of alleles ranged from two to 21 per locus and observed heterozygosity between 0.20 and 0.91. Four primer pairs also yielded reproducible results in other species of Emberizidae. These loci comprise a set of molecular markers for various applications, from moderately polymorphic loci suitable for population studies to highly polymorphic loci for pedigree analysis in Emberizidae.     

Development of microsatellite marker loci for European hazelnut (<Emphasis Type="Italic">Corylus avellana</Emphasis> L.) from ISSR fragments

Polymorphic microsatellite markers were developed for European hazelnut (Corylus avellana L.) from the sequences of inter-simple sequence repeat (ISSR) fragments and flanking regions. Twenty-five ISSR primers were used to generate fragments for cloning. Of the 520 unique sequences obtained, 41 contained long internal repeats (≥20 bp) with flanking sequences sufficient for primer design. From these, we developed 23 new polymorphic microsatellite loci. The flanking sequences were obtained for fragment ends by chromosome walking, and an additional 47 polymorphic markers were developed. Two additional polymorphic markers were developed from a GA-enriched library. The 72 new marker loci were characterized using 50 diverse hazelnut accessions. For the internal repeat loci, the number of alleles per locus ranged from 2 to 16, with a mean of 7.52. Mean values for expected heterozygosity (H_e), observed heterozygosity (H_o), and polymorphism information content (PIC) were 0.62, 0.59, and 0.58, respectively. The estimated frequency of null alleles (r) was ≥0.05 at six of the 23 loci. For the 47 marker loci developed from fragment ends, the number of alleles per locus ranged from 2 to 16, with a mean of 7.30. Mean values for H_e, H_o, and PIC were 0.62, 0.47, and 0.58, respectively. The estimated frequency of null alleles (r) was ≥0.10 at 18 of the 47 loci. Of the 70 loci developed from ISSR and flanking sequences, 50 segregated in our mapping population and were assigned to linkage groups.     

Novel tetranucleotide microsatellite markers from the Del Norte salamander (Plethodon elongatus) with application to its sister species the Siskiyou Mountain salamander (P. stormi)

Eleven tetranucleotide microsatellite loci were developed for the Del Norte salamander (Plethodon elongatus). The loci were variably polymorphic, ranging from two to 20 alleles per locus, with expected heterozygosities ranging from 0.07 to 0.86. The loci also amplified in a congener, the Siskiyou Mountain salamander (P. stormi). The microsatellite loci will be used to assess the utility of highly polymorphic markers to assay within‐ and between‐species differentiation between these two closely related species.     

Microsatellite loci in the Canary Islands endemic ground beetle Trechus flavocinctus and their applicability to cave‐dwelling related species

Six polymorphic microsatellite markers have been developed for the Canary endemic ground beetle Trechus flavocinctus, and the performance of primer pairs amplifying these loci in closely related taxa (including two cave‐obliged species) has been tested. The number of alleles in T. flavocinctus samples taken from three distant localities in two islands ranged from four to 14 per locus. Observed heterozygosities were from 0.333 to 0.933. These markers could be useful tools to study the population structure and the genetic consequences of island and cave colonization by ground beetles.     

Isolation and characterization of microsatellite loci in Fagus longipetiolata Seem. (Fagaceae)

Eleven microsatellite loci have been developed from Fagus longipetiolata and the loci were characterized for 21 individuals. All eleven loci were polymorphic, with 2–8 alleles and an average of 4.8 per locus. The observed (H _O) and expected (H _E) heterozygosities were 0.053–0.714 and 0.355–0.856, respectively. There was significant deviation from Hardy–Weinberg equilibrium at two loci. No locus pair had significant linkage disequilibrium. Cross-species amplifications of the markers were also tested in three other congeneric species.     

Nine new tetranucleotide microsatellite DNA markers for the European mudminnow Umbra krameri

We have isolated eight polymorphic and one monomorphic perfect tetra-nucleotide microsatellite markers for the rare European mudminnow Umbra krameri from genomic libraries enriched for (GACA)n and (GATA)n repeat sequences. A total of 36 individuals from two different populations (Slovakia and Hungary) were tested for these loci. The number of alleles per locus ranged from six to twelve with expected heterozygosities ranging from 0.36 to 0.83. These new developed markers should aid conservation oriented assessment studies or renaturalization programs for this threatened species.     

Isolation of polymorphic microsatellite markers in the great tit (Parus major minor)

Six microsatellite loci were developed for a passerine bird, the great tit (Parus major), using two methods. These loci were polymorphic (3–8 alleles per locus) and exhibited expected heterozygosities from 0.45 to 0.77. At one locus the genotypic frequencies deviated significantly from Hardy–Weinberg expectations.     

Isolation and Characterization of Microsatellite Loci for Striped Bass (Morone saxatilis)

Genetic variation has been difficult to detect in striped bass (Morone saxatilis). Therefore, we identified and characterized 13 microsatellite loci to provide additional genetic markers for striped bass. Microsatellites were identified by screening a striped bass genomic library or by using primers developed for European sea bass (Dicentrarchus labrax) microsatellite loci. We found that 6 of the 13 microsatellite loci were polymorphic in DNA samples obtained from wild populations of striped bass. The number of alleles per locus varied from 3 to 12, and the observed heterozygosities ranged from 0.55 to 0.78. These results indicate that microsatellite loci provide more alleles and higher heterozygosities than other genetic markers developed for striped bass. Received November 9, 1999; accepted February 11, 2000.     

Microsatellite loci in the European bee‐eater,Merops apiaster

Twelve polymorphic microsatellite markers have been developed for the European bee‐eater, Merops apiaster (Coraciiformes: Meropidae). Screening of eight individuals at these loci showed that the average allelic diversity was 5.8, with a range of two to 11 alleles per locus. The loci reported here will provide insight into the levels of extra‐pair parentage, kin selection and dispersal in this species, which has co‐operative breeding and nests in large colonies.     

Development and characterization of 11 novel microsatellite loci for the roundscale spearfish Tetrapturus georgii and their cross‐species amplification among other istiophorid species

Eleven novel polymorphic microsatellite loci were developed and characterized for the recently validated roundscale spearfish Tetrapturus georgii. Characterization of these markers, based on 35 roundscale spearfish from the western North Atlantic, revealed two to 21 alleles per locus with an average expected heterozygosity (H_E) of 0·09–0·94, and all loci conformed to Hardy–Weinberg expectations. Cross‐amplification of these 11 loci against all other eight known istiophorid species indicates promising prospects for the utility of these markers for istiophorids in general.     

Isolation and characterization of ten microsatellite loci in Blue-footed (Sula nebouxii) and Peruvian Boobies (Sula variegata)

Ten microsatellite loci were isolated and characterized from Blue-footed (Sula nebouxii) and Peruvian Boobies (S. variegata). The loci were screened in 24 Blue-footed Boobies and 27 Peruvian Boobies: 8 were polymorphic in Blue-footed Boobies with between 2 and 10 alleles per locus and 9 were polymorphic in Peruvian Boobies with between 2 and 12 alleles per locus. Observed heterozygosity ranged from 0.29 to 0.84. These loci were also tested in Brown Boobies (S. leucogaster) and were variably polymorphic. These new loci are currently being used to assess population genetic structure in Blue-footed and Peruvian Boobies and will also be used to examine hybridization between the species.     

Isolation and characterization of polymorphic microsatellites in Iris ensata (Iridaceae)

• Premise of the study: Microsatellite primers were developed in Iris ensata (Iridaceae) to provide polymorphic markers for further studies into population genetics. • Methods and Results: Thirteen polymorphic microsatellite loci were isolated from I. ensata. These loci were successfully amplified in two natural populations of I. ensata from eastern China (Longwangshan, Zhejiang Province) and northeastern China (Jinchuan, Jilin Province). There was no significant linkage disequilibrium found for any pair of loci. These loci contained between two and 12 alleles per locus across all 48 individuals of I. ensata. The number of alleles per locus varied from two to 10 at the population level and the observed and expected heterozygosities ranged from 0.167 to 0.958 and from 0.284 to 0.853, respectively. • Conclusions: These loci showed high levels of polymorphism and could be used to study the population genetic structure, genetic relationships, and phylogeography of I. ensata.     

Isolation and characterization of microsatellites in European alpine marmots (Marmota marmota)

For future investigations of the mating system of a highly social mammal (Marmota marmota), we identified 16 new microsatellites using an enrichment protocol. Five loci were revealed to be polymorphic. The polymorphism was rather low (two to six alleles among 24 individuals). However, these markers, added to the other six published microsatellites for M. marmota and Spermophilus citellus, will help to assess dispersal patterns and test for genetic monogamy in alpine marmots from the European Alps.     

Isolation and characterization of 23 microsatellite loci in the hen flea Ceratophyllus gallinae

Nineteen dinucleotide, two trinucleotide and two tetranucleotide microsatellite loci developed for the hen flea Ceratophyllus gallinae are presented. Twenty fleas were screened at each locus. Loci were polymorphic (three to nine alleles per locus). These markers will provide a system to study population segregation and diversity, gene flow, dispersal and inbreeding.     

Development and characterization of microsatellite markers for parentage analyses of the coral reef damselfish (<Emphasis Type="Italic">Pomacentrus amboinensis</Emphasis>: Pomacentridae)

Five new polymorphic microsatellite loci were developed for the coral reef damselfish Pomacentrus amboinensis. Twenty-four individuals from two Great Barrier Reef populations were genotyped at the five loci, with numbers of alleles per locus ranging from 6–23 and observed heterozygosity between 0.42–0.92. In addition, the cross-species testing of six primers developed for Stegastes partitus revealed one primer (SpGATA40) that was also polymorphic for P. amboinensis. Due to high levels of polymorphism (≥14 alleles) in at least four of the six loci and a high proportion of tetranucleotide repeats, these microsatellite markers should be useful for parentage assignment as well as other investigations of individual relatedness.     

Development of polymorphic microsatellite markers for the Steere's Liocichla (Liocichla steerii)

For the Steere's Liocichla (Liocichla steerii) we developed seven polymorphic microsatellite markers from a partial genomic library enriched for GATA, tetranucleotide repeats. Among the 27–28 individuals screened, allelic diversity ranged from six to 19 alleles per locus and the observed heterozygosity of each locus ranged from 0.250 to 0.889. All loci conformed to Hardy–Weinberg expectations except for two that appeared to be Z‐Iinked; such Z‐Iinked loci can be used in studies of male dispersal.     

Ten novel tetranucleotide microsatellite DNA markers from Asiatic black bear, Ursus thibetanus

Ten polymorphic microsatellite markers were developed for the endangered Formosan black bear (Ursus thibetanus formosanus) from a partial genomic library enriched for GAAA repeat. Polymorphism of these loci was evaluated in 27 Formosan black bear specimens of unknown relationship. The number of alleles per locus ranged from 5 to 15 and the observed heterozygosity of each locus ranged from 0.556 to 0.889. These loci should provide useful molecular tools to study conservation genetics of the Formosan black bear and other Asiatic black bears.     

Development and characterization of microsatellite markers in the fungus Antrodia cinnamomea from dbEST database

We developed and characterized 8 polymorphic microsatellite loci or simple sequence repeats from the expressed sequence tags database of the medicinal fungus, Antrodia cinnamomea. Variability was assessed in a panel of 23 strains collected from various regions of Taiwan. The number of alleles per polymorphic locus ranged from two to seven loci with an average of 4.375 alleles per locus. The observed and expected heterozygosity values ranged from 0.130 to 0.783 and from 0.122 to 0.809, respectively. No significant Hardy–Weinberg equilibrium was detected (P < 0.01) at all loci. The EST-SSR markers developed in the present study can be used for strain identification and population genetic studies in A. cinnamomea.     

Isolation and characterization of 10 polymorphic microsatellite loci from small yellow croaker (Pseudosciaena polyactis)

Small yellow croaker (Pseudosciaena polyactis) is an economically important marine fish species. About 43 microsatellite loci were isolated from two enriched genomic library of Pseudosciaena polyactis. Ten of these loci were polymorphic in a test population with alleles per locus ranging from two to six, and observed and expected heterozygosities per locus from 0.3750 to 0.8750 and from 0.3112 to 0.8121, respectively. No loci significantly deviated from Hardy–Weinberg equilibrium after Bonferroni correction and no significant linkage disequilibrium was found between pairs of loci. These polymorphic microsatellite loci would be useful for investigating genetic diversity of Pseudosciaena polyactis.     

Characterization of twenty-six polymorphic microsatellite markers for the fox squirrel (Sciurus niger) and their utility in gray squirrels (Sciurus carolinensis) and red squirrels (Tamiasciurus hudsonicus)

Twenty-six polymorphic microsatellite markers were developed for the fox squirrel (Sciurus niger). The number of alleles ranged from 2 to 11 and observed heterozygosities ranged from 0.037 to 0.815. Preliminary screening revealed that numerous loci also were polymorphic in other squirrel species: Sciurus carolinensis and Tamiasciurus hudsonicus. These loci should be useful tools for investigating the social structure, mating tactics and movement behavior of fox squirrels.