Isolation and characterization of 11 polymorphic microsatellite loci in the millipede Antichiropus variabilis Attems (Diplopoda: Polydesmida: Paradoxosomatidae)

Eleven polymorphic microsatellite loci were isolated from the Western Australian millipede Antichiropus variabilis. The number of alleles observed ranged from 2 to 12, with observed heterozygosities ranging from 0.20 to 0.80. All loci were in Hardy-Weinberg equilibrium, and no pairs of loci were in linkage disequilibrium. Many of the loci amplified successfully in eight other Antichiropus species.     

Isolation and characterization of 12 microsatellites from the black surfperch, Embiotoca jacksoni, a reef fish that lacks a pelagic larval phase

A set of 12 microsatellite markers was developed from Embiotoca jacksoni genomic DNA and tested for polymorphism using 64 individuals from two populations. All loci were polymorphic with a number of alleles ranging from two to 19 with expected heterozygosities ranging from 0.17 to 0.89. There was no evidence of linkage disequilibrium and all loci were in Hardy-Weinberg equilibrium, except for four loci in one population. High numbers of private alleles were consistent with strong population structure, and very limited dispersal. Six microsatellite markers successfully cross amplified and were polymorphic in closely related species, Embiotoca lateralis and Hyspurus caryi.     

Microsatellite analysis of the silkworm strains (Bombyx mori): high variability and potential markers for strain identification

The silkworm (Bombyx mori) is of great economic value from an industrial perspective. Casual discrimination and accumulation of genetic information from a diverse variety of silkworms are essential for practical utilization and longterm conservation. In this study, a total of 54 silkworm strains preserved in Korea were typed for nine polymorphic microsatellite loci. We determined per-locus numbers of alleles ranging from 3 to 17 with an average value of 7.5, per-locus observed heterozygosity ranging from 0.04 to 0.98, and per-locus polymorphic information content (PIC) ranging from 0.06 to 0.86, thereby indicating that some of these loci are profoundly variable. Phylogenetic analysis using the nine concatenated microsatellite loci showed no clustering on the basis of known strain characteristics and origin. A total of 17 strain-specific apomorphic alleles, which discriminate 14 among 54 silkworm strains, were obtained from eight loci. These strain-specific alleles can, therefore, be casually utilized to discriminate between applicable strains, without any further typing of other loci. Furthermore, a substantial number of homozygote strains, represented by 24 among 67 alleles in nine loci, were also detected. These results collectively implicate silkworm microsatellite DNA as useful and potentially important molecular markers for the eventual discrimination of silkworm strains, hundreds of which are currently preserved in Korea.     

Isolation and characterization of microsatellite loci in Polytremis nascens (Lepidoptera: Hesperiidae) and their cross-amplification in related species

Twelve polymorphic microsatellite loci were isolated and characterized from a Chinese endemic species Polytremis nascens Leech. The isolated loci in 53 individuals from six regional populations in China had 3–12 alleles. Analyses revealed that 53 individuals had different multilocus genotypes, with the expected heterozygosity ranging from 0.61 to 0.90 and observed heterozygosity ranging from 0.33 to 0.71. Eleven loci showed no deviation from Hardy–Weinberg proportions, and eight were amplified successfully in at least one of the eight related species. The described isolated markers will facilitate the study of population genetic structure and gene flow in P. nascens as well as in congeneric and related species.     

Population genetics of polymorphisms in Cardiff newborn. Relationship between blood group and allozyme heterozygosity and birth weight

A newborn population of Cardiff, Wales, was screened for variation at three blood group loci (ABO, Rhesus and MN) and four enzyme loci (ACP-1, PGM-1, ADA and EST-D). Birth weights were measured. There were no significant differences between mean birth weights or birth weight variances for individuals homozygous or heterozygous at the MN and the four enzyme loci. (ABO and Rhesus loci cannot be used in these tests.) There was no significant heterogeneity in contingency tables relating phenotypes at the seven loci to birth weight. There were no significant differences in mean heterozygosity per locus between babies placed in different birth weight categories, ranging from 2.5 to 4.2 kg. The genetic variation screened appears therefore to be neutral with respect to this character.     

Eight new microsatellite markers in Atlantic cod (Gadus morhua L.) derived from an enriched genomic library

One hundred and fifty random clones from an enriched genomic library of Atlantic cod were sequenced. Primer pairs were designed for 15 microsatellites containing perfect di‐, tri‐, tetra‐ and hexanucleotide repeats and characterized in 96 unrelated fish. Eight markers were successfully amplified, with the number of alleles ranging from two to nine per locus and observed heterozygosity ranging from 0.341 to 0.977. Loci Gmo‐G13 and Gmo‐G14 had a significant excess of homozygotes. 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.     

Characterization and isolation of DNA microsatellite primers in Raja clavata L. (thornback ray,Rajidae)

The thornback ray, Raja clavata, is an elasmobranch (cartilaginous fish). Since the 1950s, its stock has severely declined. In order to investigate the genetic population structure of the species, we developed microsatellite loci. The five loci reported here have eight to 48 alleles per locus and display an observed heterozygosity ranging from 0.32 to 0.98 with no deviation from the Hardy–Weinberg equilibrium. In the test panel of 122 individuals from three populations, no null alleles, band‐stuttering, large allele dropouts or linkage disequilibrium was detected.     

9号染色体短臂上7个STR基因座在基因扫描中的信息表现

为了初步探讨7个位于染色体9p区域的短串联重复序列(shorttandemrepeat,STR)基因座:D9S288、D9S157、D9S1748、D9S171、D9S161、D9S1817和D9S1805在遗传学研究及法医学应用中的意义,随机抽取225名湖南汉族无关个体,复合PCR技术扩增上述基因座,ABI377全自动测序仪进行基因分型,共检出75种等位基因,通过对基因型及等位片断频率分布的研究和数据统计分析,7个基因座基因频率分布在0.002～0.800之间,构成243种基因型。7个STR基因座基因型分布均符合Hardy Weinberg平衡定律(P>0.05),杂合度(heterozygosity,H)介于0 347～0.844之间,个体识别力(discriminationpower,DP)为0.346～0.841,非父排除率(probabilitiesofpaternityexclusion,PPE)为0.308～0.738,多态信息含量(polymorphicinformationcontent,PIC)在0.328～0.822之间。种族比较结果显示,湖南汉族与非洲黑人及欧洲白人在大多数基因座均存在显著差异(P<0.001)。研究结果丰富了中华民族基因数据库,在人类群体遗传学及法医学研究领域有重要应用价值。     

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.     

Development and characterization of eight polymorphic microsatellite loci from Pistacia lentiscus L. (Anacardiaceae)

We have developed a set of eight polymorphic nuclear microsatellite markers for the Mediterranean shrub Pistacia lentiscus by means of an enriched library method. Characterization for the eight loci was carried out on 42 individuals from two populations sampled in southern Spain. The overall number of alleles detected was 59, ranging from three to 13 per locus. Expected heterozygosity per locus and population ranged from 0.139 to 0.895. Two loci albeit only in one population (Seville) departed significantly from Hardy-Weinberg equilibrium expectations and no linkage disequilibrium between pairs of loci was detected. These markers will be used in studies of gene flow across a fragmented landscape.     

Isolation and characterization of polymorphic microsatellite loci from black snapper Sebastodes fuscescens (Houttuyn)

Black snapper Sebastodes fuscescens (Houttuyn) is an economocally important species. Fourty-five microsatellite loci were isolated from an enriched genomic library of S. fuscescens. Ten of these loci were polymorphic in a test population with alleles per locus ranging from 2 to 5, and observed and expected heterozygosities per locus from 0.21 to 1.00 and from 0.19 to 0.86, respectively. Three 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 S. fuscescens and other related species.     

Development of 19 polymorphic microsatellite loci for the intermediate horseshoe bat, Rhinolophus affinis (Rhinolophidae, Chiroptera)

The intermediate horseshoe bat (Rhinolophus affinis) is a widespread species in Southeast Asia. We developed 19 novel microsatellite loci from an enriched genomic library of the bat, and tested their polymorphism using a single population from Guangdong province, southern China. The number of alleles ranged from 3 to 15 per locus with the expected and observed heterozygosity ranging from 0.397 to 0.920 and 0.280–0.926, respectively. Three markers significantly deviated from the Hardy-Weinberg expectations after Bonferroni correction and no linkage disequilibrium was detected in any of loci. These microsatellite loci will be useful in studying the phylogeography of this species.     

Isolation and characterization of polymorphic microsatellite loci from fat greenling (Hexagrammos otakii)

Fat greenling (Hexagrammos otakii) is an economocally important marine fish species. Thirtyfive microsatellite loci were isolated from two enriched genomic library of Hexagrammos otakii. 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 0.2581 to 1.0000 and from 0.2892 to 0.7726, respectively. Four 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 Hexagrammos otakii.     

Isolation and characterization of 10 tetranucleotide microsatellite loci from the yellow-streaked greenbul (Phyllastrephus flavostriatus) and cross-species amplification in four closely related taxa

We describe the isolation of 10 tetranucleotide microsatellite loci from the yellow-streaked greenbul using an enrichment protocol. All loci were highly variable with the number of alleles ranging from 8 to 13, and observed heterozygosity ranging from 0.652 to 0.870. All loci were in Hardy-Weinberg equilibrium; however, loci Pfl12 and Pfl54 showed significant linkage disequilibrium. All 10 loci successfully amplified and were polymorphic in at least one of four related Phyllastrephus species. These loci should prove to be widely applicable to studies of phylogeography, hybridization and paternity in African greenbuls.     

Development and characterization of novel microsatellite markers from the Chinese rufous horseshoe bat (Rhinolophus sinicus) with cross-species amplification in closely related taxa

We isolated nine polymorphic microsatellite markers from the Chinese rufous horseshoe bat (Rhinolophus sinicus) using an enriched library method. We assessed genetic polymorphism at these loci in 42 individuals from a single population. We recorded high genetic diversity with four to 17 alleles per locus, and estimated expected and observed heterozygosity values ranging from 0.492 to 0.910 and from 0.462 to 0.881, respectively. No locus departed from Hardy-Weinberg equilibrium following Bonferroni correction, and no linkage disequilibrium was detected. Most loci successfully cross-amplified congeneric species. These loci will be used to characterize phylogeographical history of Rhinolophus sinicus in China.     

Characterization of 12 polymorphic microsatellite loci for the whitespotted bamboo shark (Chiloscyllium plagiosum Bennett)

Twelve polymorphic microsatellite loci without linkage disequilibrium were isolated and characterized from a microsatellite DNA-enriched DNA library for the whitespotted bamboo shark (Chiloscyllium plagiosum Bennett, 1830). These loci show polymorphism information content ranging from 0.384 to 0.885, allele number ranging from 4 to 12, effective allele number ranging from 1.686 to 9.438, and observed and expected heterozygosities from 0.229 to 1.000 and from 0.407 to 0.894 respectively. Four loci show strong Hardy-Weinberg deviations. We expect that these markers would be useful for population genetic and breeding studies of the whitespotted bamboo shark.     

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.     

Isolation and characterization of polymorphic microsatellite loci from black sea bass (Centropristis striata) and cross-species amplification

Black sea bass (Centropristis striata) is an economically important serranid species. A number of 39 microsatellite loci were isolated from two enriched genomic library of C. striata. Eleven of these loci were polymorphic in a test population with alleles per locus ranging from 3 to 8, and observed and expected heterozygosities per locus from 0.26 to 1.00 and from 0.61 to 0.84, respectively. Four loci significantly deviated from Hardy–Weinberg equilibrium after Bonferroni correction and no significant linkage disequilibrium was found between pairs of loci. 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 C. striata and other related 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.