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This research was conducted to reveal the enriched library experiences of students with a case study. Atatürk Elementary School, located in the city center of Artvin, constituted the sample of this research. The enriched library environment was observed every two weeks for a year, one year after the opening of the library, and 15 students were interviewed, the library documents were examined. Findings of the study show the following results. Students were aware of the features of the enriched library in general, showed considerable interest in the early months after the library was opened, and used the library primarily for circulation. The library also contributed to the reading habits of the students. Despite these benefits, it was clear that some problems stemming from the nature of the enriched library emerged during the process. [ABSTRACT FROM AUTHOR]

Background: Forage grasses of the African genus Urochloa (syn. Brachiaria) are the basis of Brazilian beef production, and there is a strong demand for high quality, productive and adapted forage plants. Among the approximately 100 species of the genus Urochloa, Urochloa decumbens is one of the most important tropical forage grasses used for pastures due to several of its agronomic attributes. However, the level of understanding of these attributes and the tools with which to control them at the genetic level are limited, mainly due to the apomixis and ploidy level of this species. In this context, the present study aimed to identify and characterize molecular microsatellite markers of U. decumbens and to evaluate their cross-amplification in other Urochloa species. Findings: Microsatellite loci were isolated from a previously constructed enriched library from one U. decumbens genotype. Specific primers were designed for one hundred thirteen loci, and ninety-three primer pairs successfully amplified microsatellite regions, yielding an average of 4.93 alleles per locus. The polymorphism information content (PIC) values of these loci ranged from 0.26 to 0.85 (average 0.68), and the associated discriminating power (DP) values ranged from 0.22 to 0.97 (average 0.77). Cross-amplification studies demonstrated the potential transferability of these microsatellites to four other Urochloa species. Structure analysis revealed the existence of three distinct groups, providing evidence in the allelic pool that U. decumbens is closely related to Urochloa ruziziensis and Urochloa brizantha. The genetic distance values determined using Jaccard's coefficient ranged from 0.06 to 0.76. Conclusions: The microsatellite markers identified in this study are the first set of molecular markers for U. decumbens species. Their availability will facilitate understanding the genetics of this and other Urochloa species and breeding them, and will be useful for germplasm characterization, linkage mapping and marker-assisted selection. [ABSTRACT FROM AUTHOR]

Abstract In order to study duck microsatellites, we constructed a library enriched for (CA)n, (CAG)n, (GCC)n and (TTTC)n. A total of 35 pairs of primers from these microsatellites were developed and used to detect polymorphisms in 31 unrelated Peking ducks. Twenty-eight loci were polymorphic and seven loci were monomorphic. A total of 117 alleles were observed from these polymorphic microsatellite markers, which ranged from 2 to 14 with an average of 4.18 per locus. The frequencies of the 117 alleles ranged from 0.02 to 0.98. The highest heterozygosity (0.97) was observed at the CAUD019 microsatellite locus and the lowest heterozygosity (0.04) at the CAUD008 locus, and 11 loci had heterozygosities greater than 0.50 (46.43%). The polymorphism information content (PIC) of 28 loci ranged from 0.04 to 0.88 with an average of 0.42. All the above markers were used to screen the polymorphism in other bird species. Two markers produced specific monomorphic products with the chicken DNA. Fourteen markers generated specific fragments with the goose DNA: 5 were polymorphic and 9 were monomorphic. But no specific product was detected with the peacock DNA. Based on sequence comparisons of the flanking sequence and repeat, we conclude that 2 chicken loci and 14 goose loci were true homologous loci of the duck loci. The microsatellite markers identified and characterized in the present study will contribute to the genetic map, quantitative traits mapping, and phylogenetic analysis in the duck and goose.

In this study, an (AG) n microsatellite-enriched library of Rhododendron sirnsii Planch, was constructed with FIASCO (fat Isolation by AFLP sequences containing repeats) method. Among 356 positive colonies verified by colony PCR of the (AG)n microsateHite-enriched library, 233 colonies were selected for further sequencing. According to the results, 202 colonies containing microsateHites were obtained, accounting for 86.7%. Discarding sequences with misceHaneous peaks, overlapping peaks and weak signals, 39 sequences were screened finally for subsequent primer exploitation, including 22 perfect type SSR loci, 5 imperfect type SSR loci ad 12 compound type SSR loci. The FIASCO method is efficient and feasible for constructing microsatellite-enriched libraries of RRododendron spp., which laid a solid foundation for isolation of microsateHite loci and investigation of genetic diversity ad genetic structure of RRiododendron spp. [ABSTRACT FROM AUTHOR]

We developed 31 microsatellite loci for a specialist hyperparasitoid wasp, Mesochorus cf. stigmaticus that parasitizes the parasitoid Hyposoter horticola, in Åland islands of southwestern Finland. The microsatellites were developed from 454 sequencing of the enriched libraries. The details of primers and the polymerase chain reactions for these microsatellite markers are described. All the loci tested on 24 samples were polymorphic with on average 4.1 alleles per locus (range 2-13). Genetic diversities as measured by average expected heterozygosity (H) and allelic richness (A) were generally high (H ≈ 0.10-0.80; A ≈ 1.9-11.5). These markers will be useful in population genetic studies of this parasitoid and other related taxa. [ABSTRACT FROM AUTHOR]

Egyptian clover (Trifolium alexandrinum), a multi-cut forage legume, is widely grown as an annual feed crop. Genetic improvement in this legume is largely hampered due to limited availability of genomic resources. Hence, a genomic library enriched for GA/CT motif was constructed to develop SSR primers. As a result, 667 SSR primer pairs were developed and 111 were validated in T. alexandrinum cv. Wardan s-99-1. Among these, 45 primer pairs were used for the analysis of genetic diversity among 86 Trifolium accessions, resulting in 220 alleles with an average of 4.88 alleles per locus. Polymorphic information content for these markers ranged from 0.040 to 0.819 with an average of 0.445. The observed and expected heterozygosity for individual loci ranged from 0.39 to 0.89 and 0.368 to 0.80 with an average of 0.675 and 0.659, respectively. The markers exhibited varied levels of cross-species transferability ranging from 93.33 % across T. resupinatum to 40.0 % across T. balansae. Phylogenetic relationships were analyzed using the unweighted pair group method with arithmetic averages (UPGMA)-based dendrogram which was able to establish the uniqueness of each genotype and grouped them into two major clusters. To the best of our knowledge, the SSR markers reported here are among the first to be reported in this important yet neglected forage legume. They were assessed to be highly efficient and polymorphic and found to be useful for distinguishing the closely related genotypes of Trifolium. This large resource of SSR markers would therefore facilitate new avenues for T. alexandrinum genomics. [ABSTRACT FROM AUTHOR]

Lentil ( Lens culinaris Medik.) is an economically important grain legume, yet the genetic and genomic resources remain largely uncharacterized and unexploited in this crop. Microsatellites have become markers of choice for crop improvement applications. Hence, simple sequence repeat (SSR) markers were developed for lentil through the construction of genomic library enriched for GA/CT motifs. As a result 122 functional SSR primer pairs were developed from 151 microsatellite loci and validated in L. culinaris cv. Precoz. Thirty three SSR markers were utilized for the analysis of genetic relationships between cultivated and wild species of Lens and related legumes. A total of 123 alleles were amplified at 33 loci ranging from 2-5 alleles with an average of 3.73 alleles per locus. Polymorphic information content (PIC) for all the loci ranged from 0.13 to 0.99 with an average of 0.66 per locus. Varied levels of cross genera transferability were obtained ranging from 69.70 % across Pisum sativum to 12.12 % across Vigna radiata. The UPGMA based dendrogram was able to establish the uniqueness of each genotype and grouped them into two major clusters clearly resolving the genetic relationships within lentil and related species. The new set of SSR markers reported here were efficient and highly polymorphic and would add to the existing repertoire of lentil SSR markers to be utilized in molecular breeding. Moreover, the improved knowledge about intra- and inter-specific genetic relationships would facilitate germplasm utilization for lentil improvement. [ABSTRACT FROM AUTHOR]

Limited availability of validated, polymorphic microsatellite markers in mung bean ( Vigna radiata), an important food legume of India, has been a major hurdle towards its improvement and higher yield. The present study was undertaken in order to develop a new set of microsatellite markers and utilize them for the analysis of genetic diversity within mung bean accessions from India. A GA/CT enriched library was constructed from V. radiata which resulted in 1,250 putative recombinant clones of which 850 were sequenced. SSR motifs were identified and their flanking sequences were utilized to design 328 SSR primer pairs. Of these, 48 SSR markers were employed for assessing genetic diversity among 76 mung bean accessions from various geographical locations in India. Two hundred and thirty four alleles with an average of 4.85 alleles per locus were detected at 48 loci. The polymorphic information content (PIC) per locus varied from 0.1 to 0.88 (average: 0.49 per locus). The observed and expected heterozygosities ranged from 0.40 to 0.95 and 0.40 to 0.81 respectively. Based on Jaccard's similarity matrix, a dendrogram was constructed using the unweighted pair-group method with arithmetic averages (UPGMA) analysis which revealed that one accession from Bundi, Rajasthan was clustered out separately while remaining accessions were grouped into two major clusters. The markers generated in this study will help in expanding the repertoire of the available SSR markers thereby facilitating analysis of genetic diversity, molecular mapping and ultimately broadening the scope for genetic improvement of this legume. [ABSTRACT FROM AUTHOR]

Highlights: [•] One thousand two hundred seven SSRs developed from CA and GA repeat enriched genomic libraries. [•] A total of 248 polymorphic SSRs identified with 7 accessions of J. curcas. [•] 179 and 331 polymorphic SSRs identified with parental lines of intra and inter-specific mapping population respectively. [•] Dendrogram classified all the 7 accessions of J. curcas in one cluster and the J. integerrima as out group. [•] BLASTX showed similarity of 50% with Ricinus communis followed by Populus trichocarpa (22%) and Vitis vinifera (16%). [ABSTRACT FROM AUTHOR]

Cymbidium spp. are popular flowering plants. Assessment of the genetic diversity in cultivated Cymbidium facilitates conservation of germplasm and subsequent cultivar improvement. Thus, it is important to develop more efficient polymorphic DNA markers. Although more motifs (403) were identified and more primers (206) were designed in the genomic library compared to the cDNA library, a larger number of successful primers were obtained from the cDNA library (59.9%) than from genomic DNA library (51.1%). However, higher PIC and gene diversity were identified in genomic SSRs. The average allele number per locus was also higher in genomic SSRs (7.3) than EST-SSRs (5.2), among the 24 evaluated Cymbidium accessions. AT/TA was comparatively high in EST-SSRs, while this motif was not as common in genomic SSRs. The CTT/AAG/TCT/AGA/TTC/GAA and TGC/GCA/GCT/AGC/CTG/CAG motifs were the most abundant tri-nucleotide sequences in ESTSSRs, while GTT/AAC/TGT/ACA/TTG/CAA was the most frequent in genomic SSRs. The number of repeats ranged from 3 to 12 in EST-SSRs. Currently, 52 novel polymorphic SSR markers have been evaluated, which will be useful for germplasm assessments, core set construction, evaluation of genetic diversity, and marker assisted selection (MAS) based Cymbidium breeding. [ABSTRACT FROM AUTHOR]

We identified 14 informative microsatellite loci from two taxa, Grevillea aquifolium (6) and G. infecunda (8), belonging to the Pteridifolia Group, Aspleniifolia/ Hookeriana Subgroup (Proteaceae). Loci were screened for variability among 20 individuals of their source species. The number of alleles observed per locus for G. infecunda ranged from 1 to 3. Subsequent chromosome counts revealed between 20 (normal diploid) and 30 (triploid) chromosomes in different individuals. High levels of variability were observed with the number of alleles per locus ranging from 3 to 17, with a mean of 9.8 across all loci. Nei's genetic diversity ( H) ranged from 0.399 to 0.992 with a mean of 0.783. All loci amplified in a further 9-13 taxa from the same subgroup and under the same amplification conditions. These new markers provide useful tools for evolutionary and conservation studies assessing clonality and polyploidy in this important Australian genus. [ABSTRACT FROM AUTHOR]

Microsatellites (or SSRs: simple sequence repeats) are among the most frequently used DNA markers in many areas of research. The use of microsatellite markers is limited by the difficulties involved in their de novo isolation from species for which no genomic resources are available. We describe here a high-throughput method for isolating microsatellite markers based on coupling multiplex microsatellite enrichment and next-generation sequencing on 454 GS-FLX Titanium platforms. The procedure was calibrated on a model species ( Apis mellifera) and validated on 13 other species from various taxonomic groups (animals, plants and fungi), including taxa for which severe difficulties were previously encountered using traditional methods. We obtained from 11 497 to 34 483 sequences depending on the species and the number of detected microsatellite loci ranged from 199 to 5791. We thus demonstrated that this procedure can be readily and successfully applied to a large variety of taxonomic groups, at much lower cost than would have been possible with traditional protocols. This method is expected to speed up the acquisition of high-quality genetic markers for nonmodel organisms. [ABSTRACT FROM AUTHOR]

Turbot ( Scophthalmus maximus) is a flatfish species commercially important for aquaculture. In this study, we generated a microsatellite-enriched genomic DNA library for Scophthalmus maximus, and then isolated and characterized 45 microsatellite loci by genotyping 30 individuals. The observed number of alleles ranged from 2 to 19 with an average of 6.24, while the effective number of alleles ranged from 1.30 to 11.11 with an average of 3.66. The expected heterozygosities varied from 0.235 to 0.925 4 and Polymorphie information content ranged from 0.2044 to 0.903 3, with an average of 0.622. Twelve loci deviated significantly from Hardy-Weinberg equilibrium, and no significant linkage disequilibrium was observed between any pair of loci after Bonferroni correction. In cross-species amplification, five flatfish species ( Paralichthys lethostigma, Verasper moseri, platichthys stellatus, Hippoglossoides dubius and Cynoglossus semilaevis) showed at least one Polymorphie locus. These Polymorphie microsatellite loci should prove useful for Population analysis of turbot and other related species. [ABSTRACT FROM AUTHOR]

The pomegranate ( Punica granatum L.) is a temperate climate species requiring high temperatures for proper and complete ripening. The species is consumed as a fresh fruit, but also can be used to obtain transformed products such as juice, jam, or preserve. It is a fruit tree species with a high degree of diversity, but the identification of cultivars by morphological traits is very difficult. Thus, the characterization of genotypes through molecular markers is of great value for germplasm preservation, genetic studies, and plant breeding. The number of simple sequence repeat (SSR or microsatellite) markers developed for this genus is too low, so in this work we report the development of 117 microsatellite loci from a CT/AG-enriched pomegranate genomic library. In order to check their utility, eleven accessions were analyzed. The polymorphism information content (PIC) value across all loci ranged between 0.09 and 0.71, with an average of 0.37. These markers will facilitate genetic diversity studies, mapping, and genotyping of pomegranate. [ABSTRACT FROM AUTHOR]

By combining biotin-enrichment protocol and next generation pyrosequencing, through 454 GS-FLX Titanium technology, 55 polymorphic microsatellites loci with perfect motif were isolated from the Rhone streber ( Zingel asper), a critically endangered European fish species. Eight multiplex PCR kits were optimised in order to genotype a total of 58 polymorphic loci, including three previously published loci. The level of genetic diversity was assessed for 68 Z. asper, 30 Sander lucioperca, 33 Perca fluviatilis and four Gymnocephalus schraetzer individuals. Amplification success was also assessed on Romanichthys valsanicola and Zingel streber using single individuals. These markers will be useful to investigate the population structure of the highly fragmented Rhone streber. They represent a powerful tool for conservation issues and evolutionary approaches of this endemic species. Moreover, part of our markers demonstrated applicability to other percid species, allowing for potential applications to fisheries and aquaculture management. [ABSTRACT FROM AUTHOR]

Ten tetranucleotide microsatellite loci were isolated from GATA-enriched library of rock carp ( Procypris rabaudi). The loci were tested in 25 individuals from four populations. The number of alleles per locus ranged from 3 to 6. Polymorphism information content (PIC) ranged from 0.460 to 0.788. Observed and expected heterozygosities ranged from 0.375 to 0.739 and from 0.531 to 0.833, respectively. These microsatellites will facilitate the study of population genetics of rock carp. [ABSTRACT FROM AUTHOR]

A new set of 11 microsatellite markers was isolated and characterized in the large yellow croaker (Pseudosciaena crocea Richardson) from a CA-enriched genomic library. Of 35 microsatellite markers screened, 11 microsatellite markers are highly polymorphic in one hatchery stock which contained 58 individuals. The high genetic variability was represented mainly by the number of alleles, which ranged from 7 to 16, and the observed heterozygosity, which ranged from 0.43 to 0.79. These newly isolated markers increase the available molecular resources that can be used to analyze genetic diversity and population structure of large yellow croaker. [ABSTRACT FROM AUTHOR]

Ten novel polymorphic microsatellites were isolated from the dace complex ( Leuciscus leuciscus), which is a European cyprinid species. Four multiplex polymerase chain reaction sets were optimized in order to genotype 26 polymorphic loci in all, including 16 previously published cyprinid-specific loci. The level of genetic diversity was assessed in 142 dace individuals. We also successfully applied 26 of the microsatellites to 10 related species. These primers thus will be useful to assess population structure of the dace and other cyprinid species, with application for conservation issues and phylogeographical approaches. [ABSTRACT FROM AUTHOR]

Crepidula convexa, a calyptreid gastropod with direct embryonic development, changes sex from male to female in the course of its lifetime (protandry). Under sex-allocation theory, male reproductive success should be independent from age and size (a proxy used for age). However, this may be counterbalanced by female cryptic choice or gregarious behaviour. Eleven polymorphic microsatellite loci were thus developed to examine paternity of embryos and larvae. This set of loci appears suitable to carry out paternity analyses due to the high exclusion probability of unrelated males given the maternal genotype. [ABSTRACT FROM AUTHOR]

Ten novel polymorphic microsatellites (seven with perfect motifs) were isolated from vairone species ( Telestes souffia and Telestes muticellus), which are endangered European cyprinid species. Together with 11 previously published cyprinid-specific loci, five multiplex sets were optimized, allowing the genotyping of 21 polymorphic loci. The level of genetic diversity was assessed in 97 individuals from the two species T. souffia and T. muticellus. We also successfully applied the 21 microsatellites to nine related species. These primers will thus be useful in assessing population structure of the vairone and other cyprinid species, with application for conservation issues and phylogeographical approaches. [ABSTRACT FROM AUTHOR]

We report isolation, characterization and cross-species amplification of nine microsatellite loci from the phytoparasitic nematode Xiphinema index, the vector of grapevine fanleaf virus. Levels of polymorphism were evaluated in 62 individuals from two X. index populations. The number of alleles varies between two and 10 depending on locus and population. Observed heterozygosity on loci across both populations varied from 0.32 to 0.857 (mean 0.545). The primers were tested for cross-species amplification in three other species of phytoparasitic nematodes of the Xiphinema genus. These nine microsatellite loci constitute valuable markers for population genetics and phylogeographical studies of X. index. [ABSTRACT FROM AUTHOR]

One hundred and thirty-five microsatellite markers were developed for hop Humulus lupulus L. from di- and trinucleotide-enriched libraries. Seventy-eight primers showed amplification in two tested genotypes. Twenty-four loci were further characterized on a population of 34 hop samples and the number of alleles per locus, observed heterozygosity and expected heterozygosity ranged from two to 20 (9.7 on average), from 0.0294 to 0.9412 (0.6234 on average) and from 0.0294 to 0.9170 (0.6720 on average), respectively. These microsatellite markers will be further used for studying population structures and relationships and for identifying important qualitative and quantitative loci of hop. [ABSTRACT FROM AUTHOR]

We present a new set of 15 polymorphic microsatellite primer sequences developed from Ficus carica L. The variability of specific microsatellite regions was assessed in wild population of figs from the northern Adriatic coast and all 15 primer pairs showed single-locus amplification with a total of 65 alleles and an observed heterozygosity ranging from 0.285 to 0.863. The 15 new microsatellite loci represent a significant tool for population genetic structure studies and will be further used to investigate the origin and maintenance of genetic variation within and between populations of figs along the Adriatic coastal region. [ABSTRACT FROM AUTHOR]

Simple sequence repeat-enriched library screening and expressed sequence tag database mining were adopted to develop microsatellite markers for sea cucumber ( Apostichopus japonicus). Eighty-three microsatellite loci were selected for polymorphism assessment using 48 individuals. The results showed that 45 novel loci were polymorphic. The number of alleles ranged from two to 16, and the values of observed and expected heterozygosities varied from 0 to 0.9375 and from 0.1135 to 0.9674, respectively. No significant linkage disequilibrium between pairs of loci was found and 26 loci conformed to the Hardy–Weinberg equilibrium. These markers are therefore a potential tool for studies in the population structure and linkage map construction for A. japonicus. [ABSTRACT FROM AUTHOR]

The turbot ( Scophthalmus maximus) is a flatfish species of great commercial value for aquaculture. In this study we describe the isolation and characterization of 30 novel highly polymorphic microsatellite markers in this species obtained from a genomic library enriched for seven short tandem repeated motifs. Much higher polymorphism (mean number of alleles: 13.37; mean expected heterozygosity: 0.869) and potential for parentage assignment than previously reported for microsatellites in turbot were found after the analysis of 24 wild individuals. Most loci conformed to Hardy–Weinberg expectations, excluding Sma-USC20 and Sma-USC28, which showed a high heterozygote deficit probably due to the presence of null alleles. No significant genotypic disequilibrium was observed between any pair of loci, suggesting no close linkage between them. These loci are potentially useful for kinship and population analysis in turbot. [ABSTRACT FROM AUTHOR]

An improved protocol for constructing microsatellite-enriched libraries was developed. The procedure depends on digesting genomic DNA with a restriction enzyme that generates blunt-ends, and on ligating linkers that, when dimerized, create a restriction site for a different blunt-end producing restriction enzyme. Efficient ligation of linkers to the genomic DNA fragments is achieved by including restriction enzymes in the ligation reaction that eliminate unwanted ligation products. After ligation, the reaction mixture is subjected to subtractive hybridization without purification. DNA fragments containing microsatellites are captured by biotin-labeled oligonucleotide repeats and recovered using streptavidin-coated beads. The recovered fragments are amplified by PCR using the linker sequence as primer, and cloned directly into a plasmid vector. The linker has the sequence GTTT on the 5' end. which promotes efficient adenylation of the 3' ends of the PCR products. Consequently, the amplified fragments could be cloned into vectors without purification. This procedure enables efficient enrichment and cloning of microsatellite sequences, resulting in a library with a low level of redundancy. [ABSTRACT FROM AUTHOR]

Twelve new microsatellites have been developed in olive. For that purpose, a genomic library of the olive cultivar ‘Arbequina’ was enriched for GA, GT and ACT repeats. Two methods of screening yielded 27 sequences containing microsatellites out of the 119 clones sequenced. The GA repeat seems to be the most abundant motif. Among sequences containing microsatellites, 4 (14.8%) were redundant, 1 (3.7%) was previously described in the literature and 12 (44.4%) could not be used for primers design because the repeat motifs were incomplete. Suitable primer pairs were obtained for the remaining 10 (37.0%) sequences plus an additional 14 recovered from a formerly developed library. For the 24 primer pairs designed, 4 failed to amplify, 8 produced a complex bands pattern and 12 succeeded in giving amplification products. Considering these 12 primer pairs, 10 showed single locus amplification, whereas the other 2 revealed two loci each. This was demonstrated by studying allele segregation in two olive progenies. Sixty-eight alleles were detected for the 12 microsatellites when 51 olive cultivars were analysed. The number of alleles per locus ranged from 1 to 13. The expected heterozygosity varied between 0 and 0.83. All pairs of cultivars could be distinguished using only three microsatellites due to their great discrimination power value. The data coming from genotyping the 51 olive cultivars for 7 out of the 12 new microsatellites were used for constructing a dendrogram by unweighted pair group method with arithmetic mean cluster analysis using the Dice similarity coefficient. Cultivar association according to their geographical origin was observed. [ABSTRACT FROM AUTHOR]

We developed 15 microsatellite loci in the poplar rust fungus, Melampsora larici-populina, using two enrichment protocols. Polymorphism of each locus was assessed on a panel of 30 isolates, comprising three subpanels (world, regional and local scales). Thirteen loci were polymorphic with three to eight alleles detected. The 15 loci were also tested on five related Melampsora species, M. allii-populina, M. medusae f. sp. deltoidae, M. larici-tremulae, M. rostrupii and M. pinitorqua, and partial or global cross-amplification events were detected. [ABSTRACT FROM AUTHOR]

The hydrothermal vent polychaete Branchipolynoe seepensis lives commensally inside the mantel cavity of the bivalves Bathymodiolus azoricus and Bathymodiolus puteoserpentis that form dense mussel beds around hydrothermal vents on the Mid-Atlantic Ridge. In order to study its dispersal capabilities and the way individuals develop and reproduce, nine microsatellite polymorphic markers were developed. Polymorphism was tested from three well-separated populations ranging from 14°N to 35°N along the ridge and showed significant levels of genetic differentiation. Cross-amplification tests in other polynoid species revealed that most loci could also be useful to study closely related species from both Atlantic and Pacific sites. [ABSTRACT FROM AUTHOR]

The turkey is an agriculturally important species for which, until now, there is no published genetic linkage map based on microsatellite markers — still the markers most used in the chicken and other farm animals. In order to increase the number of markers on a turkey genetic linkage map we decided to map new microsatellite sequences obtained from a GTenriched turkey genomic library. In different chicken populations more than 35-55% of microsatellites are polymorphic. In the turkey populations tested here, 43% of all turkey primers tested were found to be polymorphic, in both commercial and wild type turkeys. Twenty linkage groups (including the Z chromosome) containing 74 markers have been established, along with 37 other unassigned markers. This map will lay the foundations for further genetic mapping and the identification of genes and quantitative trait loci in this economically important species. Genome comparisons, based on genetic maps, with related species such as the chicken would then also be possible. All primer information, polymerase chain reaction (PCR) conditions, allele sizes and genetic linkage maps can be viewed at http://roslin.thearkdb.org/. The DNA is also available on request through the Roslin Institute. [ABSTRACT FROM AUTHOR]

We isolated and characterized 16 microsatellite loci from the blacktip shark, Carcharhinus limbatus, and tested cross-species amplification in 11 Carcharhinus species and five additional shark genera. Thirty-six (1.6%) and 180 (48%) colonies were positive for dinucleotide repeat motifs from unenriched and enriched libraries, respectively. Heterozygosities of polymorphic loci ranged from 0.04 to 0.96 with two to 22 alleles per locus. Amplification products were observed at nine to 13 loci (five to 11 of which where polymorphic) in 10 Carcharhinus species. Several loci were also polymorphic in each of the additional genera examined. [ABSTRACT FROM AUTHOR]

Abstract The attributes of codominance, reproducibility and high resolution have all contributed towards the current popularity of nuclear microsatellites as genetic markers in molecular ecological studies. One of their major drawbacks, however, is the development phase required to obtain working primers for a given study species. To facilitate project planning, we have reviewed the literature to quantify the workload involved in isolating nuclear microsatellites from plants. We highlight the attrition of loci at each stage in the process, and the average effort required to obtain 10 working microsatellite primer pairs. [ABSTRACT FROM AUTHOR]

Twenty-four and 12 microsatellite loci were developed in peach [ Prunus persica (L) Batsch cv. Akatsuki] by using an enriched genomic and fruit cDNA libraries, respectively. The microsatellite loci obtained from an enriched library produced 1-9 alleles per locus, 24 in total, of which 22 showed polymorphisms. The average values of observed and expected heterozygosities among the 24 loci were 0.15 and 0.68, respectively. The microsatellite loci derived from cDNA showed 1-7 alleles per locus. Eight sequences showed significant homology to the registered genes in a database. [ABSTRACT FROM AUTHOR]

We report the development of 44 microsatellite markers in three host races of the fungus Microbotryum violaceum, a sexually transmitted disease of the Caryophyllaceae. An enrichment protocol was used to isolate microsatellite loci from three strains, collected, respectively, from the plant species Gypsophila repens, Dianthus sylvestris, and Silene vulgaris. Polymorphism and cross-amplification were explored with 32 isolates of M. violaceum, collected on 12 different plant species in natural populations. [ABSTRACT FROM AUTHOR]

We report the development of microsatellites or simple sequence repeats (SSRs) in the olive tree (Olea europaea L.). Forty three positive clones obtained by the screening of a GA-enriched genomic library were sequenced and primers were designed for 13 microsatellite loci. Five primer pairs amplified polymorphic products of the expected size range. SSR polymorphism was explored in a set of 46 olive cultivars. A total of 26 alleles were detected for the five loci. Heterozygosity ranged from 0.46 to 0.71. Ninety one per cent of the cultivars had unique multilocus genotypes. Microsatellite segregation was studied in a complex population from a cross between the commercial cultivars ’Leccino’ and ’Dolce Agogia’. [ABSTRACT FROM AUTHOR]

Ten microsatellite markers were isolated and characterized for piau-verdadeiro ( Leporinus obtusidens), an important Characiform fish species in South American basins. The markers were obtained from an enriched microsatellite genomic library, and polymorphisms were evaluated in a sample of 44 individuals caught in the upper São Francisco River. The number of alleles varied from 4 to 39. The observed and expected heterozygosities ranged from 0.614 to 0.977 and from 0.541 to 0.977, respectively. All markers were in Hardy-Weinberg equilibrium and no null alleles were detected, thus making them suitable for assessing the diversity structure of the populations of this species. [ABSTRACT FROM AUTHOR]

Microsatellites are now firmly established as an informative marker system, with increasing popularity as a tool amongst molecular ecologists. We have developed a method of constructing an enriched microsatellite library for the tropical tree species Swietenia humilis Zucc. (Meliaceae). This method is based on a precloning enrichment of SSRs using synthetic oligonucleotide probes, bound to magnetic beads and hybridizing to complementary microsatellite core sequences in digested genomic DNA. Here we describe the isolation and characterization of 10 microsatellite loci that have been used to survey the genetic diversity within a natural populations of S. humilis. A total of 97 alleles were identified with an average of 9.7 alleles over all loci. Very high levels of allelic polymorphism were detected at individual loci, with 23 alleles observed at the most variable. The mean observed heterozygosity was 0.415 (range 0.038–0.815) exceeding levels of diversity detected in related species which used isozymes as the marker system. Subpopulation differentiation at a microgeographical scale was low (FST=0.036) and the values of Nm, calculated from the allelic frequencies, were greater than 1 thus reflecting the extent of gene flow occurring between individual trees. [ABSTRACT FROM AUTHOR]

Solanum tuberosum L. DNA sequences containing simple sequence repeat (SSR) motifs were extracted from the EMBL database, cDNA and selectively enriched small-insert DNA libraries. Enrichment was achieved using either triplex affinity capture or single-strand hybridisation selection. One hundred and twelve primer pairs which successfully amplified products of the correct size from potato DNA were ultimately designed and synthesised. Ninety-eight of these revealed length polymorphisms in a panel of four diploid and two tetraploid clones, in agreement with the high information content of this class of markers which has been found in other species. All of the markers were assigned a quality score of 1–5 based on their potential usefulness. Eighty-nine loci from 65 of the primer pairs were located on two genetic linkage maps of potato by segregation analysis of the amplified alleles. Fifty-two of the SSRs were clearly single locus. The maps were aligned using 23 SSR primer pairs and 13 RFLP loci mapped in both populations. The markers described constitute a class which should replace Restriction Fragment Length Polymorphisms (RFLP) as the markers of choice for future genetic studies in potato. The sequences of the primers, together with other information on these markers are provided. [ABSTRACT FROM AUTHOR]

Poecilia vivipara is a small fish found throughout the coastal lowlands of South America. We report the isolation and characterization of 25 microsatellite loci in this tropical fish. In 24 individuals from two populations, 18 loci were polymorphic. The loci had 2-12 alleles, expected heterozygosities of 0.194-0.836, observed heterozygosities of 0.208-0.792, and discriminatory powers of 0.202-0.872. Four loci showed significant deviations from Hardy-Weinberg equilibrium. The microsatellite loci will be useful for investigating genetic diversity and population genetic structure, which will contribute to the development of efficient conservation programs for coastal lowlands where P. vivipara occurs. [ABSTRACT FROM AUTHOR]

Nine microsatellite loci were isolated and characterized in the tire track eel, Mastacembelus favus by using library enriched for microsatellite region. Among 47 specimens collected from Mekong River, the number of alleles ranged from 2 to 17 while levels of observed and expected heterozygosities ranged from 0.021 to 0.873 and from 0.021 to 0.893, respectively. The primers were successfully tested in six closely related species, hence valuable for investigating population genetics and other related aspects of M. favus and its congener species. [ABSTRACT FROM AUTHOR]

Labeo fimbriatus, the fringe-lipped peninsula carp, is a commercially important fish species next to Indian major carps. We isolated and characterized polymorphic microsatellite markers to be used as a tool for delineation of genetic stock of this species. Partial genomic libraries enriched for CT and GT repeat motifs generated 103 positive clones out of which 16 loci were found with flanking regions enough for designing primers. Thirty-two individuals of L. fimbriatus collected from wild were used to characterize the polymorphism. All the 16 loci were polymorphic with allele numbers ranging from 3 to 9. The observed and expected heterozygosity ranged from 0.093 to 0.833 and from 0.146 to 0.843, respectively. Nine loci were in agreement with Hardy-Weinberg equilibrium. No significant pair wise linkage disequilibrium was found among the loci. These markers would be very useful for characterization of natural populations of this species. [ABSTRACT FROM AUTHOR]

Geelbeck ( Atractoscion aequidens) represents a valuable fishery resource in the Benguela Current region in southwestern Africa. Due to overexploitation the species is considered depleted in South Africa, and little information is available for the rest of its distribution in the region. Genetic studies of the species are essential to inform management plans for sustainable harvesting and conservation. To facilitate such studies 12 polymorphic microsatellite DNA loci were isolated from an enriched genomic library. Genotyping of 41 individuals revealed an average of 17.75 alleles per locus (range 2-37), and observed/expected heterozygosity per locus of 0.024-0.951 and 0.024-0.962, respectively. These loci constitute the first microsatellite markers isolated for geelbeck and are applicable to multiple areas of research on this species. [ABSTRACT FROM AUTHOR]

• Premise of the study: A new set of microsatellite primers was developed for Avena sativa and characterized to assess the level of genetic diversity among cultivars and wild genotypes. • Methods and Results: Using an enrichment genomic library, 14 simple sequence repeat markers were identified. The loci of these markers were characterized and found to be polymorphic in size among 48 genotypes of oat from diverse geographical locations. The number of alleles per locus ranged from two to eight, while the observed heterozygosity ranged from 0.031 to 0.75. • Conclusions: These newly identified microsatellite markers will facilitate genetic diversity studies, fingerprinting, and genetic mapping of oat. Moreover, these new primers for A. sativa will aid future studies of polyploidy and hybridization in other species in this genus. [ABSTRACT FROM AUTHOR]

Thirteen tetranucleotide microsatellite loci were isolated from tetranucleotide microsatellite enriched libraries for the Tokay gecko ( gekko gecko). The species is endemic to Southeast Asia, known to be a habitat specialist, and exposed to severe hunting pressures in the mountainous areas of Southeast Asia. These highly polymorphic markers (6-25 alleles) will facilitate the population genetic analyses of this species. In particular, they will enable estimates of gene flow between the two types of the Tokay gecko: the black-spotted Tokay gecko and the red-spotted Tokay gecko-a critical element in assessing classification between the two. [ABSTRACT FROM AUTHOR]

Lychnophora ericoides is an endangered species, endemic to cerrado rupestre. Herein we report the development and characterization of 13 polymorphic microsatellite loci for the species, using enriched genomic library. Number of alleles per locus and expected heterozygosity ranged from 06 to 13 and 0.696 to 0.894, respectively. The high combined probability of genetic identity (2.69 × 10) and probability of paternity exclusion (0.9999) showed that multilocus genotypes are likely to be unique and will allow detailed parentage studies in natural populations of L. ericoides. [ABSTRACT FROM AUTHOR]

Ten novel microsatellite loci were isolated from Arctosa sancterosae, a white dune dwelling species of wolf spider. Diversity was assessed in 273 individuals sampled from 11 populations along the Northern coast of the Gulf of Mexico. These new genetic markers will be useful for the description and conservation of these limited populations. [ABSTRACT FROM AUTHOR]

Premise of the study: Microsatellite (simple sequence repeat, SSR) primers were developed for the Chinese peony, P. lactiflora, to investigate the diversity within Chinese peony germplasm resources. • Methods and Results: Using an SSR-enriched genomic library, a set of 10 unique polymorphic genomic SSRs was developed and characterized. The primers amplified 61 alleles in all 10 loci, including di-, tri-, and tetranucleotide repeats. The primers were also effective for P. veitchii and P. obovata. • Conclusions: The new primers will be useful for genetic research of the Chinese peony and extend the repertoire of SSR markers available to study the herbaceous taxa in Paeonia. [ABSTRACT FROM AUTHOR]