Isolation and characterization of microsatellite markers for the endangered <Emphasis Type="Italic">Taxus yunnanensis</Emphasis>

Eleven polymorphic microsatellite loci were characterized for the endangered conifer Taxus yunnanensis. Eight loci were isolated through SSR-anchored PCR, one locus was developed by cross-species amplification tests, while the last two loci were obtained from cross-species microsatellite sequences available in GenBank. Variability of these markers was tested in 48 individuals collected from its main distribution range in Southwest China. The number of alleles per locus ranged from 2 to 9, and the observed and expected heterozygosity varied from 0.000 to 0.625 and from 0.062 to 0.853, respectively. Ten of these eleven loci were significantly deviated from Hardy–Weinberg expectation, except TS07 which showed a distinct heterozygote excess. The availability of these new polymorphic microsatellite markers will provide an ideal marker system for detailed population genetics studies in T. yunnanensis and potentially also for closely related species.     

Development and characteristics of microsatellite markers in Pinus merkusii

Pinus merkusii is an important industrial species that is distributed only in Southeast Asia. We isolated 10 microsatellite markers from this species using a dual‐suppression‐polymerase chain reaction technique. Of these markers, five loci were codominant and polymorphic. The number of alleles per locus ranged from three to six and the expected heterozygosity ranged from 0.389 to 0.728. These microsatellite markers will be available for analysis on population genetics and mating patterns.     

Isolation and characterization of eight polymorphic microsatellite loci for the endangered,endemic Chilean tree Gomortega keule (Gomortegaceae)

Queule (Gomortega keule) is an endangered, Chilean tree in the endemic, monotypic family Gomortegaceae. Eight polymorphic microsatellite loci were developed and characterized for this species. The number of alleles per locus ranged from four to 12 and the observed heterozygosity ranged from 0.2 to 1.0. These microsatellite markers will be valuable tools for population genetics studies and conservation planning.     

Isolation and characterization of 15 polymorphic microsatellite markers for the devastating dry rot fungus,Serpula lacrymans

Fifteen polymorphic microsatellite markers were developed from microsatellite‐enriched DNA libraries of the devastating dry rot fungus, Serpula lacrymans. The loci exhibited two to four alleles per locus and the expected heterozygosity ranged from 0.13 to 0.47. The codominant markers, described here for this fungus, will permit further studies in population genetics and phylogeography of this economically highly important species.     

Development of microsatellite markers for the dove tree, Davidia involucrata (Nyssaceae), a rare endemic from China

? Premise of the study: Microsatellite primers were developed for the endangered Davidia involucrata to assess the population genetics and infer its evolutionary history. ? Methods and Results: Using both the modified magnetic bead hybridization method and the dual-suppression PCR method, we isolated and characterized 12 polymorphic microsatellite loci using 134 individuals from five populations in southwestern China. The number of alleles per locus ranged from six to 21 (mean = 10.8). The expected heterozygosity per locus ranged from 0.404 to 0.918 and observed heterozygosity ranged from 0.015 to 0.821. ? Conclusions: All of the 12 microsatellite markers developed for D. involucrata are polymorphic, and lay a solid foundation for further studies of the population genetics of this famous tree.     

Characterization and PCR multiplexing of polymorphic microsatellite loci for the locust Locusta migratoria

Because of the scarcity of polymorphic genetic markers available in locust species, only a few population genetics studies have been carried out on this taxon. We isolated and characterized 11 polymorphic microsatellite loci in the pest locust Locusta migratoria capito, and described experimental conditions for polymerase chain reaction (PCR) multiplexing and simultaneously genotyping these loci. The number of alleles per locus ranged from six to 25, and the expected heterozygosity ranged from 0.431 to 0.957. Results of cross‐taxon amplification tests are reported in six other Locusta migratoria subspecies, six species of the Oedipodinae subfamily and two other pest locust species.     

Development and polymorphisms of microsatellite markers for hinoki (Chamaecyparis obtusa)

Fifteen microsatellite markers were developed from a microsatellite‐enriched library and characterized using 32 Chamaecyparis obtusa individuals. The number of alleles ranged from two to 27 per locus, with observed heterozygosity ranging from 0.281 to 0.906. The polymorphism information content (PIC) was also calculated for each marker and the average was 0.796 ± 0.024. These microsatellite markers will be useful for investigating population genetics, reproductive ecology, tree improvement and constructing linkage maps of the species.     

Development of microsatellite markers for Dryobalanops aromatica (Dipterocarpaceae), a tropical emergent tree in Southeast Asia

Eight polymorphic microsatellite markers were developed in Dryobalanops aromatica, an emergent tree in tropical rain forests in Southeast Asia, using an enriched library method. For the assessment of microsatellite variation, 36 individuals from a natural population were analysed. The number of alleles per locus ranged from three to 16, with observed heterozygosity of 0.056–0.833 and expected heterozygosity of 0.054–0.882. These microsatellite markers will be useful for studies of population genetics, reproductive ecology and regeneration dynamics of D. aromatica.     

Novel polymorphic microsatellite DNA markers from Malaysian giant freshwater prawn, <Emphasis Type="Italic">Macrobrachium rosenbergii</Emphasis>

Seven single locus microsatellite markers were characterized in Malaysian giant freshwater prawn, Macrobrachium rosenbergii from an enriched genomic library Primer pairs were designed to flank the repeat sequences and the loci characterized for this species. The bands resulting from the PCR amplifications of these eight microsatellite loci were polymorphic with the number of alleles ranging from 8 to 26 alleles per locus, whereas the observed heterozygosity ranged from 0.0641 to 0.6564. These newly developed microsatellite markers should prove to be useful for population studies and in the management of genetic variations in broodstocks of freshwater prawn, M. rosenbergii.     

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 of microsatellite markers in Myotis sodalis and cross-species amplification in M. gricescens, M. leibii, M. lucifugus, and M. septentrionalis

The Indiana bat (Myotis sodalis) is a highly endangered vespertilionid bat whose distribution is associated with limestone caves in the eastern United States. We present nine new polymorphic nuclear microsatellite markers for Myotis sodalis developed using an enriched library method. A total of 62 M. sodalis from two populations were used to estimate genetic diversity parameters. In M. sodalis, the number of alleles observed for each locus ranged from 17 to 48 alleles and expected heterozygosity ranged from 0.894 to 0.973. The 9 microsatellite markers were also tested on M. gricescens, M. leibii, M. lucifugus, and M. septentrionalis. These polymorphic microsatellite markers provide a valuable tool for investigating the population genetics of these species and will provide important genetic data useful for the conservation and recovery of the endangered Indiana bat.     

Microsatellite markers for population studies of the rice blast fungus,Magnaporthe grisea

We developed nine new microsatellite markers for rice blast (Magnaporthe grisea) population studies. These markers were used in addition to nine microsatellite markers previously developed by our group for mapping purpose. Altogether, the 18 markers were used in multiplex PCR (polymerase chain reaction) to characterize six populations from different geographical origins. The average number of alleles per locus across populations ranged from 1.2 to 7 and the total number of alleles detected from 2 to 19. Based on this large range of polymorphism, this set of markers is expected to be useful for different kind of population studies at different geographical scales.     

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.     

Isolation and characterization of polymorphic microsatellite markers in <Emphasis Type="Italic">Cupressus </Emphasis><Emphasis Type="Italic">chenggiana</Emphasis> S. Y. Hu (Cupressaceae)

Cupressus chenggiana S. Y. Hu (Cupressaceae) is an endemic and endangered conifer species in southwest China. In order to study the population genetics and design the effective conservation methods, we aimed to develop microsatellite primers for this species in the present study. We developed eight new microsatellite loci for this species through biotin capture method. Polymorphism of each locus was further assessed in 18 individuals from three geographically distant populations. The number of alleles per locus ranged from 6 to 11 with an average of 8.13. The observed and expected heterozygosities ranged from 0.219 to 0.296 and from 0.374 to 0.470, with averages of 0.254 and 0.417, respectively. We further found that three of nine microsatellite loci developed previously for another congeneric species showed polymorphic banding patters. We performed primer-crossing tests of these loci in the other two congeneric species which are closely related to C. chenggiana (C. gigantea and C. duclouxiana). These microsatellite markers would be effective for analyzing genetic diversity and population genetic structure of this species and its morphological differentiation with the close relatives.     

Characterization of 11 polymorphic microsatellite loci in the yellowfin seabream Acanthopagrus latus

The development and characterization of polymorphic microsatellite loci in the yellowfin seabream Acanthopagrus latus are presented. Allelic diversity was estimated in 50 samples collected from southeastern China. Eleven loci displayed polymorphism that ranged from 10 to 25 alleles per locus and the observed heterozygosity ranged from 0.48 to 0.91. These markers could be very useful for the study of population genetics of the yellowfin seabream.     

Microsatellite DNA loci from the typical halophyte <Emphasis Type="Italic">Thellungiella salsuginea</Emphasis> (Brassicaceae)

Thellungiella salsuginea (Brassiaceae) is a typical halophyte which can tolerate extreme cold, drought, and salinity. In order to understand the adaptive evolution of this species in the arid habitats, it is important to know its genetic structure. In this study, 17 polymorphic microsatellite loci were isolated and characterized from an enrichment genomic library of this species. We further assessed the polymorphisms of each locus in 18 individuals from nine geographically distant populations. The number of alleles per locus ranged from six to fourteen. The observed and expected heterozygosity ranged from 0.17 to 0.28 and 0.32 to 0.45, respectively. These markers have been crossly checked in another congeneric species, T. halophila. These microsatellite markers will be useful for investigating population genetics and adaptive evolution of this species and morphological divergence between and it and the closely related species.     

Isolation and characterization of 13 microsatellite loci in the pen shell Atrina pectinata (Bivalvia: Pinnidae)

The pen shell Atrina pectinata is a commercially important and over-exploited bivalve. In this survey, the first set of 13 polymorphic microsatellite loci for the pen shell was developed and characterized. The number of alleles ranged from 6 to 28 per locus with the observed and expected heterozygosities ranging from 0.640 to 1.000 and from 0.731 to 0.970, respectively. These informative microsatellite markers will be useful in studies of population and conservation genetics for this species.     

Characterization of nineteen microsatellite markers and development of multiplex PCRs for the wedge clam Donax trunculus (Mollusca: Bivalvia)

The wedge clam Donax trunculus is an Atlantic-Mediterranean warm-temperate species found from Senegal to the northern coast of France, including the Mediterranean and Black Sea. It is commercially exploited in several European countries and constitutes an important fishing resource due to its high economical value. To contribute to its conservation and management, nineteen microsatellite markers were isolated from two enriched genomic libraries. These loci were characterized in 30 clams from a single population from northwest Spain. The number of alleles per locus ranged from 2 to 17 and observed and expected heterozygosity varied from zero to 0.714 and from 0.078 to 0.950, respectively. Linkage disequilibrium was not detected and nine loci were in agreement with Hardy–Weinberg equilibrium. Fifteen polymorphic markers were arranged into three multiplex PCR sets to reduce both time and cost of microsatellite genotyping. This is the first time that polymorphic microsatellite markers have been reported for D. trunculus. These new markers provide a valuable resource for future population genetics studies and management and culture of this species.     

Isolation and characterization of polymorphic microsatellite loci for the invasive plant Cytisus scoparius

Scotch broom (Cytisus scoparius) is a serious weed of natural and pastoral ecosystems and is invasive in many countries worldwide. Eight polymorphic microsatellite loci were developed and characterized for this species. The number of alleles per locus ranged from five to 23. The range of observed heterozygosity was between 0.207 and 0.928. These microsatellite markers will be useful tools for studies of population genetics in the native and invasive range of this species.     

Isolation and characterization of microsatellite markers in Perilla frutescens Brit

We isolated and characterized 13 polymorphic microsatellite primers from Perilla frutescens Brit. var. frutescens by using a modified method that involves one‐way PCR amplification with single primer prior to enrichment with an ‘oligo hook’. The efficiency of this procedure for isolating unique microsatellite sequences was approximately 77%. The number of alleles per microsatellite locus ranged from three to 10 with an average of 6.5 alleles per locus while fragment size varied from 156 to 298 bp. The observed and expected heterozygosities ranged from 0.52 to 0.86 and 0.52 to 0.89, respectively. These newly isolated microsatellite markers are expected to provide valuable resources for different genetic studies currently underway in our Perilla genome research program.