Isolation and characterization of microsatellite loci from Weigela coraeensis (Caprifoliaceae)

Weigela coraeensis var. coraeensis is a deciduous shrub species distributed in Japan on the mainland, Honshu, whereas its variety W. coraeensis var. fragrans is endemic to the Izu Islands located south of Honshu. We isolated eight polymorphic microsatellite loci from the species and characterized these loci for 20 individuals from a population in Honshu. The number of alleles per locus ranged from 7 to 15 and the observed and expected heterozygosities ranged from 0.60 to 0.90 and from 0.65 to 0.90, respectively. These eight polymorphic microsatellites will be useful for examining intraspecific genetic differentiation in W. coraeensis.     

Isolation and characterization of microsatellite markers from the hosta species Hosta albomarginata (Liliaceae)

Hostas are very popular ornamental plants in gardens in the United States, Europe, and East Asia. We have developed nine microsatellite loci from an enrichment library of genomic DNA in Hosta albomarginata. We characterized these nine microsatellite loci for 20 individuals from a population of H. albomarginata. The primers developed in this study yielded an average of 12.4 alleles per locus (range five to 20) and an average expected heterozygosity of 0.86 (range 0.65 to 0.95). These markers will be powerful tools for breeding programmes of hosta cultivars, studies of population genetics, and the conservation of wild hosta species.     

Development and characterization of eight microsatellite markers in bullet tuna (Auxis rochei)

The bullet tuna (Auxis rochei) is a worldwide species of high commercial value in Spain. Nevertheless, little information is currently available about the genetic characteristics of wild A. rochei populations. In this survey, we have developed eight new microsatellites for the bullet tuna using an enriched genome library protocol. Primers were screened on a total of 78 individuals from three wild populations (Mediterranean, Atlantic, and Pacific), revealing seven to 27 alleles per locus with expected heterozygosities ranging from 0.63 to 0.97. These markers can be potentially useful tools for use in population genetic studies.     

Development and characterization of eight new microsatellite markers for the haremic sandperch,Parapercis cylindrica (family Pinguipedidae)

Eight di‐, tri‐ and tetranucleotide microsatellite markers were developed for the haremic sandperch Parapercis cylindrica using a linker‐ligated, magnetic bead enrichment protocol. Screening of at least 17 individuals showed these markers to be polymorphic with observed heterozygosity ranging from 0.381 to 1.000 (mean = 0.742) and the numbers of alleles ranging from three to 18. The average polymorphic information content for these eight loci was 0.723. These markers may be used for parentage studies aimed at exploring the complex mating strategies employed by this haremic coral reef fish and will be valuable for population genetic studies.     

Isolation and characterization of microsatellite loci in Eurycorymbus cavaleriei,a dioecious endemic tree species in China

Eleven dinucleotide microsatellites were developed and characterized for Eurycorymbus cavaleriei, a dioecious, rare and endemic tree species in China. A genomic DNA enrichment protocol was used to isolate microsatellite loci and polymorphism was explored using 30 trees from two natural populations. The observed heterozygosity (H_O) ranged from 0.379 to 0.931, with an average of 0.625. These markers provide powerful tools for the ongoing population and conservation genetics studies of E. cavaleriei.     

Development and characterization of microsatellite markers for the tropical tree species Tabebuia aurea (Bignoniaceae)

In this study, we report the development and characterization of 21 polymorphic microsatellite loci for Tabebuia aurea, using genomic library enrichment. Number of alleles per locus and expected heterozygosity ranged from nine to 26 and from 0.808 to 0.955. The high combined probability of genetic identity (1.03 × 10^?37) and probability of paternity exclusion (0.9889) showed that multilocus genotypes are likely to be unique and will allow detailed parentage studies in natural populations of T. aurea. Additionally, a high percentage of transferability was achieved for the four species of the same genus studied.     

Development of microsatellite markers for the endangered medicinal plant Launaea arborescens (Asteraceae)

• Premise of the study: Microsatellite loci were developed in Launaea arborescens, an endangered and medicinal Asteraceae species in North Africa, for further investigation of its conservation genetics. • Methods and Results: We isolated and characterized 10 polymorphic and nine monomorphic microsatellite loci from L. arborescens using the protocol of Fast Isolation by AFLP of Sequences COntaining repeats (FIASCO). For the 10 polymorphic loci, the number of alleles detected per locus varied from two to six, and the observed and expected heterozygosities ranged from 0.000 to 0.833 and 0.059 to 0.713, respectively. • Conclusions: The polymorphic markers provide a useful tool for conservation genetics studies of L. arborescens, including analysis of mating system, estimating gene flow, and identifying discrete genetic units within the species.     

Development and characterization of eight polymorphic microsatellite markers for Daphnia atkinsoni (Crustacea: Ctenodaphnia)

A microsatellite‐enriched genomic library was developed for the water flea Daphnia atkinsoni Baird, 1859, a dominant species of intermittent wetlands in Europe. Eight polymorphic microsatellite markers were successfully optimized. Characterization of 77 individuals from Belgium and Spain showed moderate (in the former) to high (in the latter) levels of polymorphism with two to 11 alleles per locus and an observed heterozygosity ranging from 0 to 0.87. Some of these microsatellite markers were successfully amplified in three other Daphnia species (D. magna n = 4, D. similis n = 6; D. obtusa n = 6).     

Isolation and characterization of microsatellite markers in the tropical tree species Dicorynia guianensis (Caesalpinaceae)

Dicorynia guianensis is a canopy tree endemic to French Guyana and the first to be exploited as timber wood. To investigate levels of diversity and dynamics of gene flow in a 40‐ha study plot, highly polymorphic microsatellite markers were developed. An enriched microsatellite library was constructed and seven microsatellite primer pairs were developed. Total genomic DNA was extracted and amplified from cambium tissue from 172 adult individuals. The level of genetic diversity was found to be low. A large heterozygote deficiency was found at one locus. This was resolved by redesigning the primer set.     

Transferability and characterization of nine microsatellite markers for the tropical tree species Tabebuia roseo-alba

Microsatellite loci that were previously developed in the tropical tree Tabebuia aurea were used for the genetic analysis of Tabebuia roseo-alba populations. Nine of 10 simple sequence repeat markers were amplified, and the polymorphism was assessed in 58 individuals sampled from two stands in southeastern Brazil. All loci were polymorphic with Mendelian inheritance. The allele numbers were high, ranging from 5 to 13 in population I and 3 to 7 in population II, with means of 8.9 and 5.5, respectively. We conclude that these markers can be efficiently used for parentage and gene-flow studies.     

Isolation and characterization of highly polymorphic microsatellite markers in Hypochaeris radicata (Asteraceae)

We developed five highly polymorphic dinucleotide microsatellite loci for the grassland species Hypochaeris radicata (Asteraceae). Polymorphism of these markers was examined in six populations in the Netherlands. All loci were polymorphic in all populations. The number of alleles per locus varied between 18 and 43. Expected heterozygosity was between 0.86 and 0.91. Cross‐species amplification was tested in six Hypochaeris species and was successful for three different loci in four species. These microsatellites are a useful tool in population genetic, dispersal and metapopulation studies or in testing levels of inbreeding.     

Testing four proposed barcoding markers for the identification of species within Ligustrum L. (Oleaceae)

DNA barcoding is a biological technique that uses short and standardized genes or DNA regions to facilitate species identification. DNA barcoding has been used successfully in several animal and plant groups. Ligustrum (Oleaceae) species occur widely throughout the world and are used as medicinal plants in China. Therefore, the accurate identification of species in this genus is necessary. Four potential DNA barcodes, namely the nuclear ribosomal internal transcribed spacer (ITS) and three chloroplast (cp) DNA regions (rbcL, matK, and trnH–psbA), were used to differentiate species within Ligustrum. BLAST, character-based method, tree-based methods and TAXONDNA analysis were used to investigate the molecular identification capabilities of the chosen markers for discriminating 92 samples representing 20 species of this genus. The results showed that the ITS sequences have the most variable information, followed by trnH–psbA, matK, and rbcL. All sequences of the four regions correctly identified the species at the genus level using BLAST alignment. At the species level, the discriminating power of rbcL, matK, trnH–psbA, and ITS based on neighbor-joining (NJ) trees was 36.8%, 38.9%, 77.8%, and 80%, respectively. Using character-based and maximum parsimony (MP) tree methods together, the discriminating ability of trnH–psbA increased to 88.9%. All species could be differentiated using ITS when combining the NJ tree method with character-based or MP tree methods. Overall, the results indicate that DNA barcoding is an effective molecular identification method for Ligustrum species. We propose the nuclear ribosomal ITS as a plant barcode for plant identification and trnH–psbA as a candidate barcode sequence.     

Development and characterization of ten new microsatellite markers in a mangrove tree species Bruguiera gymnorrhiza (L.)

Bruguiera gymnorrhiza is an ecologically and somewhat economically important mangrove tree species. We isolated 10 polymorphic microsatellite loci from B. gymnorrhiza using a dual‐suppression polymerase chain reaction (PCR) technique. These loci provided microsatellite markers with polymorphism of two to five alleles per locus within 216 individuals from nine natural populations of B. gymnorrhiza on Iriomote Island, the Sakishima Islands, Japan. The expected and observed heterozygosities ranged from 0.220 to 0.720 and from 0.104 to 0.447, respectively.     

Development and characterization of 11 microsatellite markers in a widespread Neotropical seasonally dry forest tree species,Geoffroea spinosa Jacq. (Leguminosae)

Eleven dinucleotide microsatellites were developed in Geoffroea spinosa (Leguminosae), a widespread tree of the seasonally dry Neotropical forests, and characterized on six populations from Peru, Argentina and Paraguay. Four of them amplified on the Peruvian populations only, probably because of mutations in the microsatellite flanking regions in the other populations. Ten microsatellites were found polymorphic, with within population gene diversities ranging from 0.17 to 0.95, and a number of alleles varying from seven to 19. A significant overall genetic differentiation was also found (θ = 0.212; P < 0.01).     

Characterization of microsatellite loci for a barrenwort species (Epimedium diphyllum,Berberidaceae)

Microsatellites were identified and characterized from Epimedium diphyllum, a species of barrenworts, both attractive garden plants and valuable medicinal plants. Some Japanese species of Epimedium are threatened with extinction, and are listed in the Red Data Book of plants in Japan. Natural hybrid zones also have been reported among some taxa of Japanese Epimedium. We developed eight polymorphic microsatellite primers for population genetic analyses of E. diphyllum. The numbers of alleles per locus ranged from 10 to 20, with observed levels of heterozygosity between 0.85 and 1.00. These primer sets yielded amplification in the other three Japanese Epimedium. These markers will be valuable for conservation genetics, evolutionary biology, pharmacognostic study, and horticultural study of Epimedium.     

Testing four proposed barcoding markers for the identification of species within Ligustrum L.(Oleaceae)

DNA barcoding is a biological technique that uses short and standardized genes or DNA regions to facilitate species identification. DNA barcoding has been used successfully in several animal and plant groups. Ligustrum (Oleaceae) species occur widely throughout the world and are used as medicinal plants in China. Therefore, the accurate identification of species in this genus is necessary. Four potential DNA barcodes, namely the nuclear ribosomal internal transcribed spacer (ITS) and three chloroplast (cp) DNA regions (rbcL, marK, and trnH-psbA),were used to differentiate species within Ligustrum. BLAST, character-based method, tree-based methods and TAXONDNA analysis were used to investigate the molecular identification capabilities of the chosen markers for discriminating 92 samples representing 20 species of this genus. The results showed that the ITS sequences have the most variable information, followed by trnH-psbA, matK, and rbcL. All sequences of the four regions correctly identified the species at the genus level using BLAST alignment. At the species level, the discriminating power of rbcL, matK, trnH-psbA and ITS based on neighbor-joining (NJ) trees was 36.8%, 38.9%, 77.8%, and 80%,respectively. Using character-based and maximum parsimony (MP) tree methods together, the discriminating ability of trnH-psbA increased to 88.9%. All species could be differentiated using ITS when combining the NJ tree method with character-based or MP tree methods. Overall, the results indicate that DNA barcoding is an effective molecular identification method for Ligustrum species. We propose the nuclear ribosomal ITS as a plant barcode for plant identification and trnH-psbA as a candidate barcode sequence.     

Development and characterization of microsatellite markers for the Amazonian blackwing hatchetfish,Carnegiella marthae (Teleostei,Gasteropelecidae)

The blackwing hatchetfish, Carnegiella marthae, is a small characin species distributed in forest streams of the Negro and upper Orinoco River basins in Amazonia. Freshwater hatchetfish are popular in the aquarium trade and represent an economic resource for the riverine people from middle Rio Negro, in Brazil. We isolated and characterized seven microsatellite DNA loci for the blackwing hatchetfish. Number of alleles and heterozygosity per locus in a sample of 30 fish ranged from three to 17 and from 0.19 to 0.87, respectively. These microsatellite loci provide powerful markers for studies on taxonomy, management and phylogeographic history of Amazonian hatchetfish.     

Development of 30 microsatellite markers for dab (Limanda limanda L.): a key UK marine biomonitoring species

Dab (Limanda limanda) are the principal target fish species in offshore biomonitoring programmes in the UK; however, detailed knowledge of genetic structure and connectivity among sampling locations is unavailable. Here, the isolation and characterization of 30 polymorphic microsatellite loci for dab is described. The number of alleles per locus ranged from 2 to 42, with observed heterozygosities ranging from 0.089 to 1. These loci will enable high resolution of genetic population structure and dynamics of dab around the British Isles.     

Development of polymorphic microsatellite markers in Hessian fly, Mayetiola destructor (Say)

A microsatellite library was prepared from size-selected genomic DNA of Hessian fly (Mayetiola destructor). Approximately 81% of recovered clones hybridized with microsatellite motif-specific probes. Subsequently, 2350 clones were sequenced. Sixty-two individual flies from laboratory strains were used to test for reliability and polymorphism in 50 of the microsatellites by gel electrophoresis; 18 were further tested with capillary electrophoresis. Of these, 17 behaved as a polymorphic single locus appropriate for population analysis.