Ten microsatellite loci from Solms-laubachia eurycarpa (Brassicaceae)

Solms-laubachia eurycarpa is a medicinal herb endemic to the Hengduan Mountains region of south-central China. We screened a partial genomic library enriched for microsatellites and characterized 10 polymorphic loci for S. eurycarpa. The number of alleles per locus ranged from five to 15, with an average of 9.6. The observed and expected heterozygosities ranged from 0.2 to 0.725 and from 0.585 to 0.871, respectively. Amplification in closely related taxa was successful for most loci. The results indicate significant potential for the utility of these markers in studying the population genetics of S. eurycarpa and related species.     

Characterization and cross-amplification of 13 microsatellite loci in the northern pine processionary moth, Thaumetopoea pinivora (Lepidoptera: Notodontidae)

Thirteen polymorphic microsatellite loci were developed for the northern pine processionary moth (Thaumetopoea pinivora) and tested for cross-amplification in seven other species within the Thaumetopoea family. Number of alleles ranged from two to 10 when at least 28 individuals from one population were screened and one locus, Thapin06, appears to be sex linked. Expected heterozygosity ranged from 0.094 to 0.856 and observed heterozygosity ranged from 0.097 to 0.806. Amplification success varied between sister species, with two up to seven loci being successfully amplified. The described loci will be valuable for studying the population genetic structure and dispersal behaviour of this forest pest.     

Characterization of six microsatellite markers in Trillium camschatcense using a dual‐suppression‐polymerase chain reaction technique

Trillium camschatcense is a herbaceous perennial plant distributed in Hokkaido and northern Honshu, Japan. Geographical variations in the breeding system (partial selfing or obligate outcrossing) are reported in the populations of Hokkaido. We isolated six polymorphic microsatellite loci from this species. The number of allele per locus ranged from four to 12, whereas the expected heterozygosities ranged from 0.69 to 0.83. These markers may allow further investigations to reveal the evolutionary and ecological processes of mating system in T. camschatcense.     

Microsatellite markers for the palaeotropic pioneer tree genus Macaranga (Euphorbiaceae) and their cross‐species transferability

We developed primer sequences for five polymorphic microsatellite loci in the tropical ant‐plant genus Macaranga (Euphorbiaceae). Population genetic parameters were determined on the basis of 30 individuals from each of two Macaranga species in Borneo. Allele numbers per locus ranged from three to 13. Expected and observed heterozygosities ranged from 0.160 to 0.850 and from 0.130 to 0.700, respectively. Four of the five primer pairs cross‐amplify polymorphic PCR products in a wide range of Macaranga species.     

Microsatellite markers for the cosmopolitan marine diatom Pseudo‐nitzschia pungens

Very little information is available on the genetic structure of populations of phytoplankton species. Here we report the characterization of six polymorphic microsatellite loci in the marine diatom, Pseudo‐nitzschia pungens, using clones (between 87 and 213 per locus) isolated from spring blooms in the North Sea. The number of alleles per locus ranged from five to 19 and observed heterozygosities ranged from 0.55 to 0.86.     

Isolation and characterization of 11 new microsatellite markers for Macaranga (Euphorbiaceae)

We provide primer sequences for 11 new polymorphic microsatellite markers developed in the tropical ant-plant genus Macaranga (Euphorbiaceae), after enrichment cloning of Macaranga tanarius and Macaranga hypoleuca. Allele numbers per locus ranged from two to 16 among 20 accessions of M. tanarius, and from three to 10 among 22 accessions of M. hypoleuca. Observed and expected heterozygosities ranged from 0.150 to 0.900 and from 0.375 to 0.894 in M. tanarius, and from 0.545 to 1.000 and from 0.434 to 0.870 in M. hypoleuca, respectively. Six of the 11 primer pairs successfully cross-amplified polymorphic polymerase chain reaction products in Macaranga winkleri.     

Development of new microsatellite markers from a salt-marsh sedge Carex rugulosa by compound simple sequence repeat-polymerase chain reaction

We developed 12 polymorphic microsatellite markers from a salt-marsh sedge Carex rugulosa. The number of alleles per locus ranged from two to four, with an average of 2.75. The observed and expected heterozygosities ranged from 0.067 to 0.600 and from 0.128 to 0.620, respectively. These simple sequence repeat markers will allow the identification of genets and evaluation of the genetic diversity of C. rugulosa.     

Simultaneous quantification of twelve bioactive components in San-huang-xie-xin-tang by HPLC

A simple and reliable high-performance liquid chromatographic method with ultraviolet detection (280 nm) has been developed for the simultaneous analysis of 12 bioactive components in San-huang-xie-xin-tang (SHXXT), a traditional Chinese medicine containing Rhei rhizome, Coptidis rhizome and Scutellariae radix. A relatively simple extraction procedure was employed and optimised, and separation of the components was obtained within 1 h using a reversed-phase column under gradient elution with acetonitrile and a buffer containing 0.01 M sodium 1-pentanesulphonate (pH 3). The lower limit of detection for the analytes ranged from 25 to 75 ng/mL. The correlation coefficients associated with each calibration curve were greater than 0.99. The precision and accuracy of the method ranged from 1.0 to 10.5% at low concentration levels, 0.8 to 8.7% at medium levels and 1.2 to 5.8% at high levels. In commercial products of SHXXT, baicalin and berberine were present in the highest amounts with levels up to 4.0 and 3.3%, respectively, in one sample. The HPLC method was able rapidly and efficiently to analyse constituents in crude herb and traditional Chinese medicinal preparations containing Rhei rhizome, Coptidis rhizome and Scutellariae radix.     

Isolation and characterization of microsatellite loci in the highland papaya Vasconcellea × heilbornii V. Badillo (Caricaceae)

Nine polymorphic microsatellite loci were identified in the tropical plant Vasconcellea ×heilbornii and used to estimate allelic diversity in two populations of southern Ecuador. Allelic richness ranged from two to five alleles, observed heterozygosity ranged from 0.150 to 0.947 and expected heterozygosity ranged from 0.186 to 0.701. Most of these markers also amplified microsatellite loci in two other Vasconcellea species (Vasconcellea stipulata and Vasconcellea cundinamarcensis). Hence, these markers will be useful for population genetic analysis and the evaluation of genetic diversity and gene flow in these species.     

Eight polymorphic microsatellite markers developed in the green leaf bug,Lygus lucorum Meyer‐Dür (Hemiptera: Miridae)

We developed eight polymorphic microsatellite markers for the green leaf bug, Lygus lucorum Meyer‐Dür. The number of observed alleles per locus in 48 individuals ranged from three to 13, whereas the observed and expected heterozygosities ranged from 0.133 to 0.810 and from 0.236 to 0.886, respectively. Cross‐species amplifications were conducted both within and between generas, and the amplification results indicated that these primers enriched for L. lucorum Meyer‐Dür are species‐specific.     

Isolation and characterization of polymorphic microsatellite loci in the field vole,Microtus agrestis,and their cross‐utility in the common vole,Microtus arvalis

We developed nine polymorphic microsatellite loci for the field vole, Microtus agrestis. The number of alleles ranged from five to 15 and observed heterozygosities ranged from 0.40 to 1.00. We also tested the microsatellite loci for amplification and polymorphism in the congeneric species Microtus arvalis. Five of the nine loci were successfully analysed in this species. The microsatellite markers will be employed in studies of reproductive success and fine‐scale spatial genetic structure.     

6‐Methyl 3‐chromonyl 2,4‐thiazolidinedione/2,4‐imidazolidinedione/2‐thioxo‐imidazolidine‐4‐one compounds: novel scavengers of reactive oxygen species

The benefits of antioxidants on human health are usually ascribed to their potential ability to remove reactive oxygen species providing protection against oxidative stress. In this paper the free radicals scavenging activities of nine 6‐methyl 3‐chromonyl derivatives (CMs) were evaluated for the first time by the chemiluminescence, electron paramagnetic resonance, spin trapping and 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH^?) methods. The total antioxidant capacity was also measured using a ferric‐ferrozine reagent. Compounds having a hydrogen atom at the N3‐position of the β‐ring were effective in quenching CL resulted from the KO₂/18‐crown‐6‐ether system (a source of superoxide anion radical, ) in a dose‐dependent manner over the range of 0.05–1 mmol/L [IC₅₀ ranged from 0.353 (0.04) to 0.668 (0.05) mmol/L]. The examined compounds exhibited a significant scavenging effect towards hydroxyl radicals (HO^? HO^?), produced by the Fenton reaction, and this ranged from 24.0% to 61.0%, at the concentration of 2.5 mmol/L. Furthermore, the compounds examined were also found to inhibit DPPH^? and this ranged from 51.9% to 97.4% at the same concentration. In addition, the use of the total antioxidant capacity assay confirmed that CM compounds are able to act as reductants. According to the present study, CM compounds showed effective in vitro free radical scavenging activity and may be considered as potential therapeutics to control diseases of oxidative stress‐related etiology. Copyright © 2013 John Wiley & Sons, Ltd.     

Naked Amoebae in Biofilms Collected from a Temperate Freshwater Pond

Biofilms collected on Plexiglass substrates, from a freshwater pond in northern New York State, were examined microscopically for naked amoebae densities, sizes, diversity, and estimated C‐biomass. Five samples were obtained during summer 2006 and 2007. The densities ranged from 109 to 136/cm² biofilm surface and 285 to 550/mg biofilm dry weight. Sizes ranged from 13 to 200 μm. Diversities ranged from 4.23 to 4.55. C‐biomass ranged from 64 to 543 ng C/cm² and 125 to 1,700 μg C/g dry weight. Thirty morphospecies were identified among the five samples, including very large amoebae in the range of 100–200 μm. Large amoebae (≥ 50 μm) accounted for the largest proportion of the C‐biomass.     

Isolation and characterization of 12 tetranucleotide repeat microsatellite loci from the green‐backed tit (Parus monticolus)

From a partial genomic library enriched for GATA short tandem repeats, we developed 12 polymorphic microsatellite loci from the green‐backed tit (Parus monticolus). We characterized these loci by genotyping 30 adult individuals with unknown relationship. The number of alleles ranged from four to 17 per locus (mean = 9.3 alleles) and the observed heterozygosity for each locus ranged from 0.633 to 0.933 (mean = 0.789). All loci conformed to Hardy–Weinberg expectations. Four of 66 possible pairwise comparisons between loci showed significant gametic disequilibrium.     

Characterization of polymorphic microsatellite DNA markers in the ruff (Philomachus pugnax)

Nine polymorphic microsatellite markers were isolated and characterized from the lek‐breeding ruff (Philomachus pugnax) using an enrichment cloning procedure. The number of alleles resolved per locus ranged from five to 56, and observed heterozygosity ranged from 0.311 to 0.857 in a sample of 319 ruffs from Gotland, Sweden. Two loci were located on the Z chromosome, rendering all females hemizygotes. These markers are effective in the assignment of parentage and determining male mating success.     

Development and use of anchored‐SSRs to study DNA polymorphism in bread wheat (Triticum aestivum L.)

In bread wheat, 21 anchored simple sequence repeat (SSR) primer pairs detecting SSR length polymorphism and 42 anchored SSR primers detecting microsatellite‐anchored fragment length polymorphisms (MFLPs) are reported. Eight bread wheat genotypes were used for detecting polymorphism. The number of alleles in SSR analysis ranged from two to six, with a mean of 2.9 alleles per SSR. The number of polymorphic bands in MFLP ranged from two to 40, with a mean of 12.74 polymorphic bands/primer combination, the SSRs with CT/GA motifs giving the highest level of polymorphism (a mean of 18.37 bands). The average value of polymorphic information content (PIC) was 0.473 for SSRs and 0.061 for MFLP.     

Development of 10 microsatellite loci in the ling (Molva molva)

We developed primers for two dinucleotide and eight tetranucleotide microsatellite loci in a marine fish, the ling (Molva molva). All markers were obtained from partial genomic DNA libraries and characterized in 55 unrelated individuals from one putative population. The number of alleles ranged from five to 24 (average 10.5) per locus, and the observed heterozygosity ranged from 0.218 to 0.981 (average 0.643). No loci amplified in two other gadoid species tested, the Atlantic cod (Gadus morhua) and the tusk (Brosme brosme).     

Isolation and characterization of polymorphic microsatellite markers in the white‐winged chough (Corcorax melanorhamphos)

We have isolated and characterized seven polymorphic microsatellite loci in the white‐winged chough (Corcorax melanorhamphos), a highly social, cooperatively breeding bird of Australian eucalypt woodlands. In analyses of 100 samples from 16 family groups, the number of alleles per locus ranged from four to 18, and observed heterozygosity ranged between 0.46 and 0.93. One locus appears to be sex‐linked. The primers were also tested in apostlebirds (Struthidera cinerea), the only other species in the subfamily Corcoracinae. Five loci were successfully amplified and three were polymorphic.     

Characterization of expressed sequence tag‐derived microsatellites from the kuruma prawn,Marsupenaeus japonicus

The kuruma prawn, Marsupenaeus japonicus, is a commercially important benthic marine crustacean in East Asia. Understanding the species’ population structures will be very important for its proper stock assessment and management strategy. Herein, 13 polymorphic microsatellite loci originating from expressed sequence tag libraries of M. japonicus were isolated and characterized. Number of alleles per locus ranged from two to 15, observed and expected heterozygosities ranged from 0.250 to 0.906 and from 0.310 to 0.932, respectively. The adequate level of variability within the population renders these microsatellite loci ideal markers for population genetic analysis of M. japonicus.     

Isolation of polymorphic microsatellite loci from Eurasian woodcock (Scolopax rusticola) and their cross‐utility in related species

We isolated one trinucleotide and seven tetranucleotide microsatellite loci for the Eurasian woodcock (Scolopax rusticola). We describe polymerase chain reaction conditions and primers for the successful amplification of these loci and report the results obtained from their use in 42 specimens from two populations in Europe. The number of alleles per locus ranged from three to 15, observed heterozygosity was comprised between 0.11 and 1.00 and expected heterozygosity ranged between 0.10 and 0.91. Cross‐specific amplification experiments highlighted the potential usefulness of these molecular markers for the study of three related scolopacid waders.