Isolation and characterization of nine microsatellite loci for the takin (<Emphasis Type="Italic">Budorcas taxicolor</Emphasis>)

We report on the isolation and characterization of nine microsatellite markers in the takin (Budorcas taxicolor) from genomic DNA-enriched libraries. Twenty-eight microsatellites were screened from the libraries, and nine of the screened microsatellites were polymorphic. The number of observed alleles for each locus in 28 individuals ranged from two to seven, and the expected and observed heterozygosity was 0.105–0.758 and 0.071–0.821, respectively. Four loci (TK01, TK02, TK04 and TK08) of nine deviated from Hardy-Weinberg expectation and no significant linkage association was found among all these loci. These microsatellite markers provide useful tool for population genetic studies of the takin.     

Isolation of seven polymorphic microsatellites in Ophioblennius atlanticus atlanticus (Perciformes,Blenniidae)

We isolated and characterized seven polymorphic microsatellite loci of Ophioblennius atlanticus atlanticus (Valenciennes, 1836) using an optimized protocol to construct and screen a microsatellite‐enriched genomic library. The analysis of variability was performed in 16 specimens from Faial Island (Azores, Portugal). The mean number of alleles was 8.71 ± 2.43 and the level of expected heterozygosity ranged from 0.764 to 0.903. The total exclusionary probabilities using these loci for the first and the second parent were 0.985 and 0.998, respectively, suggesting that these microsatellites are a useful tool for large‐scale parentage analysis.     

Microsatellite loci for two East African tree species, Leptonychia usambarensis (Sterculiaceae) and Sorindeia madagascariensis (Anacardiaceae)

We isolated 20 trinucleotide microsatellites from two African tree species: Sorindeia madagascariensis (nine microsatellites) and Leptonychia usambarensis (11 microsatellites). Number of alleles ranged from three to seven in Sorindeia and two to 10 in Leptonychia. Observed heterozygosity ranged from 0.025 to 0.829 for Sorindeia and from 0.226 to 0.933 for Leptonychia. Two loci from each species departed from Hardy-Weinberg equilibrium. These microsatellite markers will be used to study how forest fragmentation affects pollination and seed dispersal processes of these tree species.     

Isolation and characterization of 13 polymorphic microsatellite loci from the clam shrimp Eulimnadia texana (Crustacea: Spinicaudata)

Thirteen polymorphic microsatellite loci were isolated and characterized from the clam shrimp Eulimnadia texana. In analyses of 20–50 individuals from two populations the number of alleles ranged from two to seven with observed heterozygosity ranging between 0.00 and 0.37. The low values for heterozygosity were not unexpected for a group characterized by its unusual androdioecious mating system, in which males compete with self‐compatible hermaphrodites for offspring production. These microsatellites are likely to be useful for further evolutionary investigations of this rare mating system in these crustaceans.     

Isolation and characterization of seven tetranucleotide microsatellite loci in mungbean,Vigna radiata

A simple and rapid method for isolating tetranucleotide microsatellites in mungbean, Vigna radiata, based on the 5′‐anchored polymerase chain reaction technique, revealed 15 microsatellite sequences. We report on the characterization of seven polymorphic microsatellite loci in V. radiata. The number of alleles per locus ranged from 2 to 6, whereas the observed heterozygosity ranged from 0 to 0.561. Tetranucleotide markers are useful because they amplify fewer stutter bands thus making scoring easier. These markers will be useful for detecting genetic variation in mungbean varieties for germplasm management and development of the crop.     

Isolation and characterization of twelve novel microsatellites in yellow catfish, Pelteobagrus fulvidraco

The yellow catfish Pelteobagrus fulvidraco is a freshwater fish species. Due to overfishing and pollution of freshwater ecosystems, the wild stocks of this fish reduced substantially. We isolated and characterized 12 polymorphic microsatellites of this species. The number of alleles at the 12 microsatellite loci ranged from four to eight, with an average of 6.6/locus. The average observed heterozygosity was 0.72, whereas the expected heterozygosity ranged from 0.60 to 0.86 (average: 0.80). All 12 microsatellites conformed to Hardy–Weinberg Equilibrium and were in linkage equilibrium. These 12 novel microsatellites could facilitate studies of genetic diversity and population structure of the yellow catfish to supply necessary information of conservation of the yellow catfish.     

Characterization of microsatellite loci in Lychnis flos‐cuculi (Caryophyllaceae)

We cloned microsatellite repeats from ragged robin Lychnis flos‐cuculi (Caryophyllaceae) and developed 20 primer pairs for microsatellite marker analysis. We used 18 individuals of a large Swiss population to screen microsatellites. Seven loci were polymorphic. Between seven and 11 alleles were found per locus and the observed heterozygosity was between 0.308 and 0.813. Observed heterozygote deficits were significant in five of the seven loci, in line with the mixed mating system of L. flos‐cuculi.     

Polymorphic microsatellite loci isolated from the Atlanto‐Mediterranean colonial ascidian Pycnoclavella sp. (Ascidiacea,Tunicata)

Eight microsatellite loci were characterized in the colonial ascidian Pycnoclavella sp. from an enriched library of genomic DNA. Most microsatellites were large and compound. Allelic variation was assessed in 30 individuals collected from Cerro Gordo (SW Mediterranean Sea). The number of alleles per locus ranged from one to nine, observed heterozygosity from 0.05 to 0.29 and expected heterozygosity from 0.15 to 0.39. No significant linkage disequilibrium between pairs of loci was detected, but five loci showed significant heterozygote deficiency that may be explained either by inbreeding, population substructure or the presence of null alleles.     

Characterization of new microsatellite markers in mung bean,Vigna radiata (L.)

The present work reports the isolation and characterization of new polymorphic microsatellites in mung bean (Vigna radiata L.). Of 93 designed primer pairs, seven were found to amplify polymorphic microsatellite loci, which were then characterized using 34 mung bean accessions. The number of alleles ranged from two to five alleles per locus with an average of three alleles. Observed and expected heterozygosity values ranged from 0 to 0.088 and from 0.275 to 0.683, respectively. All seven loci showed significant deviations from Hardy–Weinberg equilibrium, whereas only one pairwise combination (GBssr‐MB77 and GBssr‐MB91) exhibited significant departure from linkage disequilibrium. These newly developed markers are currently being utilized for diversity assessment within the mung bean germplasm collection of the Korean Gene Bank.     

Novel microsatellite loci in the grass snake (Natrix natrix) and cross-amplification in the dice snake (Natrix tessellata)

Six novel polymorphic microsatellite loci are presented for the grass snake (Natrix natrix), a species with declining populations in many regions. The number of alleles per locus ranged from two to seven. Four dice snake (Natrix tessellata) microsatellites were polymorphic in the grass snake with three to four alleles. At two loci, the expected heterozygosity differed significantly from observed heterozygosity. Cross-amplification of the grass snake markers in the dice snake showed two polymorphic microsatellites with two and four alleles.     

Isolation and characterization of polymorphic microsatellite loci in nurse shark (Ginglymostoma cirratum)

We report on the development of nine polymorphic microsatellite loci in nurse shark (Ginglymostoma cirratum) using a combination of enriched and unenriched subgenomic libraries. Based on the small percentage of positive clones in the unenriched library (0.4%) it appears that microsatellites are very scarce in nurse shark genomes. Numbers of alleles at polymorphic loci ranged from two to 15; observed heterozygosity ranged from 0.17 to 0.90. We expect these loci to be useful for studies of breeding structure and paternity.     

Characterization of 18 new microsatellite loci in Atlantic cod (Gadus morhua L.)

Eighteen new microsatellite loci consisting of 10 di‐, 5 tri‐, 2 tetra‐ and 1 heptanucleotide repeats are introduced for the Atlantic cod (Gadus morhua L.). All loci were co‐amplified in two polymerase chain reactions (plus two previously published microsatellites) and all products were typed clearly. The number of alleles per locus ranged from six (PGmo130) to 45 (PGmo76) and the observed heterozygosity ranged from 0.356 (PGmo130) to 0.957 (PGmo95). All loci except one followed Hardy–Weinberg expectations. Genetic linkage disequilibrium analysis between all pairs of loci did not yield any significant values.     

Microsatellites loci isolated in the freshwater fish Brycon hilarii

We describe seven microsatellite loci, including tetra‐, tri‐ and dinucleotides, isolated from Brycon hilarii, which is a migratory fish inhabiting the Paraguay River basin (Brazil) and is highly valued for its meat quality. Three to eight alleles per locus were detected and the expected heterozygosity ranged from 0.31 to 0.81. Positive results were obtained with cross‐amplification in five other species of Brycon. These microsatellites provide a potential tool for wild population and aquaculture studies of B. hilarii and other species of the genus.     

Isolation of highly polymorphic microsatellite markers from the intertidal barnacle Chthamalus montagui Southward

This study reports the isolation and characterization of seven highly polymorphic microsatellite loci in Chthamalus montagui (Crustacea, Cirripedia). The loci were isolated from a library constructed from genomic DNA enriched for CA repeats. The markers yielded three to 43 alleles per locus (mean 16.7) in samples averaging 49 individuals. Observed heterozygosity ranged from 0.08 to 0.58 (mean 0.39). These microsatellite loci will be valuable tools for population genetic studies of this species.     

Isolation and characterization of polymorphic microsatellite loci in a traditional Chinese medicinal plant, Schisandra sphenanthera

Schisandra sphenanthera (Schisandraceae) is a traditional Chinese medicinal plant. In this study, nine polymorphic microsatellite loci were developed and characterized for the species. A genomic DNA enrichment protocol was used to isolate microsatellite loci and polymorphism was explored using 36 individuals sampled from a natural population. The observed number of alleles ranged from 2 to 10 with an average of 5.2. Expected and observed heterozygosity ranged from 0.30 to 0.82 and 0.33 to 0.97, respectively. These microsatellites have been directly applied to both population and conservation genetic studies of S. sphenanthera.     

Characterization of 35 novel microsatellite DNA markers from the duck (Anas platyrhynchos) genome and cross-amplification in other birds

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.     

Polymorphic microsatellite loci for population studies of <Emphasis Type="Italic">Bullacta exarata</Emphasis>

Nine polymorphic microsatellite loci were isolated and characterized from an AC-enriched genomic library of Bullacta exarata. They were polymorphic in 30 individuals from Qidong in Jiangsu Province, China. The number of alleles per polymorphic loci varied from 7 to 14 and the values of observed heterozygosity and expected heterozygosity ranged from 0.2593 to 0.9667 and from 0.7090 to 0.9164, respectively. Six of the nine microsatellites conformed to Hardy–Weinberg equilibrium, and inherited independently. These informative microsatellite markers will be useful in studies of population genetic structure for this species.     

Isolation and characterization of microsatellites in Sparganium emersum and cross‐species amplification in the related species S. erectum

We developed seven novel polymorphic microsatellite loci for the aquatic macrophyte Sparganium emersum (Sparganiaceae). These were characterized on 62 individuals collected from nine different populations. In this set of individuals, seven to 20 alleles per locus were detected and observed heterozygosity ranged between 0.16 and 0.95. Cross‐species amplification was tested in the related species Sparganium erectum, and was successful for five of the seven microsatellite loci.     

Isolation of polymorphic microsatellite loci from the red‐billed gull (Larus novaehollandiae scopulinus) and amplification in related species

We describe the isolation and characterization of seven dinucleotide microsatellite loci developed from the red‐billed gull (Larus novaehollandiae scopulinus). Locus‐specific primers were used to genotype individuals from 13 populations of this subspecies as well as individuals from closely related subspecies from Australia and New Caledonia. The primers were tested successfully on other species of gulls and shorebirds. The number of alleles observed within the red‐billed gull ranged from three to 17, and observed heterozygosity varied from 0.359 to 0.787. These microsatellites are likely to be useful for studies of mating systems and population genetics in a wide range of gull species.