Characterization of microsatellite loci isolated from the blacktip shark and their utility in requiem and hammerhead sharks

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.     

Isolation and characterization of microsatellite markers from Pacific white shrimp (Litopenaeus vannamei)

We report the development of 11 polymorphic microsatellite loci in pacific white shrimp (Litopenaeus vannamei) using an unenriched genomic library. The number of the alleles ranged from two to 18 and observed hererozygosity ranged from 0.0286 to 0.9429, indicating that these markers will be useful for population studies and mapping in pacific white shrimp. Seven loci were detected deviated from Hardy–Weinberg, caused by deficiency of heterozygote, suggesting population genetic structure across the sampled population. No evidence for linkage disequilibrium was found.     

Polymorphic microsatellite loci for the zebra shark Stegostoma fasciatum

We report on the development and characterization of 14 polymorphic microsatellite loci in the zebra shark (Stegostoma fasciatum). Five tetranucleotide and nine dinucleotide loci were polymorphic with heterozygosities ranging from 0.400 to 0.967 and from three to 22 alleles per locus. Cross‐species amplification of these zebra shark primers on four other species of orectolobid sharks was not successful.     

Characterization of microsatellite loci in Coenonympha hero (Lepidoptera: Nymphalidae)

Microsatellite markers are suitable tools for studying dispersal pattern among local populations. I report on the characterization of seven polymorphic microsatellite loci in the scarce heath butterfly (Coenonympha hero), from one unenriched and one enriched genomic library. Number of alleles ranged from two to 20 when 108 individuals from seven populations were screened. H_O ranged from 0.140 to 0.889. Primers were also tested on three other butterfly species. Amplification was successful for all loci in Erebia triaria, while only one gave products in Maculinea alcon and Maculinea rebeli. To my knowledge, these are the first microsatellite markers published in the Nymphalidae subfamily Satyrinae.     

Development of microsatellite markers for muskellunge (Esox masquinongy) and cross‐species amplification in two other esocids

We report the development of seven polymorphic microsatellite loci in muskellunge (Esox masquinongy) using an unenriched subgenomic library. Polymorphic loci exhibited 2–11 alleles with observed heterozygosities ranging from 0.190 to 0.917 (n = 24). All seven loci amplified by their respective primer pairs resulted in monomorphic products in northern pike (E. lucius) whereas three loci amplified monomorphic products in grass pickerel (E. americanus americanus); these results imply conservation of flanking sequence but loss of polymorphism between these closely related species. Only one of six microsatellite primers developed in a previous study in northern pike amplified polymorphic products in muskellunge.     

Assessment of multiple paternity in single litters from three species of carcharhinid sharks in Hawaii

We tested for presence or absence of multiple paternity in single litters from each of three congeneric shark species in Hawaii: the sandbar shark, Carcharhinus plumbeus, bignose shark, Carcharhinus altimus, and Galapagos shark, Carcharhinus galapagensis. Based on eight polymorphic microsatellite loci, we excluded paternity by a single sire in sandbar and bignose sharks, but could not exclude a single sire for the litter from the Galapagos shark. This study doubles the number of shark species tested for multiple paternity, and is the first demonstration of multiple paternity in sandbar and bignose sharks.     

Characterisation of 14 blue crane <Emphasis Type="Italic">Grus paradisea</Emphasis> (Gruidae,AVES) microsatellite loci for use in detecting illegal trade

Forty-two unique microsatellite loci were isolated from an unenriched and a tetranucleotide-enriched blue crane (Grus paradisea) genomic library. Fourteen polymorphic loci were characterised in 20 unrelated wild blue crane individuals from the Karoo region, South Africa, and displayed 4–27 alleles with observed heterozygosities ranging between 0.50 and 0.95. All 14 loci were also polymorphic in the grey-crowned crane (Balearica regulorum) and the wattled crane (Grus carunculatus). These markers are aimed at assisting the identification of illegal trade in the blue crane but have wider population and conservation applications for most, if not all, crane species.     

Highly polymorphic microsatellite loci in the facultatively eusocial hover wasp,Liostenogaster flavolineata and cross‐species amplification

We describe eight polymorphic microsatellite loci for facultatively eusocial hover wasps (Hymenoptera: Stenogastrinae). Loci were isolated from a partial genomic library of Liostenogaster flavolineata DNA, enriched for GA repeats and an unenriched library of Liostenogaster vechti DNA. We present our results of cross‐species amplification in three species: L. flavolineata, L. vechti and Parischnogaster alternata.     

Isolation and characterization of microsatellite loci in the whale shark (Rhincodon typus)

In preparation for a study on population structure of the whale shark (Rhincodon typus), nine species-specific polymorphic microsatellite DNA markers were developed. An initial screening of 50 individuals from Holbox Island, Mexico found all nine loci to be polymorphic, with two to 17 alleles observed per locus. Observed and expected heterozygosity per locus ranged from 0.200 to 0.826 and from 0.213 to 0.857, respectively. Neither statistically significant deviations from Hardy-Weinberg expectations nor statistically significant linkage disequilibrium between loci were observed. These microsatellite loci appear suitable for examining population structure, kinship assessment and other applications.     

Characterization and the broad cross‐species applicability of 20 anonymous nuclear loci isolated from the Taiwan Hwamei (Garrulax taewanus)

We isolated 20 anonymous nuclear loci (8556 bp in total) from the Taiwan Hwamei (Garrulax taewanus), an endemic songbird of Taiwan. A panel of nine to 15 individuals with unknown relationship was used to characterize polymorphism of these loci. We identified 46 single nucleotide polymorphic sites (SNPs) in 15 polymorphic loci. Frequency of SNPs was one per every 186 bp in average. Nucleotide diversity, θ, ranged from 0.00054 to 0.00371 per locus. We also tested cross‐species applicability of these loci on 17 species from eight different passerine families. All 20 loci could be successfully amplified (ranged from one to 16 species, mean = 7.9 species).     

Characterization of microsatellite loci in Kearney's bluestar (Amsonia kearneyana) and cross‐amplification in other Amsonia species

Kearney's bluestar (Amsonia kearneyana) is a highly endangered herbaceous perennial in the family Apocynaceae. The species is found only in the Baboquivari Mountains of southern Arizona. We report the isolation and development of 12 microsatellite loci for Kearney's bluestar. Numbers of alleles ranged from two to four and observed heterozygosities ranged from 0.20 to 0.80 in the nine loci found to be polymorphic in the test population. All loci were also tested for cross‐amplification in five other Amsonia species representing two subgenera from the southwestern United States. Some loci that were not polymorphic in the Kearney's bluestar were polymorphic in other species.     

Nine new tetranucleotide microsatellite markers for the fire‐bellied toad (Bombina bombina)

We describe nine new polymorphic tetranucleotide microsatellite loci isolated from the fire‐bellied toad (Bombina bombina). The relative yield of new loci was higher than described in previous studies in amphibians: out of 12 loci initially evaluated, nine were polymorphic and amplifying reliably. Number of alleles ranged from four to 10 and observed heterozygosities from 0.47 to 0.91. Seven loci were polymorphic also in Bombina variegata and five in Bombina orientalis. Enrichment protocols yielding long flanking regions potentially overcome difficulties (i.e, low yield of reliable loci relative to number of clones screened) which have been reported in microsatellite development in anurans.     

Isolation of microsatellite markers from the drepanosiphid aphid Tuberculatus quercicola (Homoptera,Aphididae)

We isolated five polymorphic microsatellite loci from the drepanosiphid aphid Tuberculatus quercicola (Matsumura) that is associated with the Daimyo oak, Quercus dentata Thunberg, using the magnetic particles method. The isolated loci were polymorphic, with four to 16 alleles in 40 aphids. Expected heterozygosities ranged from 0.4 to 0.82. These loci can be used to quantify seasonal changes in clonal diversity in the metapopulation and the extent of clonal mixing in the colonies.     

Characterization,multiplex conditions,and cross‐species utility of tetranucleotide microsatellite loci for Blanchard's cricket frog (Acris crepitans blanchardi)

We have isolated and characterized 17 tetranucleotide microsatellite loci for Blanchard's cricket frog (Acris crepitans blanchardi), an anuran common in the central USA. Sixteen loci were organized into four multiplex amplification reactions. These loci were highly polymorphic when screened in 55 individuals from two distant populations, with 11–48 alleles per locus (average = 24.8). Observed and expected heterozygosities ranged from 0.18 to 0.97 and from 0.17 to 0.96, respectively. Nine loci were also polymorphic in Acris crepitans crepitans, with seven polymorphic in Acris gryllus. Five loci amplified in all three taxa. These loci will be useful for population‐ and species‐level investigations of this widespread group.     

Eight new polymorphic microsatellite loci from the eastern phoebe (<Emphasis Type="Italic">Sayornis phoebe</Emphasis>)

We describe eight new polymorphic microsatellite loci (5 dinucleotide and three trinucleotide) for the eastern phoebe (Sayornis phoebe) to complement five previously published loci. The number of alleles per locus ranged from 3 to 15 and observed heterozygosities ranged from 0.33 to 0.91. Preliminary screening revealed that loci were polymorphic in other Tyrannidae: Empidonax virescens (n = 10), Tyrannus tyrannus (n = 10), Tyrannus vociferans (n = 5), and Tyrannus melancholicus (n = 28).     

Characterization of microsatellite loci In The hihi Notiomystis cincta (Notiomystidae, Aves)

We have characterized 20 polymorphic microsatellite loci in the hihi Notiomystis cincta. Loci were identified by testing loci originally isolated in other avian species and by isolating new microsatellites from a hihi genomic library. These loci were characterized in unrelated hihi from a single population on Tiritiri Matangi Island (n = 98). Each locus displayed between two and 10 alleles, and observed heterozygosities ranged between 0.29 and 0.91. Nineteen of the 20 polymorphic loci could be assigned a chromosome location in the zebra finch, Taeniopygia guttata genome based on sequence homology.     

Ten new microsatellite loci for the yellowhammer (Emberiza citrinella) and their cross‐species applicability among related taxa

We describe isolation and characterization of the first microsatellite loci specifically developed for a very common European passerine bird, the yellowhammer Emberiza citrinella. Nine out of our 10 loci are polymorphic within the species E. citrinella. Number of alleles ranged from two to 21 per locus and observed heterozygosity between 0.20 and 0.91. Four primer pairs also yielded reproducible results in other species of Emberizidae. These loci comprise a set of molecular markers for various applications, from moderately polymorphic loci suitable for population studies to highly polymorphic loci for pedigree analysis in Emberizidae.     

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.     

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.     

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.