Characterization of 28 microsatellite loci in the European roller Coracias garrulus (Coracidae,AVES)

One hundred and forty-four existing microsatellite avian loci were tested for polymorphism in two to four unrelated European rollers. This allowed the identification of 28 unique polymorphic loci that we characterized in 34 unrelated individuals from a population in Guadix, Spain. The putative chromosomal locations of all the polymorphic loci in European rollers could be assigned based on the location of their orthologues in the assembled zebra finch and chicken genomes. All loci were assigned to autosomes based on predictive mapping and/or the presence of female heterozygotes, except CAM-11 that was assigned to the Z chromosome. By using a selected subset of 15 autosomal loci, the combined non-exclusion probability for identity was 4.7E-09.     

PERMANENT GENETIC RESOURCES: Isolation, characterization and multiplex genotyping of 11 autosomal and four sex-linked microsatellite loci in the reed bunting, Emberiza schoeniclus (Emberizidae, Aves)

Fifteen highly polymorphic microsatellite loci were characterized in the reed bunting, Emberiza schoeniclus. Eleven loci were autosomal and four linked to the Z chromosome. All loci were characterized and tested in 45 unrelated reed buntings from a Swiss population. Autosomal loci displayed seven to 17 and sex-linked loci displayed four to 13 alleles with heterozygosities ranging from 0.756 to 0.933 and from 0.478 to 0.957, respectively. These loci will be used in population genetic and mating system studies of reed buntings.     

Microsatellite loci characterized in three African crane species (Gruidae, Aves)

Forty-three microsatellite loci originally isolated in Grus americana and G. japonensis were tested for polymorphism in the blue crane (G. paradisea). Amplified products were sequenced in the blue crane to aid in the design of blue crane-specific primers. When characterized in 20 unrelated blue crane individuals from South Africa, 14 loci were polymorphic, with each locus displaying between 2 and 7 alleles. Eight polymorphic loci were characterized in the grey-crowned crane (Balearica regulorum) and ten in the wattled crane (G. carunculatus).     

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.     

Characterization of Mauritius parakeet (Psittacula eques) microsatellite loci and their cross-utility in other parrots (Psittacidae, Aves)

We characterized 21 polymorphic microsatellite loci in the endangered Mauritius parakeet (Psittacula eques). Loci were isolated from a Mauritius parakeet genomic library that had been enriched separately for eight different repeat motifs. Loci were characterized in up to 43 putatively unrelated Mauritius parakeets from a single population inhabiting the Black River Gorges National Park, Mauritius. Each locus displayed between three and nine alleles, with the observed heterozygosity ranging between 0.39 and 0.96. All loci were tested in 10 other parrot species. Despite testing few individuals, between seven and 21 loci were polymorphic in each of seven species tested.     

Fourteen polymorphic microsatellite loci characterized in the house sparrow Passer domesticus (Passeridae,Aves)

We characterized 14 polymorphic microsatellite loci in the house sparrow Passer domesticus. Four loci were isolated from house sparrow genomic libraries and 10 loci were identified by testing 100 loci that had been originally isolated in other passerine species. Loci were characterized in 37–54 unrelated sparrows from British and Norwegian populations. Each locus displayed between two and 31 alleles, with the observed heterozygosity ranging between 0.30 and 0.91.     

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.     

Bovine Y chromosome microsatellite polymorphisms

Thirty-eight bovine Y chromosome (BTAY) microsatellites (MS) were assessed for polymorphisms in DNA samples obtained from 17 unrelated bulls. Thirty-three of these microsatellites are new and were used for the construction of a first generation radiation hybrid map for BTAY (Liu et al., 2002). Five MS had been previously reported and were used as positive controls. Fourteen out of 38 MS were found to be polymorphic; the remaining 24 were uninformative among the animals tested. The number of hemizygous loci per MS within individual ranged from two to over 20. Seven MS presented smear- or ladder-like bands, a unique feature for Y chromosome multi-copy hemizygous MS loci. The locus length variance, within individual, ranged from 2 to 42 bp corresponding to the MS with the minimum and maximum number of loci observed, respectively. Within the 14 polymorphic MS, the five pseudoautosomal MS, on average, were more polymorphic (35.3%) than the nine Y-specific MS (19.6%). Haplotypes resulting from combinations of these polymorphic loci will provide a powerful tool for future studies on the origin of domestic cattle and the evolution of bovid species.     

Microsatellite markers for the Indian golden silkmoth, Antheraea assama (Saturniidae: Lepidoptera)

Antheraea assama, an economically important and scientifically unexplored Indian wild silkmoth, is unique among saturniid moths. For this species, a total of 87 microsatellite markers was derived from 35 000 expressed sequence tags and a microsatellite‐enriched sub‐genomic library. Forty individuals collected from Tura and West Garo Hills region of Northeast India were screened for each of these loci. Ten loci from expressed sequence tags and one from genomic library were found to be polymorphic. These microsatellite markers will be useful resources for population genetic studies of A. assama and other closely related species of saturniids. This is the first report on development of microsatellite markers for any saturniid species.     

Isolation of sixteen autosomal loci and a sex-linked polymorphic microsatellite locus from the Milne-Edwards' sportive lemur (Lepilemur edwardsi)

We isolated 21 microsatellites from the Milne-Edwards' sportive lemur, Lepilemur edwardsi. Eighteen microsatellite sequences possessed sufficient flanking DNA for primer design. Seventeen loci amplified and were found to be polymorphic displaying two to 17 alleles in 32 unrelated individuals from a population from the National Park of Ankarafantsika in northwest Madagascar. One locus (Led-12) was found to be sex linked located on the X chromosome and can be used to sex-type 40% of female L. edwardsi lemurs. These 17 loci were characterized to investigate family structure and the phylogeography of L. edwardsi.     

Isolation and characterization of microsatellite loci in the Malaysian giant freshwater prawn, Macrobrachium rosenbergii

Eight single locus microsatellite markers were developed to characterize the Malaysian giant freshwater prawn, Macrobrachium rosenbergii. These microsatellites were isolated from an enriched genomic library contained by using a 5'-anchored polymerase chain reaction technique. Primers were designed to flank the repeat sequences and subsequently used to characterize 30 unrelated individuals of the giant freshwater prawn. The polymerase chain reaction amplification products of these eight microsatellite loci were polymorphic with the number of alleles ranging from two to 10 alleles per locus while the levels of heterozygosity ranged from 0.6333 to 0.8667.     

Twenty-two polymorphic microsatellite loci aimed at detecting illegal trade in the Cape parrot, Poicephalus robustus (Psittacidae, AVES)

Twenty-two polymorphic microsatellite loci were characterized in the Cape parrot, Poicephalus robustus. Nineteen loci were newly isolated from two Cape parrot genomic libraries, and three loci isolated from other parrot species. Loci were characterized in 40 unrelated captive Cape parrots held by aviculturalists. The loci displayed between two and 24 alleles, with the observed heterozygosities ranging between 0.10 and 0.94. This locus set is suitable for identifying clarifying parentage (parentage exclusion probabilities of P_E1 = 0.0004 and P_E2 = 0.000001). Candidate parents for any Cape parrot individual can now be genotyped to distinguish between individuals, which are truly captive bred and those suspected of being wild-caught birds. Cross-species analysis found up to 31 loci to be polymorphic across 24 additional parrot species tested.     

Identification and characterization of nine polymorphic microsatellite loci in the North American pika,Ochotona princeps

Nine polymorphic microsatellite loci were developed for the North American pika (Ochotona princeps) from di‐ and tetranucleotide repeat‐enriched genomic libraries. Polymorphism was assessed for 165 individuals from eight geographical locations in the western United States. All loci were polymorphic. The number of alleles per locus ranged from three to 14, with observed heterozygosity between 0.189 and 0.822. All loci were in Hardy–Weinberg equilibrium (< 0.05). Regional differences were evident with unique alleles at multiple loci in six of eight populations.     

New microsatellite loci isolated from the field cricket Gryllus bimaculatus characterized in two cricket species, Gryllus bimaculatus and Gryllus campestris

We have developed a new set of 27 polymorphic markers for each of two cricket species, Gryllus bimaculatus and Gryllus campestris. Initially, 14 published G. bimaculatus loci were tested in G. campestris; however, only five loci were polymorphic. Therefore, we isolated an additional 50 new microsatellite loci from G. bimaculatus and tested these in both species. In a minimum of 20 individuals, 27 of the new loci were polymorphic in G. bimaculatus and 25 in G. campestris.     

Development and optimization of microsatellite DNA primers for boreal owls (Aegolius funereus)

We developed 22 microsatellite loci for boreal owls (Aegolius funereus). We genotyped 275 unrelated boreal owls (Aegolius f. richardsoni) and 36 unrelated Tengmalm's owls (Aegolius f. funereus) using seven loci that were polymorphic and did not have detectable null alleles. Among North American and Scandinavian boreal owls, respectively, allelic diversity ranged from three to 11 alleles and from one to 11 alleles, observed heterozygosity ranged from 0.31 to 0.80 and from 0.00 to 0.81, and expected heterozygosity ranged from 0.28 to 0.81 and from 0.00 to 0.81. These markers appeared to amplify DNA in six other Strigidae species.     

Effects of topography on dispersal of black-billed magpie Pica pica sericea revealed by population genetic analysis

This study was conducted using random amplified polymorphic DNA analysis to examine whether the effects of topographical structure on the dispersal of black-billed magpie Pica pica sericea are reflected in its population genetic structure. The black-billed magpie is typically seen in the lowlands and is remarkably sedentary in Korea and Japan. The unweighted pair-group method of clustering analysis showed two main clusters: five populations in the western region of the Bekdudegan mountains (WRBM) in one cluster and five populations in the eastern region of the Bekdudegan mountains (ERBM), including the Japanese population in the other cluster. The populations in WRBM appeared to be diverged from the populations in ERBM. Analysis of molecular variance revealed that the populations in ERBM were more genetically divergent from each other than were those in WRBM. The high-rising mountains, very rugged topographical features and the sea in ERBM may have resulted in a lower dispersal rate and larger genetic variation among populations in ERBM compared to those in WRBM. These results suggest that the topographical structure may have an influence on the dispersal and population genetic structure of the black-billed magpie. Electronic Publication     

In silico whole-genome EST analysis reveals 2322 novel microsatellites for the silver-lipped pearl oyster, Pinctada maxima

Molecular stock improvement techniques such as marker assisted selection have great potential in accelerating selective breeding programmes for animal production industries. However, the discovery and application of trait/marker associations usually requires a large number of genome-wide polymorphic loci. Here, we present 2322 unique microsatellites for the silver-lipped pearl oyster, Pinctada maxima, a species of aquaculture importance throughout the Indo-Australian Archipelago for production of the highly valued South Sea pearl. More than 1.2 million Roche 454 expressed sequence tag (EST) reads were screened for microsatellite repeat motifs. A total of 12,604 sequences contained either a di, tri, tetra, penta or hexa microsatellite repeat motif (n ≥ 6), with 6435 of these sequences having sufficient flanking regions for primer development. All identified microsatellites with designed primers were condensed into 2322 unique clusters (i.e., unique loci) of which 360 were shown to be polymorphic based on multiple sequence reads with different repeat motifs. Genotyping of five microsatellite loci demonstrated that in silico evaluation of polymorphism levels was a very useful method for identification of polymorphic loci, with the variation uncovered being a lower bound. Gene Ontology annotations of sequences containing microsatellites suggest that most are derived from a diverse array of unique genes. This EST derived microsatellite database will be a valuable resource for future studies in genetic map construction, diversity analysis, quantitative trait loci analysis, association mapping and marker assisted selection, not only for P. maxima, but also closely related species within the genus Pinctada.     

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.     

Isolation and characterisation of 11 microsatellite loci in the abyssal carapine grenadier Coryphaenoides carapinus (Actinoperygii, Macrouridae) and cross-amplification in two other deep-sea macrourid species

Microsatellites represent an important tool for characterising population structure, for attributing individuals to stocks, and for revealing ecological processes taking place on population and meta-population levels. A sound knowledge of population structure is essential for sustainable management of exploited fish stocks, and helps to understand population connectivity and speciation. We developed for the first time primers for microsatellite loci in the carapine grenadier, Coryphaenoides carapinus, inhabiting the abyssal Atlantic. Eleven microsatellites were obtained from partial genomic DNA libraries enriched for tetranucleotide repeats. The loci were characterised in three unrelated individuals and nine loci were found to be polymorphic. Cross-amplification in two commercially exploited deep-sea macrourid species (Coryphaenoides rupestris and Macrourus berglax) resolved two polymorphic loci in each species.     

Isolation and cross‐species amplification of microsatellite loci from the fucoid seaweeds Fucus vesiculosus,F. serratus and Ascophyllum nodosum (Heterokontophyta,Fucaceae)

The Fucaceae is a family of brown seaweeds that dominate and frequently co‐occur on North Atlantic rocky shores. We developed nine polymorphic microsatellite markers for the fucoid seaweeds Fucus vesiculosus, F. serratus and Ascophyllum nodosum using a combined, enriched library. Six of these loci were polymorphic in at least two species, showing from two to eight alleles with heterozygosities ranging from 0.41 to 0.85. Loci were also tested on F. spiralis, revealing five polymorphic microsatellite loci in this species.