Isolation,characterization and cross‐species amplification of microsatellite DNA loci in the tropical American yam Dioscorea trifida

We developed microsatellite markers in American yam (Dioscorea trifida). A microsatellite sequence‐enriched genomic library was screened, and after sequencing, primers were designed for 20 microsatellites. Among these, eight primer pairs yielded amplification products that were both interpretable and polymorphic in 24 yam cultivars. The number of alleles per locus ranged from two to 13 and the overall expected heterozygosity was around 0.5. Six of the eight Dioscorea trifida microsatellite loci gave amplification products in other Dioscorea species.     

Polymorphic tetranucleotide microsatellite loci in the Hwamei (Garrulax canorus canorus) (Timaliidae)

Eight polymorphic tetrarepeat (GATA)_n, microsatellite loci were isolated from a babbler, Hwamei (Garrulax canorus canorus). We evaluated the polymorphism of these microsatellite loci by genotyping 36–48 individuals from the Asian mainland. The number of alleles for each locus ranged from eight to 29. The heterozygosity was between 0.587 and 0.978. Except for one locus, genotype frequencies of these microsatellites did not significantly deviate from the Hardy–Weinberg expectation. These markers should be useful for monitoring potential hybridization between different Hwamei subspecies and provide new insights into the mating system and geographical differentiation of these birds.     

Development and multiplex PCR amplification of novel microsatellite markers in the White‐tailed Sea Eagle,Haliaeetus albicilla (Aves: Falconiformes,Accipitridae)

We report the development of 14 novel polymorphic microsatellite markers cloned from the White‐tailed Sea Eagle, Haliaeetus albicilla, a formerly threatened raptor that has received much conservation attention throughout Eurasia. We also present a protocol for multiplex polymerase chain reaction (PCR) amplification of the loci. Among 40 unrelated H. albicilla individuals from southern Sweden, the markers produced two to eight alleles per locus, and average observed and expected heterozygosities were 0.463 and 0.468, respectively. We further present five microsatellite markers that appeared monomorphic in H. albicilla, but which may be of interest for use in other raptor species.     

Development of eight polymorphic microsatellites for an Australasian rainforest tree species,Cryptocarya mackinnoniana (Lauraceae)

We developed eight microsatellite markers for Cryptocarya mackinnoniana to study the spatial genetic structure and ecological correlates of parentage in secondary rainforests in Australia's Wet Tropics. The microsatellite loci were screened in 99 trees, and 623 seedlings < 0.24 m tall, at a site extending from primary rainforest into adjacent 50‐year‐old secondary rainforest. The eight loci were polymorphic, exhibiting between three and 11 alleles, and gene diversity (H_E) from 0.25 to 0.84. For the trees, no departures from Hardy–Weinberg equilibrium were detected, except at locus Cm03, which had an estimated null allele frequency of 0.0903. No locus combinations exhibited linkage disequilibrium.     

Development and characterization of microsatellite loci for Rosa odorata var. gigantea Rehder & E. H. Wilson (Rosaceae)

Eighteen microsatellite markers were developed from Rosa odorata var. gigantea (Rosaceae), including 11 new microsatellite markers and 7 modified microsatellite loci having been developed from other Rosa species. About 27 wild individuals from 3 populations were used to screen polymorphism of these 18 microsatellite makers. The average allele number of these markers was 3.9 per locus, ranging from 2 to 9. The expected and observed heterozygosities varied from 0.2711 to 0.8043 and from 0.0370 to 0.5556, respectively. Cross-species amplification in other eight Rosa species showed their potential use for congeneric species. These microsatellite primers will be used for population genetics studies, constructing genetic linkage maps or locating quantitative trait locus (QTL) of R. odorata var. gigantea and related species.     

Characterization of microsatellite loci in the brown treecreeper (Climacteris picumnus) and cross‐species amplification in the white‐throated treecreeper (Cormobates leucophaeus)

Eight polymorphic microsatellite markers were developed for the brown treecreeper, Climacteris picumnus. The number of alleles ranged from three to 25 per locus with observed heterozygosities between 0.05 and 0.76. Seven of the eight primer pairs also amplified polymorphic microsatellite loci in the white‐throated treecreeper (Cormobates leucophaeus). These markers are likely to be useful for population genetic and parentage studies in any of the Australasian treecreepers (Climacteridae) and are the first genetic markers developed for any member of this passerine family.     

Microsatellite markers for Sachalin fir (Abies sachalinensis Masters)

We developed eight polymorphic microsatellite markers from the Sachalin fir (Abies sachalinensis Masters). The number of alleles per locus ranged from eight to 31, with an average of 21.4. The observed and expected heterozygosities ranged from 0.656 to 0.937 and from 0.710 to 0.945, respectively. These markers will be available for population genetic studies and parentage analysis.     

Development, characterization and transferability of microsatellite markers for Cullen australasicum (Leguminosae)

The development of eight polymorphic microsatellite markers for use in the Australian native legume Cullen australasicum is described. The number of alleles per locus ranged from 3 to 8. Observed heterozygosity ranged from 0.208 to 0.667. The cross-species amplification of these eight markers and a ninth marker, which is monomorphic in the populations examined, but may be used to distinguish between species, was also tested in five other species. These microsatellite markers will be useful for investigating the population structure of natural populations of C. australasicum and other Cullen species which may be susceptible to genetic contamination via pollen mediated gene flow from planted pastures of C. australasicum.     

Development of microsatellite markers in a mangrove tree species Aegiceras corniculatum (Myrsinaceae)

Aegiceras corniculatum is an ecologically important mangrove tree species. We isolated 18 polymorphic microsatellite loci from this species. These loci provided microsatellite markers with polymorphism of two to eight alleles per locus. The observed and expected heterozygosities ranged from 0.050 to 0.550, and from 0.097 to 0.736, respectively. These markers will contribute to research on the conservation, genetic diversity and mating patterns of A. corniculatum.     

Isolation and characterization of microsatellite markers in Elliot's pheasant (Syrmaticus ellioti)

We isolated eight microsatellite loci from genomic DNA in Elliot's pheasant Syrmaticus ellioti using streptavidin‐coated magnetic beads. In the analyses of 53 individuals sampled, these loci displayed polymorphism varying from six to nine alleles per locus and observed heterozygosities ranging from 0.174 to 0.430. The results suggested that these novel microsatellite markers could become useful molecular tools for genetic studies of S. ellioti.     

Novel polymorphic microsatellite DNA markers from Malaysian giant freshwater prawn, <Emphasis Type="Italic">Macrobrachium rosenbergii</Emphasis>

Seven single locus microsatellite markers were characterized in Malaysian giant freshwater prawn, Macrobrachium rosenbergii from an enriched genomic library Primer pairs were designed to flank the repeat sequences and the loci characterized for this species. The bands resulting from the PCR amplifications of these eight microsatellite loci were polymorphic with the number of alleles ranging from 8 to 26 alleles per locus, whereas the observed heterozygosity ranged from 0.0641 to 0.6564. These newly developed microsatellite markers should prove to be useful for population studies and in the management of genetic variations in broodstocks of freshwater prawn, M. rosenbergii.     

Microsatellite markers for the earthworm Lumbricus rubellus

We describe eight microsatellite markers for the earthworm Lumbricus rubellus, a commonly used ‘biological sentinel’ species in ecotoxicology studies. Thirty‐four individuals from a single population were tested for polymorphism. Markers possessed between two and 15 alleles per locus, and expected heterozygosity ranged between 0.06 and 0.92 (observed heterozygosity 0.03–0.92). Unfortunately, no consistent cross‐species amplification was observed in the related congeners Lumbricus castaneus and Lumbricus terrestris.     

Isolation of polymorphic tetranucleotide microsatellite markers for the masked flowerpiercer Diglossa cyanea

We have isolated 11 polymorphic microsatellite markers for the masked flowerpiercer Diglossa cyanea from a genomic library enriched for (AAGG)_n repetitive elements. The number of alleles ranged from eight to 16 per locus with the observed heterozygosity ranging from 0.64 to 1.00. These markers will be useful for analysis of questions concerning population genetic structure and models of speciation.     

Development and characterization of polymorphic microsatellite DNA markers for Sasa senanensis (Poaceae: Bambuseae)

Sasa senanensis is a dwarf bamboo species distributed on the floors of cool temperate forests in Japan and adjacent regions. We isolated eight polymorphic microsatellite loci from this species. The number of alleles ranged from two to eight and the observed heterozygosity per locus from 0.13 to 0.74. Seven of the eight loci were also polymorphic in Sasa nipponica. Most of these markers were successfully amplified in other dwarf bamboo species. These markers will be useful for investigating clonal structure and population genetics in some dwarf bamboo species.     

Eight polymorphic microsatellite loci for the Chinese medicinal plant Artemisia annua L. (Asteraceae)

Artemisia annua is an important medicinal plant from which Artemisinin was extracted to cure malaria effectively. We developed eight microsatellite markers from the genome of A. annua using the FIASCO protocol. Polymorphism of each locus was assessed in 54 individuals from two Chinese populations. The average allele number of these microsatellites was 3.1 per locus, ranging from 2 to 6. The observed (H _O) and expected (H _E) heterozygosity were from 0.019 to 0.907 and from 0.055 to 0.793, respectively. These microsatellite markers would provide a useful tool for genetic studies of A. annua. H.-R. Huang and G. Zhou have contributed equally to this work.     

Isolation of polymorphic tetranucleotide microsatellite markers for the wedge‐billed woodcreeper Glyphorynchus spirurus

We have isolated six polymorphic microsatellite markers for the wedge‐billed woodcreeper Glyphorynchus spirurus from a genomic library enriched for (AAGG)_n repetitive elements, and characterized them in 25 individuals. The number of alleles ranged from eight to 20 per locus, with observed heterozygosity ranging from 0.43 to 0.78.     

Isolation of polymorphic tetranucleotide microsatellite markers for the skink Mabuya affinis

We isolated 10 polymorphic microsatellite markers for the skink Mabuya affinis from genomic libraries enriched for (AAGG)_n and (AAAG)_n repetitive elements. The number of alleles ranged from eight to 13 per locus with the observed heterozygosity ranging from 0.60 to 0.92. These markers will be useful for analysis of questions concerning population genetic structure and models of speciation.     

Microsatellite markers for the polygamous termite Nasutitermes corniger (Isoptera: Termitidae)

We developed eight highly variable microsatellite markers for the termite Nasutitermes corniger. Allele number per locus ranged from nine to 34, and expected heterozygosity from 0.45 to 0.94, in samples from seven sites in the former canal zone of Panama. The utility of these markers was assessed for five congeners varying in phylogenetic distance to N. corniger. The markers will be useful for fine‐scale examination of population and colony genetic structure in N. corniger and other closely related species.     

Isolation and characterization of microsatellite markers for Viola mirabilis (Violaceae) using a next‐generation sequencing platform

We isolated and characterized microsatellite loci in Viola mirabilis (Violaceae), an endangered species from South Korea. Twenty‐three polymorphic microsatellite loci were developed and tested in Korean, Chinese and Japanese populations. The number of alleles per locus varied from two to eight. The observed and expected heterozygosities within the three populations were 0.000–0.625 and 0.469–0.695, respectively. A total of six loci in the Korean population, one locus in the Chinese population and seven loci in the Japanese population deviated from Hardy–Weinberg equilibrium. We expect that these newly developed microsatellite markers will contribute to understanding the phylogeography and population genetics of V. mirabilis, which will aid in developing conservation strategies for this species.     

The development of eight microsatellite loci in the wild daffodil Narcissus triandrus (Amaryllidaceae)

We report microsatellite primer pairs for the wild tristylous daffodil, Narcissus triandrus (Amaryllidaceae). From enriched libraries, we identified 58 unique microsatellite loci. We designed primer pairs for 27 of these loci and screened genomic DNA from 38 to 40 adults from a single population. For eight polymorphic loci, the number of alleles per locus ranged from five to 17. As six primers also amplified loci in three other Narcissus species, including two horticultural varieties, we expect that some of these markers will be transferable to other Narcissus species.