Microsatellite markers for <Emphasis Type="Italic">Juglans cinerea</Emphasis> L. and their utility in other Juglandaceae species

Ten polymorphic microsatellite markers were found to amplify in butternut (Juglans cinerea; Juglandaceae). These microsatellite loci were found to amplify across most of nine other species and five hybrids examined. Loci were highly polymorphic, with 18 to 32 alleles per locus across species. These nuclear microsatellite markers will be useful in examining genetic diversity within and among populations of butternut, and in distinguishing butternut from interspecific hybrids.     

Microsatellite primers for Australian recreationally and commercially important estuarine fishes

Eight primer sets that amplify microsatellite DNA loci were developed for the estuarine fishes, dusky flathead Platycephalus fuscus and sand whiting Sillago ciliata, which are targets of recreational and commercial fishers on the east coast of Australia. Surveys across these loci revealed remarkably similar levels of genetic diversity within samples of c. 22 P. fuscus and S. ciliata from each of the two estuaries. These loci will be useful in determining the genetic stock structure of these two species.     

Isolation and characterization of microsatellite markers from Fangzheng silver crucian carp,Carassius auratus gibelio (Bloch), and cross‐amplification in the closely related species crucian carp,Carassius auratus auratus (Linnaeus)

We report the development of 20 microsatellite markers for Fangzheng silver crucian carp, Carassius auratus gibelio (Bloch). Nineteen out of 20 showed polymorphism with alleles ranging from two to 14. These loci were screened to amplify the closely related species crucian carp, Carassius auratus auratus (Linnaeus) and all of them can amplify DNA with the size similar to the former. The origin of silver crucian carp is in issue and the population genetic structure is still unclear. Microsatellite markers isolated from the silver crucian carp and their utility in the crucian carp will be useful for these researches.     

Microsatellite loci for great white herons and great blue herons (Aves,Ardeidae, Ardea herodias)

We used an enrichment technique to isolate 60 microsatellite loci in Ardea herodias. We developed primers for 17 loci, screened for variation in A. herodias and attempted to amplify these loci in three closely related species (A. alba, A. cinerea and A. cocoi). Fifteen loci were polymorphic in A. herodias. Observed heterozygosities ranged from 0.10 to 0.81. Two loci appeared to be monomorphic in A. herodias, but exhibited variation in product size among species within the genus. Our ability to amplify polymorphic products in closely related species suggests that these markers may be useful in other herons.     

Nine polymorphic microsatellite markers in Centaurea stoebe L. [subspecies C. s. stoebe and C. s. micranthos (S. G. Gmelin ex Gugler) Hayek] and C. diffusa Lam. (Asteraceae)

Centaurea stoebe (subspecies C. s. stoebe and C. s. micranthos[S. G. Gmelin ex Gugler) Hayek] and Centaurea diffusa are Eurasian plant species that have invaded much of North America. We isolated seven microsatellite loci from C. stoebe and two loci from C. diffusa. All loci described here amplify in both species and have between six and 25 alleles each. These markers will be useful in examining population structure and addressing questions regarding these invasions.     

Isolation of polymorphic microsatellite markers from <Emphasis Type="Italic">Przewalskia tangutica</Emphasis> (Solanaceae)

Microsatellite-containing regions were isolated and characterized in Przewalskia tangutica Maxim (Solanaceae), an endemic and endangered species to the Qinghai-Tibetan Plateau of China. An enrichment protocol yielded 200 positive clones. We designed primers to amplify 29 unique microsatellites, 12 of which amplified cleanly and were polymorphic. A survey of 17 individuals showed that these loci are highly variable with the number of alleles ranging from 3 to 12, and expected heterozygosity ranged from 0.2929 to 0.4947. Those markers will be useful for studies of population structure and intraspecific variation in P. tangutica.     

Novel polymorphic microsatellites for the red‐legged partridge (Alectoris rufa) and cross‐species amplification in Alectoris graeca

The red‐legged partridge, Alectoris rufa, is an endemic species of the southwestern Mediterranean, and the most popular game bird in the Iberian Peninsula. A total of 27 microsatellite loci was isolated from an enriched genomic library of A. rufa. Six perfect GT microsatellites were characterized and optimized in 45 individuals of A. rufa. All loci revealed high levels of polymorphism with a number of alleles that ranged from three to 13. Observed heterozygosity ranged from 0.2 to 0.6. Cross‐species amplification showed that all loci were also polymorphic in rock partridge, Alectoris graeca. The new markers will be useful in determining hybridization between both species of Alectoris.     

Genome‐assisted development of nuclear intergenic sequence markers for entomopathogenic fungi of the Metarhizium anisopliae species complex

Entomopathogenic fungi in the genus Metarhizium are useful for biological control programmes against economically important arthropod pests worldwide. However, understanding the true diversity and ecology of these organisms is hampered by convergent morphologies between species. The application of molecular techniques has enabled greater resolution of species than allowed by morphology alone. In particular, the commonly used biocontrol agent M. anisopliae was found to be a species complex composed of nine species. This prior work was conducted with commonly used markers in fungal phylogenetics (BTUB, RPB1, RPB2 and TEF), which likely under‐represent diversity in the M. anisopliae complex. Using sequence data from nuclear genomes of M. acridum and M. robertsii we identified regions of conserved gene synteny and developed primers to amplify intergenic regions of seven loci. Using ex‐type and authenticated tissue specimens for species in the M. anisopliae complex, we demonstrate that sequence data derived from intergenic loci is more variable and phylogenetically informative than previously available markers. These new markers will facilitate investigations at or below the species level for the M. anisopliae complex. The method of marker development employed here should be extendable to any group with sufficiently divergent genome data available.     

Polymorphic microsatellite loci from the American goldfinch (Carduelis tristis) and their cross‐amplification in a variety of passerine species

Cardueline finches have become important models in studies of sexual selection. Here I describe eight polymorphic microsatellite loci isolated from the American goldfinch (Carduelis tristis) that are useful in parentage and heterozygosity studies. The number of alleles per locus ranged from eight to 38 based on 252 goldfinches. The markers are polymorphic in at least four other Cardueline finch species, and several amplify in nonfinch passerines including red‐winged blackbird (Agelaius phoeniceus), black‐throated blue warbler (Dendroica caerulescens) and tufted titmouse (Baeolophus bicolor), indicating they should be useful in paternity and population studies of a variety of species.     

New microsatellite primers for Daphnia galeata mendotae

In order to study genetic changes in populations of Daphnia galeata mendotae, I characterized seven polymorphic microsatellite loci. Primers to amplify these loci were tested on individual eggs from the resting egg bank of Onondaga Lake, NY. Levels of polymorphism and cross‐amplification in D. g. galeata indicate that they will be useful markers for ecological genetic studies on both adults and diapausing eggs of these species.     

Isolation and characterization of microsatellite markers in Phytophthora ramorum,the causal agent of sudden oak death

We describe specific primers and conditions to amplify two dinucleotide and five trinucleotide microsatellite DNA loci isolated from the oomycete Phytophthora ramorum, the causal agent of sudden oak death. The primer sets were tested on 14–30 isolates from North America and Europe. Seven of 14 loci differentiated between A1 and A2 mating types. All seven loci successfully amplified DNA isolated from infected plant tissue. Four loci may be useful for the diagnosis of P. ramorum because they do not amplify closely related Phytophthora species.     

Characterization of six polymorphic microsatellites for the polychaete tubeworm Hydroides elegans and cross‐species amplification in the congener Hydroides hexagonus

We isolated and characterized six polymorphic microsatellite loci for the polychaete tubeworm, Hydroides elegans. Two additional loci were not reliably scorable and estimates of heterozygosity were obtained for the other six. In addition, cross‐species amplification was successful for two loci using the congener H. hexagonus. Given that few microsatellite loci are available for polychaetes, these markers will be useful in assessing dispersal and gene flow in H. elegans and probably also other polychaetes.     

Isolation and characterization of microsatellite loci in Penstemon rostriflorus (Plantaginaceae) and cross‐species amplification

Eight novel microsatellite primer pairs are presented for Penstemon rostriflorus, representing the first microsatellite markers available for this genus. Loci were characterized for 20 individuals from two populations in the Great Basin, USA. All loci are polymorphic within P. rostriflorus (seven to 13 alleles per locus; observed heterozygosity between 0.40 and 0.95), and therefore useful for population genetic studies within the species. Cross‐species transferability was tested on 40 additional species of Penstemon, and results indicate that these primers pairs will likely be useful for population genetic studies on many Penstemon species.     

Characterization of microsatellite markers in the shea tree (Vitellaria paradoxa C. F Gaertn) in Mali

In order to study the genetic diversity and structure in the population of Vitellaria paradoxa, we characterized eight polymorphic microsatellite loci. Primers to amplify these loci were tested on 169 individual trees representing a sample of the population of shea tree in Mali. The loci were all polymorphic with a number of alleles between three to nine and with observed level of heterozygosity ranging from 0.035 to 0.507. These markers will be useful for genetic and ecological studies of this species.     

Isolation and characterization of novel microsatellite loci for the short‐beaked common dolphin (Delphinus delphis) and cross‐amplification in other cetacean species

Seven tetranucleotide and three dinucleotide microsatellite loci were isolated from the short‐beaked common dolphin (Delphinus delphis). Number of alleles and observed heterozygosity ranged from three to twelve (mean 8.4) and from 0.087 to 0.978 (mean 0.741), respectively. All loci were tested in four other odontocete species and showed polymorphisms in most locus–species combinations. This panel of nuclear markers will be useful for the investigation of population ecology and social interactions in common dolphins and other cetacean species.     

Novel microsatellite loci for the study of the black‐capped vireo (Vireo atricapillus)

We identified 14 novel polymorphic microsatellite loci in the black‐capped vireo (Vireo atricapillus). We also attempted to amplify and genotype these loci in other Vireo species, including the white‐eyed vireo (Vireo griseus), red‐eyed vireo (Vireo olivaceus), and blue‐headed vireo (Vireo solitarius). In 33 genotyped black‐capped vireos from two locations, total alleles ranged from six to 20, with observed heterozygosity ranging from 0.58 to 0.91 and expected heterozygosity from 0.65 to 0.93. Two loci had detectable levels of null alleles. Many of the loci were able to be amplified in the related Vireo species.     

A set of polymorphic microsatellite loci for Vriesea gigantea and Alcantarea imperialis (Bromeliaceae) and cross‐amplification in other bromeliad species

Fifteen polymorphic microsatellite markers were isolated and characterized in two species of Bromeliaceae: Vriesea gigantea and Alcantarea imperialis. The number of alleles observed for each locus ranged from three to 16. The loci will be used for studies of the genetic structure of natural populations, reproductive biology, and evolutionary relationships among and within these genera. A cross‐amplification test in 22 taxa suggests that the markers will be useful for similar applications in numerous other bromeliad species.     

Isolation and characterization of microsatellite loci in the Pacific pleasure oyster,Crassostrea corteziensis,and their cross‐species amplification in four other oyster species

Microsatellite loci were isolated in Crassostrea corteziensis using (GT)_n, (CT)_n and (CTGT)_n‐enriched genomic libraries. Within each of 45 sequenced clones, an average of three microsatellite regions (156 total) were observed. Thirty‐three primers were designed, from which 11 microsatellite loci amplified. Ten of those were polymorphic, with a range of two to 30 alleles. Three loci were not in Hardy–Weinberg equilibrium, and linkage disequilibrium was found for six pairs of loci. These microsatellite loci will be further tested for segregation distortions and null alleles to establish a set for population genetic studies of the species in the Northwest coasts of Mexico, and for optimization of aquaculture development. Seven of the microsatellite loci cross‐amplified in Crassostrea palmula, a sympatric species, and will be useful in further genetic studies.     

Isolation and characterization of microsatellites in Leuciscus cephalus (Cypriniformes,Cyprinidae) and cross‐species amplification within the family Cyprinidae

Thirteen polymorphic microsatellites were isolated from Leuciscus cephalus, a widespread cyprinid species with great ecological tolerance. Together with the cross‐species amplification of six additional loci originally published for three cyprinid fish species, we optimized a multiplex panel for L. cephalus allowing the genotyping of 19 polymorphic loci. Number of alleles and heterozygosity per locus in a sample of 20 fish individuals ranged from two to 16 and from 0.05 to 0.90, respectively. These primers will be useful in determining the population structure of L. cephalus. In addition, successful cross‐amplification was obtained for several species of Cyprinidae.