Microsatellites from the vairone Leuciscus souffia (Pisces: Cyprinidae) and their application to closely related species

Eleven microsatellites were isolated from the vairone Leuciscus souffia (Risso 1826), an endangered fish that inhabits river systems in and around the Alps in Europe. The level of genetic diversity was assessed in 29 individuals of the subspecies L. s. souffia, and their variability was further estimated in seven individuals of a different subspecies, L. s. muticellus. Eight of these microsatellite loci were also applied to seven closely related cyprinid species. Availability of the reported microsatellite loci will facilitate the investigation of population genetic structure of these species with applications for the development of conservation strategies and phylogeographical approaches.     

Isolation and characterization of microsatellite loci in Lesquerella fendleri (Brassicaceae) and cross‐species amplification

Fifteen novel microsatellite primer pairs are presented for Lesquerella fendleri, which were developed from seven dinucleotide, five trinucleotide and three tetranucleotide microsatellite DNA loci. These loci were characterized for 40 individuals from 24 localities throughout the species range. The number of alleles observed per locus ranged from three to 16, the observed heterozygosity ranged from 0.175 to 0.750, and the polymorphic information content ranged from 0.218 to 0.889. Cross‐species transferability tested on nine species of Lesquerella and one species of the related genus Physaria indicates that these primer pairs may be useful for population genetic studies of other species in Lesquerella and possibly other closely related genera.     

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.     

Twelve microsatellites for two deep sea polychaete tubeworm species,Lamellibrachia luymesi and Seepiophila jonesi,from the Gulf of Mexico

The vestimentiferan tubeworms Lamellibrachia luymesi and Seepiophila jonesi are found at hydrocarbon seeps in the Gulf of Mexico. Primers for polymorphic microsatellite loci were developed from genomic libraries of L. luymesi (five loci) and from S. jonesi tissue (eight loci) and were used to screen individuals collected from nine northern Gulf of Mexico hydrocarbon seep sites. Loci had from four to more than 50 alleles with high expected levels of heterozygosity. Cross‐species amplification, tested on seven vestimentiferan species including both hydrothermal vent and cold seep species, was generally strong in similar species but weak in more genetically distant species.     

Isolation and characterization of the first microsatellite loci from the order Psocoptera in the economically important pest insect Liposcelis decolor (Pearman) and cross‐species amplification

A total of 11 microsatellite loci from the invasive insect pest Liposcelis decolor were isolated and characterized of which six loci were polymorphic. A population survey involving a total of 30–192 individuals per locus from five populations revealed a range of four to seven alleles per locus and moderate observed heterozygosities (0.183–0.565), highlighting the utility of these loci in further population genetic studies. Cross‐species amplifications were successful for two to 11 loci in five other Liposcelis species also of international economic importance.     

Microsatellite loci for the southern pine beetle (Dendroctonus frontalis) and cross‐species amplification in Dendroctonus

We developed microsatellite loci for the southern pine beetle (Dendroctonus frontalis). Twelve microsatellite loci were identified. Eight loci were polymorphic and sufficiently variable in 62 individuals (expected heterozygosity ranged from 0.707 to 0.880) to investigate population structure. All loci conformed to HWE except Dfr‐14, which showed heterozygote excess, and no two loci deviated from linkage equilibrium. The loci were tested for cross‐species amplification in four species of Dendroctonus (D. valens, D. terebrans, D. brevicomis, and D. ponderosae). Seven loci were polymorphic in at least one of the species tested.     

Validation of microsatellite multiplexes for parentage analysis and species discrimination in two hybridizing species of coral reef fish (Plectropomus spp., Serranidae)

Microsatellites are often considered ideal markers to investigate ecological processes in animal populations. They are regularly used as genetic barcodes to identify species, individuals, and infer familial relationships. However, such applications are highly sensitive the number and diversity of microsatellite markers, which are also prone to error. Here, we propose a novel framework to assess the suitability of microsatellite datasets for parentage analysis and species discrimination in two closely related species of coral reef fish, Plectropomus leopardus and P. maculatus (Serranidae). Coral trout are important fisheries species throughout the Indo‐Pacific region and have been shown to hybridize in parts of the Great Barrier Reef, Australia. We first describe the development of 25 microsatellite loci and their integration to three multiplex PCRs that co‐amplify in both species. Using simulations, we demonstrate that the complete suite of markers provides appropriate power to discriminate between species, detect hybrid individuals, and resolve parent–offspring relationships in natural populations, with over 99.6% accuracy in parent–offspring assignments. The markers were also tested on seven additional species within the Plectropomus genus with polymorphism in 28–96% of loci. The multiplex PCRs developed here provide a reliable and cost‐effective strategy to investigate evolutionary and ecological dynamics and will be broadly applicable in studies of wild populations and aquaculture brood stocks for these closely related fish species.     

Isolation and characterization of microsatellite loci in Lampsilis abrupta (Bivalvia: Unionidae) and cross‐species amplification within the genus

We have documented the first microsatellites isolated from a unionid and demonstrated that these markers can be useful for surveys of neutral genetic variation in several Lampsilis species. We describe the isolation and characterization of 15 polymorphic microsatellite DNA loci for the endangered unionid Lampsilis abrupta. Among individuals from five collections, allelic diversity ranged from six to 17 alleles and averaged 10.4 alleles per locus. Individual heterozygosity was observed to range from 20.0% to 86.7% and averaged 46.9%. Cross‐species amplification was investigated in nine additional Lampsilis species. A high level of flanking sequence similarity was inferred as 13 of 15 loci amplified in at least seven species.     

Microsatellite loci in the Propertius duskywing,Erynnis propertius (Lepidoptera: Hesperiidae), and related species

Fifteen polymorphic dinucleotide microsatellite loci were characterized for Erynnis propertius using an enrichment protocol. The number of alleles varied from nine to 28 for a sample of 24 individuals. Observed heterozygosities ranged from 0.25 to 0.96. Homozygote excess was detected for 10 loci. Twelve markers successfully amplified in related Erynnis species and eight loci were polymorphic in at least one other species.     

Isolation and characterization of highly polymorphic microsatellite markers from the marine invasive species Crepidula fornicata (Gastropoda: Calyptraeidae)

Many marine organisms have pelagic larvae and these are often important agents of dispersal and establishment of exotic species. The larval phase and the hermaphroditic reproductive system of the invasive marine gastropod Crepidula fornicata are crucial in determining the success of this species along the European coasts. To analyse these factors, we developed seven microsatellite loci specific to C. fornicata. On the same set of individuals, the microsatellite loci exhibited a greater level of polymorphism than previously studied allozyme markers and, thus, will permit fine‐scale analysis and larval pool studies.     

Development of microsatellite markers for Polemonium kiushianum (Polemoniaceae), a critically endangered grassland plant species in Japan

Eight microsatellite loci were isolated and characterized for Polemonium kiushianum (Polemoniaceae), an endangered perennial herb endemic to the Kyushu region in Japan. The number of individuals of this species is presumed to be about 400 individuals. The loci showed polymorphisms with two to ten alleles per locus. The expected and observed heterozygositis ranged from 0.039 to 0.744 and 0.039 to 0.784, respectively. Although significant deviations (P < 0.05) from Hardy–Weinberg equilibrium were detected for one locus, no significant deviations were detected for other seven loci. There was no evidence of significant linkage disequilibrium between any two of the loci. The markers described here will be useful to establish conservation programs based on genetic traits of P. kiushianum.     

Isolation of microsatellite loci and cross‐species amplifications in three gobiid fish of the genus Pomatoschistus

Gobiids of the genus Pomatoschistus are increasingly being investigated as models for adaptation to coastal environments and for mating system studies. Among the dozen currently analysed species, microsatellite primers have been characterized only for Pomatoschistus minutus. This paper describes seven new polymorphic loci isolated from Pomatoschistus marmoratus and Pomatoschistus microps, two species that hybridize. Cross‐species amplification was tested for these new loci, together with seven already published P. minutus loci. Systematic amplification of samples of each of the three species provided a first indication of their polymorphism.     

Characterization of microsatellite loci isolated in trumpeter swan (Cygnus buccinator)

Primers for 16 microsatellite loci were developed for the trumpeter swan (Cygnus buccinator), a species recovering from a recent population bottleneck. In a screen of 158 individuals, the 16 loci were found to have levels of variability ranging from two to seven alleles. No loci were found to be linked, although two loci repeatedly revealed significant departures from Hardy–Weinberg equilibrium. Amplification in the closely related tundra swan (Cygnus columbianus) was successful for all except one locus. These microsatellite loci will be applicable for population genetic analyses and ultimately aid in management efforts.     

Cross‐species comparison of microsatellite loci in the Culex pipiens complex and beyond

In the past, we have developed microsatellite loci from the two most common members of the Culex pipiens complex, Culex quinquefasciatus and Culex pipiens. Here we describe seven additional loci and present an extensive survey of a panel of 20 loci across most of the species and subspecies in the complex as well as in morphologically related species. Because we observed a high degree of polymorphism in the flanking regions, we designed new primers and surveyed multiple populations. We present alternate primers and discuss the cross‐species usefulness of these Culex microsatellite loci in a phylogenetic context.     

Microsatellites for use in Nothofagus cunninghamii (Nothofagaceae) and related species

Nothofagus cunninghamii (myrtle) has a widespread distribution through the temperate rainforest of southeastern Australia with some disjunct populations existing in putative glacial refugia. Polymorphic nuclear markers are required to resolve the biogeographical history of the species and will also be useful for conservation and forestry applications in this and other species of Nothofagus. Fourteen polymorphic microsatellite loci were isolated from N. cunninghamii, with four to 10 alleles amplified in 15 individuals tested. Transferability to other species of Nothofagus was successful, with six loci transferring to all eight other species tested.     

Microsatellite loci in Bactris gasipaes (Arecaceae): their isolation and characterization

We constructed a microsatellite‐enriched genomic library for Bactris gasipaes, an economically important, domesticated palm. We developed 18 polymorphic microsatellite markers, and found an average of seven alleles per locus in a sample of 14 individuals selected from a germplasm bank. Cross‐species amplification was evaluated in six other Bactris species. The loci detected will permit population studies and germplasm characterization, and can be used for genetic analyses in related species.     

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.     

Cloning and characterization of novel microsatellite DNA markers for the grass rockfish,Sebastes rastrelliger,and cross‐species amplification in 10 related Sebastes spp

Here we report development and characterization of seven polymorphic loci derived from grass rockfish (Sebastes rastrelliger) genomic DNA phagemid libraries enriched for microsatellite motifs. Within grass rockfish, average allelic diversity was 11.3 alleles per locus and average heterozygosity was 0.73. The seven loci also were surveyed in 10 related species of Sebastes, where allelic diversity ranged from highly polymorphic to monomorphic. Deviations from Hardy–Weinberg were nonsignificant in all but one species/locus combination suggesting low occurrence of null alleles. Significant linkage disequilibrium was detected among three loci, but these events were limited to a single species in each case.     

Development of novel microsatellite markers for the grassland species Lolium multiflorum,Lolium perenne and Festuca pratensis

Twelve microsatellite markers were isolated from Lolium multiflorum. Allelic variability and cross‐species amplification were assessed on 16 individuals of each of the three grassland species L. multiflorum, Lolium perenne and Festuca pratensis. Cross‐species amplification success was 100% for L. perenne and 83% for F. pratensis. The number of alleles detected ranged from one to 14 with an average of 3.4. While three microsatellite loci were polymorphic in all three species, one marker produced species‐specific alleles in all three species. These microsatellite markers provide a valuable tool for population genetic studies within and among species of the Festuca–Lolium complex.     

Isolation of microsatellite loci from spotted gum (Corymbia variegata), and cross‐species amplification in Corymbia and Eucalyptus

Corymbia variegata (spotted gum) is an important commercial hardwood timber species in Australia. Fourteen polymorphic microsatellite loci were isolated from C. variegata, with 3–5 alleles amplified in three individuals examined. Cross‐species amplification in Corymbia was successful for all primer pairs, while 10 loci (71%) were successfully transferred to at least one species in the closely related genus Eucalyptus.