Isolation of microsatellites from two species of scleractinian coral

Microsatellites were isolated from two broadcast‐spawning species of scleractinian coral (Platygyra daedalea and Goniastrea favulus) from Australia's Great Barrier Reef. We found 27 microsatellites across both species, although only five loci were polymorphic in each species. Microsatellite loci displayed a wide range of diversity levels with four to 11 alleles per locus (H_O = 0.26–0.91) in P. daedalea and two to seven alleles per locus (H_O = 0.16–0.96) in G. favulus. Most loci showed departures from Hardy–Weinberg equilibrium which may reflect nonrandom mating but may also be related to difficulties associated with coral DNA.     

Isolation and characterization of microsatellite markers in the southern emu‐wren (Stipiturus malachurus: Aves)

We isolated and characterized eight novel microsatellite loci in the southern emu‐wren (Stipiturus malachurus). We used nonradioactive polymerase chain reaction (PCR)‐based techniques to screen an enriched genomic DNA library. Based on genotypes from a single population, six loci showed no evidence of null alleles and were polymorphic (allele range = 2–9, mean heterozygosity = 0.57), and one locus was sex‐linked (N_A = 4). These loci were variable and had different allele size ranges in three other populations of southern emu‐wrens, and are therefore useful for determining levels of genetic diversity within and between populations of the species.     

Isolation and characterization of polymorphic microsatellite loci for the study of paternity and population structure in the little penguin Eudyptula minor

We isolated and characterized eight novel microsatellite loci in the little penguin Eudyptula minor, using nonradioactive polymerase chain reaction‐based techniques to screen GA and GAAA repeats from enriched genomic DNA libraries. All eight loci were polymorphic and seven were variable in our main study population (mean H_E = 0.613, mean N_A = 7.14). Cross‐amplification using a microsatellite primer developed in Spheniscus demersus (African penguin) yielded one additional polymorphic locus. This locus combined with six of the little penguin loci is suitable for paternity assignment in little penguins (exclusion probability for seven unlinked loci = 0.993).     

Isolation and characterization of variable microsatellite loci in Lilium philadelphicum (Liliaceae)

Here we describe the characterization of six polymorphic microsatellite loci for Lilium philadelphicum (Liliaceae). Polymorphism levels ranged from 7 to 30 alleles per locus with a mean number of 15 alleles per locus. Observed heterozygosity (H_O) ranged from 0.17 to 0.82 and did not deviate from Hardy–Weinberg expectations in the three western Montana populations included in the analysis. These loci are proving useful in studying gene flow between populations of this species distributed across its range in North America.     

Characterization of 14 tetranucleotide microsatellite loci derived from Pacific herring

Spawning aggregations of Pacific herring (Clupea pallasi) often exhibit significant interannual variation in allele frequencies of neutral gene markers. We isolated 14 tetranucleotide microsatellites to examine hypothetical processes that may produce this unique genetic signal. We developed and tested primer pairs for each locus and then estimated locus variability in samples (n = 60) from two populations. The number of alleles per locus ranged from five to 49. The expected heterozygosity across loci and populations ranged from 0.20 to 0.96. These microsatellites will be useful for estimating genetic variation in herring on a fine geographical scale.     

A hot topic: the genetics of adaptation to geothermal vents in Mimulus guttatus

Identifying the individual loci and mutations that underlie adaptation to extreme environments has long been a goal of evolutionary biology. However, finding the genes that underlie adaptive traits is difficult for several reasons. First, because many traits and genes evolve simultaneously as populations diverge, it is difficult to disentangle adaptation from neutral demographic processes. Second, finding the individual loci involved in any trait is challenging given the respective limitations of quantitative and population genetic methods. In this issue of Molecular Ecology, Hendrick et al. (2016) overcome these difficulties and determine the genetic basis of microgeographic adaptation between geothermal vent and nonthermal populations of Mimulus guttatus in Yellowstone National Park. The authors accomplish this by combining population and quantitative genetic techniques, a powerful, but labour‐intensive, strategy for identifying individual causative adaptive loci that few studies have used (Stinchcombe & Hoekstra 2008 ). In a previous common garden experiment (Lekberg et al. 2012), thermal M. guttatus populations were found to differ from their closely related nonthermal neighbours in various adaptive phenotypes including trichome density. Hendrick et al. (2016) combine quantitative trait loci (QTL) mapping, population genomic scans for selection and admixture mapping to identify a single genetic locus underlying differences in trichome density between thermal and nonthermal M. guttatus. The candidate gene, R2R3 MYB, is homologous to genes involved in trichome development across flowering plants. The major trichome QTL, Tr14, is also involved in trichome density differences in an independent M. guttatus population comparison (Holeski et al. 2010) making this an example of parallel genetic evolution.     

Identification and characterization of eight microsatellite loci in Aster amellus L. (Asteraceae)

Eight polymorphic microsatellites were developed in the perennial herbaceous Aster amellus L. (Asteraceae) and characterized on three populations from France and Switzerland. The number of alleles ranged between four and 30 depending on the locus, and the mean number of effective alleles was 5.8. The average gene diversity equalled 0.744 (range: 0.419–0.957) and the overall differentiation was found significant (θ = 0.092, P < 0.01). Three loci displayed significant heterozygote deficiencies, which might indicate the presence of null alleles. Amplifications were detected on eight loci in Aster alpinus L.     

Development and characterization of six new microsatellite markers for the silver‐ or gold‐lipped pearl oyster,Pinctada maxima (Pteriidae)

Six di‐, tri‐ and tetranucleotide microsatellite loci were developed for the silver‐ or gold‐lipped pearl oyster Pinctada maxima using a linker‐ligated, magnetic bead enrichment protocol. Based on a minimum of 134 Indonesian pearl oyster samples, number of alleles and observed heterozygosity at each locus ranged from six to 17 alleles and from 0.172 to 0.813 (mean = 0.448), respectively. Mean polymorphic information content for the six loci was 0.562. These loci should be very useful in DNA parentage analyses and population differentiation of P. maxima in Australia and Indonesia.     

Polymorphic microsatellite markers isolated from partially enriched genomic library of Chitala chitala

A total eight polymorphic microsatellite loci were obtained from genomic library of Indian feather back, Chitala chitala (order Osteoglossiformes, family Notopteridae) and the 46 samples were analysed to determine genetic variation. The mean number of alleles per locus ranged from 4.50 to 5.25, and expected heterozygosity ranged from 0.124 to 0.852. Deviation from Hardy–Weinberg equilibrium expectations (P < 0.002) was observed at loci Cch2, Cch9 (Bhaghirathi) and Cch9 (Brahmaputra). The identified microsatellite loci were found promising for population genetics studies of C. chitala and related species Notopterus notopterus (family Notopteridae).     

Isolation and characterization of microsatellite DNA loci in the toad‐headed lizards,Phrynocephalus przewalskii complex

To assess mechanisms of hybridization and speciation, we isolated and characterized 12 dinucleotide microsatellite DNA loci for the toad‐headed lizards, Phrynocephalus przewalskii complex. A total of 48 specimens were examined and all loci were polymorphic with seven to 25 alleles per locus. The observed and expected heterozygosities ranged from 0.282 to 0.946 and 0.400 to 0.937, respectively. These loci are therefore suitable for a wide range of population level studies within the P. przewalskii complex.     

Development and characterization of polymorphic microsatellite DNA loci for the endangered dusky gopher frog,Rana sevosa,and two closely related species,Rana capito and Rana areolata

A microsatellite library was developed using genomic DNA of the endangered dusky gopher frog, Rana sevosa. Polymerase chain reaction (PCR) primers and conditions are presented for R. sevosa (eight loci) and two sister taxa — other gopher frogs, Rana capito (seven loci) and crawfish frogs, Rana areolata (three loci). Polymorphism of each microsatellite locus was evaluated for each species. All loci have moderate to high genetic variation in terms of allelic richness (four to 10 alleles per locus), observed heterozygosity (0.595–0.946), and expected heterozygosity (0.531–0.856).     

Microsatellite loci for the critically endangered Santa Cruz long-toed salamander (Ambystoma macrodactylum croceum) and other Ambystoma taxa

The imperiled Santa Cruz long-toed salamander (Ambystoma macrodactylum croceum) is a geographically isolated lineage of the long-toed salamander species complex. Recovery of this taxon requires understanding its population genetic structure, which is necessary to identify critical habitat. To assess the population structure of A. m. croceum, I developed primers for 17 microsatellite loci and evaluated their performance. Polymorphism ranged from 3 to 10 alleles per locus, with an average of 6.3 per locus. Mean observed and expected heterozygosity were similar (H _O = 0.678; H _E = 0.708), with one locus demonstrating heterozygote deficiency. The performance of these and previously identified microsatellite loci were also characterized for two other long-toed salamander lineages, as well as two other species of Ambystoma. The utility of these markers has similar and overlapping applications for conservation and population genetics studies of other Ambystoma taxa, and will provide a basis for revising the recovery plan for A. m. croceum, particularly for designating corridors among interacting breeding populations and upland areas, and identifying areas where gene flow should be restored.     

Cross‐species amplification of microsatellite loci in Aquilegia and Semiaquilegia (Ranunculaceae)

We developed 16 microsatellite loci from an F₂ hybrid between Aquilegia formosa and Aquilegia pubescens. In samples of 28 individuals, we found an average of 14 alleles per locus from each parental species. We tested these loci for cross‐amplification in 10 additional species of Aquilegia and found that all 16 loci amplified in other North American species and 12 consistently amplified in European or Asian species. Nine loci amplified in the sister species to Aquilegia, Semiaquilegia adoxoides. The success of cross‐species amplification suggests that these microsatellites should prove useful for studies in a broad range of Aquilegia species.     

Twelve novel microsatellite markers for the Great Plains toad,Bufo cognatus

Here I report on 12 microsatellite loci designed for populations of the Great Plains toad (Bufo cognatus) in the deserts of the southwestern USA. Diversity at these loci measured for 134 individuals from four breeding aggregations was relatively high with seven to 34 alleles per locus (mean = 17.8). Observed heterozygosities ranged from 0.444 to 0.949 and expected heterozygosities ranged from 0.597 to 0.951. These markers will be useful for studies of population genetic structure, parentage and relatedness in this explosively breeding amphibian.     

Polymorphic microsatellite loci in the Chinese pond turtle (Chinemys reevesii)

Eight novel polymorphic microsatellite loci developed from a microsatellite enriched genomic library, are presented for the Chinese pond turtle (Chinemys reevesii). We screened 30 individuals from three populations and detected high levels of polymorphism for all eight loci with the number of alleles/locus ranging from 7 to 17 (average = 10.88). The values of expected and observed heterozygosities ranged from 0.703 to 0.920 and 0.321 to 0.966, respectively. These highly variable loci will provide a powerful molecular toolkit for studies of population structure, gene flow, and paternity assignment.     

Characterization of microsatellite markers for the mangrove tree Avicennia germinans L. (Avicenniaceae)

We describe 10 primers for amplification of microsatellite loci for the mangrove, Avicennia germinans. Eight loci were isolated from a DNA sample from the Pacific coast of Baja California, Mexico and two loci were isolated from a DNA sample from the Atlantic coast of Bermuda. Polymorphism was investigated in a population from the Mexican Pacific coast (n = 15) and in four samples scattered throughout the range of the species. Total number of alleles for the species ranged from two to 10. Observed heterozygosity in the Mexican Pacific coast population ranged from 0.27 to 0.60, with two loci having fixed alleles.     

Isolation,characterization and cross‐species amplification of microsatellite loci in the cycad genus Dioon (Zamiaceae). Potential utilization in population genetics studies of Dioon edule

Dioon edule (Zamiaceae) is an endemic Mexican cycad. Nineteen microsatellite loci were isolated from three enriched genomic libraries of D. edule var. angustifolium, D. tomasellii, and D. caputoi. Seven of these loci showed polymorphisms in D. edule. Levels of polymorphism were assessed using 16 individuals from each of seven populations throughout the range of this species. The number of alleles per locus ranged from two to five and the observed and expected heterozygosities ranged from 0.0 to 0.9821 and from 0.0088 to 0.6318, respectively. All loci show significant linkage disequilibrium. Three loci depart significantly from Hardy–Weinberg equilibrium.     

Characterization of microsatellite markers from the gastropod genus Melongena

Eight microsatellite loci were isolated and characterized from the Florida crown conch (Melongena spp.). Approximately 500 individuals from 20 populations encompassing all known Melongena spp. groups in Florida and Alabama were genotyped at all loci. High levels of variation were found with eight to 28 alleles per locus and observed per locus heterozygosity ranging from 0.148 to 0.779 (mean 0.624). One locus, Mco5, was monomorphic in more than half of the populations. These loci can provide useful information for a variety of studies ranging from parentage to population‐level analyses of the Florida crown conch.     

Isolation of microsatellite loci from the endangered plant Galium catalinense subspecies acrispum (Rubiaceae)

Galium catalinense subspecies acrispum (Rubiaceae) is a state‐endangered perennial shrub endemic to San Clemente Island. Eight polymorphic microsatellite loci were isolated from G. catalinense ssp. acrispum. These loci show high levels of variability, averaging 6.5 alleles per locus and an expected heterozygosity of 0.550. One locus exhibited significant deviations from Hardy–Weinberg equilibrium (P < 0.01) and one pair of loci exhibited significant linkage disequilibrium.     

New microsatellite markers for the snow crab Chionoecetes opilio (Brachyura: Majidae)

Nine novel microsatellite loci were isolated from Chionoecetes opilio by screening an enriched genomic library using nonradioactive polymerase chain reaction techniques, and the polymorphisms were examined to estimate genetic variability. The genetic variabilities varied depending on the locus. All loci were found to be polymorphic with an average of 9.7 alleles per locus (range 3–25). The observed and expected heterozygosities ranged from 0.80 to 0.98 and from 0.56 to 0.95, respectively. Five loci showed significant Hardy–Weinberg disequilibrium at the P < 0.05 level. The high variabilities revealed in this study suggest that these microsatellite loci should provide useful markers for genetic variation monitoring of C. opilio.