PERMANENT GENETIC RESOURCES: Eighteen new polymorphic microsatellite markers for the endangered Florida manatee, Trichechus manatus latirostris

Here we describe 18 polymorphic microsatellite loci for Trichechus manatus latirostris (Florida manatee), isolated using a polymerase chain reaction‐based technique. The number of alleles at each locus ranged from two to four (mean = 2.5) in specimens from southwest (n = 58) and northeast (n = 58) Florida. Expected and observed heterozygosities ranged from 0.11 to 0.67 (mean = 0.35) and from 0.02 to 0.78 (mean = 0.34), respectively. Departures from Hardy–Weinberg equilibrium occurred at two loci. There was no evidence of genotypic disequilibrium for any pair of loci. For individual identification, mean random‐mating and θ‐corrected match probabilities were 9.36 × 10^?7 and 1.95 × 10^?6, respectively.     

Development of 15 polymorphic microsatellite markers in the Atlantic tarpon (Megalops atlanticus) for capture-recapture studies

Fifteen polymorphic microsatellite DNA loci for Atlantic tarpon, Megalops atlanticus, were isolated by using PIMA, a polymerase chain reaction‐based technique. The number of alleles at each locus ranged from two to 24 (mean = 7.7) in 65 specimens from Tampa Bay, Florida. Observed and expected heterozygosities ranged from 0.27 to 0.92 (mean = 0.60) and from 0.28 to 0.95 (mean = 0.62), respectively. Genotypes at one locus deviated significantly from Hardy–Weinberg equilibrium. In exact tests for genotypic disequilibrium, there was no evidence of associations between any pair of loci. Overall, loci were well resolved and highly polymorphic, confirming their suitability for DNA fingerprinting applications and other genetic studies.     

Ten di- and trinucleotide microsatellite loci in the Caribbean spiny lobster, Panulirus argus, for studies of regional population connectivity

We describe 10 microsatellite loci for Panulirus argus (Caribbean spiny lobster). The number of alleles at each locus ranged from four to 39 (mean = 21.8) in 89 juvenile specimens collected at two different times at a recruitment site in south Florida. Levels of expected and observed heterozygosities ranged from 0.48 to 0.96 (mean = 0.83) and from 0.32 to 0.98 (mean = 0.71), respectively. Significant departures from Hardy–Weinberg equilibrium were observed at two loci. There was no evidence of genotypic disequilibrium for any pair of loci. Overall, the loci were well resolved, highly polymorphic and independently segregating, confirming their utility for population genetic studies.     

New microsatellite loci for the endangered scalloped hammerhead shark, Sphyrna lewini

We isolated 15 microsatellite markers for the scalloped hammerhead shark, Sphyrna lewini. Loci were tested on 80 specimens of S. lewini from four Eastern Pacific samples. The number of alleles per locus ranged from 6 to 31 (mean = 14). Observed and expected levels of heterozygosity per locus ranged from 0.39 to 0.91 (mean = 0.70) and from 0.54 to 0.90 (mean = 0.76), respectively. No pairs of loci were in gametic disequilibrium after Bonferroni correction of α. One locus showed significantly lower heterozygosity than expected under Hardy–Weinberg proportions in two populations, possibly caused by null alleles.     

Isolation and characterization of 23 microsatellite loci in the yellow tang, Zebrasoma flavescens (Pisces: Acanthuridae)

Twenty‐three microsatellites were isolated from the yellow tang (Zebrasoma flavescens), an ecologically and commercially important reef fish. Genetic diversity was assessed in 90 adults collected from Honokohau, Hawaii. The number of alleles per locus varied from four to 29 (mean = 13.8) and observed and expected heterozygosities ranged from 0.15 to 0.94 (mean = 0.70) and from 0.29 to 0.93 (mean = 0.81), respectively. Eight loci exhibited significant departure from Hardy–Weinberg equilibrium due to the presence of null alleles. Exact tests showed no evidence of genotypic disequilibrium between loci. Overall, loci were well resolved, easy to score and highly polymorphic.     

Microsatellite loci for the stingless bee Melipona rufiventris (Hymenoptera: Apidae)

Eight microsatellite primers were developed from ISSR (intersimple sequence repeats) markers for the stingless bee Melipona rufiventris. These primers were tested in 20 M. rufiventris workers, representing a single population from Minas Gerais state. The number of alleles per locus ranged from 2 to 5 (mean = 2.63) and the observed and expected heterozygosity values ranged from 0.00 to 0.44 (mean = 0.20) and from 0.05 to 0.68 (mean = 0.31), respectively. Several loci were also polymorphic in M. quadrifasciata, M. bicolor, M. mandacaia and Partamona helleri and should prove useful in population studies of other stingless bees.     

Development of polymorphic microsatellite markers for the sorghum plant bug, Stenotus rubrovittatus (Heteroptera: Miridae)

Nine polymorphic microsatellite DNA markers from the sorghum plant bug, Stenotus rubrovittatus, were isolated and characterized. These markers were used to analyse 22 individuals from a single field population. The number of alleles at these nine loci ranged from two to 28 (mean = 11.4) and heterozygosity ranged from 0.27 to 0.86 (mean = 0.58). Stenotus rubrovittatus has shown rapid population growth in the decades since the first report in the 1980s of serious damage to a rice crop. These microsatellite markers will be of value for studying both the population genetics and population dynamics of S. rubrovittatus.     

Isolation and characterization of 10 highly polymorphic di- and trinucleotide microsatellite markers in the mayfly Ameletus inopinatus (Ephemeroptera: Siphlonuridae)

We describe the isolation of ten polymorphic microsatellite loci from the mayfly Ameletus inopinatus. Loci had di‐ or trinucleotide repeat motifs and were highly variable with three to 17 alleles (mean = 7.15). Observed heterozygosity ranged from 0.143 to 0.905. One locus (Ami_202) showed significant deviation from Hardy–Weinberg equilibrium in one population, but no evidence for null alleles. One locus (Ami_73) was significantly linked with three other loci. The remaining nine loci should prove highly informative for population genetic studies.     

Isolation and characterization of 11 polymorphic trinucleotide microsatellite markers in the stonefly Arcynopteryx compacta (Plecoptera: Perlodidae)

We describe the isolation of 11 polymorphic trinucleotide microsatellite loci from the stonefly Arcynopteryx compacta. Loci were highly variable with 3 to 14 alleles (mean = 6.45). Observed heterozygosity ranged from 0 to 0.867. Seven loci showed significant deviation from Hardy–Weinberg equilibrium across both populations. There was no evidence for null alleles, and thus, Hardy–Weinberg departures could have resulted from genetic structure between populations or subpopulations. No linkage between loci was found. The 11 loci should prove highly informative for population genetic studies.     

Isolation and characterization of microsatellite markers from three species of swallows in the genus Tachycineta: T. albilinea, T. bicolor and T. leucorrhoa

We describe 30 microsatellite loci developed from three species of swallows in the genus Tachycineta: T. bicolor (tree swallow), T. albilinea (mangrove swallow), and T. leucorrhoa (white‐rumped swallow). These commonly studied birds nest in secondary cavities and are distributed from Alaska to Argentina. Primer pairs were designed for each species individually and tested for cross‐amplification in 40–48 individuals of all three species. Polymorphism ranged from 5 to 65 alleles per locus (mean = 19.1). These markers will allow comparative studies of extra‐pair paternity rates among members of the genus as well as the assessment of population structure.     

Isolation and characterization of microsatellite markers from the acacia-ant Crematogaster mimosae

We describe 10 microsatellite loci developed from Crematogaster mimosae, an ant species that nests mutualistically in Acacia drepanolobium trees in east Africa. Polymorphism ranged from 4 to 16 alleles per locus (mean = 7.3). Observed and expected heterozygosities ranged from 0.485 to 0.813 (mean 0.626), and from 0.502 to 0.894 (mean 0.674), respectively. These markers will foster studies of the population structure, colony structure, and reproductive strategies of these ants.     

Microsatellite DNA markers for the snow vole (Chionomys nivalis)

A total of 14 dinucleotide microsatellite loci were characterized in the snow vole (Chionomys nivalis). Allelic polymorphism across all loci and 28 individuals representing a single population in the Swiss Alps was high (mean = 10.1 alleles). No significant linkage disequilibrium between pairs of loci and no departure from Hardy–Weinberg equilibrium were found. These loci will be useful for describing mating systems and population structure and to investigate the genetic consequences of a species living in a highly fragmented habitat.     

Isolation and cross-amplification of microsatellites in pink abalone (Haliotis corrugata)

Ten novel microsatellite loci were isolated in pink abalone, Haliotis corrugata, using (GT)₁₅ and (CT)₁₅ enriched genomic libraries. Two previously reported Haliotis kamtschatkana microsatellites cross‐amplified in H. corrugata. A set of 12 polymorphic microsatellites were evaluated in a wild population sample (N = 49). The number of alleles ranged from two to 55, and the observed and expected heterozygosities ranged from 0.104 to 0.939 and from 0.213 to 0.982, respectively. Significant deviations from Hardy–Weinberg equilibrium at three loci and no linkage disequilibrium were observed. Haliotis corrugata microsatellites cross‐amplified in other abalone species, two in H. fulgens, and seven in H. rufescens.     

Development and characterization of 11 microsatellite loci for the Mona Island iguana (Cyclura cornuta stejnegeri)

We isolated and characterized 11 microsatellite loci in the Mona Island iguana (Cyclura cornuta stejnegeri). Eleven loci exhibit moderate to high allelic diversity (two to 12 alleles, mean = 4.5) and polymorphism (mean observed heterozygosity, 0.56; range, 0.26 to 0.78) in 41 adults. This marker set has low probability of identity and high parentage exclusion power and will be suitable for studies of paternity, social organization and relatedness in this species.     

Isolation and characterization of polymorphic tetranucleotide microsatellite loci in the chestnut-crowned babbler (Pomatostomus ruficeps)

We describe 18 microsatellite markers isolated in the cooperatively breeding chestnut‐crowned babbler (Pomatostomus ruficeps). The number of alleles ranged from seven to 16 per locus (mean N_a = 10.4 ± 0.54 SE) and the expected heterozygosity ranged from 0.732 to 0.889 (mean H_E = 0.836 ± 0.01 SE). Three of the 18 loci exhibited significant heterozygote deficiency, but the remaining 15 will be used to analyse population genetic structure and the mating system of this highly social species.     

Polymorphic microsatellite DNA markers for Daviesia triflora (Papilionaceae)

We developed 11 polymorphic microsatellite DNA markers for an Australian native plant, Daviesia triflora. The number of alleles per locus in 40 individuals varied from four to 19, and observed and expected heterozygosities ranged from 0.450 to 0.925 and from 0.497 to 0.899, respectively. Nine loci showed no significant deviation from Hardy–Weinberg equilibrium (P > 0.05), and null alleles appear to exist at loci DT‐A102 and DT‐B103. All loci showed independent inheritance.     

Isolation and characterization of 13 polymorphic microsatellite loci for the Florida pompano,Trachinotus carolinus

Here we describe 13 polymorphic, dinucleotide microsatellite loci for Trachinotus carolinus (Florida pompano), isolated by using PIMA, a polymerase chain reaction‐based technique. The number of alleles at each locus ranged from 3 to 29 (mean = 11.4) in 45 specimens collected from bay and nearshore waters around St Petersburg, Florida. Levels of expected and observed heterozygosities ranged from 0.15 to 0.94 (mean = 0.69) and from 0.16 to 0.98 (mean = 0.70), respectively. No significant departures from Hardy–Weinberg equilibrium expectations were observed. In exact tests for genotypic disequilibrium, there was no evidence of linkage for any pair of loci. The ability of these markers to cross‐amplify in two congeneric Trachinotus species —T. falcatus (permit) and T. goodei (palometa) — was also assessed. The loci were well‐resolved, highly polymorphic, and independently segregating in these taxa, also suggesting a general utility for intraspecific studies, species identification, and investigation of interspecific hybridization.     

Cross-species amplification of 44 microsatellite loci developed for Chaetodon trifascialis, C. lunulatus and C. vagabundus in 22 related butterflyfish species

Forty‐four microsatellite primers developed for three species of butterflyfish were cross‐tested against 22 related confamilial species. Amplification success and cross‐species transferability of these markers were moderately high. Between 24 and 37 loci were amplified successfully in each species, with a mean success rate per species of 71.7% (± 1.8 SE). Rates of amplification success were comparable among primers designed for the three source species, ranging from a mean success rate per species of 16.9 loci (± 0.8 SE) for Chaetodon trifascialis source loci to 13.7 loci (± 1.5 SE) for C. vagabundus source loci. Polymorphism rates were high (76.1%± 3.1 SE of all successfully amplified loci), and 10 loci were polymorphic in all successfully amplified species (Tri14, B11, C5, D3, D113, D6, D117, D120, D111, D118). The number of alleles per polymorphic locus ranged from 2 to 8, and the average number of alleles across all polymorphic loci and all species was 3.6 (± 0.07 SE). Polymorphism rates were higher overall in primers designed for C. vagabundus (89.9%± 3.9 SE). Overall cross‐testing success was lowest for Heniochus chrysostomus, the most phylogenetically divergent species. The significant cross‐testing reported here provides a valuable resource that will enable population genetics studies to be undertaken on a range of butterflyfishes without the need for expensive and time‐consuming de novo microsatellite development.     

Polymorphic microsatellite DNA markers for Banksia hookeriana (Proteaceae)

We developed 11 polymorphic microsatellite DNA markers for an Australian native shrub Banksia hookeriana (Proteaceae). The number of alleles per locus in 37 individuals varied from three to 17, observed and expected heterozygosities ranged from 0.297 to 0.838 and from 0.279 to 0.900, respectively. Two loci (BH‐B5 and BH‐B107) showed significant deviation from Hardy–Weinberg equilibrium (P < 0.05), and null alleles may be present at these two loci. All loci showed independent inheritance.     

Isolation and characterization of polymorphic microsatellite loci in yellowtail catfish, Pangasius pangasius (Hamilton, 1822)

A total of nine polymorphic microsatellite loci were obtained from a genomic library of Pangasius pangasius (order Siluriformes, family Pangasiidae). Samples from rivers Bhagirathi (n = 22) and Mahanadi (n = 20) were genotyped for each of the nine microsatellite loci to determine genetic variation. The mean number of alleles per locus was 5.22 in Bhagirathi and 5.78 in Mahanadi; and expected heterozygosity ranged from 0.567 (Bhagirathi) to 0.578 (Bhagirathi). Significant deviation (P < 0.003) from Hardy–Weinberg expectations was evident at three loci, Ppa2 (Bhagirathi), Ppa14 (Mahanadi) and Ppa28 (Bhagirathi and Mahanadi). The identified microsatellite loci were found to be promising for population genetics studies of P. pangasius.