Novel microsatellite loci identified from the Australian eastern small‐eyed snake (Elapidae: Rhinocephalus nigrescens) and cross species amplification in the related genus Suta

A total of 15 microsatellite primers pairs were developed for the Australian small‐eyed snake Rhinoplocephalus nigrescens. Five primers were used to screen 93 individuals of R. nigrescens and were also tested against eight species of the closely related genus Suta. Allelic diversity in R. nigrescens was high in three loci (12–27) and there was high heterozygosity (0.58–0.82). Observed heterozygosity did not deviate from Hardy–Weinberg expectations for the five loci tested. These primers will be useful in studies of population genetics and mating systems of small‐eyed snakes and related species.     

Isolation and characterization of microsatellite loci in Portunus pelagicus (Crustacea: Portunidae)

Seven dinucleotide and one tetranucleotide microsatellite loci were isolated and characterized, using routine protocols, from the blue swimmer crab, Portunus pelagicus. These loci will be useful for investigating the population structure of this broadly distributed and economically important portunid. In particular, all eight loci were polymorphic in P. pelagicus collected from throughout Australia. In addition, the genotype frequencies at each locus in each population sample were usually not significantly different from those expected under Hardy–Weinberg equilibrium conditions. Six of the eight loci also showed moderate to high levels of polymorphism in an undescribed species of Portunus from northern Australia.     

Eight polymorphic microsatellite loci for bottlenose dolphin and other cetacean species

We developed eight polymorphic microsatellite loci that can be used to elucidate population structure and aid management of bottlenose dolphins, Tursiops truncatus, in the northwest Atlantic. These loci averaged nine alleles, with no evidence for null alleles. Their cross‐species utility was examined in Cephalorhynchus commersonii, Delphinus delphis, Delphinapterus leucas, Eschrictius robustus, Globicephala macrorhynchus, Lagenorhynchus obliquidens, Orcinus orca, Steno bredanensis and Stenella clymene. On average, 75% of the loci were polymorphic in these species (range = 50–88%). Our results indicate that these loci will be useful for elucidating population structure of bottlenose dolphin as well as of other cetacean species.     

Characterization of polymorphic microsatellite loci in Castelnau's Antshrike,Thamnophilus cryptoleucus (Aves: Thamnophilidae)

In order to study the biogeography and population genetic structure of the obligate river‐island species Thamnophilus cryptoleucus (Thamnophilidae) we isolated four microsatellite loci. Number of alleles ranged from six to 11, heterozygosities from 60% to 89%, and individual populations were found in Hardy–Weinberg equilibrium. Cross‐species amplification of the loci was tested in eight species of the family Thamnophilidae and all loci successfully amplified in five or more related species.     

Microsatellite DNA primers for the endangered vermilion darter,Etheostoma chermocki,and cross‐species amplification in other darters (Percidae: Etheostoma)

The endangered vermilion darter (Etheostoma chermocki) is endemic to the Black Warrior River system in the Mobile Basin in Alabama. Restoration and conservation of this species require an understanding of its population genetic structure, which can be characterized using microsatellite DNA. Nine microsatellite loci were developed; eight loci were polymorphic. Although observed heterozygosity was lower than expected heterozygosity in most polymorphic loci, only one locus showed significant deviation from Hardy–Weinberg equilibrium. These nine markers were tested in an additional 24 species of Etheostoma and appear to have sufficient allelic variation to be useful in studies of population genetic structure.     

Identification of microsatellite loci in Collinsia verna (Veronicaceae)

We developed eight polymorphic microsatellite loci for Collinsia verna (Veronicaceae). In a sample of 18–35 individuals from a single population, we found two to 15 alleles per locus (mean 8.3). We also tested these loci for cross‐amplification in all 22 species in the tribe Collinseae. Overall, more than half the species in the tribe amplified one microsatellite while three species most closely related to C. verna (Collinsia violacea, Collinsia parviflora and Collinsia grandiflora) amplified multiple microsatellite loci. These microsatellite loci will be used in future studies of mating system in this tribe and other quantitative genetic and population genetic studies.     

Polymorphic microsatellite markers for the Florida scrub lizard (Sceloporus woodi)

The Florida scrub lizard (Sceloporus woodi) is one of a suite of species restricted to Florida scrub, a threatened ecosystem. We characterized eight microsatellite loci from scrub lizards based on screening of 75–91 individuals per locus. Polymorphism was high (8–20 alleles per locus). Observed and expected heterozygosities ranged from 0.32–0.83 and 0.77–0.91, respectively. These markers will be useful for population‐level analyses and can contribute to a genetic foundation for conservation strategies for this endemic species.     

Characterization of microsatellite DNA markers for the alligator snapping turtle,Macrochelys temminckii

Two trinucleotide and seven tetranucleotide microsatellite loci were isolated from an alligator snapping turtle Macrochelys temminckii. To assess the degree of variability in these nine microsatellite loci, we genotyped 174 individuals collected from eight river drainage basins in the southeastern USA. These markers revealed a moderate degree of allelic diversity (six to 16 alleles per locus) and observed heterozygosity (0.166–0.686). These polymorphic microsatellite loci provide powerful tools for population genetic studies for a species that is afforded some level of conservation protection in every state in which it occurs.     

Development and characterization of 10 novel microsatellite markers from Chital deer (Cervus axis) and their cross–amplification in other related species

Ten polymorphic microsatellite loci were isolated and characterized from Chital deer (Cervus axis). These loci show high levels of allelic diversity with four to eight alleles per locus in the 22 individuals of the free‐ranging population of Chital deer in Nehru Zoological Park, Hyderabad. In addition, we found that all the loci show cross–amplification in closely related as well as distantly related deer species. The amplification of these markers in different genera further indicates that these can be applied to a wide range of endangered deer species for their population genetics studies and conservation management.     

Microsatellite markers characterized in the barn owl (Tyto alba) and of high utility in other owls (Strigiformes: AVES)

We have identified 15 polymorphic microsatellite loci for the barn owl (Tyto alba), five from testing published owl loci and 10 from testing non‐owl loci, including loci known to be of high utility in passerines and shorebirds. All 15 loci were sequenced in barn owl, and new primer sets were designed for eight loci. The 15 polymorphic loci displayed two to 26 alleles in 56–58 barn owls. When tested in 10 other owl species (n = 1–6 individuals), between four and nine loci were polymorphic per species. These loci are suitable for studies of population structure and parentage in owls.     

Isolation and characterization of microsatellite loci in the bee Ceratina flavipes

Polymorphic microsatellite markers can provide essential information on genetic analyses of sociality and mating behaviour of insects. We developed eight microsatellite loci in the small carpenter bee, Ceratina flavipes, using a magnetic bead hybridization enrichment protocol. These loci showed two to eight alleles with expected heterozygosity of 0.21–0.87, and also seemed useful to such congeneric species as Ceratina okinawana distributed in southern Japan. By using these loci, the sociality and mating systems in Ceratina species, that are still controversial, are possible to be analysed.     

Polymorphic microsatellite markers for studies of the conservation and reproductive genetics of imperilled sand tiger sharks (Carcharias taurus)

We report on the isolation of eight microsatellites from the sand tiger shark, Carcharias taurus, using an enrichment protocol. All loci, with the exception of Cta45–183, were in Hardy–Weinberg equilibrium. Loci exhibited three to 15 alleles, and observed and expected heterozygosities of 0.095–1.000 and 0.284–0.924, respectively. An additional marker (Iox‐12) developed from a shortfin mako library was variable in sand tigers. These markers will be used to examine population genetics and mating patterns of this imperilled species.     

Isolation and characterization of 10 microsatellite loci in the Ponerinae ant Pachycondyla luteipes (Hymenoptera; Formicidae)

We developed 10 polymorphic microsatellite loci for the Ponerinae ant Pachycondyla luteipes using the magnetic particle method. Nine of the 10 loci tested were highly polymorphic, having four to eight alleles in P. luteipes with expected heterozygosity of 0.38–0.73, and also appeared to be applicable to other related species such as Pachycondyla chinensis. These loci can be used to test the hypotheses on the population genetic structures, such as if this ant is unicolonial or not.     

Isolation and characterization of polymorphic microsatellite loci from Wilsonia backhousei (Convolvulaceae)

Wilsonia backhousei is a clonal saltmarsh plant restricted to the southern latitudes of Australasia and threatened in New South Wales. We have identified eight informative microsatellite loci in the species from (AG)_n‐ and (AC)_n‐enriched libraries. In 48 samples from two populations we detected an average of five alleles per locus (range 2–8, average H_E = 0.45), of which 72% were unique to one population or the other. Six of the eight loci were also amplifiable in Wilsonia rotundifolia under the same reaction conditions. The markers will be excellent tools for use in the management and conservation of both species.     

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 loci in Pseudoplatystoma corruscans (Siluriformes: Pimelodidae) and cross‐species amplification

A total of five polymorphic microsatellites loci from Pseudoplatystoma corruscans were isolated and characterized. A population survey involving 43 specimens resolved a large number of alleles (range seven to eight among loci) and high observed heterozygosity (0.500–0.615), indicating its usefulness in population genetics studies. Cross‐species amplification was successful in four other Pimelodidae species.     

Isolation and characterization of eight highly variable microsatellite markers in the Pacific jumping mouse (Zapus trinotatus)

In patchily distributed populations it is possible to investigate gene flow and relatedness at multiple scales. Using an enrichment procedure, I developed a suite of eight microsatellite markers in such a heterogeneously distributed species, the Pacific jumping mouse (Zapus trinotatus). The identified loci were highly polymorphic (10–36 alleles per locus) and will be valuable for investigating this species at multiple levels, from that of parentage and breeding structure within a subpopulation, to gene‐flow and population structure across populations.     

Polymorphic microsatellite markers for the gliding marsupials Petaurus australis and Petaurus breviceps

Habitat destruction is causing population decline of many hollow dependent species such as gliding marsupials of the Family Petauridae. Three petaurid species are now listed in some Australian states as either threatened, rare or vulnerable, precipitating a need for information on their basic biology and population structure. We isolated and characterized three polymorphic microsatellite loci from the yellow‐bellied glider (Petaurus australis) and six polymorphic microsatellite loci from the sugar glider (P. breviceps). Per‐locus heterozygosities range from 42%–92%, and cross‐species amplification studies show that between five and seven loci are polymorphic in the two target species as well as a related species P. norfolcensis.     

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).     

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.