Polymorphic microsatellite markers for endangered spotted suslik,Spermophilus suslicus

Spotted suslik Spermophilus suslicus is one of the most endangered mammal species in Poland. Over the last 50 years, it lost more than 90% of its populations and about 70% of its individuals. In order to establish a conservation and reintroduction program, the knowledge of population structure of the species is crucial. We have developed nine polymorphic microsatellite loci to analyse the population structure. Six of the primer sets also amplify polymorphic markers in the European suslik Spermophilus citellus.     

Semen cryopreservation in the Indian rhinoceros (Rhinoceros unicornis)

The objective was to identify an extender and cryoprotectant combination for Indian rhinoceros (Rhinoceros unicornis) sperm that yielded high post-thaw sperm quality. Male Indian rhinoceroses (n = 6; 7.5-34 yr old) were anesthetized and subjected to a regimented electroejaculation procedure (75-100 mAmps; 4-10 volts; 7-150 stimuli; total of 10 electroejaculation procedures). High quality semen fractions from each ejaculate were divided into four aliquots and a 2 x 2 factorial design used to compare the effect of two sperm extenders (standard equine [EQ] and skim milk-egg-yolk-sugar [SMEY]), and two cryoprotectants (glycerol and dimethylsulfoxide [DMSO]). Cyropreserved samples were thawed and assessed for motility, viability and acrosome integrity over time. Electroejaculate fractions processed for cryopreservation had high sperm concentration (516 × 10⁶/mL) and motility (79%). Post-thaw sperm characteristics were higher (P < 0.05) when semen was cryopreserved in EQ versus SMEY. Post-thaw motility of sperm cyropreserved in EQ averaged 50-55% compared to 22-37% in SMEY, with no significant differences in sperm characteristics of samples cyropreserved in glycerol and DMSO. In conclusion, sperm collected from Indian rhinoceroses via electroejaculation were cryopreserved using EQ extender with either glycerol or DMSO; post-thaw quality was adequate for use in assisted reproductive procedures.     

Polymorphic microsatellite markers in the painted dragon lizard,Ctenophorus pictus

We isolated eight polymorphic microsatellite markers from the painted dragon, Ctenophorus pictus, from a genomic library simultaneously enriched for a suite of di‐, tri‐ and tetranucleotide repeat units. Genetic variation was assessed in 40 individuals from populations in Victoria and South Australia. The number of alleles per locus ranged from four to 45 with observed heterozygosity ranging from 0.23 to 0.95. These markers will be useful for studies of natural and sexual selection and the effects of habitat fragmentation in semiarid Australia.     

Polymorphic tetranucleotide microsatellite markers in the Caribbean spiny lobster,Panulirus argus

Ten tetranucleotide microsatellite loci are described for the Caribbean spiny lobster Panulirus argus. Loci were polymorphic (4–15 alleles per locus) and exhibited high levels of expected (0.553–0.921) and observed heterozygosity (0.469–0.906) from samples caught off Belize and Puerto Rico coasts. No significant departure from Hardy–Weinberg equilibrium conditions were observed for any locus. All microsatellite loci should be useful for assessing population discrimination for this valuable marine animal currently subjected to excessive fishing efforts.     

Polymorphic tetranucleotide microsatellite loci in the Hwamei (Garrulax canorus canorus) (Timaliidae)

Eight polymorphic tetrarepeat (GATA)_n, microsatellite loci were isolated from a babbler, Hwamei (Garrulax canorus canorus). We evaluated the polymorphism of these microsatellite loci by genotyping 36–48 individuals from the Asian mainland. The number of alleles for each locus ranged from eight to 29. The heterozygosity was between 0.587 and 0.978. Except for one locus, genotype frequencies of these microsatellites did not significantly deviate from the Hardy–Weinberg expectation. These markers should be useful for monitoring potential hybridization between different Hwamei subspecies and provide new insights into the mating system and geographical differentiation of these birds.     

Development and characterization of microsatellite loci in endangered Astrophytum asterias (Cactaceae)

We report the development of polymorphic microsatellite markers for the endangered North American cactus Astrophytum asterias (Cactaceae). Six loci, averaging 8.5 alleles per locus, were found to amplify genomic DNA consistently in 94 individuals from four geographically defined demes in South Texas. These markers will permit the generation of appropriate data for estimating population genetic parameters, population structure and the degree of inbreeding in the small, fragmented populations of A. asterias that currently exist. These are the first microsatellites reported for the genus Astrophytum and for the tribe Cacteae.     

Characterization of microsatellite loci in the Kaka,Nestor meridionalis

Eight microsatellite loci were characterized from the Kaka (Nestor meridionalis), a New Zealand parrot, using a polymerase chain reaction‐based isolation technique. Locus‐specific primers were used to genotype nine Kaka populations and tested on 25 other species of parrot. The number of alleles observed within Kaka ranged from one to 16 in the 12–126 individuals screened and two loci exhibited greater than 60% heterozygosity. Furthermore, these primers are likely to be useful in population‐level studies of two other New Zealand parrots.     

Fifteen microsatellite loci for the jungle perch, Kuhlia rupestris

We developed and optimized 15 polymorphic microsatellite loci in the jungle perch, Kuhlia rupestris. Loci were screened in a single population (n = 24) from Fraser Island, Queensland, Australia. Number of alleles per locus ranged from 3 to 19 and observed heterozygosity from 0.25 to 1. No significant linkage disequilibrium was detected between any pair of loci. Genotype proportions for these loci in the population sampled were in Hardy–Weinberg equilibrium.     

Polymorphic microsatellite loci in Plantago lanceolata

The genus Plantago is particularly interesting for evolutionary studies because of its wide range of mating systems. We have developed primers for five highly polymorphic microsatellite loci isolated from P. lanceolata. All five loci amplified and were polymorphic in the two populations examined, Lowsteads Beach in the United Kingdom and Duke in the United States. These new markers will allow a comparison of population structure between the outcrossing species P. lanceolata, and the highly selfing species P. major.     

Polymorphic microsatellite markers isolated from the neptune whelk Neptunea arthritica

Eight polymorphic microsatellite DNA loci were isolated from the neptune whelk Neptunea arthritica, which is an important fishery resource in northern Japan. The number of alleles at the loci ranged from two to six, with observed and expected heterozygosities of 0.192-0.807 and 0.233-0.738, respectively. The observed variations suggest that these loci can be used as markers for population and kinship analyses in this species.     

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.     

Polymorphic microsatellite DNA markers for the Patagonian squid,Loligo gahi (Cephalopoda)

Squid populations are being put under rapidly increasing commercial fishing pressure worldwide. The same species are known to be susceptible to extreme population fluctuations, so detailed knowledge of population substructuring and genetic diversity is essential for rational management. We present a set of microsatellite DNA loci suitable for population genetic analysis of Loligo gahi, the squid species subject to the most detailed monitoring and fishery control (around the Falkland Islands), with the future aim of generating management‐related information to aid conservation of this valuable natural resource.     

Cross-species amplification of Eucalyptus microsatellite loci

This study examined the interspecific amplification of nuclear microsatellite loci developed mainly for eucalypts in the subgenus Symphyomyrtus across five species within the second most speciose subgenus, subgenus Eucalyptus. A set of eight to 10 loci, depending on taxon, have been identified that are highly variable and easily scored. The successful transfer of microsatellite loci to these eucalypt species sidesteps the expensive and time-consuming development of species-specific microsatellite libraries. This primer set will enable the examination and cross-species comparison of the genetic resources of commercially and ecologically important members of the subgenus Eucalyptus.     

Development and characterization of 16 polymorphic microsatellite loci in the Tibet endemic fish Glyptosternum maculatum

Sixteen novel polymorphic microsatellite loci were developed and characterized in Glyptosternum maculatum, an endemic fish inhabiting the Yarlung Zangbo River valley, Tibet, China. These loci were developed after the enrichment of microsatellite-containing genomic DNA fragments with a subtractive hybridization protocol. The polymorphism of these loci was analysed with 36 individuals representing a geographical population. The number of alleles found at these loci ranged from 3 to 10. The observed heterozygosity and expected heterozygosity ranged from 0 to 1 and from 0.31 to 0.75, respectively. These microsatellite loci will certainly facilitate the determination of the genetic structure of G. maculatum and the conservation of its genetic resource.     

Genetic diversity, phylogeny and conservation of the Javan rhinoceros (Rhinoceros sondaicus)

With a total population of less than 60 individuals limited to two locations, the Javan rhinoceros is perhaps the most endangered large mammal on earth. Although species specific information is crucial to its conservation, its precarious status, habitat inaccessibility, and behavioral adaptations pose major obstacles to its study. Here we report on the first genetic analysis of the two extant populations, in Ujung Kulon, Indonesia, and Cat Tien, Vietnam, and discuss their conservation. As its critically endangered status precluded invasive sampling, we extracted DNA from dung, amplifying and sequencing segments of the mtDNA 12S rRNA gene and the non-coding D-loop. Divergence between Javan rhinos from Ujung Kulon and Cat Tien was similar to that between recognized subspecies of African rhinos, and exceeded that between Sumatran rhinos. The Ujung Kulon and Cat Tien populations represent separate Evolutionary Significant Units, advocating independent management. However, given the precariousness of the Cat Tien population, demographic considerations may override genetic issues in the short term. Genetic diversity of Javan rhinos was low and population expansion in the immediate future will be critical for its survival.     

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.     

Polymorphic microsatellite loci in Linum usitatissimum

We report the characterization of 28 polymorphic microsatellite markers in Linum usitatissimum that allow distinguishing almost all cultivars of both flax and linseed. Polymorphism was low, ranging from two to 10 alleles per locus in the 93 cultivars screened. Linkage disequilibrium was found at about a third of the pairs of loci likely due to self‐fertilization and strong selection by breeders. We tested these loci for cross‐amplification in nine additional species of Linum and found that three species amplified a majority of loci.     

Isolation and characterization of microsatellite loci from rock shell, Thais clavigera

Imposex (superimposition of male genital organs on female phenotype) of the rock shell or whelk, Thais clavigera, shows typical evidence of endocrine disruption by organotin compounds within inter-tidal zones polluted by such compounds. It will be informative to see how low fertility caused by imposex of this species finally affects the genetic diversity of polluted populations. For future use in population genetic research of the rock shell, we report the isolation and characteristics of 11 highly polymorphic microsatellite markers with a mean of 11.7 alleles per locus. We observed heterozygosities of these sequences ranging from 0.43 to 0.95. These markers will be useful for future ecological genetic studies of rock shell.     

Isolation and characterization of 49 polymorphic microsatellite loci in the ayu,Plecoglossus altivelis

A total of 49 novel polymorphic microsatellite markers in the ayu fish, Plecoglossus altivelis, were successfully isolated and characterized. The number of alleles per locus, observed heterozygosity, and expected heterozygosity ranged from two to 25 (11.3 on average), from 0.125 to 1.000 (0.648 on average), and from 0.123 to 0.957 (0.751 on average), respectively. These microsatellite markers will be useful for both studying population structure and identifying quantitative trait loci that control evolutionarily and economically important traits of ayu.