Isolation of microsatellite loci in the pollinating fig wasp of Ficus hispida, Ceratosolen solmsi

Microsatellite loci were isolated for Ceratosolen solmsi, pollinator of the dioecious Ficus hispida. We developed nine polymorphic microsatellite loci based on the method of polymerase chain reaction isolation of microsatellite arrays （PIMA）. Enrichment of genomic libraries was performed by random amplified polymorphic DNA （RAPD）. A subset of 38 positive clones was sequenced; 15 clones showed microsatellite loci. We tested 15 designed primer pairs and nine of them produced polymorphic amplification in 48 individual wasps collected from different fruits of the dioecious host fig Ficus hispida in China. Among the 48 individuals, 49 alleles were obtained at the nine loci. The observed heterozygosity ranged between 0.357 and 0.634.

Isolation and Characterization of 17 Microsatellite DNA Loci for Odorrana margaretae(Anura: Ranidae)

We isolated and characterized 17 microsatellite DNA loci for the odorous frog Odorrana margaretae from its transcriptome sequence data. These loci were screened with 24 individuals from Mt. Emei. All loci were polymorphic, with the number of alleles ranging from 2 to 8. The observed and expected heterozygosity, polymorphism information content, ranged from 0.04 to 1, 0.04 to 0.81, and 0.040 to 0.763, respectively. All loci were in linkage equilibrium and six loci were significantly deviated from Hardy-Weinberg equilibrium after sequential Bonferroni corrections. Cross- species amplification test was conducted for ten odorous frog species, and 12 loci were amplifiable in most species. With the high cross-species amplification rates, thesc markers will provide useful molecular tools for conservation genetic and phylogeographic studies on the genus Odorrana and Bamburana.

Development of novel microsatellite markers for Holothurian scabra (Holothuriidae), Apostichopusjaponicas (Stichopodidae) and cross-species testing in other sea cucumbers

Thirty-five new microsatellite loci from the sea cucumbers H olothurian scabra(Jaeger, 1833) and Apostichopus japonicas(Selenka, 1867) were screened and characterized using the method of magnetic bead enrichment. Of the twenty-four polymorphic loci tested, eighteen were consistent with Hardy-Weinberg equilibrium after a modified false discovery rate(B-Y FDR) correction, whereas six showed statistically significant deviations(CHS2 and CHS11: P <0.014 790; FCS1, FCS6, FCS8 and FCS14: P <0.015 377). Furthermore, four species of plesiomorphous and related sea cucumbers(Holothurian scabra, Holothuria leucospilota, Stichopus horrens and Apostichopus japonicas) were tested for mutual cross-amplification using a total of ninety microsatellite loci. Although transferability and universality of all loci were generally low, the results of the cross-species study showed that the markers can be applied to identify individuals to species according to the presence or absence of specific microsatellite alleles. The microsatellite markers reported here will contribute to the study of genetic diversity, assisted breeding, and population conservation in sea cucumbers, as well as allow for the identification of individuals to closely related species.

Development and Characterization of Fifteen Polymorphic M icosatellites for Extremely Endangered Salamander, Echinotriton chinhaiensis

Echinotriton chinhaiensis is a critically endangered salamander and its distribution is restricted to Ningbo, Zhejiang Province of China. In this study, we developed and characterized fifteen polymorphic microsatellite loci for E. ehinhaiensis from dinucleotide- and tetranucleotide- enriched library. The number of alleles ranges from 4 to 12 with an average of 7.27 alleles per locus. The observed and expected heterozygosities values were from 0.250 to 0.844 and 0.511 to 0.872 with an average value of 0.596 and 0.722; respectively. The polymorphic microsatellite loci described in this paper are useful in the further study on genetic diversity and gene flow, which would be helpful to formulate effective conservation strategies for the E. chinhaiensis.

Patterns of geographic variation between mitochondrial and nuclear markers in Heaviside’s(Benguela)dolphins(Cephalorhynchus heavisidii)

The Heaviside’s(or Benguela)dolphin(Cephalorhynchus heavisidii)is endemic to the west coast of southern Africa.The present study investigated the population genetic structure across a large portion of the species dis­tribution using mitochondrial control region and nuclear(microsatellite)markers.A total of 395 biopsy skin samples were analyzed;they were collected from free-ranging Heaviside’s dolphins in 7 locations along 1650 km of coast between Table Bay,South Africa and Walvis Bay,Namibia.Both genetic markers rejected the hy­pothesis of 1 homogenous population but revealed contrasting results in the genetic structuring of putative pop­ulations.Mitochondrial DNA suggested either 2 populations or a fine-scale division with 6(sub)populations,while microsatellite markers were indicative of 2 widespread populations with measurable gene flow between them.Neutrality tests and mismatch distribution of the mitochondrial sequences indicated a departure from mu­tation-drift equilibrium due to a population expansion at the 2 extremes of the geographic range,but not to­wards the middle of the distribution.These results highlight the importance of evaluating multiple genetic mark­ers to gain reliable insights into population processes and structure.