Development of microsatellite markers for <Emphasis Type="Italic">Metrosideros boninensis</Emphasis> (Myrtaceae), an endangered endemic plant species from the Bonin Islands,Japan

Metrosideros boninensis is an endangered endemic plant species from the Bonin Islands. We isolated and characterized nine microsatellite loci in this species. The expected heterozygosities of these nine markers ranged from 0.127 to 0.768. These markers described here will be useful for investigating the genetic diversity, genetic structure and gene flow, and planning for conservation of M. boninensis.     

Development of microsatellite markers for <Emphasis Type="Italic">Echinops setifer</Emphasis> (Asteraceae), an endangered grassland plant species in Japan

Echinops setifer is an endangered grassland plant species in Japan. We isolated and characterized nine microsatellite loci in this species. The number of alleles ranged from 3 to 7 and expected heterozygosities from 0.108 to 0.761. These markers described here will be useful for investigating the genetic diversity, genetic structure and gene flow, and planning for conservation of E. setifer.     

Isolation and characterization of microsatellite loci in Shortia rotundifolia (Diapensiaceae), an endangered relict plant on the Ryukyu Islands and Taiwan

Ten microsatellite loci were isolated and characterized for Shortia rotundifolia, a relict temperate plant on the subtropical Ryukyu Islands and Taiwan. The number of alleles ranged from 2 to 21. The expected (H _E) and observed (H _O) heterozygosities were 0.027–0.877 and 0.027–0.622, respectively, from 74 individuals on Iriomotejima Island of Ryukyus. One locus exhibited significantly fewer heterozygotes than expected under Hardy–Weinberg equilibrium (P < 0.05). These primers amplifying microsatellites in this species may provide a useful tool for population genetics to establish conservation strategy.     

Development and characterization of microsatellite loci in the endangered Tricyrtis ishiiana (Convallariaceae), a local endemic plant in Japan

Nine microsatellite loci were isolated and characterized for the endangered and tertiary relict perennial herb, Tricyrtis ishiiana. The number of alleles ranged from 2 to 33. The expected (H _E) and observed (H _O) heterozygosities were 0.207–0.944 and 0.215–0.813, respectively, from 96 individuals on one population. Five loci exhibited significantly fewer heterozygotes than expected under Hardy–Weinberg equilibrium (P < 0.05). These primers amplifying microsatellites in this species may provide a useful tool for population genetics to establish conservation strategy.     

Isolation and characterization of microsatellite markers in Dysosma versipellis (Berberidaceae), a rare endemic from China

Dysosma versipellis (Berberidaceae) is an important threatened medicinal plant (TMP) species. Here we isolated nine polymorphic microsatellite loci from D. versipellis using a modified biotin-capture method. Our isolated loci provided SSR markers with polymorphism of 2–7 alleles per locus. The expected and observed heterozygosities ranged from 0.507 to 0.864 and from 0.360 to 0.720, respectively. These markers would be the useful tools for analyzing questions concerning population genetic structure and mating system of D. versipellis.     

Isolation and characterization of microsatellite markers for Cycas hainanensis C. J. Chen (Cycadaceae)

Cycas hainanensis is an endangered cycad species endemic to Hainan Island, China. To enrich our scientific conservation for this species, we developed eight microsatellite markers using repetitive DNA enriched libraries. The number of alleles per microsatellite locus varied from 8 to 17. The expected (H _E ) and observed (H _O ) heterozygosities varied from 0.4754 to 0.8846 and from 0.3636 to 0.9600, respectively. These markers will be employed to determine whether the ex situ C. hainanensis individuals in Sourh China Botanical Garden capture a representative portion of genetic diversity of the wild populations.     

Development and characterisation of nine polymorphic microsatellite markers for Tephrosia calophylla Bedd. (Fabaceae)

Tephrosia calophylla Bedd. (Fabaceae) is an endangered tropical plant endemic to southwestern Ghats, India. The objective of this study was to contribute to the characterisation of the diversity of this rare species, which is necessary for its future conservation. Accordingly, microsatellite markers were designed, and their ability to detect polymorphisms was determined. Nine microsatellite markers were developed using genomic libraries, and all of the markers were successfully amplified in 42 individuals. Three to nine alleles per locus were observed, and the heterozygosity of the loci ranged from 0.381 to 0.905. The nine newly developed polymorphic markers recognise a sufficient number of varying loci to perform further studies on the conservation and breeding of this medicinal cultivar.     

Isolation and characterization of polymorphic microsatellite markers for the endangered tree <Emphasis Type="Italic">Handeliodendron bodinieri</Emphasis> (Sapindaceae)

Nine polymorphic microsatellite loci were developed and characterized for Handeliodendron bodinieri, an endangered tree endemic to karst forest in southwest China. Each locus displayed between 4 and 11 alleles. The observed heterozygosity ranged from 0.345 to 0.931 and expected heterozygosity varied from 0.414 to 0.824. The combined probabilities of paternity exclusion over all loci were 0.931 for P _r(Ex₁) and 0.993 for P _r(Ex₂), respectively. These loci represent a powerful tool in investigating gene flow and population structure in natural populations of H. bodinieri.     

Development of microsatellite DNA loci for <Emphasis Type="Italic">Pugionium dolabratum</Emphasis> (Brassicaceae), an endangered psammophyte

Pugionium (Brassicaceae) is a small Central-Asian endemic genus with four tentative species described before. All of them grow in the desert habitat and have been considered as important psammophytes for evolutionary studies and ecological restorations. However, the wild resources of these species have been declined because of overexploitation. In this study, 12 polymorphic microsatellite loci have been developed for P. dolabratum Maxim, and the performance of primer pairs amplifying these loci in related three species has also been tested. The number of alleles in 18 individuals from 13 geographically distant populations ranged from three to seven per locus. The observed and expected heterozygosities varied from 0.17 to 0.30 and from 0.32 to 0.48, respectively. We further performed cross-priming tests in the remaining three species of this genus, P. cornutum, P. calcaratum and P. cristatum. These newly recovered microsatellite loci will be useful for studying the population genetics and adaptive evolution of these endangered psammophytes in the desert.     

Isolation and characterisation of microsatellite loci in Calystegia soldanella (Convolvulaceae), an endangered coastal plant isolated in Lake Biwa,Japan

Eight microsatellite loci of Calystegia soldanella useful for comparisons of the genetic structure of isolated populations in the ancient Lake Biwa and coastal populations in Japan were isolated and characterised. The number of alleles ranged from 2 to 5. The expected (H _E) and observed (H _O) heterozygosities were 0.097–0.583 and 0.000–0.380, respectively, from 100 individuals from Lake Biwa and coastal populations. Seven of the eight loci exhibited significantly fewer heterozygotes than expected based on the Hardy–Weinberg equilibrium (P < 0.05). These primers amplifying microsatellites in C. soldanella may provide a population genetics tool useful in the establishment of a conservation strategy.     

Isolation of polymorphic microsatellite loci in Livistona chinensis (Jacq.) R.Br. ex Mart. var. boninensis Becc., an endemic palm species of the oceanic Bonin Islands, Japan

A set of microsatellites markers were developed for Livistona chinensis var. boninensis, an endemic palm tree of the Bonin Islands. We obtained 123 sequences containing unique microsatellites from an enriched library. Twelve loci were screened for their feasibility using 32 trees. They showed polymorphisms with two to nine alleles per locus. No significant deviation from Hardy–Weinberg equilibrium was observed for 11 loci. No genotypic disequilibrium was detected between any two of the loci. Total exclusionary powers for the first and the second parents were 0.978774 and 0.998987, respectively. These markers will allow us to investigate the gene flow within/among populations of the species.     

Development of microsatellite markers for Pasania edulis (Makino) Makino, one of the dominant species of lucidphyllous forests in southern Kyushu, Japan

Pasania edulis (Makino) Makino is a Fagaceous tree species endemic to Japan and one of the dominant species in lucidphyllous forests in southern Kyushu, Japan. Recently, P. edulis was attacked by ambrosia beetles, Platypus quereivorus (Murayama) and mass mortality occurred in several areas in southern Kyushu. We isolated and characterized ten microsatellite loci in this species to reveal the genetic structure and gene flow. The number of alleles ranged from 3 to 11 and expected heterozygosities from 0.3761 to 0.8346. The markers described here will be useful for investigating the genetic diversity, genetic structure and gene flow, and planning for conservation of viable population of P. edulis.     

Development and characterization of microsatellite markers for Hibiscus glaber Matsum. ex Nakai, an endemic tree species of the oceanic Bonin Islands, Japan

Polymorphic microsatellite markers were developed for Hibiscus glaber, an endemic tree of the Bonin Islands. Eighty-seven of the 208 sequences from an enriched library were unique and containing microsatellites. Ten loci were proved to be highly polymorphic among 78 individuals from the Nishi-jima Island. Total exclusionary powers for the first and the second parents were 99.989% and 99.999%, respectively. Nine loci also amplified single fragment from genomic DNA of H. tiliaceus, a related and widespread congener. Our markers can be reliably used for the estimation of current gene flow within/among populations of the two woody Hibiscus species.     

Isolation and characterization of microsatellite loci in <Emphasis Type="Italic">Melicope quadrilocularis</Emphasis> (Rutaceae), an endemic plant species of the Bonin Islands,Japan, and cross-species amplification in closely related taxa

Twelve microsatellite loci were isolated and characterized for Melicope quadrilocularis, an insular endemic tree species of the Bonin Islands. The observed number of alleles at each locus ranged from 1 to 18. The range of expected heterozygosity was 0.0000–0.9445. The inter-specific applicability of these loci was evaluated by analyzing two other endemic species and one endemic variety of Melicope that are also distributed on the Bonin Islands. All primer pairs for the 12 loci tested successfully amplified the loci in all taxa, except for primers for four loci in M. nishimurae.     

Population genetic characterization of the endangered dung beetle Copris tripartitus (Coleoptera: Scarabaeidae) using novel microsatellite markers

The dung beetle Copris tripartitus (Coleoptera: Scarabaeidae) has long been considered an endangered insect in South Korea; the detection of recent population increases leaves its endangered status uncertain. Population genetic analysis subsequent to development of molecular markers is essential for establishing proper conservation strategies. In this study, we developed ten microsatellite markers specifically for C. tripartitus. Sixty-eight individuals of C. tripartitus collected from six South Korean localities were genotyped to validate these markers and preliminarily assess population genetic characteristics. Per-locus observed number of alleles, observed heterozygosity (H_O), and expected heterozygosity (H_E) ranged from 5–12, 0.499–0.958, and 0.54–0.743, respectively. All populations showed higher H_O than H_E, negative values of inbreeding coefficient, and, overall, no sign of recent population bottlenecks (excluding one population, Seosan). This suggests that C. tripartitus did not suffer from genetic drift and inbreeding, which are typically severe in small, isolated populations. Nevertheless, detection of only one of the two gene pools in some populations and resultant genetic subdivision into two population groups may suggest that the population size is not enough to cover both gene pools. Thus, a more extended period of protection may be required to ensure higher genetic diversity of widespread populations and achieve the long-term conservation goal.     

Two new species of the genusAmblyeleotris (Pisces: Gobiidae) from the Ryukyu Islands,Japan

Two new shrimp-associated gobies,Amblyeleotris yanoi sp. nov. andA. masuii sp. nov. are described on the basis of specimens from Iriomote-jima Island and Okinawa-jima Islands, Okinawa Prefecture, Japan.A. yanoi is distinguished from other members of the genus by the combination of the following characters: 14 anal fin soft rays, 19 pectoral fin rays, 97–103 longitudinal scales, a candle flame-shaped marking on the caudal fin, a very low membrane connecting the pelvic fins and absence of a ventral frenum.A. masuii differs from all other congeners by having 92–97 longitudinal scales, the length of the interpelvic connecting membrane relative to the longest pelvic fin ray (0.43–0.66), black blotches on the sides of the chin, and blue spots on the opercle and preopercle.     

Isolation and characterization of microsatellite loci in endangered Cycas changjiangensis (Cycadaceae)

Eight microsatellite loci were isolated from repetitive DNA enriched libraries for Cycas changjiangensis, an endangered endemic species in Hainan Island, China. The number of allele ranged from three to seven. The observed (H _O) and expected (H _E) heterozygosities ranged from 0.0000 to 0.8750 and from 0.2359 to 0.7582, respectively. These microsatellite loci will enrich our scientific understanding for C. changjiangensis conservation.     

Development and characterization of microsatellite markers for the endangered anchialine squat lobster Munidopsis polymorpha

Species of the genus Munidopsis are typically distributed in bathyal and abyssal zones, but the anchialine species Munidopsis polymorpha is an exception. It inhabits a volcanic tube on Lanzarote Island (Canary Islands, NE Atlantic) and is currently listed as endangered due to its highly restricted distribution and degree of endemism. Microsatellite loci were isolated from partial genomic libraries that had been enriched for AC, ACAG, GATA, AAAC and AAG repeat sequences. Eight loci were polymorphic in a sample of 24 individuals. The number of alleles per locus ranged from 2 to 4 with observed and expected heterozygosities ranging from 0.083 to 0.875 and from 0.080 to 0.681, respectively. These markers will be used to evaluate levels of genetic diversity and inbreeding, providing essential information for the development of a management and conservation strategy for this species.     

Genetic architecture in bur oak,Quercus macrocarpa (Fagaceae), inferred by means of spatial autocorrelation analysis

Allozyme variation at seven putative polymorphic gene loci (Fle, Idh, Mdh-2, Mdh-3, 6 Pgd-2, Pgi-2, Pgm) was quantified in an isolatedQuercus macrocarpa population in northwestern Ontario, Canada. The bur oaks studied were slightly less genetically variable (H _e = 0.196,A = 3.43) than theQuercus subg.Lepidobalanus average. To assess the genetic differentiation within the topodeme,Moran's spatial autocorrelation coefficients were used for the nearest-neighbor map and correlograms. Results indicate that most alleles were randomly distributed in space. However, significant genetic patches were detected for twoFle-alleles. Patch width was inferred to be less than 10m. This spatial substructure may be due to limited gene flow and/or demographic factors. Spatially varying selection can be probably excluded as a spatial evolutionary force, since site conditions were very homogeneous.     

A phenetic study of the taxonomic delimitation of Quercus acutifolia and Q. conspersa (Fagaceae)

A phenetic study ofQuercus acutifolia andQ. conspersa (Fagaceae) is carried out. The study was based on morphological characters, and included cluster analysis and principal component analysis. The results revealed that the characters employed so far to separateQuercus acutifolia andQ. conspersa seem to be quite unreliable. Nevertheless, both species can still be recognized as taxonomically different entities, although mostly based on characters of leaf margins. We propose that the individuals that have leaves with dentate-aristate margins correspond toQ. acutifolia, whereas those that have leaves with entire and not dentate-aristate margins belong toQ. conspersa.