Isolation and characterization of polymorphic microsatellite markers in <Emphasis Type="Italic">Cupressus </Emphasis><Emphasis Type="Italic">chenggiana</Emphasis> S. Y. Hu (Cupressaceae)

Cupressus chenggiana S. Y. Hu (Cupressaceae) is an endemic and endangered conifer species in southwest China. In order to study the population genetics and design the effective conservation methods, we aimed to develop microsatellite primers for this species in the present study. We developed eight new microsatellite loci for this species through biotin capture method. Polymorphism of each locus was further assessed in 18 individuals from three geographically distant populations. The number of alleles per locus ranged from 6 to 11 with an average of 8.13. The observed and expected heterozygosities ranged from 0.219 to 0.296 and from 0.374 to 0.470, with averages of 0.254 and 0.417, respectively. We further found that three of nine microsatellite loci developed previously for another congeneric species showed polymorphic banding patters. We performed primer-crossing tests of these loci in the other two congeneric species which are closely related to C. chenggiana (C. gigantea and C. duclouxiana). These microsatellite markers would be effective for analyzing genetic diversity and population genetic structure of this species and its morphological differentiation with the close relatives.     

Isolation of polymorphic microsatellite loci from the endangered mudflat crab Chasmagnathus convexus (Crustacea: Decapoda: Varunidae)

Chasmagnathus convexus is a mudflat crab that is distributed throughout East Asia. However, most populations of this species in Korea are endangered due to continuing seashore developments and habitat fragmentations. In this study, we isolated and characterized nine microsatellite loci from C. convexus. The number of alleles per locus ranged from 3 to 23. The expected heterozygosities ranged from 0.145 to 0.974 and the observed heterozygosities ranged from 0.150 to 0.947. Cross-species amplifications for 10 other Varunid crab species were performed. These microsatellite markers could be useful for planning conservation strategies for C. convexus and related species.     

Development and characterization of microsatellite DNA markers for the Alpine plant species Campanula thyrsoides

We isolated and characterized eight polymorphic microsatellite markers for the Alpine plant species Campanula thyrsoides (Campanulaceae). Number of alleles per locus ranged from six to 12 and the observed heterozygosity was between 0.529 and 0.900. Observed vs. expected heterozygote deficits were significantly different in one out of eight loci following Bonferroni's correction for multiple tests. We did not find evidence for linkage disequilibrium between locus pairs. The microsatellite markers are being used for the study of genetic variation and gene flow within and among populations of C. thyrsoides in the Swiss Alps.     

Identification and characterization of microsatellite DNA markers developed in ide Leuciscus idus and Siberian roach Rutilus rutilus

The first five microsatellite markers for the ide, Leuciscus idus, and four microsatellite markers for the Siberian roach, Rutilus rutilus, were designed. Cross‐amplification of ide markers was examined in Siberian roach and vice versa. The number of alleles per locus ranged from three to 13 in ide and from two to eight in roach. The expected heterozygosity ranged from 0.313 to 0.909 in ide and from 0.119 to 0.775 in roach. These markers could be used to evaluate the genetic population structure of these species and other fish from the Cyprinidae family.     

Development and characterization of microsatellite loci for Rosa odorata var. gigantea Rehder & E. H. Wilson (Rosaceae)

Eighteen microsatellite markers were developed from Rosa odorata var. gigantea (Rosaceae), including 11 new microsatellite markers and 7 modified microsatellite loci having been developed from other Rosa species. About 27 wild individuals from 3 populations were used to screen polymorphism of these 18 microsatellite makers. The average allele number of these markers was 3.9 per locus, ranging from 2 to 9. The expected and observed heterozygosities varied from 0.2711 to 0.8043 and from 0.0370 to 0.5556, respectively. Cross-species amplification in other eight Rosa species showed their potential use for congeneric species. These microsatellite primers will be used for population genetics studies, constructing genetic linkage maps or locating quantitative trait locus (QTL) of R. odorata var. gigantea and related species.     

Ten new microsatellite loci for analysis of genetic diversity in isolated populations of the Alpine land snail <Emphasis Type="Italic">Cylindrus obtusus</Emphasis>

In this study we characterized 10 polymorphic microsatellite markers for the land snail Cylindrus obtusus, an endemism of the Austrian Alps with a distribution in isolated populations above approximately 1,600 m. The microsatellite loci were analyzed in 44 individuals from two populations. Number of alleles per locus ranged between two and eight. Observed heterozygosity ranged between 0.00 and 1.00, and expected heterozygosity between 0.09 and 0.72. No significant linkage disequilibrium was found between pairs of loci. One of the sampled populations (Dachstein) showed no deviation from Hardy–Weinberg equilibrium and no presence of null alleles, whereas the other one (Schneeberg) did. These diverging results probably reflect differences in population structure rather than characteristics of the microsatellite loci and underline the usefulness of these markers for studying genetic diversity, population structure and differentiation in C. obtusus.     

Polymorphic microsatellites for the study of newly established populations of the gastropod Cyclope neritea

Both human‐mediated introductions and climatic changes may promote the settlement of species in new areas outside of their natural geographical range. To investigate the settlement of recently established populations of the neogastropod Cyclope neritea, we developed eight microsatellite markers. Their usefulness was studied in two native populations previously found to be monomorphic with mitochondrial markers. The eight loci were found to be polymorphic in both populations, with two to 18 alleles per locus. This result shows promise for these loci in studies of recently founded populations of C. neritea.     

Development and characterisation of microsatellite markers for <Emphasis Type="Italic">Cordia verbenacea</Emphasis> (Boraginaceae), an important medicinal species from the Brazilian coast

Cordia verbenacea DC (Boraginaceae) is a Brazilian coastal shrub species extensively utilized in the folk medicine and also by the pharmaceutical industry. Despite its enormous economic and pharmacological potential, little is known about its reproductive biology, genetic structure and genetic diversity of natural populations. Due to the rapid degradation of its natural habitat, this knowledge became imperative. We developed a set of eight highly polymorphic microsatellite markers for the study of genetic structure and diversity of C. verbenacea natural populations and germplasm collections. Six of them revealed a multibanded pattern typical of polyploid species. The total number of bands per locus ranged from 2 to 12 and the number of bands per locus per individual ranged from 1 to 8. The primer pairs presented here will be used to generate information to guide conservation programs and rational exploitation of C. verbenacea.     

Isolation and characterization of microsatellite loci in the highland papaya Vasconcellea × heilbornii V. Badillo (Caricaceae)

Nine polymorphic microsatellite loci were identified in the tropical plant Vasconcellea ×heilbornii and used to estimate allelic diversity in two populations of southern Ecuador. Allelic richness ranged from two to five alleles, observed heterozygosity ranged from 0.150 to 0.947 and expected heterozygosity ranged from 0.186 to 0.701. Most of these markers also amplified microsatellite loci in two other Vasconcellea species (Vasconcellea stipulata and Vasconcellea cundinamarcensis). Hence, these markers will be useful for population genetic analysis and the evaluation of genetic diversity and gene flow in these species.     

Development and characterization of polymorphic microsatellite DNA markers for Sasa senanensis (Poaceae: Bambuseae)

Sasa senanensis is a dwarf bamboo species distributed on the floors of cool temperate forests in Japan and adjacent regions. We isolated eight polymorphic microsatellite loci from this species. The number of alleles ranged from two to eight and the observed heterozygosity per locus from 0.13 to 0.74. Seven of the eight loci were also polymorphic in Sasa nipponica. Most of these markers were successfully amplified in other dwarf bamboo species. These markers will be useful for investigating clonal structure and population genetics in some dwarf bamboo species.     

Characterization of microsatellite loci in two closely related Lissotriton newt species

We have developed eight di- and tetranucleotide Lissotriton microsatellite markers. Eight loci were polymorphic in the palmate newt Lissotriton helveticus and six were polymorphic in the smooth newt L. vulgaris. Polymorphism detected in 33 and 37 individuals per species ranged from 3 to 15 alleles. These markers are suitable for the investigation of population structure, genetic variation and taxonomic identification in the two focal species, and may also be of use in other Lissotriton–Triturus species.     

Development of microsatellite markers for the endangered grassland perennial herb Vincetoxicum atratum (Apocynaceae–Asclepiadoideae)

Eight microsatellite markers were developed for the endangered grassland perennial herb Vincetoxicum atratum. The number of alleles ranged from 4 to 14, and the expected heterozygosities were from 0.575 to 0.933 in a population of V. atratum. Five of the eight loci did not significantly deviated from the Hardy–Weinberg equilibrium. All eight loci were tested for cross-species amplification in five other species of Vincetoxicum in Japan. These microsatellite loci will be useful for conservation genetics of V. atratum and other species of Vincetoxicum.     

Development of polymorphic microsatellite markers for the fungal tree pathogen Cryphonectria eucalypti

Polymorphic microsatellite DNA markers were developed from a single spore isolate of Cryphonectria eucalypti collected from a Eucalyptus stem canker in South Africa. Markers were obtained using the enrichment technique known as fast isolation by AFLPs of sequences containing repeats (FIASCO). Ten polymorphic markers were isolated, of which, two were discarded due to their high polymorphism in the flanking region. The mean number of alleles produced by the remaining eight markers from 20 isolates was 7.25, and alleles per locus ranged from four to 12. The markers will be used to study populations of C. eucalypti.     

Isolation and characterization of microsatellite loci from Corydalis ambigua (Papaveraceae)

Primers for seven unlinked and highly polymorphic microsatellite loci were developed for Corydalis ambigua to investigate its mating system and population genetics. This species can be used to explore the impact of an alien bumblebee on plant reproduction. Genetic diversity and other population genetic parameters were estimated in two populations with and without the alien bumblebee. The number of alleles per locus ranged from 4 to 20, and observed heterozygosities ranged from 0.355 to 0.969. These markers can be applied to study mating systems and population genetics in C. ambigua.     

Isolation and characterization of eight polymorphic microsatellite loci for the critically endangered Arenaria nevadensis (Caryophyllaceae)

We report on the isolation and characterization of eight microsatellite markers from enriched libraries for Arenaria nevadensis, one of the most critically endangered plant species in the Iberian Peninsula. These are the first microsatellite loci reported for Arenaria species. The number of alleles ranged from two to eight, and the expected heterozygosity from 0.067 to 0.873. These markers will be useful for characterizing the genetic diversity in A. nevadensis and understanding its population structure, and will provide important genetic data for the conservation and recovery of this species.     

Identification and characterization of nuclear microsatellite loci in Abies alba Mill.

Eleven polymorphic nuclear microsatellite markers for Abies alba Mill. were developed from an enriched genomic library. An average of 5.2 alleles per locus and a mean expected heterozygosity of 0.532 were found in a sample of 24 Abies alba individuals from different populations within Europe. These loci can be used in studies of genetic diversity for parentage analysis and for estimation of gene flow in silver fir populations. Moreover, successful amplifications were obtained for eight other Mediterranean Abies species, suggesting that these loci may be useful for similar applications in other fir species.     

Isolation and characterization of eight microsatellite loci from the house finch (Carpodacus mexicanus)

I isolated the first set of polymorphic microsatellite markers from the house finch, Carpodacus mexicanus, a well‐studied North American bird species, as part of an effort to compare levels of genetic diversity in introduced and native populations. Here, I describe eight independently assorting microsatellite loci screened for polymorphism using 40 house finches. Polymorphism levels ranged from six to 14 alleles (mean = 10.6), making these markers a powerful tool for paternity and population level analyses of this widely distributed North American species.     

基于SSR标记探讨三种金花茶植物的遗传多样性和遗传结构

薄叶金花茶、小花金花茶和小瓣金花茶是三种濒危金花茶植物,为了解珍稀濒危植物遗传多样性和遗传结构,该研究利用微卫星标记对他们的7个种群共184个个体进行了遗传多样性和遗传结构分析。结果表明:11个位点共检测到等位基因92个。在物种水平上,小瓣金花茶平均等位基因数(N_A)为3.9、有效等位基因数(N_E)为2.328、观测杂合度(H_o)为0.520、期望杂合度(H_e)为0.501,高于薄叶金花茶和小花金花茶。在种群水平上,有效等位基因数(N_E)在1.788～2.466之间,期望杂合度(H_e)在0.379～0.543之间;种群间遗传分化系数(FST)在0.143 7～0.453 3之间,种群间基因流(N_m)在0.301 5～1.488 9之间。AMOVA分子变异分析显示65.72%的变异存在于种群内。三种金花茶具有较低水平的遗传多样性和高水平的种群间遗传分化。STRUCTURE和PCoA种群遗传结构分析结果将取样种群分为2组,即薄叶金花茶和小花金花茶大部分个体分为一组,小瓣金花茶大部分个体分为一组。现存所有种群应根据实际情况尽快采取就地保护或迁地保护措施。     

Isolation and characterization of eight microsatellite loci from <Emphasis Type="Italic">Lycorma delicatula</Emphasis> (White) (Hemiptera: Fulgoridae) for population genetic analysis in Korea

Lycorma delicatula (White) is native to China but is becoming an important insect pest in Korea. Polymorphic DNA markers like microsatellites are widely used for characterizing dispersal patterns and capacity of invasive insect pests which can contribute to designing effective management of the species. To facilitate such population genetic studies of L. delicatula in Korea, we isolated and characterized eight microsatellite loci for L. delicatula using a hybridization-biotin enrichment method. We further used these novel microsatellite loci to determine population genetic parameters for 33 L. delicatula specimens collected from Cheonan, South Korea where outbreaks of this species were first reported in Korea. The number of alleles per locus ranged from three to ten, with an average of 6.25. The mean expected (H _E) and observed heterozygosities (H _O) were 0.575 and 0.626, respectively. The eight loci showed no deviation from Hardy–Weinberg equilibrium according to the adjusted significance threshold (P = 0.00625), and there was no linkage disequilibrium between each pair of these eight markers. Bayesian cluster analysis using the program structure revealed no evidence of genetic structuring in L. delicatula samples from Cheonan. These new microsatellite markers will be widely applicable to future ecological genetic studies of L. delicatula, including assessment of the level of gene flow and genetic connectivity among populations that are necessary for effective management and monitoring of the species.     

A set of polymorphic microsatellite loci for Vriesea gigantea and Alcantarea imperialis (Bromeliaceae) and cross‐amplification in other bromeliad species

Fifteen polymorphic microsatellite markers were isolated and characterized in two species of Bromeliaceae: Vriesea gigantea and Alcantarea imperialis. The number of alleles observed for each locus ranged from three to 16. The loci will be used for studies of the genetic structure of natural populations, reproductive biology, and evolutionary relationships among and within these genera. A cross‐amplification test in 22 taxa suggests that the markers will be useful for similar applications in numerous other bromeliad species.