Microsatellite loci from the ectomycorrhizal basidiomycete Suillus pictus associated with the genus Pinus subgenus Strobus

Seven microsatellite loci were isolated and characterized from ectomycorrhizal basidiomycete Suillus pictus associated with soft pine species (Pinus subgenus Strobus) using a dual‐suppression‐PCR technique. Microsatellite variation was assessed using 38 isolates of S. pictus sampled from three different local populations in Japan. The number of alleles per locus ranged from three to 12 within populations. These microsatellite loci can be used for studying the allelic variation and genetic structure in/among local populations of S. pictus.     

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.     

GENETIC DIVERSITY AND POPULATION STRUCTURE IN CAMELLIA JAPONICA L. (THEACEAE)

Camellia japonica is a widespread and morphologically diverse tree native to parts of Japan and adjacent islands. Starch gel electrophoresis was used to score allelic variation at 20 loci in seeds collected from 60 populations distributed throughout the species range. In comparison with other plant species, the level of genetic diversity within C. japonica populations is very high: 66.2% of loci were polymorphic on average per population, with a mean number of 2.16 alleles per locus; the mean observed and panmictic heterozygosities were 0.230 and 0.265, respectively. Genotypic proportions at most loci in most populations fit Hardy-Weinberg expectations. However, small heterozygote deficiencies were commonly observed (mean population fixation index = 0.129). It is suggested that the most likely cause of the observed deficiencies is population subdivision into genetically divergent subpopulations. The overall level of population differentiation is greater than is typically observed in out-breeders: The mean genetic distance and identity (Nei's D and I) between pairs of populations were 0.073 and 0.930, respectively, and Wright's Fst was 0.144. Differences among populations appeared to be manifested as variation in gene frequencies at many loci rather than variation in allelic composition per se. However, the patterns of variation were not random. Reciprocal clinal variation of gene frequencies was observed for allele pairs at six loci. In addition, principal components analysis revealed that populations tended to genetically cluster into four regions representing the geographic areas Kyushu, Shikoku, western Honshu, and eastern Honshu. There was a significant relationship between genetic and geographic distance (r = 0.61; P < 0.01). Analysis of variance on allozyme frequencies showed that there was approximately four times as much differentiation among populations within regions, as among regions. It is likely that the observed patterns of population relationships result from the balance between genetic drift in small subpopulations and gene flow between them.     

Microsatellite identification of individual sockeye salmon in Barkley Sound, British Columbia

Population structure of sockeye salmon Oncorhynchus nerka in Barkley Sound, British Columbia, Canada was examined by analysis of microsatellite variation at 14 loci in three populations sampled in each of 3 years. The mean F _ST over all 14 loci was 0·063. Differences among populations accounted for 12 times the variation observed among years within populations. The number of alleles present at a locus was related to the power of the locus to provide accurate identification of individuals to population. The more alleles that were present at a locus, the greater was the power of the locus for individual identification. Individuals were correctly classified to one of three lakes of origin at a rate of 89%, and to one of two river drainages at a rate of 96%.     

COVARIATION OF SELFING RATES WITH PARENTAL GENE FIXATION INDICES WITHIN POPULATIONS OF MIMULUS GUTTATUS

Wright's gene fixation index F and two single-locus effective selfing rates—the selfing rate at loci with fixed alleles, and the selfing rate at loci without fixed alleles—were estimated in five populations of Mimulus guttatus. These two effective selfing rates describe the inbreeding observed at a single locus when both uniparental and biparental inbreeding are practiced. Estimates were made using progeny arrays assayed for six allozyme loci and two morphological loci exhibiting dominance. The average of the two selfing rates computed for subpopulations (ca. 10 m diameter) ranged from 24% to 59%, with a mean of 37%. When computed for populations (ca. 1 km diameter), average selfing rates were about 10% higher. In four populations, the selfing rate at loci with fixed alleles was higher than the selfing rate at loci without fixed alleles. Thus, the covariance of selfing with parental gene fixation was positive. In one of the populations, estimates for individual plants sampled along a transect gave positive correlations for selfing rates and for gene-fixation indices between adjacent plants. A highly positive correlation between selfing rate and gene fixation of individual plants was also observed. In another population, the covariance of selfing with gene fixation was higher for a locus causing leaf spots than for allozyme loci. This covariance is partially caused by 1) variation in homozygosity among neighborhoods and 2) biparental inbreeding within neighborhoods. The consequences of this covariance are discussed.     

Genetic diversity of Ustilago maydis strains

Thirty wild isolates belonging to five different locations in Mexico plus two laboratory strains of Ustilago maydis were characterized by restriction fragment length polymorphism (RFLP) analysis using 23 different clones as probes derived from a PstI library and two restriction enzymes. All loci analysed presented a high level of polymorphism, including one locus with thirty one different alleles. Geographical grouping of the populations was based on Nei's genetic distance and there was no correlation between genetic and geographic distances among these isolates. Our results suggest that DNA fingerprinting is a useful method for detecting genetic variation in populations of U. maydis. This work demonstrated that considerable genetic variation may be present within field populations of U. maydis.     

Genomic variation in cline shape across a hybrid zone

Hybrid zones are unique biological interfaces that reveal both population level and species level evolutionary processes. A genome‐scale approach to assess gene flow across hybrid zones is vital, and now possible. In Mexican towhees (genus Pipilo), several morphological hybrid gradients exist. We completed a genome survey across one such gradient (9 populations, 140 birds) using mitochondrial DNA, 28 isozyme, and 377 AFLP markers. To assess variation in introgression among loci, cline parameters (i.e., width, center) for the 61 clinally varying loci were estimated and compiled into genomic distributions for tests against three empirical models spanning the range of observed cline shape. No single model accounts for observed variation in cline shape among loci. Numerous backcross individuals near the gradient center confirm a hybrid origin for these populations, contrary to a previous hypothesis based on social mimicry and character displacement. In addition, the observed variation does not bin into well‐defined categories of locus types (e.g., neutral vs. highly selected). Our multi‐locus analysis reveals cross‐genomic variation in selective constraints on gene flow and locus‐specific flexibility in the permeability of the interspecies membrane.     

Genetic variation and differentiation in Nordic populations of Elymus alaskanus (Scrib. ex Merr.) Löve (Poaceae)

 To gain information on the extent and nature of genetic variation in Elymus alaskanus, levels and distribution of genetic variation were assessed within and among 13 populations originating from Iceland, Norway, Sweden and Russia using allozymes. The results showed that four (30.7%) of the 13 loci were polymorphic within the species, while the mean percentage of polymorphic loci within the populations was 1.9%. The mean number of alleles per locus for the species was 1.8 and 1.02 across the populations. Genetic diversity at the species level was low (H _es=0.135), and mean population diversity was notably lower (H _ep=0.005). A high degree of genetic differentiation was observed among populations. The salient points emerging from this study are: (1) statistically significant differences were found in allele frequencies among populations for every polymorphic locus (P<0.001), (2) the high mean coefficient of gene differentiation (G _ST) showed that 95% of the total allozyme variation was attributable to differences among populations, and (3) relatively high genetic distances between the populations were obtained (mean D=0.16). The Norwegian populations had the highest genetic diversity as compared with the other populations. Geographical comparisons revealed three different groups of populations clearly differentiated, i.e. Scandinavia (Norway and Sweden), Iceland and Russia. Cluster and principal coordinates analyses revealed the same genetic patterns of relationships among populations. Generally, this study indicates that E. alaskanus contains low allozymic variation in its populations. The implications of these results for the conservation of the species are discussed. Received: 23 October 1998 / Accepted: 19 December 1998     

Genetic analysis of differentiation among breeding ponds reveals a candidate gene for local adaptation in Rana arvalis

One of the main questions in evolutionary and conservation biology is how geographical and environmental features of the landscape shape neutral and adaptive genetic variation in natural populations. The identification of genomic polymorphisms that account for adaptive variation can aid in finding candidate loci for local adaptation. Consequently, a comparison of spatial patterns in neutral markers and loci under selection may help disentangle the effects of gene flow, genetic drift and selection at the landscape scale. Many amphibians breed in wetlands, which differ in environmental conditions and in the degree of isolation, enhancing the potential for local adaptation. We used microsatellite markers to measure genetic differentiation among 17 local populations of Rana arvalis breeding in a network of wetlands. We found that locus RC08604 deviated from neutral expectations, suggesting that it is a good candidate for directional selection. We used a genetic network analysis to show that the allele distribution in this locus is correlated with habitat characteristics, whereas this was not the case at neutral markers that displayed a different allele distribution and population network in the study area. The graph approach illustrated the genomic heterogeneity (neutral loci vs. the candidate locus for directional selection) of gene exchange and genetic divergence among populations under directional selection. Limited gene flow between wetlands was only observed at the candidate genomic region under directional selection. RC08604 is partially located inside an up‐regulated thyroid‐hormone receptor (TRβ) gene coordinating the expression of other genes during metamorphosis and appears to be linked with variation in larval life‐history traits found among R. arvalis populations. We suggest that directional selection on genes coding larval life‐history traits is strong enough to maintain the divergence in these genomic regions, reducing the effective recombination of locally adapted alleles but not in other regions of the genome. Integrating this knowledge into conservation plans at the landscape scale will improve the design of management strategies to preserve adaptive genetic diversity in wetland networks.     

GENETIC VARIATION IN BROMUS TECTORUM (POACEAE): POPULATION DIFFERENTIATION IN ITS NORTH AMERICAN RANGE

Allelic variation in seedlings from 60 North American populations of the alien annual grass Bromus tectorum was determined at 25 loci using starch gel electrophoresis. Populations were collected from four regions; east of the Rocky Mountains, Nevada and California, the Intermountain West, and British Columbia. Compared to other diploid seed plants, genetic variation within these populations of B. tectorum is low: 4.60% of loci are polymorphic per population, with an average of 1.05 alleles per locus and a mean expected heterozygosity of 0.012. Although 2,141 individuals were analyzed, no heterozygous individuals were detected, and consequently, mean observed heterozygosity is 0.000. Extensive deviations from Hardy-Weinberg expectations were observed at every polymorphic locus due to heterozygote deficiencies. The mean genetic identity (Nei's I) between population pairs was 0.980 and indicates a high level of overall genetic similarity among populations. The among-population component of the total gene diversity is high (G_ST ⁼ 0.478), indicating substantial genetic differentiation among populations. These results are consistent with previous reports for highly self-pollinating plants of low genetic variation and substantial genetic differentiation among populations. Despite the lack of genetic variation as measured by enzyme electrophoresis, this weedy grass has become exceedingly abundant in a diverse array of arid environments throughout much of western North America, perhaps due to phenotypic plasticity.     

Allozyme variations in six natural populations of scots pine (<Emphasis Type="Italic">Pinus sylvestris</Emphasis>) in Turkey

Genetic variation in six natural populations of Scots pine (Pinus sylvestris L.) was determined with isoenzyme analyses. For this purpose, haploid female gametophytes of seeds and horizontal starch gel electrophoresis technique were used. A total of 17 loci and 58 alleles were observed in studying 10 enzyme systems. The average proportion of polymorphic loci for populations ranged from 58.8% to 70.6%. The average number of alleles per locus per population was 2.65. The mean estimated expected heterozygosity (He) of populations was 0.294. A rather high proportion of genetic diversity (96.4%) was due to within-population variation and the remaining (3.6%) was due to variation among populations. The level of gene flow (N_em) was found to be 6.69 per generation. Nei’s genetic distance coefficient ranged from 0.006 to 0.027 (mean 0.017) among all possible population pairs. The mean value of Nei’s genetic distance is similar to the values reported for other European Scots pine populations. The low mean value of Nei’s genetic distance among populations is enough to explain low interpopulation variation. According to genetic variation parameters, three out of six populations (Akdagmadeni-Yozgat, Refahiye-Erzincan and Vezirkopru-Samsun) appear to be preferable populations for genetic conservation and forest tree breeding programs.     

Allozyme variation among biotypes of the brown planthopperNilaparvata lugens in the Philippines

Allozyme variation was studied in threeNilaparvata lugens biotypes infesting specific rice varieties and a biotype infesting a weed grass,Leersia hexandra. Of the 20 enzymes inN. lugens for which activity was noted, 9 were polymorphic. Eleven enzyme loci were monomorphic for the same allele in all biotype populations; the rest were polymorphic for two or more alleles. The mean number of alleles per polymorphic locus was 2.3, while the mean number of alleles per locus was 1.5; heterozygosity ranged from 0.02 to 0.06 (biotype 1 > biotype 3 >Leersia-infesting biotype > biotype 2). Allelic frequency differences were observed in five loci among the four biotypes. However, the coefficient of genetic identity (I) of 0.99+ showed that the four biotype populations were genetically close relatives or merely populations ofN. lugens undergoing genetic differentiation. This work was partly supported by a financial grant received from the Directorate for Technical Cooperation and Humanitarian Aid, Switzerland.     

POPULATION GENETIC STRUCTURE IN DIPLOID BLUEBERRY VACCINIUM SECTION CYANOCOCCUS (ERICACEAE)

Vaccinium section Cyanococcus comprises diploid, tetraploid, and hexaploid species (x = 12) all of which are highly self-sterile. In order to assess the distribution of genetic variation within and among the diploids, population genetic analyses of allozyme data were conducted on three species: V. elliottii, V. myrtilloides, and V. tenellum. Populations were located throughout the range of these taxa in eastern North America. Data were collected at 20 loci, 12 of which are mendelian based on formal genetic analyses. Consistent with expectations for out-crossing taxa, these species exhibited high levels of variation within populations. Eighteen loci were polymorphic with the number and frequency of alleles varying among taxa. Mean values for the number of alleles per polymorphic locus, proportion of polymorphic loci, and expected heterozygosity were 2.8, 48.2, and 0.148, respectively. High infraspecific genetic identities exceeding 0.9 indicated that these taxa are homogeneous. All populations were in Hardy-Weinberg equilibrium with slight heterozygote excess observed in V. myrtilloides and V. tenellum. Although total genetic diversity was lower than that observed for other predominantly out-crossing species, it was apportioned similarly. The majority could be attributed to differences among individuals within populations. Conspecific populations were relatively undifferentiated with genetic differentiation similar to other self-incompatible species.     

Characterization of microsatellite markers from the gastropod genus Melongena

Eight microsatellite loci were isolated and characterized from the Florida crown conch (Melongena spp.). Approximately 500 individuals from 20 populations encompassing all known Melongena spp. groups in Florida and Alabama were genotyped at all loci. High levels of variation were found with eight to 28 alleles per locus and observed per locus heterozygosity ranging from 0.148 to 0.779 (mean 0.624). One locus, Mco5, was monomorphic in more than half of the populations. These loci can provide useful information for a variety of studies ranging from parentage to population‐level analyses of the Florida crown conch.     

Identification of microsatellite markers in <Emphasis Type="Italic">Hieracium pilosella</Emphasis> L.

In order to investigate the genetic diversity and structure of Hieracium pilosella L., we developed eleven pairs of primers for nuclear microsatellites through screening of three genomic DNA enriched libraries. About 127 individuals from six locations of the Trentino region (Italy) were analysed. These loci were all polymorphic and displayed 5–33 alleles per locus. These microsatellite markers constitute an efficient tool in investigating the genetic patterns of H. pilosella L. populations.     

Development and characterization of microsatellite markers to study population genetic diversity of Przewalski's naked carp Gymnocypris przewalskii in China

Two populations of Przewalski's naked carp Gymnocypris przewalskii, 30 individuals per population, were screened for 10 microsatellite loci. Moderate allele variation was found in these loci with two to eight alleles per locus. The expected and observed heterozygosity ranged from 0·019 to 0·805 and from 0·160 to 0·575, respectively.     

Genetic Variation and Differentiation among Populations of Chum Salmon Oncorhynchus ketaWalbaum from Southern Russian Far East

Allozyme variation of populations of chum salmon Oncorhynchus ketafrom southern Russian Far East was examined. Of 55 loci screened, 31 were polymorphic. Within-population variation accounted for most of the allele diversity; F _STaveraged over loci was 0.052. Linkage disequilibrium was found in less than 5% of locus pairs in the chum population examined. Analysis of within- and among-population variance components of linkage disequilibrium using D-statistics (Ohta, 1982) showed that most genetic variation was distributed among populations.     

GENETIC CONSEQUENCES OF RARITY IN ASTER FURCATUS (ASTERACEAE), A THREATENED,SELF-INCOMPATIBLE PLANT

Aster furcatus is a rare, self-incompatible plant with fewer than 50 known populations throughout its range. We verified self-incompatibility in A. furcatus by conducting experimental self- and cross-pollinations and by examining seed set in a small population comprised of a single clonal genet. We examined variation at 22 electrophoretic loci in 23 populations of A. furcatus from across its range in Wisconsin, Illinois, Indiana, and Missouri. Except for two rare alleles found in single individuals in three populations, all loci but one of those examined were fixed for single alleles. The only variable locus (triosephosphate isomerase, TPI-1) tended to exhibit genotype frequencies in Hardy-Weinberg equilibrium or with a slight excess of heterozygotes. Although overall gene diversity was extremely low, TPI genotype frequencies were indicative of an outcrossing plant. We examined the subpopulation genetic structure among clonal plants within one Wisconsin population in greater detail. F statistics indicated that much of the genetic variation at the polymorphic TPI locus was due to differentiation among populations. We discuss the implications of self-incompatibility and low levels of genetic variation for the evolution and conservation of Aster furcatus and other rare plants with similar breeding systems.     

种间转移扩增筛选仙湖苏铁微卫星位点及其遗传多样性研究

运用刺叶苏铁、葫芦苏铁、海南苏铁的SSR引物,在仙湖苏铁中进行种间转移扩增,筛选得到7对引物能扩增出清晰的特异带,其中3对引物的扩增产物具有多态性.为验证微卫星的真实性,扩增产物切胶回收后克隆测序.结果表明：重复单元的数目变化是扩增片段长度多态性的主要来源.运用筛选出的3对 SSR标记对4个仙湖苏铁野生种群进行遗传结构研究,等位基因数从2~5,杂合度从0．000~0．667,期望杂合度为0．000~0．610.种群两两遗传分化系数从0~0．382.总体上仙湖苏铁遗传多样性水平低,而种群间遗传分化显著.STRUCTURE分析结果表明,4个野生种群可被分配到3个假想的遗传簇.BOTTLENECK 分析结果表明种群近期没有遭遇瓶颈效应.     

Discriminating coho salmon (Oncorhynchus kisutch) populations within the Fraser River, British Columbia, using microsatellite DNA markers

Three microsatellite loci were used to examine genetic variation among 16 coho salmon ( Oncorhynchus kisutch ) populations within the Fraser River drainage system, in British Columbia, Canada. Each locus was highly polymorphic with 30 alleles at the Ots 101 locus, 15 alleles at the Ots 3 locus and 38 alleles at the Ots 103 locus. Average observed heterozygosities were 86.1%, 70%, and 56.1%, respectively. With the exception of the Dunn and Lemieux River populations, Chi-square tests and F _ST values indicated that all populations had significantly different allele frequencies. Two distinct population groups within the Fraser River drainage were observed. Lower Fraser River populations were strongly differentiated from populations spawning in the upper Fraser River, which includes the Thompson River (a tributary flowing into the upper Fraser) and the portion of the Fraser River beyond the precipitous Fraser River canyon. This regional population structure may have resulted from colonization of the upper and lower Fraser River regions by different founder populations following Pleistocene glaciation, and be maintained by adaptive differences between the two groups of coho salmon. Coho salmon populations in the upper Fraser and Thompson River drainages form an evolutionarily significant unit (ESU) of importance for conservation of biodiversity in coho salmon. Microsatellite DNA loci show promise as technically simple and highly informative genetic markers for coho salmon population management.