IDENTICAL MAJOR GENE LOCI FOR HEAVY METAL TOLERANCES THAT HAVE INDEPENDENTLY EVOLVED IN DIFFERENT LOCAL POPULATIONS AND SUBSPECIES OF SILENE VULGARIS

Heavy metal tolerant Silene vulgaris plants, originating from different metalliferous sites in Germany and one in Ireland, were crossed to each other and to nontolerant plants from a nonmetalliferous site in The Netherlands. Analysis of the crosses suggested that there were two distinct major gene loci for zinc tolerance among a total of five tolerant populations. The tolerance loci for zinc, copper, and cadmium in the Irish plants were shown to be identical with those in the German populations. It is argued that the occurrence of common major genes for tolerance among different geographically isolated populations must have resulted from independent parallel evolution in local nontolerant ancestral populations. Each of the tolerances studied seems to be controlled by only a few specific major genes.     

Genetic variation of chloroplast and nuclear markers in natural populations of hazelnut (Corylus avellana L.) in Germany

Corylus avellana L. (hazel) is a long-lived, monoecious and wind-pollinated shrub species, widespread all over Europe. In Germany, hazel is intensively traded and planted, and thus is of central interest from a nature conservancy point of view. To assess the within- and between-population differentiation of hazel, 20 natural populations (18 from Germany, one from Italy and one from Hungary) were investigated genetically. Seven isozyme systems comprising 11 gene loci were analysed in up to 100 samples (average 92.6) per population, amplified fragment length polymorphisms (AFLP) were analysed in up to 50 samples (average 47.4) and nine cpDNA-SSR markers were assessed in 20 samples per population. Results for overall isozyme variability with Na 2.46 alleles per locus, allelic diversity (Ne) 1.39, expected heterozygosity He 21 % and 79 % polymorphic loci were in accordance with the findings of previous studies. The respective values for AFLPs were lower, but both marker systems revealed the same level of about 3.5 % differentiation between populations. For cpSSR only the Italian sample showed within-population variation and the two haplotypes were completely differentiated from all other populations expressing a unique genetic structure with one single haplotype. Among the three marker systems AFLPs showed the best ability to differentiate between populations. While only one isozyme locus revealed significant differentiation, 41 AFLP loci showed highly significant differentiation between all populations, but 26 loci when only German populations were considered. Consequently geographic differentiation analyses focused mainly on molecular markers. Mantel tests showed significant correlations between genetic and geographic distance, but in the unweighted pair-group method with arithmetic mean analyses, adjacent populations did not always form clusters. While chloroplast markers were able to clearly distinguish only the Hungarian population, the nuclear markers revealed clear spatial genetic structures. The correlations between geographic and genetic distance was high for AFLPs. The correlograms illustrate this effect for all populations as well as for the German populations.     

欧洲中部刺槐种源群体等位酶变异研究

从欧洲中部及美国收集了18个刺槐种源种子,以2年生苗木为材料,采用水平切片淀粉凝胶电泳和聚丙烯酰胺凝胶电泳对11个酶系统进行了检测,共发现20个酶位点,其中14个为多态位点,多态性位点百分数为70％。在参加计算的7个酶系统的12个多态性位点中,平均每个位点的等位基因数(A)和平均每位点等位基因有效数(Ae)为2.733和1．794,平均每个位点的实际杂合度(Ho)和预期杂合度(He)为0．368和0．400,固定指数(F)为0．080,绝大多数位点固定指数为正值。各基因位点遗传参数与12个位点平均值进行相关分析表明,Fe-6和Lap-a的大部分群体遗传参数(A、Ae、Ho、He等)与平均值存在紧密的相关关系,其他位点各项遗传参数和总平均值相关不明显,这些位点的重要性可能比相关性较好的位点更大。德国8个种源群体间遗传距离变化在0．09-0．26之间,来自匈牙利的6个种源之间遗传距离很小,绝大部分均在0．11以下;德国的8个种源与匈牙利和斯洛伐克的8个种源之间的遗传距离变化在0．09-0．24之间;来自美国的2个种源,与欧洲的种源间的遗传距离在0．09-0．23之间,这一距离没有超出欧洲种源之间的差异。匈牙利和斯洛伐克的8个种源群体的各项遗传参数均高于德国种源,表明这两个国家的遗传多样性水平较高。同时来自这两个国家的种源群体间变异系数变化在2．92％-9．97％,说明群体之间的遗传差别很小。德国的8个种源群体各项遗传参数相对较低,但群体间变异系数较高,群体间遗传差别较大。以各个国家为群体单位,4个群体间各项遗传参数的变异均较小,变异系数变化在3．861％-5．139％,表明刺槐种源间地理变异模式不明显。     

Genetic variability and differentiation of wild and cultured tench populations inferred from microsatellite loci

Nine species-specific microsatellites were used to characterize 792 tench, Tinca tinca (L.), from 21 wild and cultured populations. Seven loci were polymorphic expressing four to 22 alleles. A Spanish cultured strain was homozygous at all loci for all individuals studied. Low variability was also observed in a wild population from Sapanca Lake, Turkey and a Chinese cultured strain. In contrast, the highest variabilities were found in wild tench from lake Felchowsee (average number of alleles), and the cultured strain from Königswartha (average heterozygosity), both from Germany. Genetic differentiation between populations was moderate to high. The smallest genetic distances were found between the geographically most distant populations. A Neighbor-Joining tree showed only two major clades consisting of 4 and 17 populations, respectively. Within the smaller clade the Turkish wild and Spanish and Chinese cultured tench formed a sub-cluster with 100% bootstrap support. Possible reasons for the latter unexpected grouping are discussed.     

Biochemical and DNA markers yield strikingly different results regarding variability and differentiation of roe deer (Capreolus capreolus, Artiodactyla: Cervidae) populations from northern Germany

Three mainland and two island roe deer ( Capreolus capreolus ) populations with a total sample size of 105 individuals from Schleswig–Holstein, northern Germany, were analysed with regard to genetic variability within and differentiation among populations as revealed by eight allozyme loci known to be polymorphic in roe deer, eight microsatellite loci and 404 bp of the mitochondrial control region. Surprisingly, the allozymes were completely monomorphic, but microsatellite and control region variability were high. Hypotheses as to demographic reasons for the variability patterns found, including bottlenecks, founder effects and translocations, are put forward. There were no statistically significant differences between the island and the mainland populations in terms of genetic variability as measured by expected heterozygosity, inbreeding coefficient and allelic richness. The correlations of the various variability indices were not statistically significant after Bonferroni correction. Nevertheless, there was a clear tendency for differentiation indices to yield concordant results for microsatellite and mitochondrial markers.     

T-Specific DNA Polymorphisms among Wild Mice from Israel and Spain

Lehrach and his coworkers have isolated a series of DNA probes that specifically hybridize with different regions of mouse chromosome 17 within the t complex. The probes display restriction fragment length polymorphisms, RFLPs, which are specific for the t haplotypes in all laboratory mouse strains tested thus far. Some of these probes have been used to test wild mice populations for these t-associated DNA forms. It is demonstrated that populations from Germany, Switzerland, Italy, Greece, Yugoslavia, Australia, Costa Rica, and Venezuela contain chromosomes in which all the tested DNA loci display the t-specific polymorphisms. The frequency of mice carrying these chromosomes is as high as 31%. Wild mice from Israel and Spain, on the other hand, carry chromosomes displaying t-specific DNA forms only at one or two of the probed loci, while the other loci carry the wild-type (+) forms. These chromosomes thus resemble the partial t haplotypes known from the study of laboratory mice. One possible interpretation of these findings is that these DNA polymorphisms contributed to the assembly of the complete t haplotypes and that these haplotypes may have originated in the Middle East.     

Postglacial colonisation of western Central Europe by Polyommatus coridon (Poda 1761) (Lepidoptera: Lycaenidae): evidence from population genetics

The genetic population structure of Polyommatus coridon (Poda 1761) over large regions of France, Italy and Germany was studied by allozyme electrophoresis. The genetic diversity within populations was high for all parameters analysed (number of alleles 2.72; observed and expected heterozygosity 19.6% and 20.3%, respectively; percentage of polymorphic loci: total: 76.4% and, with polymorphism if the frequency of the commonest allele is below 95%: 53.1%), whereas genetic differentiation between populations was comparatively low (FST = 0.021 +/- 0.002). The mean number of alleles declined significantly from southern to northern populations (r = -0.53, P = 0.0005). Similar effects were found also for other parameters of genetic diversity. This is interpreted as a loss of genetic diversity during postglacial expansion. However, samples from France and Italy had similar patterns of genetic diversity indicating no significant loss in this region. Populations from southern Germany were genetically uniform, well differentiated from French populations and showed a significant loss of genetic diversity. Probably, this is due to a bottleneck during passing through the Burgundian Gap, which is a migration corridor from north-eastern France to southern Germany. In contrast to southern German populations, western German populations were not well differentiated from French populations. Nevertheless, they were genetically impoverished, probably as a result from local bottlenecks and post-expansion phenomena.     

Development of microsatellite markers for noninvasive DNA samples of the eastern green lizard Lacerta viridis

We report the isolation and characterization of microsatellite markers for the eastern green lizard (Lacerta viridis) usable for noninvasive samples. Ten polymorphic loci were obtained by screening 3000 recombinant clones and tested on 39 noninvasive DNA samples of individuals from different locations of the Danube area, Germany. Allelic richness ranged from three to nine alleles, the observed heterozygosity from 0.33 to 0.80, and the expected heterozygosity from 0.36 to 0.85. Our 10 loci, along with another 12 loci previously described, deliver effective analytical tools to analyse the genetic variability and to assess the social structure of eastern green lizard populations.     

Isolation of ten tetranucleotide microsatellite loci in the Northern Wheatear (Oenanthe oenanthe)

We isolated 10 polymorphic microsatellite DNA loci from the Northern Wheatear (Oenanthe oenanthe) and optimized these for future studies in population genetics and behavioural ecology. The loci were screened for polymorphism using 107 individuals from one population in Germany. The primers amplified loci with high numbers of alleles ranging from two to 20 alleles per locus.     

Low diversity but high differentiation: the population genetics of Aglaope infausta (Zygaenidae: Lepidoptera)

Aim Aglaope infausta is a thermophilous Zygaenid of Atlanto-Mediterranean origin, distributed in Portugal, Spain, France and north-western Italy reaching its north-eastern distribution limit in western Germany. The local, regional and inter-regional genetic structure of this species is studied in this analysis. Location and methods The allozymes of 456 individuals from 12 populations (11 from western Germany and one from southern Portugal) were studied by electrophoresis. Results Six of the 19 loci analysed were monomorphic. Genetic differentiation between populations was high (F_ST: 0.404), while the mean genetic diversity was low (H_e: 3.4%). Most (96.5%) of the genetic variance between populations was between the Portuguese and the German samples, but also the differentiation within Germany was considerable (F_SR: 0.101). In Germany, A. infausta occurs in two major regions (middle Rhine and Nahe) that are geographically separated, and 55.5% of the genetic variance was found between these two regions. The populations of both areas do not differ in their genetic diversity, but those of the middle Rhine have significantly higher genetic distances among them than the Nahe populations (0.020 and 0.015, respectively). F_ST was also higher in the middle Rhine region than in the Nahe region (0.089 and 0.045, respectively). Main conclusions Aglaope infausta shows a very low level of genetic heterogeneity for a lepidopteran species. However, this genetic poverty is not affecting the species’ viability. During the ice ages, differentiation into two genetic lineages occurred, most probably in a south-western and a south-eastern differentiation centre in Iberia. The gap in the distribution range in Germany is clearly reflected in the genetic structure. This differentiation must have developed relatively quickly because western Germany most probably was colonized during the climatic optimum 6000 years ago.     

Genetic differentiation of Anopheles claviger s.s. in France and neighbouring countries

An investigation of polymorphism of 11 autosomal and one sex-linked allozyme loci was made on 18 samples of Anopheles claviger Meigen (Diptera: Culicidae) from localities across France and neighbouring sites in Germany and Switzerland, plus one sample of Anopheles petragnani Del Vecchio from the French Pyrénées. Genetic differentiation between these two sibling species was confirmed (Nei genetic distance 0.33-0.44) and two genetically distinct groups of populations were identified within An. claviger. These two forms of An. claviger showed contiguous geographical distributions, Group I found across western and Central France, Group II in eastern France and nearby parts of Germany and Switzerland. The two groups were in contact in a region near the Rhone Valley where two intermediate samples were found. The taxonomic significance of this finding is discussed in the context of the recent climatic history of Europe and in relation to the vector potential of each member of the An. claviger complex.     

Genetic Variation Within and Among Metal-Tolerant and Non-Tolerant Populations of Armeria maritima (Mill.) Willd. s.l. (Plumbaginaceae) in Central and Northeast Germany

Abstract: Six metal-tolerant populations and sub-populations of Armeria maritima ssp. halleri, ssp. hornburgensis, and ssp. bottendorfensis and two non-tolerant populations of ssp. elongata in Central and Northeast Germany have been analysed using RAPD markers. The populations show very strong genetic differentiation (Φ_ST = 0.46), corresponding gene flow between them is low (N_em = 0.29). A moderate positive correlation between the matrices of genetic and geographical distances was found between the seven populations and sub-populations of central Germany (r = 0.68, p < 0.001). Calculated parameters of genetic variability are molecular variance, percentage of heterozygosity and percentage of polymorphic loci. A significant correlation between population size and parameters of genetic variability was not recognisable. Genetic structure was investigated by an analysis of molecular variance (AMOVA). The studied populations show strong genetic differentiation. Genetic variation within populations ("normal" as well as metalliferous) is higher (53.9 %) than among them (46.1 %). Six hypotheses of possible genetic relatedness between the studied populations have been tested by AMOVA. A data set structure above the populational level is hardly recognisable. It was impossible to combine the populations to edaphic (tolerant and "non-tolerant") or taxonomic groups. A. maritima ssp. halleri of the north Harz mountains and ssp. hornburgensis are clearly separated from a geographical group containing all other populations (across taxonomic and edaphic boundaries). These results are a further indication for a polyphyletic origin of metal-tolerant populations of A. maritima s.l. by multiple colonizations of metalliferous sites from neighbouring populations on non-metalliferous soil.     

Lack of reproductive isolation between the Western and Eastern phylogroups of the tench

The Eurasian range of the tench distribution is subdivided into deeply divergent Western and Eastern phylogroups evidenced by nuclear and mitochondrial DNA sequence markers. A broad zone of overlap exists in central and western Europe, suggesting post-glacial contact with limited hybridisation. We conducted a population genetic test of this indication that the two phylogroups may represent distinct species. We analysed variation at introns of nuclear genes, microsatellites, allozymes and mitochondrial DNA in populations from two postglacial lakes within the contact zone in Germany. The test is based on the expectation that in the presence of strong barriers to reproduction, a hybrid population will show genome-wide associations among alleles and genotypes from each phylogroup even after hundreds of generations of interbreeding. In contrast to this expectation, no consistent significant deviations from linkage and Hardy–Weinberg equilibria were found. Samples from both lakes did show significant disequilibria but they were limited to individual loci and were not concordant between populations, and were not robust to the method used. The single consistent association can be attributed to physical linkage between two microsatellite loci. Thus, results of our study support the hypothesis of free interbreeding between the two phylogroups of tench. Therefore, although the phylogroups may be considered as separate phylogenetic species, the present data suggest that they are a single species under the biological species concept.     

Extent of linkage disequilibrium between the androgen receptor gene CAG and GGC repeats in human populations: implications for prostate cancer risk

While studies have implicated alleles at the CAG and GGC trinucleotide repeats of the androgen receptor gene with high-grade, aggressive prostate cancer disease, little is known about the normal range of variation for these two loci, which are separated by about 1.1 kb. More importantly, few data exist on the extent of linkage disequilibrium (LD) between the two loci in different human populations. Here we present data on CAG and GGC allelic variation and LD in six diverse populations. Alleles at the CAG and GGC repeat loci of the androgen receptor were typed in over 1000 chromosomes from Africa, Asia, and North America. Levels of linkage disequilibrium between the two loci were compared between populations. Haplotype variation and diversity were estimated for each population. Our results reveal that populations of African descent possess significantly shorter alleles for the two loci than non-African populations (P<0.0001). Allelic diversity for both markers was higher among African Americans than any other population, including indigenous Africans from Sierra Leone and Nigeria. Analysis of molecular variance revealed that approx. 20% of CAG and GGC repeat variance could be attributed to differences between the populations. All non-African populations possessed the same common haplotype while the three populations of African descent possessed three divergent common haplotypes. Significant LD was observed in our sample of healthy African Americans. The LD observed in the African American population may be due to several reasons; recent migration of African Americans from diverse rural communities following urbanization, recurrent gene flow from diverse West African populations, and admixture with European Americans. This study represents the largest genotyping effort to be performed on the two androgen receptor trinucleotide repeat loci in diverse human populations.     

RFLP diversity and relationships among traditional European maize populations

Given the large extent of hybrid cultivation, the importance of conserving the diversity of crop genetic resources has given birth to numerous collections of old races. In the present paper, we conduct a molecular characterisation of a large collection of 488 European maize populations using the bulk RFLP analysis. The analysis of 23 RFLP loci showed a high allelic richness of 11.5 alleles per locus. Populations from eastern Europe (Poland, Austria, Germany, etc.) showed the lowest genetic diversity, a lower number of unique alleles and a higher percentage of fixed loci than populations from southern Europe. In fact, genetic diversity appeared higher in Southern regions where the first maize populations are thought to have been introduced. Molecular classification based on Rogers' distance (i.e. alleles frequencies) allowed us to distinguish three main clusters which were highly consistent with geographic origins. A Northeastern cluster grouped together early or intermediate populations from Northeastern countries and the Balkans, a southeastern cluster joined late and partially dent populations from Greece and Italy, and, a southwestern cluster was made up of early flint populations from northern Spain, Portugal and the Pyrenees. A correlation between allelic frequencies at some loci and latitude and/or longitude was observed. Such tendencies may reflect the direction of gene flow between different races of maize: for instance, North American (Northern flint) and Caribbean populations were introduced, respectively, to northern and southern Europe, in the past.     

Microsatellites and allozymes as the genetic memory of habitat fragmentation and defragmentation in populations of the ground beetle Carabus auronitens (Col., Carabidae)

Aim This paper has three aims: (1) to reconstruct the population history of a flightless silvicolous (forest) ground beetle in a region where strong anthropogenic activity has altered the connectivity of the landscape; (2) to estimate the effects of both contemporary and historical landscape structure on the genetics of populations; and (3) to investigate the reasons for clinal variation in one gene locus found in an earlier study carried out in the same geographical location. Location Münster (Westphalia), north‐west Germany. Methods We investigated 26 populations of the ground beetle Carabus auronitens Fabricius, 1792 by analysing seven polymorphic microsatellite loci and an allozyme locus. Samples of at least 16 individuals per site were studied. These were obtained from dry pitfall traps placed at 23 sites and from three additional (refuge) populations. We used regression and correlation analyses to investigate the effects of both historical and contemporary landscape structure on the allele frequency distributions of the investigated loci. Spatial autocorrelation analysis was used to study possible clinal variations, and admixture rates were calculated in order to assess the genetic influence of populations from possible refuges. Possible reasons for the development of the cline were examined using simulation models. Results The allele frequency distributions at the investigated loci could not be explained by selection or isolation by distance. We found clinal variation in 50% of the investigated loci and our simulations indicated that this was unlikely to have developed by chance. Admixture rates show significant influences of the investigated refuge populations on the populations under study. Main conclusions The findings strongly suggest that the clinal variation is secondary. It results from recolonization of the area by C. auronitens from multiple refuges after anthropogenic landscape changes caused forest fragmentation and led to species isolation.     

Isolation and characterization of 16 novel microsatellite loci in two inbred strains of the Chinese hamster (Cricetulus griseus)

The Shanyi inbred A and E strains of the Chinese hamster are widely used in biomedical research, but detailed genetic characterization has been lacking. We developed microsatellite markers that could be used for genetic diversity analysis and linkage map construction. We isolated and characterized 16 novel microsatellite loci from a microsatellite-enriched genomic DNA library. These loci were genotyped in 48 animals from the two strains, and the polymorphic information content was determined. In the Shanyi A and E populations, 14 and 15 loci were found to be polymorphic, respectively, with polymorphic information content ranging from 0.1393 to 0.8082 and from 0.1109 to 0.7397, respectively. A total of 115 alleles were found for the 16 microsatellite loci in the two populations; the mean observed heterozygosity (H(O)) was 0.5191 and 0.4333 for the A and E populations, respectively, indicating marked genetic variation within the two populations. Correspondingly, the F(ST) values ranged from 0.002 to 0.9253, with an overall mean of 0.1935, indicating significant genetic difference between the two strains. The population differentiation levels were substantiated by Nei's genetic distance and full Bayesian analyses computed with STRUCTURE. Despite the genetic diversity and differentiation within and between the two inbred populations, the 48 individuals were correctly allocated into their original populations with high statistical confidence based on these 16 microsatellite loci. These novel microsatellite loci should be useful genetic markers for these two Chinese hamster inbred strains.     

Isolation and characterization of microsatellite markers in the caddisfly Drusus discolor (Trichoptera: Limnephilidae)

We describe the development of and amplification conditions for microsatellite primers isolated from the caddisfly Drusus discolor. Eight polymorphic microsatellite loci were developed and screened for variability using 37 individuals from two populations from central Europe. The primers yielded an average of 8.6 alleles per loci. No linkage disequilibrium between loci was detected, while three loci showed deviations from Hardy–Weinberg equilibrium in one of the two tested populations.     

Microsatellite loci for genetic mapping in the turkey (Meleagris gallopavo)

New microsatellite loci for the turkey (Meleagris gallopavo) were developed from two small insert DNA libraries. Polymorphism at these new loci was examined in domestic birds and two resource populations designed for genetic linkage mapping. The majority of loci (152 of 168) was polymorphic in domestic turkeys and informative in two mapping resource populations and thus will be useful for genetic linkage mapping.     

Microsatellite markers isolated from barn swallows (Hirundo rustica)

Fifteen polymorphic microsatellite DNA loci were isolated from barn swallow (Hirundo rustica) and optimized for future studies of radiation‐induced mutations in populations from Ukraine. The loci were screened for variability among 25 individuals from two populations. The primers amplified loci with relatively high numbers of alleles ranging from five to 32 alleles per locus and polymorphic information content from 0.481 to 0.951. Observed heterozygosity varied from 0.458 to 0.960. None of the loci showed deviations from Hardy–Weinberg equilibrium in either population.