Molecular markers reveal differentiation among isolates of Coccidioides immitis from California, Arizona and Texas

Coccidioides immitis causes coccidioidomycosis, a fungal disease of both immunocompromised and otherwise healthy people; it is capable of causing large epidemics and the disease is often refractory to chemotherapy. To quantify the magnitude of population differentiation and estimate levels of gene flow in C. immitis , multilocus genotypes were scored for 20–25 clinical isolates from each of Bakersfield (California), Tucson (Arizona), and San Antonio (Texas). The molecular markers used were PCR products with polymorphic restriction endonuclease sites, found and characterized in a previous study of the Tucson population. The data show very highly significant differences in allele frequencies between all three populations, and suggest very low levels of migration between populations. One isolate in the San Antonio sample was an outlier, showing the California-specific allele at all four of the loci distinguishing the two populations, and subsequent inquiries indicated that the infection had indeed been acquired in California. Thus, genetic information can be used to infer the geographical origin of a fungal infection.     

The estimation of population differentiation with microsatellite markers

Microsatellite markers are routinely used to investigate the genetic structuring of natural populations. The knowledge of how genetic variation is partitioned among populations may have important implications not only in evolutionary biology and ecology, but also in conservation biology. Hence, reliable estimates of population differentiation are crucial to understand the connectivity among populations and represent important tools to develop conservation strategies. The estimation of differentiation is c from Wright's FST and/or Slatkin's RST, an FST -analogue assuming a stepwise mutation model. Both these statistics have their drawbacks. Furthermore, there is no clear consensus over their relative accuracy. In this review, we first discuss the consequences of different temporal and spatial sampling strategies on differentiation estimation. Then, we move to statistical problems directly associated with the estimation of population structuring itself, with particular emphasis on the effects of high mutation rates and mutation patterns of microsatellite loci. Finally, we discuss the biological interpretation of population structuring estimates.     

Does natural selection promote population divergence? A comparative analysis of population structure using amplified fragment length polymorphism markers and quantitative traits

Divergent natural selection is considered an important force in plant evolution leading to phenotypic differentiation between populations exploiting different environments. Extending an earlier greenhouse study of population differentiation in the selfing annual plant Senecio vulgaris, we estimated the degree of population divergence in several quantitative traits related to growth and life history and compared these estimates with those based on presumably neutral molecular markers (amplified fragment length polymorphisms; AFLPs). This approach allowed us to disentangle the effects of divergent selection from that of other evolutionary forces (e.g. genetic drift). Five populations were examined from each of two habitat types (ruderal and agricultural habitats). We found a high proportion of total genetic variance to be among populations, both for AFLP markers (phiST = 0.49) and for quantitative traits (range of QST: 0.26-0.77). There was a strong correlation between molecular and quantitative genetic differentiation between pairs of populations (Mantel's r = 0.59). However, estimates of population differentiation in several quantitative traits exceeded the neutral expectation (estimated from AFLP data), suggesting that divergent selection contributed to phenotypic differentiation, especially between populations from ruderal and agricultural habitats. Estimates of within-population variation in AFLP markers and quantitative genetic were poorly correlated, indicating that molecular marker data may be of limited value to predict the evolutionary potential of populations of S. vulgaris.     

梯棱羊肚菌全基因组SNP/Indel分析及基于Indel标记的遗传连锁图谱构建

基于梯棱羊肚菌Morchella importuna两个子囊孢子培养物的全基因组测序数据,对全基因组范围内的变异位点进行分析。共鉴定到18 438个变异位点,平均每Mbp的变异位点数量为361个;变异位点以单核苷酸多态性SNP为主,共计17 104个,基因组中SNP的频率为335SNPs/Mbp;Indel多态性位点1 334个,以2-10bp的插入缺失为主;73.4% SNP/Indel位于基因间隔区域,外显子区域共检测到3 042个变异位点,占总数的16.50%;对基因功能产生确定影响的移码突变有1 088个,占5.90%,错义突变916个,占比4.97%;不同Scaffold上的SNP/Indel出现频率不同,SNP频率最大的为Scaffold80,平均每Mbp包含2 856个SNP,频率最低为Scaffold60和Scaffold75,分别为16SNPs/Mbp和30SNPs/Mbp;对≥11bp的Indel变异位点进行标记开发和多态性群体分析,成功开发出75对Indel标记。采用原生质体单细胞分离技术,获得了梯棱羊肚菌M04的两个可亲和的同核体菌株M04P01和M04P40,同时采用来自M04子囊果的58个单孢菌株作为作图群体,初步构建了包含75个Indel标记和1个交配型基因的梯棱羊肚菌遗传连锁图谱,共获得12个连锁群,连锁群总长度273.7cM。     

Stability of genetic structure and effective population size inferred from temporal changes of microsatellite DNA polymorphisms in the land snail Helix aspersa (Gastropoda: Helicidae)

Temporal evolution of genetic variability may have far-reaching consequences for a diverse array of evolutionary processes. Within the polders of the Bay of Mont-Saint-Michel (France), populations of the land snail Helix aspersa are characterized by a metapopulation structure with occasional extinction processes resulting from farming practices. A temporal survey of genetic structure in H . aspersa was carried out using variability at four microsatellite loci, in ten populations sampled two years apart. Levels of within-population genetic variation, as measured by allelic richness, H _e or F _is , did not change over time and similar levels of population differentiation were demonstrated for both sampling years. The extent of genetic differentiation between temporal samples of the same population established (i) a stable structure for six populations, and (ii) substantial genetic changes for four populations. Using classical F -statistics and a maximum likelihood method, estimates of the effective population size ( N _e) illustrated a mixture of stable populations with high N _e, and unstable populations characterized by very small N _e estimates (of 5–11 individuals). Owing to human disturbances, intermittent gene flow and genetic drift are likely to be the predominant evolutionary processes shaping the observed genetic structure. However, the practice of multiple matings and sperm storage is likely to provide a reservoir of variability, minimizing the eroding genetic effects of population size reduction and increasing the effective population size.  © 2004 The Linnean Society of London, Biological Journal of the Linnean Society , 2004, 82 , 89–102.     

Genetic structure of the shrub Daphne laureola across the Baetic Ranges, a Mediterranean glacial refugium and biodiversity hotspot

Distribution margins constitute areas particularly prone to random and/or adaptive intraspecific differentiation in plants. This trend may be particularly marked in species discontinuously distributed across mountain ranges, where sharp geographic isolation gradients and habitat boundaries will enhance genetic isolation among populations. In this study, we analysed the level of neutral genetic differentiation among populations of the long-lived shrub Daphne laureola (Thymelaeaceae) across the Baetic Ranges, a glacial refugium and biodiversity hotspot in the western Mediterranean Basin. Within this area, core and marginal populations of D. laureola were compared with regard to their spatial isolation, size, genetic diversity and differentiation. A spatially explicit analysis conducted on the vast majority of the species' known populations in the study area (N = 111) showed that marginal populations (western and eastern) present larger spatial isolation than core populations, but are not smaller. We compared genetic diversity and differentiation between core and marginal populations using a subsample of 15 populations and 225 amplified fragment length polymorphism (AFLP) markers. Core and marginal populations did not differ in genetic diversity, probably because of the occurrence of large populations on the local margins. Western populations were strongly differentiated from the other populations. In addition, spatial and genetic differentiation among populations was larger on the western margin. Eastern populations constituted a genetically homogeneous group closely related to core populations, despite their greater spatial isolation. Results suggest that studies on phenotypic differentiation between core and marginal populations of D. laureola, and presumably other species having discontinuous distributions across the Baetic ranges, should take into account geographical differences in levels of genetic differentiation between the different distribution borders.     

TETRASAT: a program for the population analysis of allotetraploid microsatellite data

Microsatellite markers are quite popular due to their degree of polymorphism and efficiency; however, the utility of such markers for analysing allotetraploid species is often hampered by an inability to determine allele copy number for partial heterozygotes. tetrasat is a program that uses an iterative substitution process to account for all probable combinations of allele copy numbers in populations with partial heterozygote samples. The program subsequently calculates allele frequencies, and mean Hardy–Weinberg expected heterozygosity (H_E), Shannon–Weiner Diversity Index (H′) and Nei's measure of population differentiation (G_ST) are reported for each locus and population. Of equal importance is the calculation of statistical variability generated by the missing data and allele substitution process, which allows for assessment of the strength of conclusions drawn from the statistics.     

Enzyme polymorphism inDrosophila melanogaster populations collected in two different habitats in Hungary

The level of enzyme polymorphism was compared in tenDrosophila melanogaster populations collected in farmyards and distilleries in two regions of Hungary. The total genetic diversity was partitioned into between-and within-population components at each investigated locus using Wright's F-statistics. Population differentiation was studied in two different ways. Genetic distances between pairs of populations were calculated and a hierarchical analysis of gene diversity was performed. Based on the F values gene flow was estimated among the populations at different levels of the hierarchy. The results indicated that our farmyard populations collected within a region could be considered as parallel samples from a panmictic population rather than samples of distinct populations. In distilleries, the flies might be influenced by two different evolutionary forces: (i) selection due to the extremely high concentration of ethanol in the fermenting mash and (ii) genetic drift due to the combination of repeated founder effects and fluctuating population size. Our results suggested that distillery populations could not be regarded as real populations either. They could be considered as peculiar cases: founder individuals taken from the total population (region) established special populations which survived in the distilleries for many generations. Thus the dominating force acting on the distillery populations was genetic drift.     

基于微卫星标记的印度中部Madhya Pradesh地区疟疾强化控制和非强化控制区的斯氏按蚊基因流和种群遗传结构分析

[目的]斯氏按蚊Anopheles stephensi是亚洲东南部城市人体疟疾的主要媒介,印度12％的疟疾病例由其引起.本实验研究了印度中部Madhya Pradesh地区东北部的疟疾强化控制(EMCP)区和非强化控制(非EMCP)区斯氏按蚊的基因流.在EMCP区,由于采用了各种疟疾防控措施因而疟疾病例首先降低,但是很快回升,说明总的疟疾风险维持稳定.[方法]应用7个微卫星位点,对印度中部Madhya Pradesh地区东北部的4个EMCP区和非EMCP区采集的斯氏按蚊进行基因分型,以分析各种群参数.[结果]发现各标记在所有种群中表现出高度的多态性.在两区间未发现很大的遗传多样性.观察到EMCP区的东部种群(FST=0.0485,RST =0.1112)比非EMCP区的北部种群(FST=0.020,RST =0.0145)具有较高的遗传分化,在EMCP区和非EMCP区之间观察到较高的基因流(12.90,6.16,5.06和2.38).RST的灵敏度高于FST,说明分化可能是由于突变而非遗传漂变引起的.[结论]本研究表明,在EMCP区和非EMCP区内以及EMCP区和非EMCP区之间存在很高的基因流.基因流水平高以及抗虫性的发展似乎是EMCP区和非EMCP区疟疾病例发生增加的重要原因.     

The efficiency of purifying selection in Mammals vs. Drosophila for metabolic genes

The nearly neutral theory of molecular evolution states that the efficiency of natural selection depends on the effective population size. By using a wide range of multispecies data on nucleotide polymorphism, we have tried to ascertain whether there are any differences in the level of selective constraints of metabolic process genes between Mammals and Drosophila species. The results are consistent with a higher selective constraint in Drosophila than in Mammals, according to the expected under the nearly neutral model: purifying selection seems to be more efficient in species with a larger effective population size.