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用串联重复 DNA 探针研究香蒲(香蒲科)的遗传多样性及其对污染物的影响。

Genetic diversity of Typha latifolia (Typhaceae) and the impact of pollutants examined with tandem-repetitive DNA probes.

机构信息

Department of Biological Sciences ML6, University of Cincinnati, Cincinnati, Ohio 45221-0006;

出版信息

Am J Bot. 1999 Sep;86(9):1226-38.

Abstract

Genetic diversity at variable-number-tandem-repeat (VNTR) loci was examined in the common cattail, Typha latifolia (Typhaceae), using three synthetic DNA probes composed of tandemly repeated "core" sequences (GACA, GATA, and GCAC). The principal objectives of this investigation were to determine whether: (1) the previously reported almost complete lack of polymorphism at allozyme loci in this species was indicative of a reduced amount of genetic diversity at VNTR loci as well; (2) VNTR markers were informative about possible clonal propagation; and (3) significant differences in genetic structure of sampling sites were associated with differences in environmental levels of pollutants at those sites. Previously, widespread sampling across the eastern United States, surveying across ten allozyme loci, has detected only two genotypes, involving a difference at a single locus, among 104 populations. In this study, the amount of genetic diversity detected at VNTR loci: (1) among ramets (N = 40; 40 genotypes detected) collected at ∼8-km intervals along a 320-km transect; (2) among ramets (N = 220; 117 genotypes detected) from five study sites separated by 50-3000 m; and (3) even among ramets within each study site [N = 44 per site; from 13 to 34 genotypes detected per site (270 m(2))] exceeds that previously found in those more geographically widespread allozyme surveys. Among the 260 ramets analyzed here, the mean number of bands scored per individual was 48.61 (SD = 2.80). Mean genetic similarity among ramets collected along the 320-km transect was 0.91, which was within the range of mean genetic similarity within the five study sites (range: 0.89-0.95). Among the five study sites, 61% of the samples analyzed appeared to be clonal ramets, with up to 12 clones detected for 44 ramets sampled within a site. Clones grew intermingled and ranged up to 39 m in extent. Permutation tests of genetic similarity revealed significant genetic differentiation between each of the five study sites. Consistent with the previous allozyme studies, T. latifolia was characterized by extremely low genetic variation relative to levels of polymorphism detected at VNTR loci in other plant species. Estimated heterozygosity among ramets along the 320-km transect ranged from 0.11 to 0.13, while that within the five study sites ranged from 0.05 to 0.12. Estimates of F(st) (0.32-0.41) also indicated considerable genetic subdivision among these stands. Significantly higher genetic diversity was detected at the two study sites that chemistry and toxicity data indicate to be the most severely impacted by pollutants. Although this correlation does not establish cause and effect, the results of this study indicate that the analysis of genetic diversity at VNTR loci may be a useful tool for monitoring anthropogenic-induced changes in the genetic structure of natural populations of plants.

摘要

使用由串联重复“核心”序列(GACA、GATA 和 GCAC)组成的三个合成 DNA 探针,研究了普通香蒲(Typha latifolia,香蒲科)在可变数目串联重复(VNTR)位点的遗传多样性。本研究的主要目的是确定:(1)以前在该物种等位酶位点报告的几乎完全缺乏多态性是否表明 VNTR 位点的遗传多样性也减少了;(2)VNTR 标记是否可以提供有关可能克隆繁殖的信息;(3)采样地点的遗传结构差异是否与这些地点的污染物环境水平差异有关。在此之前,在美国东部进行了广泛的采样,在十个等位酶位点进行了调查,仅在 104 个种群中发现了两种基因型,涉及一个位点的差异。在这项研究中,在 VNTR 位点检测到的遗传多样性:(1)在沿 320 公里长的样带上以 8 公里的间隔收集的 40 个(N = 40;检测到 40 种基因型)配子体之间;(2)在五个研究地点之间收集的 220 个配子体(N = 220;检测到 117 种基因型),相距 50-3000 米;(3)即使在每个研究地点的配子体之间[每个地点 N = 44;每个地点检测到 13-34 种基因型(270 m2)],这也超过了以前在那些地理分布更广泛的等位酶调查中发现的水平。在分析的 260 个配子体中,每个个体评分的带数平均值为 48.61(SD = 2.80)。沿 320 公里长的样带收集的配子体之间的平均遗传相似性为 0.91,这在五个研究地点的平均遗传相似性范围内(范围:0.89-0.95)。在五个研究地点中,分析的 61%的样本似乎是克隆配子体,在一个地点内对 44 个采样的配子体检测到多达 12 个克隆。克隆体相互交织,范围可达 39 米。遗传相似性的置换检验显示五个研究地点之间存在显著的遗传分化。与以前的等位酶研究一致,与其他植物物种在 VNTR 位点检测到的多态性水平相比,香蒲的遗传变异极低。沿 320 公里长的样带的配子体之间的估计杂合度范围为 0.11 至 0.13,而在五个研究地点内的范围为 0.05 至 0.12。Fst 的估计值(0.32-0.41)也表明这些种群之间存在相当大的遗传划分。在两个研究地点检测到的遗传多样性明显更高,化学和毒性数据表明这些地点受到污染物的影响最严重。尽管这种相关性不能确定因果关系,但本研究的结果表明,VNTR 位点遗传多样性的分析可能是监测自然种群遗传结构人为诱导变化的有用工具。

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