Field K G, Gordon D, Wright T, Rappé M, Urback E, Vergin K, Giovannoni S J
Department of Microbiology, Oregon State University, Corvallis 97331, USA.
Appl Environ Microbiol. 1997 Jan;63(1):63-70. doi: 10.1128/aem.63.1.63-70.1997.
Small-subunit (SSU) ribosomal DNA (rDNA) gene clusters are phylogenetically related sets of SSU rRNA genes, commonly encountered in genes amplified from natural populations. Genetic variability in gene clusters could result from artifacts (polymerase error or PCR chimera formation), microevolution (variation among rrn copies within strains), or macroevolution (genetic divergence correlated with long-term evolutionary divergence). To better understand gene clusters this study assessed genetic diversity and distribution of a single environmental SSU rDNA gene cluster, the SAR11 cluster. SAR11 cluster genes, from an uncultured group of the alpha subclass of the class Proteobacteria, have been recovered from coastal and midoceanic waters of the North Atlantic and Pacific. We cloned and bidirectionally sequenced 23 new SAR11 cluster 16S rRNA genes, from 80 and 250 m in the Sargasso Sea and from surface coastal waters of the Atlantic and Pacific, and analyzed them with previously published sequences. Two SAR11 genes were obviously PCR chimeras, but the biological (nonchimeric) origins of most subgroups within the cluster were confirmed by independent recovery from separate gene libraries. Using group-specific oligonucleotide probes, we analyzed depth profiles of nucleic acids, targeting both amplified rDNAs and bulk RNAs. Two subgroups within the SAR11 cluster showed different highly depth-specific distributions. We conclude that some of the genetic diversity within the SAR11 gene cluster represents macroevolutionary divergence correlated with niche specialization. Furthermore, we demonstrate the utility for marine microbial ecology of oligonucleotide probes based on gene sequences amplified from natural populations and show that a detailed knowledge of sequence variability may be needed to effectively design these probes.
小亚基(SSU)核糖体DNA(rDNA)基因簇是SSU rRNA基因的系统发育相关集,常见于从自然群体中扩增的基因中。基因簇中的遗传变异可能源于人为因素(聚合酶错误或PCR嵌合体形成)、微观进化(菌株内rrn拷贝之间的变异)或宏观进化(与长期进化分歧相关的遗传分化)。为了更好地理解基因簇,本研究评估了单个环境SSU rDNA基因簇——SAR11簇的遗传多样性和分布。SAR11簇基因来自变形菌纲α亚类的一个未培养群体,已从北大西洋和太平洋的沿海水域和大洋中部水域中获得。我们克隆并双向测序了23个新的SAR11簇16S rRNA基因,这些基因分别来自马尾藻海80米和250米深处以及大西洋和太平洋的沿海表层水域,并将它们与先前发表的序列进行了分析。有两个SAR11基因明显是PCR嵌合体,但该簇中大多数亚群的生物学(非嵌合)起源通过从单独的基因文库中独立获得而得到证实。我们使用组特异性寡核苷酸探针分析了核酸的深度分布,同时针对扩增的rDNA和总RNA。SAR11簇中的两个亚群显示出不同的高度深度特异性分布。我们得出结论,SAR11基因簇内的一些遗传多样性代表了与生态位特化相关的宏观进化分歧。此外,我们展示了基于从自然群体中扩增的基因序列的寡核苷酸探针在海洋微生物生态学中的实用性,并表明可能需要详细了解序列变异性才能有效地设计这些探针。