Application of sequence-specific labeled 16S rRNA gene oligonucleotide probes for genetic profiling of cyanobacterial abundance and diversity by array hybridization
Application of sequence-specific labeled 16S rRNA gene oligonucleotide probes for genetic profiling of cyanobacterial abundance and diversity by array hybridization
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DOI：
10.1128/aem.66.9.4004-4011.2000
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发表时间：
2000-09-01
影响因子：
4.4
通讯作者：
Jakobsen, KS
中科院分区：
文献类型：
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作者：
Rudi, K;Skulberg, OM;Jakobsen, KS
利用从蓝藻培养物中获得的小亚基rRNA基因(16S RDNA)的DNA序列信息，研究了蓝藻在自然生境中的存在及其丰度。本研究选取了8个浮游生物群落，分别发育在藻类生物量相对较低的湖泊(中营养化)和生物量相对较高的湖泊(富营养化)。对水样的有机成分进行了遗传分析，方法是对针对16S rDNA的寡核苷酸探针进行多重序列特异性标记，然后将标记的探针与其各自的补体杂交到固体载体(DNA阵列)上。建立了10个探针，以确定在选定的湖泊中遇到的可辨别蓝藻的相对丰度，这些探针通常是针对其目标的，通过克隆培养分析确定。在随后的天然蓝藻群落分析中，为所调查的湖泊建立了可重复的丰度剖面。然后将遗传分析的结果与标准水生生物和水化学分析获得的信息进行比较。在定性上，不同湖泊中发现的生物群(念珠藻、微囊藻和Planktothrix种)之间存在较好的相关性。定量数据的相关性水平较低。然而，这可能是由于样品处理技术的差异。这些比较的结论是，遗传丰度分布可能为分离和量化自然栖息地中遗传上截然不同的蓝藻群体提供基础。
DNA sequence information for the small-subunit rRNA gene (16S rDNA) obtained from cyanobacterial cultures was used to investigate the presence of cyanobacteria and their abundance in natural habitats. Eight planktonic communities developing in lakes characterized by relatively low algal biomass (mesotrophic) and in lakes with correspondingly high biomass (eutrophic) were selected for the study. The organismal compositions of the water samples were analyzed genetically, using multiplex sequence-specific labeling of oligonucleotide probes targeted to 16S rDNA and subsequent hybridization of the labeled probes to their respective complements spotted onto a solid support (DNA array). Ten probes were established to determine the relative abundances of the discernible cyanobacteria encountered in the selected lakes, The probes were generally specific for their targets, as determined through analyses of clone cultures. Reproducible abundance profiles were established for the lakes investigated in the subsequent analyses of natural cyanobacterial communities. The results from the genetic analyses were then compared with information obtained from standard hydrobiological and hydrochemical analyses. Qualitatively, there were relatively good correlations among the groups of organisms (Nostoc, Microcystis, and Planktothrix species) found in the different lakes. The levels of correlation were lower for the quantitative data. This may, however, be due to differences in sample processing technique. The conclusions from these comparisons are that the genetic abundance profiles may provide a foundation for separating and quantifying genetically distinct groups of cyanobacteria in their natural habitats.