Department of Biology, Babeş-Bolyai University, 1 Kogălniceanu Street, Cluj-Napoca, Romania.
Antonie Van Leeuwenhoek. 2011 Feb;99(2):271-81. doi: 10.1007/s10482-010-9486-5. Epub 2010 Jul 28.
The biodiversity of a specific cyanobacterial mat associated to a thermomineral spring from the Western Plain of Romania was investigated. Light and electron microscopy, together with molecular tools (denaturing gradient gel electrophoresis-DGGE, automated ribosomal intergenic spacer analysis-ARISA and amplified ribosomal DNA restriction analysis-ARDRA), based on 16S rDNA and 16S-23S internal transcribed spacer markers were used. Based on the partial 16S rRNA fragments sequenced, eight cyanobacterial taxons were identified, all belonging to the Oscillatoriales order, Phormidium and Leptolyngbya being dominant. A significant difference was observed, in comparison with the morphological approach. In certain conditions, DGGE can provide misleading information due to multiple melting domains in the same sequence, to multiple rrn operons in the same genome and due to unspecific hybridization among closely related sequences. This can lead to an overestimated species abundance which can cause incorrect description of the microbial community investigated. Additional techniques, such as ARISA and ARDRA, can improve the microbial biodiversity studies, thus providing optimal results.
对罗马尼亚西部平原一处热矿泉相关的特定蓝藻席微生物多样性进行了调查。采用了光学显微镜和电子显微镜，以及分子工具（变性梯度凝胶电泳-DGGE、自动核糖体基因间隔区分析-ARISA 和扩增核糖体 DNA 限制分析-ARDRA），基于 16S rDNA 和 16S-23S 内部转录间隔区标记物。根据测序的部分 16S rRNA 片段，鉴定出了 8 种蓝藻分类群，均属于颤藻目，优势种为 Phormidium 和 Leptolyngbya。与形态学方法相比，观察到了显著差异。在某些条件下，由于同一序列中存在多个融解域、同一基因组中存在多个 rrn 操纵子以及密切相关序列之间的非特异性杂交，DGGE 可能会提供误导性信息。这可能导致物种丰度被高估，从而导致对所研究微生物群落的不正确描述。其他技术，如 ARISA 和 ARDRA，可以改进微生物多样性研究，从而提供最佳结果。
