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Abstract：
The vast diversity of microalgae imposes the challenge of identifying them through the most common and economical identification method, morphological identification, or through using the more recent molecular-level identification tools. Here we report an approach combining enrichment and metagenomic molecular techniques to enhance microalgae identification and identify microalgae diversity from environmental water samples. From this perspective, we aimed to identify the most suitable culturing media and molecular approach (using different primer sets and reference databases) for detecting microalgae diversity. Using this approach, we have analyzed three water samples collected from the River Nile on several enrichment media. A total of 37 microalgae were identified morphologically to the genus level. While sequencing the three-primer sets (16S rRNA V1-V3 and V4-V5 and 18S rRNA V4 region) and aligning them to three reference databases (GG, SILVA, and PR2), a total of 87 microalgae were identified to the genus level. The highest eukaryotic microalgae diversity was identified using the 18S rRNA V4 region and alignment to the SILVA database (43 genera). The two 16S rRNA regions sequenced added to the eukaryotic microalgae identification, 26 eukaryotic microalgae. Cyanobacteria were identified through the two sequenced 16S rRNA regions. Alignment to the SILVA database served to identify 14 cyanobacteria to the genera level, followed by Greengenes, 11 cyanobacteria genera. Our multiple-media, primer, and reference database approach revealed a high microalgae diversity that would have been overlooked if a single approach had been used over the other.
摘要：
微藻的巨大多样性带来了通过最常见和最经济的鉴定方法来鉴定它们的挑战。形态学鉴定，或通过使用最新的分子水平鉴定工具。在这里，我们报告了一种将富集和宏基因组分子技术相结合的方法，以增强微藻的鉴定并从环境水样中鉴定微藻的多样性。从这个角度来看,我们的目的是确定用于检测微藻多样性的最合适的培养基和分子方法（使用不同的引物集和参考数据库）。使用这种方法，我们分析了在几种富集介质上从尼罗河收集的三个水样。在形态上总共鉴定了37种微藻。在对三引物组（16SrRNAV1-V3和V4-V5和18SrRNAV4区域）进行测序并将其与三个参考数据库（GG，SILVA,andPR2),共有87种微藻类被鉴定为属级。使用18SrRNAV4区域并与SILVA数据库（43属）进行比对，鉴定了最高的真核微藻多样性。测序的两个16SrRNA区域添加到真核微藻鉴定中，26个真核微藻。通过两个测序的16SrRNA区域鉴定蓝细菌。与SILVA数据库的比对有助于鉴定14个蓝细菌的属水平，紧随其后的是格林基因，11个蓝藻属。我们的多媒体，底漆,参考数据库方法揭示了高度的微藻多样性，如果使用一种方法代替另一种方法，则会被忽略。
