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Environmental Sequencing Provides Reasonable Estimates of the Relative Abundance of Specific Picoeukaryotes

AuthorsGiner, Caterina R.; Forn, Irene ; Romac, Sarah; Logares, Ramiro ; Vargas, Colomban de ; Massana, Ramon
Issue DateAug-2016
PublisherAmerican Society for Microbiology
CitationApplied and Environmental Microbiology 82(15): 4757-4766 (2016)
AbstractHigh-throughput sequencing (HTS) is revolutionizing environmental surveys of microbial diversity in the three domains of life by providing detailed information on which taxa are present in microbial assemblages. However, it is still unclear how the relative abundance of specific taxa gathered by HTS correlates with cell abundances. Here, we quantified the relative cell abundance of 6 picoeukaryotic taxa in 13 planktonic samples from 6 European coastal sites using epifluorescence microscopy on tyramide signal amplification-fluorescence in situ hybridization preparations. These relative abundance values were then compared with HTS data obtained in three separate molecular surveys: 454 sequencing of the V4 region of the 18S ribosomal DNA (rDNA) using DNA and RNA extracts (DNA-V4 and cDNA-V4) and Illumina sequencing of the V9 region (cDNA-V9). The microscopic and molecular signals were generally correlated, indicating that a relative increase in specific 18S rDNA was the result of a large proportion of cells in the given taxa. Despite these positive correlations, the slopes often deviated from 1, precluding a direct translation of sequences to cells. Our data highlighted clear differences depending on the nucleic acid template or the 18S rDNA region targeted. Thus, the molecular signal obtained using cDNA templates was always closer to relative cell abundances, while the V4 and V9 regions gave better results depending on the taxa. Our data support the quantitative use of HTS data but warn about considering it as a direct proxy of cell abundances
Description10 pages, 4 figures, 3 tables, supplemental material https://dx.doi.org/10.1128/AEM.00560-16
Publisher version (URL)https://dx.doi.org/10.1128/AEM.00560-16
Identifiersdoi: 10.1128/AEM.00560-16
issn: 0099-2240
e-issn: 1098-5336
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