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Primer selection impacts specific population abundances but not community dynamics in a monthly time‐series 16S rRNA gene amplicon analysis of coastal marine bacterioplankton
Author(s) -
Wear Emma K.,
Wilbanks Elizabeth G.,
Nelson Craig E.,
Carlson Craig A.
Publication year - 2018
Publication title -
environmental microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.954
H-Index - 188
eISSN - 1462-2920
pISSN - 1462-2912
DOI - 10.1111/1462-2920.14091
Subject(s) - biology , roseobacter , primer (cosmetics) , amplicon , thaumarchaeota , 16s ribosomal rna , population , bacterioplankton , ribosomal rna , phylogenetic tree , ecology , evolutionary biology , genetics , clade , gene , polymerase chain reaction , phytoplankton , chemistry , demography , organic chemistry , sociology , nutrient
Summary Primers targeting the 16S small subunit ribosomal RNA marker gene, used to characterize bacterial and archaeal communities, have recently been re‐evaluated for marine planktonic habitats. To investigate whether primer selection affects the ecological interpretation of bacterioplankton populations and community dynamics, amplicon sequencing with four primer sets targeting several hypervariable regions of the 16S rRNA gene was conducted on both mock communities constructed from cloned 16S rRNA genes and a time‐series of DNA samples from the temperate coastal Santa Barbara Channel. Ecological interpretations of community structure (delineation of depth and seasonality, correlations with environmental factors) were similar across primer sets, while population dynamics varied. We observed substantial differences in relative abundances of taxa known to be poorly resolved by some primer sets, such as Thaumarchaeota and SAR11, and unexpected taxa including Roseobacter clades. Though the magnitude of relative abundances of common OTUs differed between primer sets, the relative abundances of the OTUs were nonetheless strongly correlated. We do not endorse one primer set but rather enumerate strengths and weaknesses to facilitate selection appropriate to a system or experimental goal. While 16S rRNA gene primer bias suggests caution in assessing quantitative population dynamics, community dynamics appear robust across studies using different primers.