Methods to estimate microbial diversity have developed rapidly in an effort to understand the distribution and diversity of microorganisms in natural environments. For bacterial communities, the 16S rRNA gene is the phylogenetic marker gene of choice, but most studies select only a specific region of the 16S rRNA to estimate bacterial diversity. Whereas biases derived from from DNA extraction, primer choice and PCR amplification are well documented, we here address how the choice of variable region can influence a wide range of standard ecological metrics, such as species richness, phylogenetic diversity, β-diversity and rank-abundance distributions. We have used Illumina paired-end sequencing to estimate the bacterial diversity of 20 natural lakes across Switzerland derived from three trimmed variable 16S rRNA regions (V3, V4, V5). Species richness, phylogenetic diversity, community composition, β-diversity, and rank-abundance distributions differed significantly between 16S rRNA regions. Overall, patterns of diversity quantified by the V3 and V5 regions were more similar to one another than those assessed by the V4 region. Similar results were obtained when analyzing the datasets with different sequence similarity thresholds used during sequences clustering and when the same analysis was used on a reference dataset of sequences from the Greengenes database. In addition we also measured species richness from the same lake samples using ARISA Fingerprinting, but did not find a strong relationship between species richness estimated by Illumina and ARISA. We conclude that the selection of 16S rRNA region significantly influences the estimation of bacterial diversity and species distributions and that caution is warranted when comparing data from different variable regions as well as when using different sequencing techniques.
Lakes in the Alps represent a considerable fraction of nutrient-poor lakes in Central Europe, with unique biodiversity and ecosystem properties. Although some individual lakes are well studied, less knowledge is available on large-scale patterns essential to general understanding of their functioning. Here, we aimed to describe crustacean zooplankton communities (Cladocera, Copepoda) and identify their environmental drivers in the pelagic zone of 54 oligotrophic lakes in the montane region of the Alps (400–1200 m) in Austria, Germany, and Switzerland, covering a spatial scale of 650 km. Moreover, we aimed to provide data on the distribution and ecological requirements of the North American invader Bythotrephes longimanus in its Central European native range. Communities were mainly dominated by widespread species typical of lowland habitats, and only a few true specialists of oligotrophic alpine lakes were present. The most frequent taxa were the Daphnia longispina complex and Eudiaptomus gracilis, with 48 and 45 occurrences, respectively. Species richness decreased with altitude and increased with lake area. The main structuring factors of community composition were chlorophyll a concentration and depth, which drove an apparent separation of mesotrophic and oligotrophic communities. Bythotrephes had 13 occurrences, showing a preference for deep oligotrophic lakes. Its presence was not coupled with lower crustacean species richness, as was repeatedly observed in North America. Additionally, it frequently co-occurred with the other large predatory cladoceran, Leptodora kindtii. B. longimanus might be considered a truly montane species in Central Europe, given its absence in lowland and alpine lakes.
The fate of dissolved organic carbon (DOC) is partly determined by its availability to microbial degradation. Organisms at upper trophic levels could influence the bioavailability of DOC via cascading effects on primary producers and bacteria. Here we experimentally tested whether the presence of fish in aquatic food webs can indirectly affect the composition of the DOC pool. We found that fish had strong positive effects on phytoplankton biomass that affected the dynamics of DOC composition. Specifically, fish increased protein‐like, algae‐derived DOC mid‐experiment, concurrent with the strongest fish‐induced increase in phytoplankton biomass. Fish also increased bacterial abundance, altered the community composition and diversity of bacteria, and temporarily increased DOC compounds with fluorescence properties indicative of microbially‐reprocessed organic matter. Overall, our experiment revealed that fish can positively influence the substrate (algae‐produced DOC) and the key players (bacteria) of the microbial carbon pump. Consequently, fish could contribute to carbon sequestration by stimulating both the production of bioavailable DOC and the microbial degradation of bioavailable to persistent DOC. We propose this as a novel mechanism whereby the loss of predators from global ecosystems could alter carbon cycling.
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