The ISME Journal (2017) 11, 405–414 © 2017 International Society for Microbial Ecology All rights reserved 1751-7362/17 www.nature.com/ismej ORIGINAL ARTICLE Trade-offs between microbiome diversity and productivity in a stratified microbial mat Hans C Bernstein1,2,3, Colin Brislawn2, Ryan S Renslow2,3, Karl Dana1, Beau Morton2, Stephen R Lindemann2, Hyun-Seob Song2, Erhan Atci3, Haluk Beyenal3, James K Fredrickson2, Janet K Jansson2 and James J Moran1 1Chemical and Biological Signature Science, Pacific Northwest National Laboratory, Richland, WA, USA; 2Biological Sciences Division, Pacific Northwest National Laboratory, Richland, WA, USA and 3The Gene and Linda Voiland School of Chemical Engineering and Bioengineering, Washington State University, Pullman, WA, USA Productivity is a major determinant of ecosystem diversity. Microbial ecosystems are the most diverse on the planet yet very few relationships between diversity and productivity have been reported as compared with macro-ecological studies. Here we evaluated the spatial relationships of productivity and microbiome diversity in a laboratory-cultivated photosynthetic mat. The goal was to determine how spatial diversification of microorganisms drives localized carbon and energy acquisition rates. We measured sub-millimeter depth profiles of net primary productivity and gross oxygenic photosynthesis in the context of the localized microenvironment and community structure, and observed negative correlations between species richness and productivity within the energy- replete, photic zone. Variations between localized community structures were associated with distinct taxa as well as environmental profiles describing a continuum of biological niches. Spatial regions in the photic zone corresponding to high primary productivity and photosynthesis rates had relatively low-species richness and high evenness. Hence, this system exhibited negative species– productivity and species–energy relationships. These negative relationships may be indicative of stratified, light-driven microbial ecosystems that are able to be the most productive with a relatively smaller, even distributions of species that specialize within photic zones. The ISME Journal (2017) 11, 405–414; doi:10.1038/ismej.2016.133; published online 1 November 2016 Introduction Pinkerton, 1955). Organisms growing as members of communities have been observed to have higher The majority of primary producers on the planet are power, that is, rates of energy acquisition, compared powered by oxygenic photosynthesis and their rates with members grown alone (DeLong, 2008). In of carbon fixation typically set the upper bound for relation, the species-energy theory proposes a posi- ecosystem productivity (Field et al., 1998). Commu- tive correlation between biological energy capture nities exhibiting the highest net rates of primary and a system’s species richness (May, 1975; Wright, productivity should produce the most biomass. 1983; Hurlbert and Stegen, 2014). This collective However, the question remains as to whether theoretical framework suggests that, within photo- higher-species diversity results in greater rates of autotrophically driven ecosystems, the rates of gross biological energy and carbon acquisition. Ecological oxygenic photosynthesis and net primary productiv- theories, such as the maximum power principle and ity should drive a positive relationship with species species-energy theory, address the relationships diversity. between the rate of biological energy capture and Although the relationship between community species-level fitness and richness. The maximum productivity and species diversity has been frequently power principle postulates that the fitness of investigated, a unifying consensus on the shape of the self-organizing systems increases when they are relationship is lacking (Rajaniemi, 2003; Bienhold designed to maximize power intake and subsequent et al., 2012). Similar ecosystems show different energy transformation (Lotka, 1922; Odum and productivity–diversity relationships depending on the spatial scale of observation (Chase and Leibold, Correspondence: HC Bernstein or JJ Moran, Chemical and 2002; Grace et al., 2016). Spatial gradients of produc- Biological Signature Science, Pacific Northwest National Labora- tivity are inherent to photoautotrophically driven tory, PO Box 999, MS-IN: P7-50, Richland, WA 99352, USA. E-mail: [email protected] or [email protected] ecosystems in which light is attenuated at a character- Received 28 April 2016; revised 26 July 2016; accepted 5 August istic distance related to the physical structure of the 2016; published online 1 November 2016 ecosystem. Benthic microbial mats and biofilms are of Productivity and diversity gradients HC Bernstein et al 406 particular interest in this respect, as tight physical renewal of fresh media and mitigation of batch associations between cells and extracellular polymeric effects. Incubations were performed in experi- substances strongly attenuate light over short dis- mental duplicate each containing four flow cells tances. These compact microbial ecosystems exhibit operated in parallel. Every measurement presented stratified biological organization created and con- in this study was performed in a minimum of three trolled by steep photo- and geochemical gradients on biological replicates. The flow cell module consisted sub-millimeter spatial scales (Jørgensen et al., 1979; of a black aluminum housing containing four Ley et al.,2006;Jensenet al., 2011). 14.9 × 3.8 × 3.3 cm chambers covered by gasket lids In this study, we evaluated the spatial relation- exposing a 2.5 × 9 cm area (centered borosilicate ships between productivity and diversity in a glass window) that directed incident irradiance to photosynthetic microbial mat. The goal was to the mat surface. Media was delivered at very low, determine how vertical gradients in community laminar flows (1.7 ml min − 1per chamber) to mini- composition and associated micro-environmental mize shear. The overlaying liquid depth was con- conditions translate to productivity. The experimen- trolled (1 cm) and mat samples were incubated on a tal microcosm was derived from a benthic microbial 2 mm bed of sterile (autoclaved), native sediment. mat native to epsomitic Hot Lake; located in Sterile media entered a pre-mixing continuous Washington State, USA (Anderson, 1958; Linde- stirred tank reactor upstream of the flow cell that mann et al., 2013; Zachara et al., 2016). Productivity was used to control temperature (22 °C), pH 7.5 and was assessed by two spatially resolved, complemen- gas composition of the media during the incubation tary metrics: the gross rate of oxygenic photosynth- phase. Media delivery to the system was performed esis and %C replacement of biomass over time. The by two distinct protocols. First, during an acclima- measured gross rate of oxygenic photosynthesis tion phase (14 days of flow) the continuous stirred was used as an upper bound of photosynthetic tank reactor was sparged at 1 l min − 1 air enriched energy acquisition. We evaluated the net rate with 1% CO2. The second, experimental phase, was of primary productivity at similar spatial scales performed over the last 24 h diel cycle to measure 13 − using laser ablation isotope ratio mass spectrometry net primary productivity; C labeled HCO3 was that enabled quantification of accumulation of 13C, introduced by replacing the continuous stirred tank − derived from labeled HCO3, into microbial biomass reactor input for a carboy containing pH-adjusted (Moran et al., 2014). Both photosynthesis and net (7.5) sterile media with 20 mM NaHCO3 at a 4:1 ratio primary productivity were powered by absorption of of natural isotope abundance and 13C-labled sub- light energy, which was inherently constrained to strate. The labeled media was primed into the flow the photic zone, and regions of highest producti- cell at high flow rates (~170 ml min− 1) to mitigate vity exhibited relatively low-species richness but dilution effects upon introduction. The 13C-labled high evenness. Hence, this system exhibited media was introduced at the beginning of a 12 h dark − negative-productivity richness and energy-richness cycle to enable labeled HCO3 permeation through relationships. the mat in the absence of photosynthetic activity corresponding to negligible carbon fixation. Materials and methods Micro-profiling Cultures and media Depth profiles of δ13C(‰) were sampled and Microbial mats were sampled from the previously measured in triplicate from cross sections of mat described Hot Lake field site (Lindemann et al., excised over the incubation time course. As pre- 2013). Whole-mat samples and underlying sediment viously described, lyophilized cross sections of these – were transferred under dark, cool (4 10 °C) condi- samples were analyzed using laser ablation sampling tions. Each microcosm was maintained under (Cetac LSX-500 laser ablation system, Omaha, 12:12 h light:dark cycles, illuminated at 150 μmol NE, USA) coupled to isotope ratio mass spectrometry − 2 − 1 photons m sec (see spectrum Supplementary (Thermo Delta V plus isotope ratio mass spectro- Figure S6) and fully submerged in a modified BG-11 meter, Waltham, MA, USA) at sub-millimeter resolu- media containing (mM): 120 Na2SO4, 30 KCl, 0.187 tion (50–300 μm); the most common step size was ∙ NH4Cl, 600 MgSO4 7H2O, 0.0057 K2HPO4, 0.245 250 μm (Moran et al., 2011, 2014). Porosity profiles ∙ CaCl2 2H2O, 0.0028 disodium EDTA and supple- were measured in excised mat