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DOE OSTI2023

maranasgroup/k-sacce306

The parameterization of kinetic models requires measurement of fluxes and/or metabolite levels for a base strain and a few genetic perturbations thereof. Unlike stoichiometric models that are mostly invariant to the specific strain, it remains unclear whether kinetic models constructed for different strains of the same species have similar or significantly different kinetic parameters. This important question underpins the applicability range and prediction limits of kinetic reconstructions. To this end, herein we parameterize two separate large-scale kinetic models using K-FIT with genome-wide coverage corresponding to two distinct strains of Saccharomyces cerevisiae: CEN.PK 113-7D strain (model k-sacce306-CENPK), and growth-deficient BY4741 (isogenic to S288c; model k-sacce306-BY4741). The metabolic network for each model contains 306 reactions, 230 metabolites, and 119 substrate-level regulatory interactions. The two models (for CEN.PK and BY4741) recapitulate, within one standard deviation, 77% and 75% of the fitted dataset fluxes, respectively, determined by 13C metabolic flux analysis for wild-type and eight single-gene knockout mutants of each strain. Strain-specific kinetic parameterization results indicate that key enzymes in the TCA cycle, glycolysis, and arginine and proline metabolism drive the metabolic differences between these two strains of S. cerevisiae. Our results suggest that although kinetic models cannot be readily used across strains as stoichiometric models, they can capture species-specific information through the kinetic parameterization process.

Dinh, Hoang↗

DOE OSTI2022

Data for Glucose Feeds the Tricarboxylic Acid Cycle via Excreted Ethanol in Fermenting Yeast

Ethanol and lactate are typical waste products of glucose fermentation. In mammals, glucose is catabolized by glycolysis into circulating lactate, which is broadly used throughout the body as a carbohydrate fuel. Individual cells can both uptake and excrete lactate, uncoupling glycolysis from glucose oxidation. Here we show that similar uncoupling occurs in budding yeast batch cultures of Saccharomyces cerevisiae and Issatchenkia orientalis . Even in fermenting S. cerevisiae that is net releasing ethanol, media 13C-ethanol rapidly enters and is oxidized to acetaldehyde and acetyl-CoA. This is evident in exogenous ethanol being a major source of both cytosolic and mitochondrial acetyl units. 2H-tracing reveals that ethanol is also a major source of both NADH and NADPH high-energy electrons, and this role is augmented under oxidative stress conditions. Thus, uncoupling of glycolysis from the oxidation of glucose-derived carbon via rapidly reversible reactions is a conserved feature of eukaryotic metabolism.

Conversion↗

DOE OSTI2021

Data from Raczka et al., Interactions between microbial diversity and substrate chemistry determine the fate of carbon in soil, from Elizabeth Woods in Morgantown, West Virginia, USA

File types include: Soil respiration as 13C cumulative respiration from soil incubation experiment, qSIP (quantitative stable isotope probing) data that is "18S apes wide" which is the fungal atom percent excess or how much the fungi took up of the labeled litter and "16S apes wide" which is the bacterial atom percent excess or how much the bacteria took up of the labeled litter, metabolomic data from the incubated soil samples called "Metab Soil Unique Filtered", which is the metabolomic data that had an initial filter from the raw data, and lipidomic data from the incubated soil samples called "All lipids soil". These files were used according to methods in Raczka et al. 2021 Scientific Reports. Headers show soil type (Oak, Poplar) and litter type (also called substrate here, which is labeled as no litter (control), poplar, and oak). This research was done to understand interactions between ecosystem-level processes and microbial diversity that impacts the composition of decomposition products that can form stable soil organic matter.

54 ENVIRONMENTAL SCIENCES↗

DOE OSTI2023

Arctic shrub size and leaf traits, northern Alaska, summer 2017 (Version 2.0)

This data package contains leaf and size trait and environmental data collected from 170 plots of rapidly expanding shrub genera (Alnus, Betula, and Salix) and a widespread sedge (Eriophorum vaginatum) along a latitudinal and climate gradient in northern Alaska. The trait data were collected in summer 2017 and include leaf area, specific leaf area, leaf nitrogen concentration, leaf delta 15N, leaf delta 13C, shrub height, aboveground biomass, and the ratio of root biomass to aboveground biomass. These raw data support a submitted manuscript that examines the intraspecific variations of size, leaf and root traits of shrubs across the graminoid-dominated nutrient-poor arctic tundra and reveals the environmental drivers of deciduous shrub traits in tundra ecosystems (Fraterrigo et al., submitted). Data are presented by site (n=5) and patch (shrub or sedge plot) in separate csv files. The location data are provided in the “plot coordinates” file; all other files contain the data in the file title. Detailed methods are in the submitted manuscript. A companion data package contains root trait data collected simultaneously from the same plots (Fraterrigo and Chen, 2020). See the "Related references" section for more information.2023/07/20 Update: The latest version of this dataset publication is version 2.0. The latest version of the data package was updated to correct the leaf size trait.csv file.

54 ENVIRONMENTAL SCIENCES↗

DOE OSTI2022

C fluxes from incubations of California upland soil with and without synthetic iron (hydr)oxide associations

There is a growing understanding that interactions with soil mineral phases contribute to the accumulation and retention of otherwise degradable organic matter in soils and sediments. However, the bioavailability of organic compounds in mineral-organic-associations, especially under varying environmental conditions is not well known. To assess the impact of mineral association and warming on the decomposition of an easily respirable organic substrate (glucose), we conducted a series of laboratory incubations at different temperatures with field-collected soils from 10 to 20 cm, 50–60 cm, and 80–90 cm depth at the Blodgett Forest Research Station. We added 13C-labeled glucose either directly to native soil or sorbed to one of two synthetic iron (hydr)oxide phases (goethite and ferrihydrite) that differ in crystallinity and affinity for sorbing glucose. We incubated the soils at 25 deg C. This dataset comprises the added glucose C and the cumulative fluxes over 72 and 1896 hours. We found that: (1) association with the Fe (hydr)oxide minerals reduced the decomposition rate of glucose by > 99.5% relative to rate of decomposition for free glucose in soil; (2) the respiration rate per gram carbon did not differ appreciably with depth, suggesting a similar degree of decomposability for native C across depths and that under the incubation conditions total carbon availability represents the principal limitation on respiration under these conditions as opposed to reduced abundance of decomposers or moisture and oxygen limitations; (3) addition of free glucose enhanced native carbon respiration at all soil depths with the largest effect at 50–60 cm; (4) in general respiration of the organo-mineral complex (glucose and iron-(hydr)oxide) was less temperature sensitive than was respiration of native carbon; (5) the addition of organic free mineral decreased the rate of soil respiration in the intermediate 50–60 cm depth soil. The results emphasize the key role of MOAs in regulating the fluxes of carbon from soils to the atmosphere and in turn the stocks of soil carbon.This dataset lists the glucose additions made to soils with and without added geothite or ferrihidrite and the C respired over the timespan of the incubations.

54 ENVIRONMENTAL SCIENCES↗

DOE OSTI2022

Soil C stock and CO2 production data for Soong et al. 2021: Effects of five years of soil warming at Blodgett Forest, CA.

Subsoils below 20 cm are an important reservoir in the global carbon cycle, but little is known about their vulnerability under climate change. We measured subsoil carbon in warmed vs control plots of a conifer forest after 4.5 years of whole-soil warming (4°C). We also measured soil CO2 efflux at the surface and soil CO2 concentrations along the soil profile. Over the 4.5 years we observed that warming resulted in a loss of subsoil carbon stocks (−33 ± 11%), primarily from unprotected particulate organic matter. Warming also stimulated a sustained 30 ± 4% increase in soil CO2 efflux due to increased CO2 production through the whole-soil profile. The observed in situ decline in subsoil carbon stocks with warming is now definitive evidence of a positive soil carbon-climate feedback, which could not be concluded based on increases in CO2 effluxes alone. This dataset comprises (1) 13C and C recovery in 2014, 2016, and 2018 1m soil cores sampled in 10cm increments. (2) Monthly surface CO2 fluxes from 2014 to 2018. (3) Monthly gas well sample [CO2] and production data. And (4) density fraction data for three depths of cores recovered in 2017.

54 ENVIRONMENTAL SCIENCES↗

DOE OSTI2022

Influence of root- vs needle-derived litter on soil C and N stabilization in a California coniferous forest from 2001 to 2011

How plant inputs from above- versus below-ground affect long term carbon (C) and nitrogen (N) retention and stabilization in soils is not well known. We present results of a decade-long field study that traced the decomposition of 13C- and 15N-labeled Pinus ponderosa needle and fine root litter placed in O or A soil horizons of a sandy Alfisol under a coniferous forest. We measured the retention of litter-derived C and N in particulate (>2 mm) and bulk soil (<2 mm) fractions, as well as in density-separated free light and three mineral-associated fractions. After 10 years, the influence of slower initial mineralization of root litter compared to needle litter was still evident: almost twice as much root litter (44% of C) was retained than needle litter (22–28% of C). After 10 years, the O horizon retained more litter in coarse particulate matter implying the crucial comminution step was slower than in the A horizon, while the A horizon retained more litter in the finer bulk soil, where it was recovered in organo-mineral associations. Retention in these A horizon mineral-associated fractions was similar for roots and needles. Nearly 5% of the applied litter C (and almost 15% of the applied N) was in organo-mineral associations, which had centennial residence times and potential for long-term stabilization. Vertical movement of litter-derived C was minimal after a decade, but N was significantly more mobile. Overall, the legacy of initial litter quality influences total SOM retention; however, the potential for and mechanisms of long-term SOM stabilization are influenced not by litter type but by soil horizon.This dataset comprises (1) litter recovery for each of 5 timepoints in >2mm, <2mm, and density fractions, (2) density fractionation results for the 10-year timepoint, and (3) vertical movement of applied litter.

54 ENVIRONMENTAL SCIENCES↗

DOE OSTI2024

Organic carbon feedbacks to soil structure in Profundihumic and Haplic Ferralsols in Campinas, Sao Paulo, Brazil

Increasing soil organic matter content coincides with increasing soil porosity and decreasing bulk density, but the mechanisms that lead to changes in soil structure are unclear. We quantified and compared soil characteristics across two contrasting soils (heavily weathered ferralsols in Brazil) to clarify the effects of organic matter content on soil porosity. We found that high organic matter content fosters higher biologic activity, including root growth and animal burrowing, which in turn increase soil porosity and decreases bulk density.This dataset includes soil physical, morophological, and chemical variables from 6 soil profiles were sampled in Campinas, Sa ̃o Paulo state, southeastern Brazil to 1 m depth. Sites are regenerated mixed savanna and rainforest in the protected area of Viracopos Airport in Campinas, lightly grazed by cattle.This dataset includes soil color, bulk density, texture (percent sand, silt, clay, and coarse fragments as well as class), soil pH, cation exchange capacity, C stock, C concentration, N stock, N concentration, delta 13C, Delta 14C, and soil porosity in Excel and csv format.

54 ENVIRONMENTAL SCIENCES↗

DOE OSTI2024

Soil Carbon Dynamics Following Land Use Changes and Conversion to Oil Palm Plantations in Tropical Lowlands Inferred From Radiocarbon

We measured the 14C and 13C isotopic values of soil organic carbon in mineral soil from lowland tropical forests to provide insight into how quickly carbon is turning over in the soil following conversion of primary forests to oil palm plantations. In addition to areas converted to oil palm plantations in Peru, Indonesia, and Cameroon, we examine pastures and secondary forests in Peru as a comparison to the carbon cycling processes operating in the oil palm plantations.This dataset includes radiocarbon (Δ14C) and stable carbon (δ13C) isotopes of soil organic carbon in mineral soils from natural lowland forests and oil palm plantations in Peru, Indonesia, and Cameroon. We additionally examine plots of secondary forests following agricultural use and pastures on cleared natural forest in Peru. In addition to isotopic data, this dataset includes soil carbon and nitrogen concentrations and stock, soil texture (percent sand, silt, and clay), pH, ECEC (effective cation exchange capacity), base saturation, and bulk density. Soils were sampled in 4 depth increments to 100 cm depth.This dataset supports the publication Finstad et al., 2020. Finstad, K., van Straaten, O., Veldkamp, E., & McFarlane, K. (2020). Soil carbon dynamics following land use changes and conversion to oil palm plantations in tropical lowlands inferred from radiocarbon. Global Biogeochemical Cycles, 34, e2019GB006461. https://doi.org/10.1029/2019GB006461

54 ENVIRONMENTAL SCIENCES↗

DOE OSTI2022

A radioisotope - enabled reactive transport model for deep vadose zone carbon

In mountainous regions, which constitute the principle source of recharge to major rivers and regional aquifers, infiltration occurs through fractured, partially saturated, weathered bedrock that acts as a boundary layer between saturated aquifers and surface soil. Commonly this deep vadose zone (DVZ) is many meters thick, and yet its role in regulating the generation, retention and mobility of reactive solutes, including nutrients, contaminants and weathering products, is largely unknown. In particular, many of the key reactions that drive the formation of the weathered DVZ and the quality of water moving through it are redox processes, regulated by the availability of organic carbon and oxygen below the soil layer. The role of the DVZ is thus also poorly constrained in the context of carbon stocks and mobility, particularly in lithologies that are naturally high in organic carbon, such as shales. The overarching hypothesis of this study is that upland regions developed in geologic settings with abundant petrogenic carbon store and actively cycle carbon in the weathered DVZ below the soil and above the water table at rates that are significant and currently unconstrained. In order to quantify this cycling, the current study combines novel instrumentation techniques allowing new direct sampling of DVZ systems with advanced numerical reactive transport simulations of carbon transport and transformation. Critically, these simulations will explicitly treat the three isotopes of carbon (the abundant 12C, the stable rare 13C and the radioactive 14C) in a unified framework, thus clearly parsing between the contributions of modern surface derived carbon and lithologic carbon sources in integrated measurements of fluid and gas phase fluxes. This novel model capability will be applied to test the role of DVZ carbon cycling as a regulator of water quality and geological weathering in two complementary field sites both located in organic carbon rich shale lithologies. The first is the Eel River Critical Zone Observatory (ERCZO) in Mendocino County, California, and the second is the Lawrence Berkeley National Laboratory Watershed Function Scientific Focus Area (SFA) in the East River watershed, near Crested Butte, Colorado. At the ERCZO, a novel vadose zone monitoring system has been installed in a 20 m thick, partially saturated, weathered shale hillslope, and preliminary data already indicate substantial CO2 flux generated many meters below the soil surface. At the SFA field site, an instrumented hillslope transect indicates a more complex multi-dimensional fluid and solute transport regime, which will serve as a key test of the calibrated models. Collectively, this project will advance understanding of the cycling of carbon belowground and in relation to transport pathways across the poorly constrained DVZ characteristic of primary water recharge areas. The key product of this work will be enhanced isotope simulation capabilities that are robust and publicly available for application across a broad diversity of systems.

58 GEOSCIENCES↗

DOE OSTI2022

Syntrophic Co-Cultures of Clostridium Organisms to Produce Higher Alcohols & Other C6-C8 Metabolites (Final Report)

The goal of this project was to advance the systems biology understanding and predictive modeling of synthetic & syntrophic Clostridium microbial consortia, focusing on elucidation of metabolic networks and environmental signals in the consortia. The project has direct applicability to lignocellulosic-biomass based production of higher alcohols as advanced biofuels and C6-C8 metabolites, that can be used as chemicals or serve as biofuel precursors. This project studied and optimized three synthetic syntrophic systems. It examined the population dynamics using flow-cytometry, time-lapse microscopy and PCR analysis. 13C-based tracer analyses was used to examine the metabolite exchange between the syntrophic cell populations and the impact of those interactions on the transcriptome of the individual populations. To enhance our analytical and predictive capabilities, genome-scale models (GSMs) for these syntrophies was developed. RNAseq data for these syntrophic coculture systems were acquired to enable a molecular level understanding of the syntrophies aiming to identify the genetic networks of each organism in the co-culture and compare those against the networks of pure cultures.

09 BIOMASS FUELS↗

DOE OSTI2020

Creating multifunctional synthetic lichen platforms for sustainable biosynthesis of biofuel precursors

In this project, we were creating a sustainable platform for biofuel production, utilizing carbon-fixing autotrophs to supply oxygen and organic substrates to heterotrophic partners, which in turn produce carbon dioxide to feed the autotrophs. This symbiotic lichen community could lower the input cost, optimize metabolic exchanges and improve the generation of biofuel precursors through multi-omics driven genetic engineering. The cyanobacteria Synechococcus elongatus (S. elongatus) was used as the primary autotroph to provide oxygen and organic substrates, especially sucrose, to a co-culture system. The strain with overexpression of sucrose transporter cscB demonstrated a significant increase in sucrose production under salt stress as what we expected. We also implemented 13C metabolic flux analysis on the sucrose secreting strain S. elongatus cscB-NaCl. Next, transporters proteins like glutamate exporter mscCG from Corynebacterium glutamicum was overexpressed in S. elongatus to improve metabolite exchange. We provided sucrose and glutamate to filamentous fungi to enable their growth and production of biochemicals using substrates from the cyanobacterium. Two fungi ( Aspergillus nidulans and Aspergillus niger ) and two yeast strains ( Rhodotorula toruloides and Lipomyces starkeyi ) served as heterotrophs in this system. They were co-cultured with S.elongatus under different pH condition, and growth on different carbon source to understand the symbiotic system and optimize the parameters for production. The two fungi strained grew much better at pH 7, 8 and 9 with Yeast Nitrogen Base (YNB ) supplementation. Two yeast strains grew well on pH 7 and 8 with YNB supplementation. All four species achieved the highest biomass level when utilizing sucrose as the main carbon sources, which demonstrates a great pairing with S.elongatus . In addition to investigate the metabolite exchange and growth condition of the co-culture system, we incorporated an Aspergillus strain expressing three genes (PAND, BAPAT and HPDH). Expression of these three genes enabled production of 3-hydroxypropionic acid (3HP) with a titer of 3-6 g/L regardless of pH and other nutrient conditions. Moreover, we developed a computational model with different heterotrophic fungi symbiosing with S.elongatus to evaluate the exchange of hundreds of metabolites. The accuracy and sensitivity had been optimized by high-throughput phenotyping assays. This project demonstrated ways to enhance a synthetic lichen platform’s efficiency and scalability to create a robust system for sustainable bioproduct synthesis. The project has enabled us to explore the technological and commercial potential of sustainable lichen co-culture. The project has also trained the next generation of scientists to address sustainability, carbon fixation, and design solutions to contemporary global challenges. The lichen platform represented a novel approach to sustainably producing bioproducts with a reduced carbon footprint and lower costs.

09 BIOMASS FUELS↗

DOE OSTI2024

Creating multifunctional synthetic lichen platforms for sustainable biosynthesis of biofuel precursors

Betenbaugh, Michael↗

DOE OSTI2024

Cellulose crystallinity index (CrI) of switchgrass measured by solid-state NMR

Here we present a curated dataset of switchgrass cellulose crystallinity index (CrI) measured using solid state nuclear magnetic resonance (NMR) spectroscopy. Seventy-two topline switchgrass lines grown in greenhouse were collected and ground to -20/+80 mesh. The switchgrass was then Soxhlet-extracted with toluene/ethanol for 24 h to remove extractives. The extractives-free switchgrass was holopulped by using peracetic acid at 5 g loading per g biomass and the solution consistency was adjusted to 5% with DI water. Holopulping was conducted at room temperature for 24 h with stirring. The obtained holocellulose was washed excessively with DI water and air-dried at room temperature for 24 h. The dried holocellulose was treated with hydrochloric acid (2.5 M) for 2 h to remove hemicellulose. The isolated cellulose was collected by filtration, rinsed with DI water, and used to analyze cellulose crystallinity by solid-state NMR. The NMR samples were prepared by packing the moisturized cellulose into 4-mm cylindrical Zirconia MAS rotors. Cross polarization/magic angle spinning (CP/MAS) NMR analysis of cellulose was carried out on a Bruker Avance III 400-MHz spectrometer operating at 100.59 MHz for 13C in a Bruker double-resonance MAS probe head at spinning speeds of 8 kHz. The CP/MAS experiments utilized a 5 ms (90°) proton pulse, 1.5 ms contact pulse, 4 s recycle delay and 4000 scans. The cellulose crystallinity index was determined from the areas of the cellulose crystalline C4 signal (δ 86-92 ppm) over the entire C4 regions (δ 79-92 ppm) in the NMR spectra.

Cellulose, crystallinity index, switchgrass, solid↗

DOE OSTI2024

Proton NMR spectra of lignin isolated from field grown transgenic poplar

Here we present a curated dataset of a series of 1H nuclear magnetic resonance (NMR) spectra of lignin isolated from transgenic monolignol 4-O-methyltransferase (MOMT4) engineered poplar. The transgenic poplar was collected from a 2-year-old rotation trees within a three-year field trial experiment. Two replicates were collected for each transgenic poplar for the 1H NMR analysis. The poplar samples were Soxhlet-extracted with toluene/ethanol to remove the extractives and the extractives-free poplar was then ball-milled in a Retsch PM100 planetary ball mill using a porcelain jar with ceramic balls at 600 rpm for 2 h. The ball-milled materials were subjected to enzymatic hydrolysis for 48 h followed by centrifugation and washing with deionized water. The solid residue was extracted twice with 96:4 (v/v) 1,4-dioxane/water mixture at room temperature overnight. The extracts were combined, rotary evaporated, and freeze-dried to recover lignin. The dry lignin samples were dissolved in deuterated dimethyl sulfoxide and transferred into a 5 mm NMR tube. 1H NMR experiments were performed in a Bruker Avance III HD 500 MHz NMR spectrometer operating at a frequency of 125.12 MHz for the 13C nucleus using a standard Bruker pulse sequence (zg) on a Prodigy platform cryoprobe. The NMR spectra were acquired with 16 ppm spectra width, 32k data points, 3s pulse delay, and 16 scans. All the data was processed using the Bruker’s TopSpin 3.6 software. Additional meta data is embedded in the raw spectra files.

1H NMR, lignin, poplar, field trial, MOMT4, CBI↗

DOE OSTI2021

Sensing Techniques for Organochlorides through Intermolecular Interaction with Bicyclic Amidines

Toxic organochloride molecules are widely used in industry for various purposes. With their high volatility, the direct detection of organochlorides in environmental samples is challenging. Here, a new organochloride detection mechanism using 1,5-diazabicyclo[4.3.0]non-5-ene (DBN) is introduced to simplify a sensing method with higher detection sensitivity. Three types of organochloride compounds-trichloroethylene (TCE), dichloromethane (DCM), and dichlorodiphenyltrichloroethane (DDT)—were targeted to understand DCM conjugation chemistry by using nuclear magnetic resonance (NMR) and liquid chromatography with a mass spectrometer (LC-MS). 13C-NMR spectra and LC-MS data indicated that DBN can be labeled on these organochloride compounds by chlorine–nitrogen interaction. Furthermore, to demonstrate the organochloride sensing capability, the labeling yield and limit of detection were determined by a colorimetric assay as well as micellar electrokinetic chromatography (MEKC). The interaction with DBN was most appreciable for TCE, among other organochlorides. TCE was detected at picomolar levels, which is two orders of magnitude lower than the maximum contaminant level set by the United States Environmental Protection Agency. MEKC, in conjunction with this DBN-labeling method, enables us to develop a field-deployable sensing platform for detecting toxic organochlorides with high sensitivity.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

DOE OSTI2023

Suppression of Stacking Order with Doping in 1T-TaS 2–x Se x

In 1T-TaS 2–x Se x , the charge density wave (CDW) state features a star of David lattice that expands across layers as the system becomes commensurate upon cooling. The layers can also order along the c-axis, and different stacking orders have been proposed. For this work, using neutron scattering on powder samples, we compared the stacking order previously observed in 1T-TaS 2 when the system is doped with Se. While at low temperature, a 13c layer sequence stacking was observed in TaS 2 ; this type of ordering was not evident with doping. Doping with Se results in a metallic state in which the Mott transition is suppressed, which may be linked to the absence of layer stacking.

75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND↗

DOE OSTI2022

Microbial Community Structure and Ecological Networks during Simulation of Diatom Sinking

Microbial-mediated utilization of particulate organic matter (POM) during its downward transport from the surface to the deep ocean constitutes a critical component of the global ocean carbon cycle. However, it remains unclear as to how high hydrostatic pressure (HHP) and low temperature (LT) with the sinking particles affects community structure and network interactions of the particle-attached microorganisms (PAM) and those free-living microorganisms (FLM) in the surrounding water. In this study, we investigated microbial succession and network interactions in experiments simulating POM sinking in the ocean. Diatom-derived 13C- and 12C-labeled POM were used to incubate surface water microbial communities from the East China Sea (ECS) under pressure (temperature) of 0.1 (25 °C), 20 (4 °C), and 40 (4 °C) MPa (megapascal). Our results show that the diversity and species richness of the PAM and FLM communities decreased significantly with HHP and LT. Microbial community analysis indicated an increase in the relative abundance of Bacteroidetes at high pressure (40 MPa), mostly at the expense of Gammaproteobacteria, Alphaproteobacteria, and Gracilibacteria at atmospheric pressure. Hydrostatic pressure and temperature affected lifestyle preferences between particle-attached (PA) and free-living (FL) microbes. Ecological network analysis showed that HHP and LT enhanced microbial network interactions and resulted in higher vulnerability to networks of the PAM communities and more resilience of those of the FLM communities. Most interestingly, the PAM communities occupied most of the module hubs of the networks, whereas the FLM communities mainly served as connectors of the modules, suggesting their different ecological roles of the two groups of microbes. These results provided novel insights into how HHP and LT affected microbial community dynamics, ecological networks during POM sinking, and the implications for carbon cycling in the ocean.

59 BASIC BIOLOGICAL SCIENCES↗