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Divergent responses of soil microorganisms to throughfall exclusion across tropical forest soils driven by soil fertility and climate history

Model projections predict tropical forests will experience longer periods of drought and more intense precipitation cycles under a changing climate. Such transitions have implications for structure-function relationships within microbial communities. We examine how throughfall exclusion might reshape prokaryotic and fungal communities across four lowland forests in Panama with a wide variation in mean annual precipitation and soil fertility. Four sites were established across a 1000 mm span in Mean Annual Precipitation (MAP: 2335–3421 mm). We expected microbial communities at sites with lower MAP to be less sensitive to throughfall exclusion than sites with higher MAP and fungal communities to be more resistant to disturbance than prokaryotes. At each location, partial throughfall exclusion structures were established over 10 × 10 m plots to reduce direct precipitation input. After short-term (~3–9 months) throughfall exclusion, prokaryotic communities showed no change in composition. However, prolonged (12–18 months) throughfall exclusion resulted in divergent prokaryotic community responses, reflecting MAP and soil fertility. We observed the emergence of a “drought microbiome” within infertile sites, whereby the community structure of the experimental throughfall exclusion plots at the lower MAP sites diverged from their respective control sites and converged towards overlapping assemblages. Furthermore, under throughfall exclusion, taxa increasing in relative abundance at the wettest site reflected that endemic to control plots at the lowest MAP site, suggesting a shift toward communities with lifehistory traits selected for under a lower MAP. By contrast, fungal community composition across sites was resilient to throughfall exclusion; however, biomass diverged in response to throughfall exclusion, increasing at two sites while decreasing in the other two. Broadly, our results suggest that microbial communities’ sensitivity to frequent drying and rewetting periods in tropical forest soils will depend on climate history and soil fertility, with infertile sites likely to respond readily to changes in precipitation.

54 ENVIRONMENTAL SCIENCES↗

Panama Rainforest Changes with Experimental Drought (PaRChED): Initial Effects of Partial Throughfall Exclusion on Soil Dynamics in Lowland Forests Across Variation in Rainfall and Soil Fertility

Changes in rainfall are predicted across tropical regions, with effects on nutrient, water, and carbon cycling. This chapter summarizes results from the first two years of a throughfall exclusion experiment in four lowland Panamanian forests that span a 1,000-mm change in rainfall and variation in soil fertility. Soil respiration (i.e., soil carbon dioxide [CO2 ] flux) declined with throughfall exclusion, with a site*season interaction, and the radiocarbon age of respired carbon was older in exclusion versus control plots. The decline in soil CO2 flux could be related to reduced fine root production and soil microbial biomass. Microbial community composition also changed with throughfall exclusion in infertile soils, and soil nutrients accumulated more in exclusion versus control plots during the dry season. The net effects on soil carbon storage will depend on the relative strengths of these effects over time. Continued research could improve predictions of tropical forest-climate feedbacks with changes in precipitation.

54 ENVIRONMENTAL SCIENCES↗

Soil Respiration Responses to Throughfall Exclusion Are Decoupled From Changes in Soil Moisture for Four Tropical Forests, Suggesting Processes for Ecosystem Models

Abstract Climatic drying is predicted for many tropical forests yet models remain poorly parameterized for these ecosystems, hampering predictions of forest‐climate interactions. We applied an integrated model–experiment approach, parameterizing an ecosystem model with tropical forest observational data and comparing model predictions to a field drying manipulation. We hypothesized that drying suppresses soil CO 2 fluxes (i.e., respiration) in relatively dry tropical forests but increases CO 2 fluxes in wetter tropical forests by alleviating anaerobiosis. We measured soil CO 2 fluxes during wet‐dry cycles from 2015 to 2022 in four Panamanian forests that vary in rainfall and soil fertility. Measured soil CO 2 fluxes declined in the dry season and peaked in the early wet season ahead of peak soil moisture, resulting in lower soil moisture optima for respiration than previously modeled. We then parameterized the model using field data and the new moisture‐respiration response functions. The updated model predicted increased soil CO 2 fluxes with drying in wetter and fertile forests and suppressed fluxes in drier, infertile forests. In contrast to model predictions, a chronic throughfall exclusion experiment initially suppressed soil respiration across forests, with sustained suppression for four years in the wettest forest only (−28% ± 4% during the dry season). In the fertile forest, drying eventually elevated CO 2 fluxes over this period (+75% ± 28% during the late wet season). The unexpected negative drying effect in the wettest, infertile forest could have resulted from reduced vertical flushing of nutrients into soils. Including hydro‐nutrient interactions in ecosystem models could improve predictions of tropical forest‐climate feedbacks.

54 ENVIRONMENTAL SCIENCES↗

Throughfall exclusion and fertilization effects on tropical dry forest tree plantations, a large-scale experiment

Abstract. Across tropical ecosystems, global environmental change is causing drier climatic conditions and increased nutrient deposition. Such changes represent large uncertainties due to unknown interactions between drought and nutrient availability in controlling ecosystem net primary productivity (NPP). Using a large-scale manipulative experiment, we studied for 4 years whether nutrient availability affects the individual and integrated responses of aboveground and belowground ecosystem processes to throughfall exclusion in 30-year-old mixed plantations of tropical dry forest tree species in Guanacaste, Costa Rica. We used a factorial design with four treatments: control, fertilization (F), drought (D), and drought + fertilization (D + F). While we found that a 13 %–15 % reduction in soil moisture only led to weak effects in the studied ecosystem processes, NPP increased as a function of F and D + F. The relative contribution of each biomass flux to NPP varied depending on the treatment, with woody biomass being more important for F and root biomass for D + F and D. Moreover, the F treatment showed modest increases in maximum canopy cover. Plant functional type (i.e., N fixation or deciduousness) and not the experimental manipulations was the main source of variation in tree growth. Belowground processes also responded to experimental treatments, as we found a decrease in nodulation for F plots and an increase in microbial carbon use efficiency for F and D plots. Our results emphasize that nutrient availability, more so than modest reductions in soil moisture, limits ecosystem processes in tropical dry forests and that soil fertility interactions with other aspects of drought intensity (e.g., vapor pressure deficit) are yet to be explored.

54 ENVIRONMENTAL SCIENCES↗

Data from a throughfall exclusion experiment: Fine root dynamics, morphology, chemistry, and AMF colonization across four lowland Panamanian forests

Fine roots regulate forest nutrient, carbon, and water cycling, yet their variation within and among tropical forests remains under-characterized. We quantified root productivity, disappearance, and stocks to 1 m using minirhizotron imaging, and we measured morphology, elemental composition [root carbon (C), root nitrogen (N), root phosphorus (P)], and arbuscular mycorrhizal fungi (AMF) colonization to 20 cm using ingrowth cores and sequential coring. Sampling took place in four distinct lowland Panamanian forests (32 plots; 8 per forest) from 2018 through 2022 under control and throughfall-exclusion (drought) treatments in the Panama Rainforest Changes with Experimental Drying (PARCHED) experiment.The dataset is presented as an Excel workbook with six tabs. The first tab is the data dictionary. Tab S1 contains ingrowth-core production and mortality, morphology and soil moisture. Tab S2 contains sequential-coring standing stocks with associated morphology and soil moisture. Tab S3 contains minirhizotron row data records to 1 m depth, including per-frame root length and diameter, normalized length metrics, and session timing. Tab S4 contains AMF colonization. Tab S5 contains fine-root chemistry at 0–10 cm, reporting %P, %C, %N, and C:N for samples collected via ingrowth cores and sequential-coring standing stocks. CSV mirrors for each tab are provided, and a KML file supplies coordinates for all 32 plots.Key variables span live and dead fine-root biomass (and coarse fractions where applicable), specific root length (SRL) and area (SRA), diameter, root tissue density (RTD), soil moisture, AMF colonization, root %N, %C, %P, and C:N, along with minirhizotron root length and diameter. Depth, season, treatment, and plot/site identifiers are included to support cross-tab integration and analysis from 0–100 cm (minirhizotron) and 0–20 cm (cores).Units are reported in-column and missing values are coded as NA. No special software is required to open or use the files (Excel, CSV, and KML compatible).

54 ENVIRONMENTAL SCIENCES↗

Microbial community data from throughfall exclusion experiment: Metadata, SI, community composition, LefSe, and FunGuilR data tables from PARCHED Panama tropical forest soils, 2024-2025

Soil contains more carbon (C) than terrestrial vegetation and the atmosphere combined, with some of the largest terrestrial C stocks in tropical rainforests. Soil microbes decompose organic matter, playing a vital role in the storage or loss of soil C. With climate change, drought conditions are predicted to increase in many tropical regions, including both chronic drying and extended drought, potentially influencing these processes. This project explored the effects of chronic and seasonal drying on soil microbial communities across four distinct tropical forests in a long-term drying experiment. We investigated the effects of a chronic drying manipulation on soil microbial community abundance and variation across different forests and seasons. We also compared findings with previously published data from these forests after short-term drying. This project used soils from a long-term drying experiment established in 2018 across four seasonal lowland forests in Panama. Soils were collected from 0 – 10 cm depths during three seasonal periods in control and drying plots in 2024 and 2025 from a total of 32 plots (n = 4 per forest per treatment). The forests varied in baseline rainfall and soil fertility. We calculated alpha and beta diversity indices and compared taxonomic community composition. We found significant biogeographic variation in microbial diversity and taxonomy, with significant differences across the forests and significant effects of the drying treatment. Metadata and sample IDs are within Metadata_16S.csv and Metadata_ITS.csv. Relative abundance tables of every sample at every season are shown in the Excel workbooks 16S Relative Abundance.xlsx and ITS Relative Abundance.xlsx. They are then also shown in CSV files by each taxonomic level. Linear discriminant analysis effect size (LefSe) tables are shown for the full 16S and ITS datasets (n = 96), subsets for every site at every season (n = 8), and then for the forests with each plot merged by season (n = 8). FunGuildR data table of ITS data is uploaded.

Bacteria↗

16S and ITS Amplicon Sequencing Fastq files and metadata from PARCHED Panama Tropical Forest soils, 2019-2020,

Model projections predict tropical forests will experience longer periods of drought and more intense precipitation cycles under a changing climate. Such transitions have implications for structure-function relationships within microbial communities. We examine how chronic drying might reshape prokaryotic and fungal communities across four lowland forests in Panama with a wide variation in mean annual precipitation and soil fertility. Four sites were established across a 1000 mm span in mean annual precipitation (2335 to 3300 mm). We expected microbial communities at sites with lower MAP to be less sensitive to chronic drying than sites with higher MAP; while fungal communities to be more resistant to disturbance than prokaryotes. At each location, partial throughfall exclusion structures were established over 10 x 10 m plots to reduce direct precipitation input. Raw demultiplexed sequences (bacteria, archaea, fungal) from soil samples taken from PARCHED Panama Tropical Forest throughfall exclusion experiments. Files that contain 16S are sequences from prokaryotes, ITS indicates sequences from fungi. Compressed fastq files are contained in Field_PARCHED_ITS_fastq_2020.zip, Field_PARCHED_ITS_fastq_2019.zip, Field_PARCHED_16S_fastq_2020.zip, Field_PARCHED_16S_fastq_2019.zip.There are four sites across the isthmus of Panama, each with 4 control and 4 exclusion plots. Throughfall exclusion shelters were built to intercept 50% of throughfall precipitation that hits the soil. Samples were taken on May 2019 and Jan 2020 from 0-10 cm and 10-20 cm depth approximately 9 and 18 months after shelter installation. Metadata and sample IDs for fastq files for 2019 sampling are within Field_PARCHED_Metadata_2019_ITS.csv and Field_PARCHED_Metadata_2019_16S.csv. The metadata for both 16S and ITS fastq files for January 2020 is included in Field_PARCHED_Metadata_2020.csv. No data processing or QA/QC was done on the raw data. Data processing example provided in R notebook file.

54 ENVIRONMENTAL SCIENCES↗

Effects of experimental and seasonal drying on soil microbial biomass and nutrient cycling in four lowland tropical forests

Changes in precipitation represent a major effect of climate change on tropical forests, which contain some of the earth’s largest terrestrial carbon (C) stocks. Such changes are expected to influence microbes, nutrients, and the fate of C in tropical forest soils. To explore this, we assessed soil microbial biomass, potential extracellular enzyme activities, and nutrient availability in a partial throughfall exclusion experiment in four seasonal lowland tropical humid forests in Panama with wide variation in precipitation and soil fertility. We hypothesized that throughfall exclusion would reduce microbial biomass and activity and accentuate dry season soil nutrient accumulation, with larger effects in wetter, less droughtresistant forests. We observed a baseline seasonal pattern of decreased microbial biomass and increased extractable dissolved organic C (DOC), total dissolved nitrogen (TDN), nitrate (NO 3 – ), and resinextractable phosphorus (P) in the dry season, with the strongest patterns for nitrogen (N). However, potential enzyme activities showed no consistent seasonality. In line with seasonal drying, throughfall exclusion decreased soil microbial biomass in the wet season and increased TDN and NO 3 – , especially in the dry season. In contrast to seasonal drying, throughfall exclusion decreased DOC and did not affect resinextractable P, but slightly decreased potential phosphatase activities. Potential enzyme activities varied among sites and sampling times, but did not explain much variation in microbial biomass or substrate availability. We conclude that reduced rainfall in tropical forests might accentuate some dry season patterns, like reductions in microbial biomass and accumulation of extractable nutrients. However, our data also suggest new patterns, like reduced inputs of DOC to soils with drying, which could have cascading effects on soil ecological function and C storage.

54 ENVIRONMENTAL SCIENCES↗

Tropical Tree Species Identity and Diameter at Breast Height in a Throughfall-Reduction Drying Experiment in Four Lowland Panamanian Forests

Objectives: Climatic drying is predicted for many tropical forests, yet effects on soil properties across moisture and soil gradients within tropical forests remain poorly characterized, hampering predictions of forest-climate feedbacks. We hypothesized that drying would suppress soil CO2 fluxes (i.e., respiration) in already-drier tropical forests by further reductions in soil moisture, but increases CO2 fluxes in wetter tropical forests by alleviating anaerobiosis and soil saturation. We measured soil CO2 fluxes, soil moisture, soil temperature, and forest floor biomass during wet-dry cycles (2015 – 2022) in four Panamanian forests that vary in rainfall and soil fertility. We also surveyed all tree species and identified to species in 2018 and 2019.Results: We found that soil moisture peaked in the wet season and declined in the dry season. Measured soil CO2 fluxes declined in the dry season and peaked in the early wet season ahead of peak soil moisture, resulting in a lower soil moisture optimum for respiration than previously modeled. Chronic throughfall exclusion also suppressed soil moisture across the four forests to 20cm depths, and also initially suppressed soil CO2 fluxes across forests. There was sustained suppression of soil CO2 fluxes after four years in the wettest forest only (-28 ± 4% during the dry season), but elevated soil CO2 fluxes in a fertile forest after four years (+75 ± 28% during the late wet season). The unexpected negative drying effect in the wettest, most infertile forest could have resulted from reduced vertical flushing of nutrients into soils, as the drying effect increased with time. Including hydro-nutrient interactions in ecosystem models could improve predictions of tropical forest-climate feedbacks (results presented in Cusack et al. 2023). Datasets included: Datasets included here include .csv and .xls files for tree species identity and diameter at breast height (dbh) in the study plots. There is also a .kml file that includes coordinates for all 32 plots included in the study of four forests (n = 4 throughfall reduction and n = 4 control plots per site). No special software is needed to open these files.

54 ENVIRONMENTAL SCIENCES↗

Throughfall-Reduction Drying Effects on Forest Floor Plant Tissue Biomass (Oi+Oe soil horizons) and Moisture Content in Four Lowland Panamanian Forests

Objectives: Climatic drying is predicted for many tropical forests, yet effects on soil properties across moisture and soil gradients within tropical forests remain poorly characterized, hampering predictions of forest-climate feedbacks. We hypothesized that drying would suppress soil CO2 fluxes (i.e., respiration) in already-drier tropical forests by further reductions in soil moisture, but increases CO2 fluxes in wetter tropical forests by alleviating anaerobiosis and soil saturation. We measured soil CO2 fluxes, soil moisture, soil temperature, and forest floor biomass during wet-dry cycles (2015 – 2022) in four Panamanian forests that vary in rainfall and soil fertility. We also surveyed all tree species and identified to species in 2018 and 2019.Results: We found that soil moisture peaked in the wet season and declined in the dry season. Measured soil CO2 fluxes declined in the dry season and peaked in the early wet season ahead of peak soil moisture, resulting in a lower soil moisture optimum for respiration than previously modeled. Forest floor biomass peaked in the dry season, in contrast. Chronic throughfall exclusion also suppressed soil moisture across the four forests to 20cm depths, and also initially suppressed soil CO2 fluxes across forests. There was sustained suppression of soil CO2 fluxes after four years in the wettest forest only (-28 ± 4% during the dry season), but elevated soil CO2 fluxes in a fertile forest after four years (+75 ± 28% during the late wet season). The unexpected negative drying effect in the wettest, most infertile forest could have resulted from reduced vertical flushing of nutrients into soils, as the drying effect increased with time. Including hydro-nutrient interactions in ecosystem models could improve predictions of tropical forest-climate feedbacks (results presented in Cusack et al. 2023). Datasets included: Datasets included here include .csv and .xls files for forest floor biomass (equivalent to the Oi+Oe soil horizons), and a subset include forest floor biomass moisture content (weight/weight) in the study plots. There is also a .kml file that includes coordinates for all 32 plots included in the study of four forests (n = 4 throughfall reduction and n = 4 control plots per site). No special software is needed to open these files.

54 ENVIRONMENTAL SCIENCES↗

Throughfall-Reduction Drying Effects on Soil Moisture and Soil Temperature at 30-Minute Intervals Four Lowland Panamanian Forests

Objectives: Climatic drying is predicted for many tropical forests, yet effects on soil properties across moisture and soil gradients within tropical forests remain poorly characterized, hampering predictions of forest-climate feedbacks. We hypothesized that drying would suppress soil CO2 fluxes (i.e., respiration) in already-drier tropical forests by further reductions in soil moisture, but increases CO2 fluxes in wetter tropical forests by alleviating anaerobiosis and soil saturation. We measured soil CO2 fluxes, soil moisture, soil temperature, and forest floor biomass during wet-dry cycles (2015 – 2022) in four Panamanian forests that vary in rainfall and soil fertility. We also surveyed all tree species and identified to species in 2018 and 2019.Results: We found that soil moisture peaked in the wet season and declined in the dry season. Measured soil CO2 fluxes declined in the dry season and peaked in the early wet season ahead of peak soil moisture, resulting in a lower soil moisture optimum for respiration than previously modeled. Chronic throughfall exclusion also suppressed soil moisture across the four forests, and also initially suppressed soil CO2 fluxes across forests. There was sustained suppression of soil CO2 fluxes after four years in the wettest forest only (-28 ± 4% during the dry season), but elevated soil CO2 fluxes in a fertile forest after four years (+75 ± 28% during the late wet season). The unexpected negative drying effect in the wettest, most infertile forest could have resulted from reduced vertical flushing of nutrients into soils, as the drying effect increased with time. Including hydro-nutrient interactions in ecosystem models could improve predictions of tropical forest-climate feedbacks (Cusack et al. 2023). Datasets included: Datasets here include .csv and .xls files for volumetric soil moisture (volume/volume) and soil temperature (°C). Data were taken at 30-minute intervals at 5cm and 20cm depths using permanent probes and data loggers (see methods). Data presented are from 2018-2019. There is also a .kml file that includes coordinates for all 32 plots included in the study of four forests (n = 4 throughfall reduction and n = 4 control plots per site). No special software is needed to open these files.

54 ENVIRONMENTAL SCIENCES↗

Throughfall-Reduction Drying Effects on Soil Respiration, Moisture, and Temperature in Four Distinct Lowland Panamanian Forests from 2015-2022

Objectives: Climatic drying is predicted for many tropical forests, yet effects on soil properties across moisture and soil gradients within tropical forests remain poorly characterized, hampering predictions of forest-climate feedbacks. We hypothesized that throughfall reduction (i.e. chronic drying) would suppress soil CO2 fluxes (i.e., respiration) in already-drier tropical forests by further reductions in soil moisture, but increases CO2 fluxes in wetter tropical forests by alleviating anaerobiosis and soil saturation. We conducted a field study in four distinct lowland Panamanian forests that vary in background rainfall and soil fertility. In each forest we constructed throughfall reduction structures that reduce moisture inputs to plots by ~50% (see methods). In drying and paired control plots we measured soil CO2 fluxes, soil moisture (volume/volume), soil temperature (°C), air temperature (°C), and forest floor biomass during wet-dry cycles (2015 – 2022). We also collected litterfall biomass and surveyed all tree species for diameter at breast height (dbh), and identified tree species in 2018 and 2019.Results:We found that soil moisture peaked in the wet season and declined in the dry season. Measured soil CO2 fluxes declined in the dry season and peaked in the early wet season ahead of peak soil moisture, resulting in a lower soil moisture optimum for respiration than previously modeled. Chronic throughfall exclusion also suppressed soil moisture across the four forests, and also initially suppressed soil CO2 fluxes across forests. There was sustained suppression of soil CO2 fluxes after four years in the wettest forest only (-28 ± 4% during the dry season), but elevated soil CO2 fluxes in a fertile forest after four years (+75 ± 28% during the late wet season). The unexpected negative drying effect in the wettest, most infertile forest could have resulted from reduced vertical flushing of nutrients into soils, as the drying effect increased with time. Including hydro-nutrient interactions in ecosystem models could improve predictions of tropical forest-climate feedbacks. Datasets included: Datasets included here include .csv and .xls files for soil respiration (carbon dioxide flux) collected using the LiCor (see methods), air temperature °C , soil moisture (vl/vl), and soil temperature °C. Soil moisture and soil temperature were collected at 0-10cm depths using hand-held probes (see methods). Data are on ~3 week timescales from 2015-2022 with some gaps. There is also a .kml file that includes coordinates for all 32 plots included in the study of four forests (n = 4 throughfall reduction and n = 4 control plots per site). No special software is needed to open these files.

54 ENVIRONMENTAL SCIENCES↗

Throughfall-Reduction Drying Effects on Gravimetric Soil Moisture at Two Depths in Four Lowland Panamanian Forests from 2015-2022

Objectives: Climatic drying is predicted for many tropical forests, yet effects on soil properties across moisture and soil gradients within tropical forests remain poorly characterized, hampering predictions of forest-climate feedbacks. We hypothesized that drying would suppress soil CO2 fluxes (i.e., respiration) in already-drier tropical forests by further reductions in soil moisture, but increases CO2 fluxes in wetter tropical forests by alleviating anaerobiosis and soil saturation. We measured soil CO2 fluxes, soil moisture, soil temperature, and forest floor biomass during wet-dry cycles (2015 – 2022) in four Panamanian forests that vary in rainfall and soil fertility. We also surveyed all tree species and identified to species in 2018 and 2019.Results: We found that soil moisture peaked in the wet season and declined in the dry season. Measured soil CO2 fluxes declined in the dry season and peaked in the early wet season ahead of peak soil moisture, resulting in a lower soil moisture optimum for respiration than previously modeled. Chronic throughfall exclusion also suppressed soil moisture across the four forests to 20cm depths, and also initially suppressed soil CO2 fluxes across forests. There was sustained suppression of soil CO2 fluxes after four years in the wettest forest only (-28 ± 4% during the dry season), but elevated soil CO2 fluxes in a fertile forest after four years (+75 ± 28% during the late wet season). The unexpected negative drying effect in the wettest, most infertile forest could have resulted from reduced vertical flushing of nutrients into soils, as the drying effect increased with time. Including hydro-nutrient interactions in ecosystem models could improve predictions of tropical forest-climate feedbacks (results presented in Cusack et al. 2023). Datasets included: Datasets included here include .csv and .xls files for gravimetric soil moisture (weight/weight). Soil moisture was collected on a quarterly basis from 0-10 cm and 10-20 cm depths using hand-held constant-volume soil corers. Data are on ~3 month timescales from 2015-2022 with some gaps. There is also a .kml file that includes coordinates for all 32 plots included in the study of four forests (n = 4 throughfall reduction and n = 4 control plots per site). No special software is needed to open these files.

54 ENVIRONMENTAL SCIENCES↗

Carbon Organisms Rhizosphere and Protection in Soil Environment model script and input data for soil moisture-respiration responses in tropical forests

Objectives: Climatic drying is predicted for many tropical forests, yet models remain poorly parameterized for tropical forests, hampering predictions of forest-climate feedbacks. We applied an integrated model–experiment approach, parameterizing an ecosystem model Carbon Organisms Rhizosphere and Protection in the Soil Environment (CORPSE) with tropical forest observational data, and comparing model predictions with a field drying manipulation. We hypothesized that drying would suppress soil CO2 fluxes (i.e., respiration) in already-drier tropical forests, but increases CO2 fluxes in wetter tropical forests by alleviating anaerobiosis. We measured soil CO2 fluxes, soil moisture, soil temperature, and forest floor biomass during wet-dry cycles (2015 – 2022) in four Panamanian forests that vary in rainfall and soil fertility. We used the field data to parameterize and run tests in the model.Results: Measured CO2 fluxes declined in the dry season and peaked in the early wet season ahead of peak soil moisture, resulting in a lower soil moisture optimum for respiration than previously modeled. We used this data to parameterize the model, which then predicted increased soil CO2 fluxes in wetter and fertile forests with drying, and decreased fluxes in drier, infertile forests. In contrast to model predictions, a chronic throughfall exclusion experiment in the forests initially suppressed soil CO2 fluxes across forests, with sustained suppression after four years in the wettest forest only (-28 ± 4% during the dry season), but elevated soil CO2 fluxes in a fertile forest after four years (+75 ± 28% during the late wet season), as predicted by the model. The unexpected negative drying effect in the wettest, most infertile forest could have resulted from reduced vertical flushing of nutrients into soils. Including hydro-nutrient interactions in ecosystem models could improve predictions of tropical forest-climate feedbacks (results presented in Cusack et al. 2023). Datasets included: Code files:CORPSE_array.py: Defines the equations of the CORPSE modelCORPSE_solvers: Functions for running the CORPSE model using either iterative or ordinary differential equation (ODE) solversrun_Panama_sims.py: Read in datasets and run the model simulations for this studyInput data:PanamaGradientEcosystemChem_BT_CPools_20152016CO2_DC_20190615.xlsx: Plot characteristics used in running model simulationsLiCor compiled surface flux only to 2020_03 DC_20200825.xlsx: Surface gas exchange fluxes used in model-data comparisonsPARCHED litterfall data for Ben Sulman LD 20200902.xlsx: Litterfall data used to drive model simulationsInitialization data:state_500y_20190823.csv: Initial state of model pools based on previous spinup runsOutput data:Outputs/prev_moisture_response.csv: Simulations of multiple sites using original model moisture response function.Outputs/updated_moisture_response.csv: Simulations of multiple sites using updated model moisture response function.Outputs/dry15_prev_moisture_response.csv: Simulations with soil moisture reduced by 15%, using original moisture response function.Outputs/dry15_updated_moisture_response.csv: Simulations with soil moisture reduced by 15%, using updated moisture response function.Outputs/dry30_prev_moisture_response.csv: Simulations with soil moisture reduced by 30%, using original moisture response function.Outputs/dry30_updated_moisture_response.csv: Simulations with soil moisture reduced by 30%, using updated moisture response function.Outputs/latestart_prev_moisture_response.csv: Simulations with extended dry season, using original moisture response function.Outputs/latestart_updated_moisture_response.csv: Simulations with extended dry season, using updated moisture response function.Outputs/[site name]_oneyear.csv: One-year simulation for each site in expanded site list using original moisture response function.Outputs/[site name]_oneyear_dried.csv: One-year simulation for each site in expanded site list using original moisture response function, with soil moisture reduced by 25%.Outputs/[site name]_oneyear_updated_moisture_response.csv: One-year simulation for each site in expanded site list using updated moisture response function.Outputs/[site name]_oneyear_updated_moisture_response_dried.csv: One-year simulation for each site in expanded site list using updated moisture response function, with soil moisture reduced by 25%.Field plot location data:There is also a .kml file that includes coordinates for all 32 plots included in the study of four forests (n = 4 throughfall reduction and n = 4 control plots per site).

54 ENVIRONMENTAL SCIENCES↗

Differential effects of drought on nonstructural carbohydrate storage in seedlings and mature trees of four species in a subtropical forest

Nonstructural carbohydrates (NSC) play important roles in forest vulnerability to climate change, especially under increasing drought intensity and frequency. Understanding NSC dynamics is essential for accurately predicting the resistance and resilience of forests in response to drought. However, our knowledge of NSC re-sponses to drought is still limited due to the lack of research in trees of di?erent life stages. In this study, we conducted a throughfall exclusion experiment (TFE) with four subtropical evergreen tree species to examine drought e?ects on NSC in mature trees and understory seedlings. Our results showed the differential effects of drought on NSC dynamics of understory seedlings and mature trees. In the TFE experiment, mature trees of all four species were relatively homoeostatic with the insigni?cantly changed NSC pools, photosynthesis, and growth under the drought treatment compared to the control. In contrast, understory seedlings displayed sig-ni?cant decreases in total NSC and soluble sugars (-14.70 ± 3.66% and -16.93 ± 3.85%, respectively) with the exception of Lithocarpus glaber (L. glaber). The seedlings of L. glaber with the highest hydraulic resistance maintained or slightly increased NSC and its components in response to drought. Our study highlights the im-portance of life stage in assessing drought e?ects of trees on NSC storage and then forest C cycling, which could be incorporated into the dynamic global vegetation models (DGVMs) to better understand drought effects on forest C balance in the future.

Zhang, Peipei↗

The effects of mixed-species root zones on the resistance of soil bacteria and fungi to long-term experimental and natural reductions in soil moisture

Mixed forest stands tend to be more resistant to drought than species-specific stands partially due to complementarity in root ecology and physiology. For this work, we asked whether complementary differences in the drought resistance of soil microbiomes might contribute to this phenomenon. We experimented on the effects of reduced soil moisture on bacterial and fungal community composition in species-specific (single species) and mixed-species root zones of Norway spruce and European beech forests in a 5-year-old throughfall-exclusion experiment and across seasonal (spring-summer-fall) and latitudinal moisture gradients. Bacteria were most responsive to changes in soil moisture, especially members of Rhizobiales, while fungi were largely unaffected, including ectomycorrhizal fungi (EMF). Community resistance was higher in spruce relative to beech root zones, corresponding with the proportions of drought-favored (more in spruce) and drought-sensitive bacterial taxa (more in beech). The spruce soil microbiome also exhibited greater resistance to seasonal changes between spring (wettest) and fall (driest). Mixed-species root zones contained a hybrid of beech- and spruce-associated microbiomes. Several bacterial populations exhibited either enhanced resistance or greater susceptibility to drought in mixed root zones. Overall, patterns in the relative abundances of soil bacteria closely tracked moisture in seasonal and latitudinal precipitation gradients and were more predictive of soil water content than other environmental variables. We conclude that complementary differences in the drought resistance of soil microbiomes can occur and the likeliest form of complementarity in mixed-root zones coincides with the enrichment of drought-tolerant bacteria associated with spruce and the sustenance of EMF by beech.

54 ENVIRONMENTAL SCIENCES↗

Experimental warming and drying increase older carbon contributions to soil respiration in lowland tropical forests

Tropical forests account for over 50% of the global terrestrial carbon sink, but climate change threatens to alter the carbon balance of these ecosystems. We show that warming and drying of tropical forest soils may increase soil carbon vulnerability, by increasing degradation of older carbon. In situ whole-profile heating by 4 °C and 50% throughfall exclusion each increased the average radiocarbon age of soil CO 2 efflux by ~2–3 years, but the mechanisms underlying this shift differed. Warming accelerated decomposition of older carbon as increased CO 2 emissions depleted newer carbon. Drying suppressed decomposition of newer carbon inputs and decreased soil CO 2 emissions, thereby increasing contributions of older carbon to CO 2 efflux. These findings imply that both warming and drying, by accelerating the loss of older soil carbon or reducing the incorporation of fresh carbon inputs, will exacerbate soil carbon losses and negatively impact carbon storage in tropical forests under climate change.

54 ENVIRONMENTAL SCIENCES↗

Radiocarbon of soil CO2 efflux and soils with experimental soil warming and drying in lowland tropical forests in Panama

In 2019, we measured the Δ14C and δ13C of soil respired carbon dioxide (CO2) in Panamanian forests that are subject to either in situ experimental soil warming (4C above ambient temperature to 1.2 m depth) or in situ experimental drying (50% throughfall exclusion). The warming site and one drying site are both within the Barro Colorado Nature Monument in nearby and similar forests on similar soils, enabling direct comparison of warming and drying effects on soil CO2 efflux. A second drying experiment is on the northern side of the Panama Isthmus on infertile soils where mean annual precipitation is greater, representative of a broad geographic area of the tropics. Given the seasonality of these forests, we performed measurements at stages of the seasonal cycle for which we expected the largest variation in CO2 efflux between control and experimental plots based on previous studies – the wet season (October-December) and dry season (March/April) or dry-to-wet season transition (May). This dataset includes Δ14C and δ13C of in situ soil surface CO2 flux as well as CO2 flux rates, volumetric soil moisture, soil temperature, and calculated partitioning of the fraction of total soil respiration from heterotrophs vs roots at the time of isotope sampling in AllSites_SoilResp_14C_data.xlsx. This dataset also includes Δ14C and δ13C of bulk soil, density fractions, and CO2 respired during laboratory incubations in AllSites_bulk_soil14C.xlsx. Datafiles are also available in csv format.

54 ENVIRONMENTAL SCIENCES↗