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Suspended sediment transport in montane headwaters is important to water quality and nutrient balances. However, predictions of sediment source and transport can be difficult, in part, because of a changing climate and increasing frequencies of disturbances. We used observations from 10 headwater streams in water year (WY; starting on 1st October ending on 30th September) 2007–2009 and 2013–2018 to determine the potential impacts of climate and forest management on suspended sediment delivery. We analysed hysteretic responses of suspended sediment for 76 events in five headwater catchments within a snow-dominated site and another five within a lower-elevation, rain-snow transition site, in the mixed-conifer zone of California's Sierra Nevada. Hysteresis patterns were predominantly clockwise at both sites, suggesting localized sediment sources such as streambeds and banks. The warmer, transition site exhibited a lower proportion of clockwise-loop events, faster transport speed and higher peak sediment concentrations than the snow-dominated site. This suggests extended sediment sources and increases in transport can occur as currently snow-dominated areas become rain-snow transitional. Over the nine water years, we observed similar hysteresis effects amongst years under drought, near-average, and extremely wet conditions. Hence, fluctuations in precipitation amounts across years may not influence sediment source area substantially. Furthermore, we compared hysteresis metrics between the control, thin only, burn only and thin combined with burn catchments during the posttreatment period (WY 2013– 2018). Hysteresis effects remained unchanged amongst treatments, which may be attributed to the combinations of low-intensity operations implemented with best management practises combined with a four-year drought (WY 2013–2016). Taken together, sediment sources in small headwater catchments will probably remain localized with changing precipitation levels and low-intensity management operations, but it may be extended and potentially lead to higher sediment yields as the main hydrologic input shifts from primarily snow to a mix of rain and snow.

Increasing wildfire severity, which is common throughout the western United States, can have deleterious effects on plant regeneration and large impacts on carbon (C) and nitrogen (N) cycling rates. Soil microbes are pivotal in facilitating these elemental cycles, so understanding the impact of increasing fire severity on soil microbial communities is critical. Here, we assess the long-term impact of high-severity fires on the soil microbiome. We find that high-severity wildfires result in a multi-decadal (>25 y) recovery of the soil microbiome mediated by concomitant differences in aboveground vegetation, soil chemistry, and microbial assembly processes. Our results depict a distinct taxonomic and functional successional pattern of increasing selection in post-fire soil microbial communities. Changes in microbiome composition corresponded with changes in microbial functional potential, specifically altered C metabolism and enhanced N cycling potential, which related to rates of potential decomposition and inorganic N availability, respectively. Based on metagenome-assembled genomes, we show that bacterial genomes enriched in our earliest site (4 y since fire) harbor distinct traits such as a robust stress response and a high potential to degrade pyrogenic, polyaromatic C that allow them to thrive in post-fire environments. Taken together, these results provide a biological basis for previously reported process rate measurements and explain the temporal dynamics of post-fire biogeochemistry, which ultimately constrains ecosystem recovery.

Freshwater lakes are important but poorly constrained sources of methane (CH4) to the atmosphere, reflecting high but variable rates of CH4 production as well as limited and inconsistent measuremen...

Understanding the transport of dissolved organic carbon (DOC) and nitrogen (N) as water flows through headwater basins is important for predicting downstream water quality. With increased recognition of climatic impact on nutrient transport, more studies are needed in headwater basins experiencing a Mediterranean-type climate, such as those of the Sierra Nevada, California. We analyzed water samples collected over 5 years from eight low-order and mixed-conifer watersheds to elucidate the temporal variation of water chemistry and evaluate their responses to prolonged drought and low-intensity forest thinning. We observed higher stream DOC concentrations in October compared to other months within water years prior to drought and thinning, suggesting the importance of antecedent moisture conditions on seasonal C export. In unthinned watersheds, stream DOC concentrations were lower (62%) and DOC aromaticity was higher (68 and 92%, depending on the index used) during drought compared to non-drought years. In thinned watersheds during drought years, stream water had higher DOC concentrations (66–94% in three consecutive years following thinning) and dissolved inorganic N (24%, in the third year following thinning) compared to unthinned watersheds during drought. Additionally, lower stream DOC concentrations were found in watersheds with higher elevations and lower drainage densities in the year with near-average precipitation; however, these correlations were not significant in years with greater or extremely low precipitation. Taken together, our results suggest that stream concentrations of DOC and dissolved N in Mediterranean headwater basins are extremely variable over time due to the high temporal climatic variabilities and periodic management practices.

As the largest biogeochemically active terrestrial reserve of carbon (C), soils have the potential to either mitigate or amplify rates of climate change. Ecosystems with large C stocks and high rates of soil C sequestration, in particular, may have outsized impacts on regional and global C cycles. Montane meadows have large soil C stocks relative to surrounding ecosystems. However, anthropogenic disturbances in many meadows may have altered the balance of C inputs and outputs, potentially converting these soils from net C sinks to net sources of C to the atmosphere. Here, we quantified ecosystem-level C inputs and outputs to estimate the annual net soil C flux from 13 montane meadows spanning a range of conditions throughout the California Sierra Nevada. Our results suggest that meadow soils can be either large net C sinks (577.6 ± 250.5 g C m−2 y−1) or sources of C to the atmosphere (− 391.6 ± 154.2 g C m−2 y−1). Variation in the direction and magnitude of net soil C flux appears to be driven by belowground C inputs. Vegetation species and functional group composition were not associated with the direction of net C flux, but climate and watershed characteristics were. Our results demonstrate that, per unit area, montane meadows hold a greater potential for C sequestration than the surrounding forest. However, legacies of disturbance have converted some meadows to strong net C sources. Accurate quantification of ecosystem-level C fluxes is critical for the development of regional C budgets and achieving global emissions goals.

Giant sequoia (Sequoiadendron giganteum) is an iconic conifer that lives in relict populations on the western slopes of the California Sierra Nevada. In these settings, it is unusual among the dominant trees in that it associates with arbuscular mycorrhizal fungi rather than ectomycorrhizal fungi. However, it is unclear whether differences in microbial associations extend more broadly to nonmycorrhizal components of the soil microbial community. To address this question, we used next‐generation amplicon sequencing to characterize bacterial/archaeal and fungal microbiomes in bulk soil (0–5 cm) beneath giant sequoia and co‐occurring sugar pine (Pinus lambertiana) individuals. We did this across two groves with distinct parent material in Yosemite National Park, USA. We found tree‐associated differences were apparent despite a strong grove effect. Bacterial/archaeal richness was greater beneath giant sequoia than sugar pine, with a core community double the size. The tree species also harbored compositionally distinct fungal communities. This pattern depended on grove but was associated with a consistently elevated relative abundance of Hygrocybe species beneath giant sequoia. Compositional differences between host trees correlated with soil pH and soil moisture. We conclude that the effects of giant sequoia extend beyond mycorrhizal mutualists to include the broader community and that some but not all host tree differences are grove‐dependent., Giant sequoia (Sequoiadendron giganteum) is an iconic conifer that lives in relict populations on the western slopes of the California Sierra Nevada. In these settings, it is unusual among the dominant trees in that it associates with arbuscular mycorrhizal fungi rather than ectomycorrhizal fungi. We found that the effects of giant sequoia extend beyond mycorrhizal mutualists to include the broader community and that some but not all host tree differences are grove‐dependent.

Chemical forms of phosphorus (P) in airborne particulate matter (PM) are poorly known and do not correlate with solubility or extraction measurements commonly used to infer speciation. We used P X-ray absorption near-edge structure (XANES) and 31P nuclear magnetic resonance (NMR) spectroscopies to determine P species in PM collected at four mountain sites (Colorado and California). Organic P species dominated samples from high elevations, with organic P estimated at 65-100% of total P in bulk samples by XANES and 79-88% in extracted fractions (62-84% of total P) by NMR regardless of particle size (≥10 or 1-10 μm). Phosphorus monoester and diester organic species were dominant and present in about equal proportions, with low fractions of inorganic P species. By comparison, PM from low elevation contained mixtures of organic and inorganic P, with organic P estimated at 30-60% of total P. Intercontinental PM transport determined from radiogenic lead (Pb) isotopes varied from 0 to 59% (mean 37%) Asian-sourced Pb at high elevation, whereas stronger regional PM inputs were found at low elevation. Airborne flux of bioavailable P to high-elevation ecosystems may be twice as high as estimated by global models, which will disproportionately affect net primary productivity.

Tobacco and cannabis product use can result in debris (a.k.a. litter) on the landscape, with implications to soil and water quality and thus potential impacts to ecosystems. More information is needed regarding how much debris exists on the landscape, such that the magnitude of potential associated environment problems can be quantified. Such information can raise awareness in the public about environmental hazards from tobacco and cannabis product use. The goal of this protocol is to quantify, via a timed survey approach, the tobacco and cannabis debris load (i.e. cigarette butts, cannabis or e-cigarette waste) at study sites. Multiple sites in a geographical location may be of interest for comparative analysis, for example sites used regularly by humans (e.g. trails, parking lots, around buildings, garbage cans) versus other sites that are suspected to have low human use. In any case, the same survey data sheet would be used (included with this protocol). Each survey will take 15 minutes per site to conduct. In the process of surveying, debris is collected and retained for later sorting, counting, and photographic documentation. The time to sort and count the collected debris will depend on the quantity and diversity of debris collected.

Subsoil microbiomes play important roles in soil carbon and nutrient cycling, yet our understanding of the controls on subsoil microbial communities is limited. Here, we investigated the direct (mean annual temperature and precipitation) and indirect (soil chemistry) effects of climate on microbiome composition and extracellular enzyme activity throughout the soil profile across two elevation-bioclimatic gradients in central California, USA. We found that microbiome composition changes and activity decreases with depth. Across these sites, the direct influence of climate on microbiome composition and activity was relatively lower at depth. Furthermore, we found that certain microbial taxa change in relative abundance over large temperature and precipitation gradients only in specific soil horizons, highlighting the depth dependence of the climatic controls on microbiome composition. Our finding that the direct impacts of climate are muted at depth suggests that deep soil microbiomes may lag in their acclimation to new temperatures with a changing climate.

Increasing wildfire severity, which is common throughout the western United States, can have deleterious effects on plant regeneration and large impacts on carbon (C) and nitrogen (N) cycling rates. Soil microbes are pivotal in facilitating these elemental cycles, so understanding the impact of increasing fire severity on soil microbial communities is critical. Here, we assess the long-term impact of high-severity fires on the soil microbiome. We find that high-severity wildfires result in a multi-decadal (25 y) recovery of the soil microbiome mediated by concomitant differences in aboveground vegetation, soil chemistry, and microbial assembly processes. Our results depict a distinct taxonomic and functional successional pattern of increasing selection in post-fire soil microbial communities. Changes in microbiome composition corresponded with changes in microbial functional potential, specifically altered C metabolism and enhanced N cycling potential, which related to rates of potential decomposition and inorganic N availability, respectively. Based on metagenome-assembled genomes, we show that bacterial genomes enriched in our earliest site (4 y since fire) harbor distinct traits such as a robust stress response and a high potential to degrade pyrogenic, polyaromatic C that allow them to thrive in post-fire environments. Taken together, these results provide a biological basis for previously reported process rate measurements and explain the temporal dynamics of post-fire biogeochemistry, which ultimately constrains ecosystem recovery.

The effects of organic matter amendments (OMA) on soil fertility in permanent cropping systems like orchards is under-studied compared to annual cropping systems. We evaluated experimentally the impact of OMAs on soil fertility in almond (Prunus dulcis) orchards over a two-year period with annual applications. Two OMAs, derived from composted green waste (GWC) or composted manure wood chips (MWC), were applied as surface mulch and compared to a control at two sites with different soil textures (sandy loam and loamy sand). OMAs increased soil moisture content (0–0.1 m depth) at both sites by 27–37%. Both amendments increased soil inorganic N at the sandy loam (GWC: 194%; MWC: 114%) and loamy sand (GWC: 277%; MWC: 114%) sites the month following application, but soil inorganic N concentrations quickly decreased to values similar to those of control plots. After two-years, the GWC and the MWC amendments increased the soil cation exchange capacity (CEC) by 112% and 29%, respectively, in the sandy loam site, but no change was observed in the loamy sand site. The greatest increase in soil extractable K occurred in the GWC-amended plots at the sandy loam site even though the initial K concentration of MWC was higher. Both OMAs increased soil organic carbon (SOC) after two years, but the SOC increase in the GWC-amended plots was greater. Our results suggest that OMAs can significantly improve soil fertility after one or two annual applications, and that fertility gains appear to be dependent on soil texture than the nutrient concentrations of the OMA.

Unlike plants and vertebrates, the ecological preferences, and potential vulnerabilities of soil invertebrates to environmental change, remain poorly understood in terrestrial ecosystems globally. We conducted a cross-biome survey including 83 locations across six continents to advance our understanding of the ecological preferences and vulnerabilities of the diversity of dominant and functionally important soil invertebrate taxa, including nematodes, arachnids and rotifers. The diversity of invertebrates was analyzed through amplicon sequencing. Vegetation and climate drove the diversity and dominant taxa of soil invertebrates. Our results suggest that declines in forest cover and plant diversity, and reductions in plant production associated with increases in aridity, can result in reductions of the diversity of soil invertebrates in a drier and more managed world. We further developed global atlases of the diversity of these important soil invertebrates, which were cross-validated using an independent database. Our study advances the current knowledge of the ecological preferences and vulnerabilities of the diversity and presence of functionally important soil invertebrates in soils from across the globe. This information is fundamental for improving and prioritizing conservation efforts of soil genetic resources and management policies.

Belowground organisms play critical roles in maintaining multiple ecosystem processes, including plant productivity, decomposition, and nutrient cycling. Despite their importance, however, we have a limited understanding of how and why belowground biodiversity (bacteria, fungi, protists, and invertebrates) may change as soils develop over centuries to millennia (pedogenesis). Moreover, it is unclear whether belowground biodiversity changes during pedogenesis are similar to the patterns observed for aboveground plant diversity. Here we evaluated the roles of resource availability, nutrient stoichiometry, and soil abiotic factors in driving belowground biodiversity across 16 soil chronosequences (from centuries to millennia) spanning a wide range of globally distributed ecosystem types. Changes in belowground biodiversity during pedogenesis followed two main patterns. In lower-productivity ecosystems (i.e., drier and colder), increases in belowground biodiversity tracked increases in plant cover. In more productive ecosystems (i.e., wetter and warmer), increased acidification during pedogenesis was associated with declines in belowground biodiversity. Changes in the diversity of bacteria, fungi, protists, and invertebrates with pedogenesis were strongly and positively correlated worldwide, highlighting that belowground biodiversity shares similar ecological drivers as soils and ecosystems develop. In general, temporal changes in aboveground plant diversity and belowground biodiversity were not correlated, challenging the common perception that belowground biodiversity should follow similar patterns to those of plant diversity during ecosystem development. Taken together, our findings provide evidence that ecological patterns in belowground biodiversity are predictable across major globally distributed ecosystem types and suggest that shifts in plant cover and soil acidification during ecosystem development are associated with changes in belowground biodiversity over centuries to millennia. European Union's Horizon 2020 research and innovation program under Marie Sklodowska-Curie Grant [702057]; US National Science FoundationNational Science Foundation (NSF) [EAR1331828, DEB 1556090] This project received funding from the European Union's Horizon 2020 research and innovation program under Marie Sklodowska-Curie Grant Agreement 702057. N.F. was supported through grants from the US National Science Foundation (EAR1331828, DEB 1556090). Any use of trade, product, or firm names is for descriptive purposes only and does not imply endorsement by the US Government. An extended version of the acknowledgments is provided in SI Appendix. Public domain – authored by a U.S. government employee

Increasing global temperatures are predicted to stimulate soil microbial respiration. The direct and indirect impacts of warming on soil microbes, nevertheless, remain unclear. This is particularly true for understudied subsoil microbes. Here, we show that 4.5 years of whole-profile soil warming in a temperate mixed forest results in altered microbial community composition and metabolism in surface soils, partly due to carbon limitation. However, microbial communities in the subsoil responded differently to warming than in the surface. Throughout the soil profile—but to a greater extent in the subsoil—physiologic and genomic measurements show that phylogenetically different microbes could utilize complex organic compounds, dampening the effect of altered resource availability induced by warming. We find subsoil microbes had 20% lower carbon use efficiencies and 47% lower growth rates compared to surface soils, which constrain microbial communities. Collectively, our results show that unlike in surface soils, elevated microbial respiration in subsoils may continue without microbial community change in the near-term., There is much uncertainty on the response of soil microbial communities to warming, particularly in the subsoil. Here, the authors investigate microbial community and metabolism response to 4.5 years of whole-profile soil warming, finding depth-dependent effects and elevated subsoil microbial respiration.

The rapidly increasing global population and anthropogenic climate change have created intense pressure on agricultural systems to produce increasingly more food under steadily challenging environmental conditions. Simultaneously, industrial agriculture is negatively affecting natural and agricultural ecosystems because of intensive irrigation and fertilization to fully utilize the potential of high-yielding cultivars. Growth-promoting microbes that increase stress tolerance and crop yield could be a useful tool for helping mitigate these problems. We investigated if commercially grown almonds might be a resource for plant colonizing bacteria with growth promotional traits that could be used to foster more productive and sustainable agricultural ecosystems. We isolated an endophytic bacterium from almond leaves that promotes growth of the model plant Arabidopsis thaliana. Genome sequencing revealed a novel Erwinia gerundensis strain (A4) that exhibits the ability to increase access to plant nutrients and to produce the stress-mitigating polyamine spermidine. Because E. gerundensis is known to be able to colonize diverse plant species including cereals and fruit trees, A4 may have the potential to be applied to a wide variety of crop systems.

Riding the global waves of decriminalization, medical or recreational use of cannabis (Cannabis sativa spp.) is now legal in more than 50 countries and U.S. states. As governments regulate this formerly illegal crop, there is an urgent need to understand how cannabis may impact the environment. Due to the challenges of researching quasi-legal commodities, peer-reviewed studies documenting environmental impacts of cannabis are limited, slowing the development of policies and agricultural extension guidelines needed to minimize adverse environmental outcomes. Here we review peer-reviewed research on relationships between cannabis and environmental outcomes, and identify six documented impact pathways from cannabis cultivation (land-cover change, water use, pesticide use, energy use, and air pollution) and consumption (water pollution). On the basis of reviewed findings, we suggest policy directions for these pathways. We further highlight the need to formalize existing traditional and gray literature knowledge, expand research partnerships with cannabis cultivators, and ease research restrictions on cannabis. Finally, we discuss how science might contribute to minimize environmental risks and inform the development of regulations for a growing global cannabis industry.

During the past century, systematic wildfire suppression has decreased fire frequency and increased fire severity in the western United States of America. While this has resulted in large ecological changes aboveground such as altered tree species composition and increased forest density, little is known about the long-term, belowground implications of altered, ecologically novel, fire regimes, especially on soil biological processes. To better understand the long-term implications of ecologically novel, high-severity fire, we used a 44-yr high-severity fire chronosequence in the Sierra Nevada where forests were historically adapted to frequent, low-severity fire, but were fire suppressed for at least 70 yr. High-severity fire in the Sierra Nevada resulted in a long-term (44 +yr) decrease (50%, P 0.05) in soil extracellular enzyme activities, basal microbial respiration (56-72%, P 0.05), and organic carbon (50%, P 0.05) in the upper 5 cm compared to sites that had not been burned for at least 115 yr. However, nitrogen (N) processes were only affected in the most recent fire site (4 yr post-fire). Net nitrification increased by over 600% in the most recent fire site (P 0.001), but returned to similar levels as the unburned control in the 13-yr site. Contrary to previous studies, we did not find a consistent effect of plant cover type on soil biogeochemical processes in mid-successional (10-50 yr) forest soils. Rather, the 44-yr reduction in soil organic carbon (C) quantity correlated positively with dampened C cycling processes. Our results show the drastic and long-term implication of ecologically novel, high-severity fire on soil biogeochemistry and underscore the need for long-term fire ecological experiments.

The role of soil biodiversity in regulating multiple ecosystem functions is poorly understood, limiting our ability to predict how soil biodiversity loss might affect human wellbeing and ecosystem sustainability. Here, combining a global observational study with an experimental microcosm study, we provide evidence that soil biodiversity (bacteria, fungi, protists and invertebrates) is significantly and positively associated with multiple ecosystem functions. These functions include nutrient cycling, decomposition, plant production, and reduced potential for pathogenicity and belowground biological warfare. Our findings also reveal the context dependency of such relationships and the importance of the connectedness, biodiversity and nature of the globally distributed dominant phylotypes within the soil network in maintaining multiple functions. Moreover, our results suggest that the positive association between plant diversity and multifunctionality across biomes is indirectly driven by soil biodiversity. Together, our results provide insights into the importance of soil biodiversity for maintaining soil functionality locally and across biomes, as well as providing strong support for the inclusion of soil biodiversity in conservation and management programmes. Combining field data from 83 sites on five continents, together with microcosm experiments, the authors show that nutrient cycling, decomposition, plant production and other ecosystem functions are positively associated with a higher diversity of a wide range of soil organisms. Marie Sklodowska-Curie We thank N. Fierer, M. Gebert, J. Henley, V. Ochoa, F. T. Maestre and B. Gozalo for their help with laboratory analyses; O. Sala, C. Siebe, C. Currier, M. A. Bowker, V. Parry, H. Lambers, P. Vitousek, V. M. Pena-Ramirez, L. Riedel, J. Larson, K. Waechter, W. Williams, S. Williams, B. Sulman, D. Buckner and B. Anacker for their help with soil sampling in Colorado, Hawaii, Iceland, New Mexico, Arizona, Mexico and Australia; the City of Boulder Open Space and Mountain Parks for allowing us to conduct these samplings; C. Cano-Diaz for her advice about R analyses; S. K. Travers for her help with mapping. This project has received funding from the European Union's Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie grant agreement no. 702057. M.D.-B. is supported by the Spanish Government under a Ramon y Cajal contract RYC2018-025483-I. This research is supported by the Australian Research Council projects (DP170104634; DP190103714). S.A. and F.D.A. are funded by FONDECYT 1170995, IAI-CRN 3005, PFB-23 (from CONICYT) and P05-002 (from Millennium Scientific Initiative). N.A.C. acknowledges support from Churchill College, University of Cambridge; and M.A.W. from the Wilderness State Park, Michigan for access to sample soil and conduct ecosystem survey. B.K.S. acknowledges a research award from the Humboldt Foundation. J.-Z.H. acknowledges support from the Australia Research Council (project DP170103628); and A.G. from the Spanish Ministry (project CGL2017-88124-R). F.B. thanks the Spanish Ministry and FEDER funds for the CICYT project AGL2017-85755-R, the CSIC project 201740I008 and funds from 'Fundacion Seneca' from Murcia Province (19896/GERM/15). P.T. thanks K. Little for her help with laboratory analyses. S.C.R. was supported by the US Geological Survey Ecosystems Mission Area. Any use of trade, firm or product names is for descriptive purposes only and does not imply endorsement by the US Government. S.N. was funded by the Austrian Science Fund (grant Y801-B16). Public domain – authored by a U.S. government employee

Sequential chemical extraction has been widely used to study soil phosphorus (P) dynamics and inform nutrient management, but its efficacy for assigning P into biologically meaningful pools remains unknown. Here, we evaluated the accuracy of the modified Hedley extraction scheme using P K-edge X-ray absorption near-edge structure (XANES) spectroscopy for nine carbonate-free soil samples with diverse chemical and mineralogical properties resulting from different degrees of soil development. For most samples, the extraction markedly overestimated the pool size of calcium-bound P (Ca-P, extracted by 1 M HCl) due to (1) P redistribution during the alkaline extractions (0.5 M NaHCO3 and then 0.1 M NaOH), creating new Ca-P via formation of Ca phosphates between NaOH-desorbed phosphate and exchangeable Ca2+ and/or (2) dissolution of poorly crystalline Fe and Al oxides by 1 M HCl, releasing P occluded by these oxides into solution. The first mechanism may occur in soils rich in well-crystallized minerals and exchangeable Ca2+ regardless of the presence or absence of CaCO3, whereas the second mechanism likely operates in soils rich in poorly crystalline Fe and Al minerals. The overestimation of Ca-P simultaneously caused underestimation of the pools extracted by the alkaline solutions. Our findings identify key edaphic parameters that remarkably influenced the extractions, which will strengthen our understanding of soil P dynamics using this widely accepted procedure.

Endophytes have been defined as microorganisms living inside plant tissues without causing negative effects on their hosts. Endophytic microbes have been extensively studied for their plant growth-promoting traits. However, analyses of endophytes require complete removal of epiphytic microorganisms. We found that the established tests to evaluate surface sterility, polymerase chain reaction, and leaf imprints, are unreliable. Therefore, we used scanning electron microscopy (SEM) as an additional assessment of epiphyte removal. We used a diverse suite of sterilization protocols to remove epiphytic microorganisms from the leaves of a gymnosperm and an angiosperm tree to test the influence of leaf morphology on the efficacy of these methods. Additionally, leaf tissue damage was also evaluated by SEM, as damaging the leaves might have an impact on endophytes and could lead to inaccurate assessment of endophytic communities. Our study indicates, that complete removal of the leaf cuticle by the sterilization technique assures loss of epiphytic microbes, and that leaves of different tree species may require different sterilization protocols. Furthermore, our study demonstrates the importance of choosing the appropriate sterilization protocol to prevent erroneous interpretation of host-endophyte interactions. Moreover, it shows the utility of SEM for evaluating the effectiveness of surface sterilization methods and their impact on leaf tissue integrity.

The importance of soil age as an ecosystem driver across biomes remains largely unresolved. By combining a cross-biome global field survey, including data for 32 soil, plant, and microbial properties in 16 soil chronosequences, with a global meta-analysis, we show that soil age is a significant ecosystem driver, but only accounts for a relatively small proportion of the cross-biome variation in multiple ecosystem properties. Parent material, climate, vegetation and topography predict, collectively, 24 times more variation in ecosystem properties than soil age alone. Soil age is an important local-scale ecosystem driver; however, environmental context, rather than soil age, determines the rates and trajectories of ecosystem development in structure and function across biomes. Our work provides insights into the natural history of terrestrial ecosystems. We propose that, regardless of soil age, changes in the environmental context, such as those associated with global climatic and land-use changes, will have important long-term impacts on the structure and function of terrestrial ecosystems across biomes., This project received funding from the European Union’s Horizon 2020 research and innovation program under the Marie Sklodowska-Curie Grant Agreement No. 702057 (CLIMIFUN). M.D.-B. is supported by a Ramón y Cajal grant from the Spanish Ministry of Science and Innovation (RYC2018-025483-I), and by the BES Grant Agreement No. LRB17\1019 (MUSGONET). F.B. is grateful to the Spanish Ministry and FEDER funds for the project AGL2017–85755-R, the i-LINK+2018 (LINKA20069) from CSIC, and received funds from “Fundación Séneca” from Murcia Province (19896/GERM/15). S.R. was supported by the US Geological Survey Ecosystems Mission Area. C.P. acknowledges support from the Spanish State Plan for Scientific and Technical Research and Innovation (2013–2016), award ref. AGL201675762-R (AEI/FEDER, UE). A.G. acknowledges support from the Spanish Ministry of Science (CGL2017-88124-R). F.A. is supported by FONDECYT 11180538 and S.A. by FONDECYT 1170995. We would like to thank Peter Vitousek for his comments on a previous draft of this paper. Moreover, we thank Matt Gebert, Jessica Henley, Fernando T. Maestre, Victoria Ochoa, and Beatriz Gozalo for their help with lab analyses, and Emilio Guirado for his advice with topographic analyses. We also want to thank Osvaldo Sala, Matthew A. Bowker, Peter Vitousek, Courtney Currier, Martin Kirchmair, Victor M. Peña-Ramírez, Lynn Riedel, Julie Larson, Katy Waechter, David Buckner, and Brian Anacker for their help with soil sampling, and to the City of Boulder Open Space and Mountain Parks for allowing us to conduct these samplings. We are also grateful to the Division of Forestry and Wildlife of the State of Hawai’i and Koke’e State Park for their logistical assistance and for allowing us access to the HA sites. The Arizona research sites were established with the support of an EPA‐STAR Graduate Fellowship (U‐916251), a Merriam‐Powell Center for Environmental Research Graduate Fellowship, an Achievement Rewards for College Scientists (ARCS) Foundation of Arizona Scholarship, and McIntire‐Stennis appropriations to Northern Arizona University and the State of Arizona. Any use of trade, product, or firm names is for descriptive purposes only and does not imply endorsement by the U.S. Government.

The release of water during snowmelt orchestrates a variety of important belowground biogeochemical processes in seasonally snow-covered ecosystems, including the production and consumption of greenhouse gases (GHGs) by soil microorganisms. Snowmelt timing is advancing rapidly in these ecosystems, but there is still a need to isolate the effects of earlier snowmelt on soil GHG fluxes. For an improved mechanistic understanding of the biogeochemical effects of snowmelt timing during the snow-free period, we manipulated a high-elevation forest that typically receives over two meters of snowfall but little summer precipitation to influence legacy effects of snowmelt timing. We altered snowmelt rates for two years using black sand to accelerate snowmelt and white fabric to postpone snowmelt, thus creating a two- to three-week disparity in snowmelt timing. Soil microclimate and fluxes of carbon dioxide (CO2 ), methane (CH4 ), and nitrous oxide (N2 O) were monitored weekly to monthly during the snow-free period. Microbial abundances were estimated by potential assays near the end of each snow-free period. Although earlier snowmelt caused soil drying, we found no statistically significant effects (p

Large uncertainty remains in the spatial distribution of deep soil organic carbon (OC) storage and how climate controls belowground OC. This research aims to quantify OC stocks, characterize soil OC age and chemical composition, and evaluate climatic impacts on OC storage from the soil surface through the deep critical zone to bedrock. These objectives were carried out at four sites along a bio-climosequence in the Sierra Nevada, California. On average, 74% of OC was stored below the A horizon, and up to 30% of OC was stored in saprock (friable weakly weathered bedrock). Radiocarbon, spectroscopic, and isotopic analyses revealed the coexistence of very old organic matter (OM) (mean radiocarbon age = 20 300 years) with relatively recent OM (mean radiocarbon age = 4800 years) and highly decomposed organic compounds with relatively less decomposed material in deep soil and saprock. This co-mingling of OM suggests OC is prone to both active cycling and long-term protection from degradation. In addition to having direct effects on OC cycling, climate indirectly controls deep OC storage through its impact on the degree of regolith weathering (e.g. thickening). Although deep OC concentrations are low relative to soil, thick saprock represents a large, previously unrealized OC pool.

While the impacts of cigarette smoking on human health are widely known, a less recognized impact of tobacco product use and disposal is environmental pollution. This review discusses the current literature related to cigarette and e-cigarette contamination in the context of environmental sources and impacts, with a focus on the documented influences on biota, ranging from bacteria to mammals. Cigarette butts and electronic cigarette components can leach contaminants into soil, water, and air. Cellulose acetate cigarette filters comprising the butts are minimally degradable and are a source of bulk plastic and microplastic pollution, especially in aquatic ecosystems where they tend to accumulate. Cigarette combustion and aerosol production during e-cigarette use result in air contamination from sidestream, exhaled, and thirdhand pathways. The chemical byproducts of tobacco product use contaminate wastewater effluents, landfill leachates, and urban storm drains. The widespread detection of nicotine and cotinine in the environment illustrates the potential for large-scale environmental impacts of tobacco product waste. Studies show that cigarette butt leachate and nicotine are toxic to microbes, plants, benthic organisms, bivalves, zooplankton, fish, and mammals; however, there remain critical knowledge gaps related to the environmental impacts of tobacco product waste on environmental health and ecosystem functioning.

The generation and transport of mineral dust is strongly related to climate on seasonal, year-to-year, and glacial-interglacial timescales. The modern dust cycle is influenced by soil moisture, which is partly a function of drought duration and severity. The production and transport of dust can therefore be amplified by global and regional droughts, thereby moderating ecosystem vulnerability to disturbance through the influence of dust on nutrient delivery to ecosystems. In this work, we use strontium and neodymium isotopes in combination with trace element concentrations in modern dust samples collected in 2015 to quantify the role of regionally versus globally supplied dust in nutrient delivery to a montane ecosystem. The study sites lie along an elevational transect in the southern Sierra Nevada, USA, with samples spanning the dry seasons of 2014 (Aciego et al., 2017) and 2015 (this study), when the region was experiencing a historic drought. The goal of our research was to quantify the spatial and temporal variability and sensitivity of the dust cycle to short term changes at nutrient-limited sites. We find that, during the dry season of 2015, Asian sources contributed between 10 and 40% of dust to sites located along this elevational transect, and importantly increased in importance during the summer growing season compared to regional dust sources. These changes are likely related to the prolonged drought in Asia in 2015, highlighting both the sensitivity of dust production and transport to drought and the teleconnections of dust transport in terrestrial ecosystems.

OBJECTIVE: To analyze HIV drug resistance among men who have sex with men (MSM) recruited for participation in the HPTN 078 study, which evaluated methods for achieving and maintaining viral suppression in HIV-infected MSM. METHODS: Individuals were recruited at four study sites in the United States (Atlanta, Georgia; Baltimore, Maryland; Birmingham, Alabama; and Boston, Massachusetts; 2016–2017). HIV genotyping was performed using samples collected at study screening or enrollment. HIV drug resistance was evaluated using the Stanford v8.7 algorithm. A multi-assay algorithm was used to identify individuals with recent HIV infection. Clustering of HIV sequences was evaluated using phylogenetic methods. RESULTS: High-level HIV drug resistance was detected in 44 (31%) of 142 individuals (Atlanta: 21%, Baltimore: 29%, Birmingham: 53%, Boston: 26%); 12% had multi-class resistance, 16% had resistance to tenofovir or emtricitabine, and 8% had resistance to integrase strand transfer inhibitors (INSTIs); 3% had intermediate-level resistance to second-generation INSTIs. In a multivariate model, self-report of ever having been on antiretroviral therapy (ART) was associated with resistance (p=0.005). One of six recently-infected individuals had drug resistance. Phylogenetic analysis identified five clusters of study sequences; two clusters had shared resistance mutations. CONCLUSIONS: High prevalence of drug resistance was observed among MSM. Some had multi-class resistance, resistance to drugs used for pre-exposure prophylaxis (PrEP), and INSTI resistance. These findings highlight the need for improved HIV care in this high-risk population, identification of alternative regimens for PrEP, and inclusion of integrase resistance testing when selecting ART regimens for MSM in the US.

Giant sequoia (Sequoiadendron giganteum) is an iconic conifer that lives in relic populations on the western slopes of the California Sierra Nevada. In these settings it is unusual among the dominant trees in that it associates with arbuscular mycorrhizal fungi rather than ectomycorrhizal fungi. However, it is unclear whether differences in microbial associations extends more broadly to non-mycorrhizal components of the soil microbial community. To address this question we characterized microbiomes associated with giant sequoia and co-occurring sugar pine (Pinus lambertiana) by sequencing 16S and ITS1 of the bulk soil community at two groves with distinct parent material. We found tree-associated differences were apparent despite a strong grove effect. Bacterial/archaeal richness was greater beneath giant sequoia than sugar pine, with a unique core community that was double the size. The tree species also harbored compositionally distinct fungal communities. This pattern depended on grove but was associated with a consistently elevated relative abundance ofHygrocybespecies beneath giant sequoia. Compositional differences between host trees correlated with soil pH, calcium availability, and soil moisture. We conclude that the effects of giant sequoia extend beyond mycorrhizal mutualists to include the broader community, and that some but not all host tree differences are grove-dependent.

Soil profiles are rarely homogeneous. Resource availability and microbial abundances typically decrease with soil depth, but microbes found in deeper horizons are still important components of terrestrial ecosystems. By studying 20 soil profiles across the United States, we documented consistent changes in soil bacterial and archaeal communities with depth. Deeper soils harbored communities distinct from those of the more commonly studied surface horizons. Most notably, we found that the candidate phylum Dormibacteraeota (formerly AD3) was often dominant in subsurface soils, and we used genomes from uncultivated members of this group to identify why these taxa are able to thrive in such resource-limited environments. Simply digging deeper into soil can reveal a surprising number of novel microbes with unique adaptations to oligotrophic subsurface conditions., While most bacterial and archaeal taxa living in surface soils remain undescribed, this problem is exacerbated in deeper soils, owing to the unique oligotrophic conditions found in the subsurface. Additionally, previous studies of soil microbiomes have focused almost exclusively on surface soils, even though the microbes living in deeper soils also play critical roles in a wide range of biogeochemical processes. We examined soils collected from 20 distinct profiles across the United States to characterize the bacterial and archaeal communities that live in subsurface soils and to determine whether there are consistent changes in soil microbial communities with depth across a wide range of soil and environmental conditions. We found that bacterial and archaeal diversity generally decreased with depth, as did the degree of similarity of microbial communities to those found in surface horizons. We observed five phyla that consistently increased in relative abundance with depth across our soil profiles: Chloroflexi, Nitrospirae, Euryarchaeota, and candidate phyla GAL15 and Dormibacteraeota (formerly AD3). Leveraging the unusually high abundance of Dormibacteraeota at depth, we assembled genomes representative of this candidate phylum and identified traits that are likely to be beneficial in low-nutrient environments, including the synthesis and storage of carbohydrates, the potential to use carbon monoxide (CO) as a supplemental energy source, and the ability to form spores. Together these attributes likely allow members of the candidate phylum Dormibacteraeota to flourish in deeper soils and provide insight into the survival and growth strategies employed by the microbes that thrive in oligotrophic soil environments.

Identifying the global drivers of soil priming is essential to understanding C cycling in terrestrial ecosystems. We conducted a survey of soils across 86 globally-distributed locations, spanning a wide range of climates, biotic communities, and soil conditions, and evaluated the apparent soil priming effect using 13C-glucose labeling. Here we show that the magnitude of the positive apparent priming effect (increase in CO2 release through accelerated microbial biomass turnover) was negatively associated with SOC content and microbial respiration rates. Our statistical modeling suggests that apparent priming effects tend to be negative in more mesic sites associated with higher SOC contents. In contrast, a single-input of labile C causes positive apparent priming effects in more arid locations with low SOC contents. Our results provide solid evidence that SOC content plays a critical role in regulating apparent priming effects, with important implications for the improvement of C cycling models under global change scenarios., The global ecological predictors of soil priming remain unclear. Here the authors conducted a global survey of soils from 86 global locations using an isotopic approach and find that in more mesic sites with high SOC concentrations, soil priming effects are more likely to be negative.

Plants frequently exhibit tradeoffs between reproduction and growth when resources are limited, and often change these allocation patterns in response to stress. Shorter-lived plants such as annuals tend to allocate relatively more resources toward reproduction when stressed, while longer-lived plants tend to invest more heavily in survival and stress defense. However, severe stress may affect the fitness implications of allocating relatively more resources to reproduction versus stress defense. Increased drought intensity and duration have led to widespread mortality events in coniferous forests. In this review, we ask how potential tradeoffs between reproduction and survival influence the likelihood of drought-induced mortality and species persistence. We propose that trees may exhibit what we call ‘fight or flight’ behaviors under stress. ‘Fight’ behaviors involve greater resource allocation toward survival (e.g., growth, drought-resistant xylem and pest defense). ‘Flight’ consists of higher relative allocation of resources to reproduction, potentially increasing both offspring production and mortality risk for the adult. We hypothesize that flight behaviors increase as drought stress escalates the likelihood of mortality in a given location.

Understanding how altered soil organic carbon (SOC) availability affects microbial communities and their function is imperative in predicting impacts of global change on soil carbon (C) storage and ecosystem function. However, the response of soil microbial communities and their function to depleted C availability insitu is unclear. We evaluated the role of soil C inputs in controlling microbial biomass, community composition, physiology, and function by (1) experimentally excluding plant C inputs insitu for 9 yr in four temperate forest ecosystems along a productivity gradient in Oregon, USA; and (2) integrating these findings with published data from similar C-exclusion studies into a global meta-analysis. Excluding plant C inputs for 9 yr resulted in a 13% decrease in SOC across the four Oregon sites and an overall shift in the microbial community composition, with a 45% decrease in the fungal:bacterial ratio and a 13% increase in Gram-positive:Gram-negative bacterial ratio. Although gross N mineralization decreased under C exclusion, decreases in gross N immobilization were greater, resulting in increased net N mineralization rates in all but the lowest-productivity site. Microbial biomass showed a variable response to C exclusion that was method dependent; however, we detected a 29% decrease in C-use efficiency across the sites, with greater declines occurring in less-productive sites. Although extracellular enzyme activity increased with C exclusion, C exclusion resulted in a 31% decrease in microbial respiration across all sites. Our meta-analyses of published data with similar C-exclusion treatments were largely consistent with our experimental results, showing decreased SOC, fungal:bacterial ratios, and microbial respiration, and increased Gram-positive:Gram-negative bacterial ratio following exclusion of C inputs to soil. Effect sizes of SOC and respiration correlated negatively with the duration of C exclusion; however, there were immediate effects of C exclusion on microbial community composition and biomass that were unaltered by duration of treatment. Our field-based experimental results and analyses demonstrate unequivocally the dominant control of C availability on soil microbial biomass, community composition, and function, and provide additional insight into the mechanisms for these effects in forest ecosystems.

BackgroundPersons who inject drugs (PWID) have high HIV incidence and prevalence, and may have limited access to antiretroviral therapy (ART) in some settings. We evaluated HIV drug resistance in PWID in a randomized clinical trial (HPTN 074). The study intervention included ART at any CD4 cell count with enhanced support for ART and substance use treatment.MethodsHPTN 074 enrolled HIV-infected PWID (index participants) with viral loads ≥1,000 copies/mL and their HIV-uninfected injection-network partners in Indonesia, Ukraine, and Vietnam; the study limited enrollment of people who reported being on ART. HIV drug resistance testing and antiretroviral (ARV) drug testing were performed using samples collected from index participants at study enrollment.ResultsFifty-four (12.0%) of 449 participants had HIV drug resistance; 29 (53.7%) of the 54 participants had multi-class resistance. Prevalence of resistance varied by study site and was associated with self-report of prior or current ART, detection of ARV drugs, and a history of incarceration. Resistance was detected in 10 (5.6%) of 177 newly diagnosed participants. Participants with resistance at enrollment were less likely to be virally suppressed after 52 weeks of follow-up, independent of study arm.ConclusionsIn HPTN 074, many of the enrolled index participants had HIV drug resistance and more than half of those had multi-class resistance. Some newly-diagnosed participants had resistance, suggesting that they may have been infected with drug-resistant HIV strains. Behavioral and geographic factors were associated with baseline resistance. Baseline resistance was associated with reduced viral suppression during study follow-up. These findings indicate the need for enhanced HIV care in this high-risk population to achieve sustained viral suppression on ART.