Soil warming-induced reduction in water content enhanced methane uptake at different soil depths in a subtropical forest.

Global warming can significantly impact soil CH4 uptake in subtropical forests due to changes in soil moisture, temperature sensitivity of methane-oxidizing bacteria (MOB), and shifts in microbial communities. However, the specific effects of climate warming and the underlying mechanisms on soil CH4 uptake at different soil depths remain poorly understood. To address this knowledge gap, we conducted a soil warming experiment (+4 °C) in a natural forest. From August 2020 to October 2021, we measured soil temperature, soil moisture, and CH4 uptake rates at four different soil depths: 0-10 cm, 10-20 cm, 20-40 cm, and 40-60 cm. Additionally, we assessed the soil MOB community structure and pmoA gene (with qPCR) at the 0-10 and 10-20 cm depths. Our findings revealed that warming significantly enhanced soil net CH4 uptake rate by 12.28 %, 29.51 %, and 61.05 % in the 0-10, 20-40, and 40-60 cm soil layers, respectively. The warming also led to reduced soil moisture levels, with more pronounced reductions observed at the 20-40 cm depth compared to the 0-20 cm depth. At the 0-10 cm depth, warming increased the relative abundance of upland soil cluster α (a type of MOB) and decreased the relative abundance of Methylocystis, but it did not significantly increase the pmoA gene copies. Our structural equation model analysis indicated that warming directly regulated soil CH4 uptake rate through the decrease in soil moisture, rather than through changes in the pmoA gene and MOB community structure at the 0-20 cm depth. In summary, our results demonstrate that warming enhances soil CH4 uptake at different depths, with soil moisture playing a crucial role in this process. Under warming conditions, the drier soil pores allow for better CH4 penetration, thereby promoting more efficient activity of MOB.

Assembly and succession of the phyllosphere microbiome and nutrient-cycling genes during plant community development in a glacier foreland.

The phyllosphere, particularly the leaf surface of plants, harbors a diverse range of microbiomes that play a vital role in the functioning of terrestrial ecosystems. However, our understanding of microbial successions and their impact on functional genes during plant community development is limited. In this study, considering core and satellite microbial taxa, we characterized the phyllosphere microbiome and functional genes in various microhabitats (i.e., leaf litter, moss and plant leaves) across the succession of a plant community in a low-altitude glacier foreland. Our findings indicate that phyllosphere microbiomes and associated ecosystem stability increase during the succession of the plant community. The abundance of core taxa increased with plant community succession and was primarily governed by deterministic processes. In contrast, satellite taxa abundance decreased during plant community succession and was mainly governed by stochastic processes. The abundance of microbial functional genes (such as C, N, and P hydrolysis and fixation) in plant leaves generally increased during the plant community succession. However, in leaf litter and moss leaves, only a subset of functional genes (e.g., C fixation and degradation, and P mineralization) showed a tendency to increase with plant community succession. Ultimately, the community of both core and satellite taxa collaboratively influenced the characteristics of phyllosphere nutrient-cycling genes, leading to the diverse profiles and fluctuating abundance of various functional genes during plant community succession. These findings offer valuable insights into the phyllosphere microbiome and plant-microbe interactions during plant community development, advancing our understanding of the succession and functional significance of the phyllosphere microbial community.

Spatial variation and controls of soil microbial necromass carbon in a tropical montane rainforest.

Soil microbial necromass carbon is an important component of the soil organic carbon (SOC) pool which helps to improve soil fertility and texture. However, the spatial pattern and variation mechanisms of fungal- and bacterial-derived necromass carbon at local scales in tropical rainforests are uncertain. This study showed that microbial necromass carbon and its proportion in SOC in tropical montane rainforest exhibited large spatial variation and significant autocorrelation, with significant high-high and low-low clustering patterns. Microbial necromass carbon accounted for approximately one-third of SOC, and the fungal-derived microbial necromass carbon and its proportion in SOC were, on average, approximately five times greater than those of bacterial-derived necromass. Structural equation models indicated that soil properties (SOC, total nitrogen, total phosphorus) and topographic features (elevation, convexity, and aspect) had significant positive effects on microbial necromass carbon concentrations, but negative effects on its proportions in SOC (especially the carbon:nitrogen ratio). Plant biomass also had significant negative effects on the proportion of microbial necromass carbon in SOC, but was not correlated with its concentration. The different spatial variation mechanisms of microbial necromass carbon and their proportions in SOC are possibly related to a slower accumulation rate of microbial necromass carbon than of plant-derived organic carbon. Geographic spatial correlations can significantly improve the microbial necromass carbon model fit, and low sampling resolution may lead to large uncertainties in estimating soil carbon dynamics at specific sites. Our work will be valuable for understanding microbial necromass carbon variation in tropical forests and soil carbon prediction model construction with microbial participation.

Cross-scale spatial variability and associations of carbon pools provide insight into regulating carbon sequestration in tropical montane rainforests.

The spatial distribution of plant, soil, and microbial carbon pools, along with their intricate interactions, presents a great challenge for the current carbon cycle research. However, it is not clear what are the characteristics of the spatial variability of these carbon pools, particularly their cross-scale relationships. We investigated the cross-scale spatial variability of microbial necromass carbon (MNC), soil organic carbon (SOC) and plant biomass (PB), as well as their correlation in a tropical montane rainforest using multifractal analysis. The results showed multifractal spatial variations of MNC, SOC, and PB, demonstrating their adherence to power-law scaling. MNC, especially low MNC, exhibited stronger spatial heterogeneity and weaker evenness compared with SOC and PB. The cross-scale correlation between MNC and SOC was stronger than their correlations at the measurement scale. Furthermore, the cross-scale spatial variability of MNC and SOC exhibited stronger and more stable correlations than those with PB. Additionally, this research suggests that when SOC and PB are both low, it is advisable for reforestations to potentiate MNC formation, whereas when both SOC and PB are high some thinning can be advisable to favour MNC formation. Thus, these results support the utilization of management measures such as reforestation or thinning as nature-based solutions to regulate carbon sequestration capacity of tropical forests by affecting the correlations among various carbon pools.

Multi-ecosystem services differently affected by over-canopy and understory nitrogen additions in a typical subtropical forest.

Obtaining a holistic understanding of the impacts of atmospheric nitrogen deposition on multiple ecosystem services of forest is essential for developing comprehensive and sustainable strategies, particularly in heavy N deposition regions such as subtropical China. However, such impacts remain incompletely understood, with most previous studies focus on individual ecosystem function or service via understory N addition experiments. To address this knowledge gap, we quantified the effects of over-canopy and understory N additions on multiple ecosystem services based on a 7-year large-scale field experiment in a typical subtropical forest. Our results showed continued over-canopy N addition with 50 kg ha-1 year-1 over a period of 4-7 years significantly increased plant nutrient retention, but did not affect the services of soil nutrient accumulation, water yield, C sequestration (in plants and soil), or oxygen release. There were trade-offs between the soil and plant on providing the services of nutrient accumulation/retention and C sequestration under over-canopy N addition. However, without uptake and retention of tree canopy, the trade-off between soil and plant were more weaken under the understory N addition with 50 kg ha-1 year-1 , and their relationships were even synergetic under the understory N addition with 25 kg ha-1 year-1 . The results suggest that understory N addition cannot accurately simulate the effects of atmospheric N deposition on multiple services, along with mutual relationships. Interestingly, the services of plant N, P retention, and C sequestration exhibited a synergetic increase under the over-canopy N addition but a decrease under the understory N addition. Our results also found tree layer plays a primary role in providing plant nutrient retention service and is sensitive to atmospheric N deposition. Further studies are needed to investigate the generalized effects of forest canopy processes on alleviating the threaten of global change factors in different forest ecosystems.

Net greenhouse gas balance in U.S. croplands: How can soils be part of the climate solution?

Agricultural soils play a dual role in regulating the Earth's climate by releasing or sequestering carbon dioxide (CO2 ) in soil organic carbon (SOC) and emitting non-CO2 greenhouse gases (GHGs) such as nitrous oxide (N2 O) and methane (CH4 ). To understand how agricultural soils can play a role in climate solutions requires a comprehensive assessment of net soil GHG balance (i.e., sum of SOC-sequestered CO2 and non-CO2 GHG emissions) and the underlying controls. Herein, we used a model-data integration approach to understand and quantify how natural and anthropogenic factors have affected the magnitude and spatiotemporal variations of the net soil GHG balance in U.S. croplands during 1960-2018. Specifically, we used the dynamic land ecosystem model for regional simulations and used field observations of SOC sequestration rates and N2 O and CH4 emissions to calibrate, validate, and corroborate model simulations. Results show that U.S. agricultural soils sequestered 13.2 ± 1.16 $$ 13.2\pm 1.16 $$ Tg CO2 -C year-1 in SOC (at a depth of 3.5 m) during 1960-2018 and emitted 0.39 ± 0.02 $$ 0.39\pm 0.02 $$ Tg N2 O-N year-1 and 0.21 ± 0.01 $$ 0.21\pm 0.01 $$ Tg CH4 -C year-1 , respectively. Based on the GWP100 metric (global warming potential on a 100-year time horizon), the estimated national net GHG emission rate from agricultural soils was 122.3 ± 11.46 $$ 122.3\pm 11.46 $$ Tg CO2 -eq year-1 , with the largest contribution from N2 O emissions. The sequestered SOC offset ~28% of the climate-warming effects resulting from non-CO2 GHG emissions, and this offsetting effect increased over time. Increased nitrogen fertilizer use was the dominant factor contributing to the increase in net GHG emissions during 1960-2018, explaining ~47% of total changes. In contrast, reduced cropland area, the adoption of agricultural conservation practices (e.g., reduced tillage), and rising atmospheric CO2 levels attenuated net GHG emissions from U.S. croplands. Improving management practices to mitigate N2 O emissions represents the biggest opportunity for achieving net-zero emissions in U.S. croplands. Our study highlights the importance of concurrently quantifying SOC-sequestered CO2 and non-CO2 GHG emissions for developing effective agricultural climate change mitigation measures.

Microbial controls over soil priming effects under chronic nitrogen and phosphorus additions in subtropical forests.

The soil priming effect (PE), defined as the modification of soil organic matter decomposition by labile carbon (C) inputs, is known to influence C storage in terrestrial ecosystems. However, how chronic nutrient addition, particularly in leguminous and non-leguminous forests, will affect PE through interaction with nutrient (e.g., nitrogen and phosphorus) availability is still unclear. Therefore, we collected soils from leguminous and non-leguminous subtropical plantations across a suite of historical nutrient addition regimes. We added 13C-labeled glucose to investigate how background soil nutrient conditions and microbial communities affect priming and its potential microbial mechanisms. Glucose addition increased soil organic matter decomposition and prompted positive priming in all soils, regardless of dominant overstory tree species or fertilizer treatment. In non-leguminous soil, only combined nitrogen and phosphorus addition led to a higher positive priming than the control. Conversely, soils beneath N-fixing leguminous plants responded positively to P addition alone, as well as to joint NP addition compared to control. Using DNA stable-isotope probing, high-throughput quantitative PCR, enzyme assays and microbial C substrate utilization, we found that positive PE was associated with increased microbial C utilization, accompanied by an increase in microbial community activity, nutrient-related gene abundance, and enzyme activities. Our findings suggest that the balance between soil available N and P effects on the PE,  was dependent on rhizosphere microbial community composition. Furthermore, these findings highlight the roles of the interaction between plants and their symbiotic microbial communities in affecting soil priming and improve our understanding of the potential microbial pathways underlying soil PEs.

Soil acidification suppresses phosphorus supply through enhancing organomineral association.

It has long been assumed that soil acidification increases reactive iron and/or aluminum (Fe/Al) oxides and promotes Pi sorption onto mineral surfaces, resulting in a decrease in Pi. However, this assumption has seldom been tested in long-term field experiments. Using a 12-year acid addition experiment in a tropical forest, we demonstrated that soil acidification increased the content of noncrystalline Fe and Al oxides by 16.3 % and 27.7 %, respectively; whereas it did not alter the absorbed Pi pool and Pi sorption capacity. Furthermore, soil acidification increased the Fe/Al-bound organic matter content by 82.5 %, causing a 54.9 % reduction in Pi desorption, a 42.3 % decrease in soluble Pi content, and a 9.2 % increase in occluded Pi content. Our findings demonstrate that soil acidification reduces Pi bioavailability by repressing Pi desorption rather than enhancing Pi sorption. These results could be attributed to the enhanced organomineral association, which competes for sorption sites with Pi and promotes the Pi occlusion. However, the interactions between organomineral-Pi have not been incorporated into global land models, which may overestimate ecosystem productivity under future acid rain scenarios.

Spatial variation of soil seed banks along a gradient of anthropogenic disturbances in tropical forests on coral islands.

Poor regeneration of natural vegetation is a major factor contributing to the degradation of tropical coral islands. Soil seed banks (SSB) are important for maintaining the resilience of plant communities. However, the community characteristics and spatial distribution of SSBs and the controlling factors along human disturbance on coral islands are unclear. To fill this gap, we measured the community structure and spatial distributions of forest SSBs on three coral islands in the South China Sea, with varying degrees of human disturbance. The results showed that strong human disturbance increased the diversity, richness, and density of SSBs, as well as increased the richness of invasive species. With increased human disturbance, the heterogeneity pattern of SSBs spatial distribution changed from difference between forest east and west to forest center and edge. The similarity between the SSBs and above-ground vegetation also increased, and the distribution of invasive species extended from the edge to the central area of the forests, demonstrating that human disturbance limited the outward dispersal of seeds of resident species but increased the inward dispersal of seeds of invasive species. Interaction between soil properties, plant characteristics, and human disturbance explained 23-45% of the spatial variation of forest SSBs on the coral islands. However, human disturbance reduced the correlations of plant communities and spatial distribution of SSBs with soil factors (i.e., available phosphorus and total nitrogen) and increased the correlations of the community characteristics of SSB with landscape heterogeneity index, road distance, and shrub and litter cover. Resident seed dispersal on tropical coral islands might be enhanced by reducing building height, constructing buildings in down-wind locations, and preserving corridors that support animal movement among forest fragments.

Divergent accumulation of amino sugars and lignins mediated by soil functional carbon pools under tropical forest conversion.

Tropical primary forests are being destroyed at an alarming rate and converted for other land uses which is expected to greatly influence soil carbon (C) cycling. However, our understanding of how tropical forest conversions affect the accumulation of compounds in soil functional C pools remains unclear. Here, we collected soils from primary forests (PF), secondary forests (SF), oil-palm (OP), and rubber plantations (RP), and assessed the accumulation of plant- and microbial-derived compounds within soil organic carbon (SOC), particulate (POC) and mineral-associated (MAOC) organic C. PF conversion to RP greatly decreased SOC, POC, and MAOC concentrations, whereas conversion to SF increased POC concentrations and decreased MAOC concentrations, and conversion to OP only increased POC concentrations. PF conversion to RP decreased lignin concentrations and increased amino sugar concentrations in SOC pools which increased the stability of SOC, whereas conversion to SF only increased the lignin concentrations in POC, and conversion to OP just increased lignin concentrations in POC and decreased it in MAOC. We observed divergent dynamics of amino sugars (decrease) and lignin (increase) in SOC with increasing SOC. Only lignin concentrations increased in POC with increasing POC and amino sugars concentrations decreased in MAOC with increasing MAOC. Conversion to RP significantly decreased soil enzyme activities and microbial biomasses. Lignin accumulation was associated with microbial properties, whereas amino sugar accumulation was mainly associated with soil nutrients and stoichiometries. These results suggest that the divergent accumulation of plant- and microbial-derived C in SOC was delivered by the distribution and original composition of functional C pools under forest conversions. Forest conversions changed the formation and stabilization processes of SOC in the long run which was associated with converted plantations and management. The important roles of soil nutrients and stoichiometry also provide a natural-based solution to enhance SOC sequestration via nutrient management in tropical forests.

Depth-driven responses of microbial residual carbon to nitrogen addition approaches in a tropical forest: Canopy addition versus understory addition.

Canopies play an important role in nitrogen (N) redistribution in forest ecosystems, and ignoring the canopy's role might bias estimates of the ecological consequences of anthropogenic atmospheric N deposition. We investigated the effects of the approach of N addition (Canopy addition vs. Understory addition) and level of N addition (25 kg N ha-1yr-1 vs. 50 kg N ha-1yr-1) on microbial residual carbon (MRC) accumulation in topsoil and subsoil. We found that the response of MRC to both approach and level of N addition varied greatly with soil depth in a tropical forest over eight years of continuous N addition. Specifically, N addition enhanced the accumulation of fungal and total MRC and their contribution to soil organic C (SOC) pools in the topsoil, whereas it decreased the contribution of fungal and total MRC to SOC in the subsoil. The contrasting effects of N addition on MRC contribution at varying soil depths were associated with the distinct response of microbial residues production. Understory N addition showed overall greater effects on MRC accumulation than canopy N addition did. Our results suggest that the canopy plays an important role in buffering the impacts of anthropogenic atmospheric N deposition on soil C cycling in tropical forests. The depth-dependent response of microbial residues to N addition also highlights the urgent need for further studies of different response mechanisms at different soil depths.

Overcompensation of ecosystem productivity following sustained extreme drought in a semiarid grassland.

Drought events are projected to be more extreme and frequent in the future and have profound influences on the structure and functions of terrestrial ecosystems. Thus, better understanding the mechanisms of recovery is critical for predicting the future dynamics of terrestrial ecosystems. We performed a 7-year field precipitation experiment to examine recovery of a grassland ecosystem from different magnitudes of sustained drought, from slight to extreme. The ecosystem was exposed to precipitation treatments in the first 3 years (2010-2012) and recovered during the last 4 years (2013-2016) without precipitation treatments. Overall, large reductions of aboveground net primary productivity (ANPP, -43.3%) and perennial forb biomass (-83.1%) were observed in the third year (2012) of extreme drought only. Nevertheless, ANPP fully recovered within 1 year after the drought treatments were terminated, and the rapid recovery was mainly due to increased soil total nitrogen and root biomass allocation after drought. Surprisingly, large increases of ANPP under the extreme drought treatment occurred during the recovery periods from 2013 to 2015 (+74.1, +88.5, and +119.8 g m-2 year-1 ) compared to the control. The overcompensation offset the extreme drought-induced reduction of ANPP in the treatment years and was primarily ascribed to the enhanced biomass of perennial grasses (PG). Higher resistance to drought and fast resource acquisition strategy might drive the rapid recovery and expansion of PG. Our findings revealed the rapid recovery of grasslands and the critical role of community overcompensation in maintaining grassland ecosystem function and stability under future climate change scenarios.

Differential responses and mechanistic controls of soil phosphorus transformation in Eucalyptus plantations with N fertilization and introduced N2 -fixing tree species.

Introducing N2 -fixing tree species into Eucalyptus plantations could replace nitrogen (N) fertilization to maintain high levels of N consumption and productivity. However, N enrichment may exacerbate phosphorus (P) limitation as Eucalyptus robusta Smith is extensively planted in P-poor tropical and subtropical soils. We conducted a field experiment in a pure plantation of Eucalyptus urophylla × grandis to investigate the impacts of N fertilization and introduced an N2 -fixing tree of Dalbergia odorifera T. Chen on soil P transformation. Nitrogen fertilization significantly enhanced soil occluded P pool and reduced the other P pools due to acidification-induced pH-sensitive geochemical processes, lowering Eucalyptus leaf P concentration with higher N : P ratio. By contrast, introduced N2 -fixing tree species did not change soil pH, labile inorganic P pool, and Eucalyptus leaf N : P ratio, even enhanced organic P pools and reduced occluded P pool probably due to altering microbial community composition particularly stimulating arbuscular mycorrhiza fungal abundance. Our results revealed differential responses and mechanistic controls of soil P transformation in Eucalyptus plantations with N fertilization and introduced N2 -fixing tree species. The dissolution of occluded P pool along with organic P accumulation observed in the mixed plantations may represent a promising future to better manage soil P availability.

Significant effects of precipitation frequency on soil respiration and its components-A global synthesis.

Global warming intensifies the hydrological cycle, which results in changes in precipitation regime (frequency and amount), and will likely have significant impacts on soil respiration (Rs ). Although the responses of Rs to changes in precipitation amount have been extensively studied, there is little consensus on how Rs will be affected by changes in precipitation frequency (PF) across the globe. Here, we synthesized the field observations from 296 published papers to quantify the effects of PF on Rs and its components using meta-analysis. Our results indicated that the effects of PF on Rs decreased with an increase in background mean annual precipitation. When the data were grouped by climate conditions, increased PF showed positive effects on Rs under the arid condition but not under the semi-humid or humid conditions, whereas decreased PF suppressed Rs across all the climate conditions. The positive effects of increased PF mainly resulted from the positive response of heterotrophic respiration under the arid condition while the negative effects of decreased PF were mainly attributed to the reductions in root biomass and respiration. Overall, our global synthesis provided for the first time a comprehensive analysis of the divergent effects of PF on Rs and its components across climate regions. This study also provided a framework for understanding and modeling responses of ecosystem carbon cycling to global precipitation change.

Dynamics of soil microbial communities involved in carbon cycling along three successional forests in southern China.

Dynamics of plant communities during forest succession have been received great attention in the past decades, yet information about soil microbial communities that are involved in carbon cycling remains limited. Here we investigated soil microbial community composition and carbohydrate degradation potential using metagenomic analysis and examined their influencing factors in three successional subtropical forests in southern China. Results showed that the abundances of soil bacteria and fungi increased (p ≤ 0.05 for both) with forest succession in relation to both soil and litter characteristics, whereas the bacterial diversity did not change (p > 0.05) and the fungal diversity of Shannon-Wiener index even decreased (p ≤ 0.05). The abundances of microbial carbohydrate degradation functional genes of cellulase, hemicellulase, and pectinase also increased with forest succession (p ≤ 0.05 for all). However, the chitinase gene abundance did not change with forest succession (p > 0.05) and the amylase gene abundance decreased firstly in middle-succession forest and then increased in late-succession forest. Further analysis indicated that changes of functional gene abundance in cellulase, hemicellulase, and pectinase were primarily affected by soil organic carbon, soil total nitrogen, and soil moisture, whereas the variation of amylase gene abundance was well explained by soil phosphorus and litterfall. Overall, we created a metagenome profile of soil microbes in subtropical forest succession and fostered our understanding of microbially-mediated soil carbon cycling.

Zero Discharge of Nutrient Solution to the Environment in a Soilless Greenhouse Cucumber Production System.

With the development of the economy and society, more attention is being paid to energy costs and the potential environmental pollution caused by vegetable cultivation. The aim of this study was to investigate the impacts of zero discharge of nutrient solutions on cucumber growth, leaf photosynthesis, and the yield and quality of cucumber under greenhouse conditions. The results show that zero discharge treatment did not change plant height, stem diameter, internode length, leaf area, net photosynthetic rate (Pn), stomatal conductance (Gs), intercellular CO2 concentration (Ci), transpiration rate (Tr), and leaf relative chlorophyll content on the most measurement days. Only Pn and relative chlorophyll content were significantly reduced after 16 days of treatment but soon recovered over time. Cucumber plants can adapt to treatment circumstances over the course of days. Leaf mineral element contents showed significant differences on some treatment days compared to the control, and trace elements of Fe, Mn, Cu, and Mo can be appropriately supplied during the treatment days. The cucumber yield and fruit quality in the zero discharge treatment did not change during the whole experimental period. This study confirmed that the irrigation method of a nutrient solution with zero discharge can be applied in cucumber cultivation practices. The strict management of irrigation strategy, plant growth, and greenhouse climate are very important for zero discharge cultivation. The cultivation method with zero discharge of nutrient solution can reduce the energy costs of disinfection, save water and fertilizers, and reduce the environmental pollution in cucumber cultivation.

Nitrogen budgets of a lower subtropical forest as affected by 6 years of over-canopy and understory nitrogen additions.

Although tropical and subtropical regions have replaced temperate regions as the global-change hotspots for increased atmosphere nitrogen (N) deposition, whether the regional forests reach N saturation is still unclear. Understory or floor N addition has been commonly used in N-deposition studies, but the results of such studies have recently been challenged because they fail to account for canopy interception, assimilation, and leaching processes. Here, we conducted a field experiment to quantify the effects of over-canopy and understory N addition on N budgets in a lower subtropical monsoon evergreen broadleaved (LSMEB) forest. We found that the LSMEB forest was not N saturated after receiving additional N at 25 and 50 kg ha-1 yr-1 for 6 years. Plants were able to absorb the added N by increasing the N concentrations in their organs, with 120-412 % increasing trend of plant N pools under N-addition treatments. Canopy absorption of N resulting from over-canopy N addition led to increases in N concentrations in tree organs but not to increases in tree biomass. Understory N addition could underestimate the effects of N deposition in forests due to neglecting canopy N interception and canopy effects on N redistribution. Additional experiments using over-canopy N addition are needed to assess the true effects of N deposition on different forest ecosystems in different climate zones.

Increased precipitation and nitrogen addition accelerate the temporal increase in soil respiration during 8-year old-field grassland succession.

Ecological succession after disturbance plays a vital role in influencing ecosystem structure and functioning. However, how global change factors regulate ecosystem carbon (C) cycling in successional plant communities remains largely elusive. As part of an 8-year (2012-2019) manipulative experiment, this study was designed to examine the responses of soil respiration and its heterotrophic component to simulated increases in precipitation and atmospheric nitrogen (N) deposition in an old-field grassland undergoing secondary succession. Over the 8-year experimental period, increased precipitation stimulated soil respiration by 11.6%, but did not affect soil heterotrophic respiration. Nitrogen addition increased both soil respiration (5.1%) and heterotrophic respiration (6.2%). Soil respiration and heterotrophic respiration linearly increased with time in the control plots, resulting from changes in soil moisture and shifts of plant community composition from grass-forb codominance to grass dominance in this old-field grassland. Compared to the control, increased precipitation significantly strengthened the temporal increase in soil respiration through stimulating belowground net primary productivity. By contrast, N addition accelerated temporal increases in both soil respiration and its heterotrophic component by driving plant community shifts and thus stimulating soil organic C. Our findings indicate that increases in water and N availabilities may accelerate soil C release during old-field grassland succession and reduce their potential positive impacts on soil C accumulation under future climate change scenarios.

Mycorrhizal fungi alleviate acidification-induced phosphorus limitation: Evidence from a decade-long field experiment of simulated acid deposition in a tropical forest in south China.

South China has been experiencing very high rate of acid deposition and severe soil acidification in recent decades, which has been proposed to exacerbate the regional ecosystem phosphorus (P) limitation. We conducted a 10-year field experiment of simulated acid deposition to examine how acidification impacts seasonal changes of different soil P fractions in a tropical forest with highly acidic soils in south China. As expected, acid addition significantly increased occluded P pool but reduced the other more labile P pools in the dry season. In the wet season, however, acid addition did not change microbial P, soluble P and labile organic P pools. Acid addition significantly increased exchangeable Al3+ and Fe3+ and the activation of Fe oxides in both seasons. Different from the decline of microbial abundance in the dry season, acid addition increased ectomycorrhizal fungi and its ratio to arbuscular mycorrhiza fungi in the wet season, which significantly stimulated phosphomonoesterase activities and likely promoted the dissolution of occluded P. Our results suggest that, even in already highly acidic soils, the acidification-induced P limitation could be alleviated by stimulating ectomycorrhizal fungi and phosphomonoesterase activities. The differential responses and microbial controls of seasonal soil P transformation revealed here should be implemented into ecosystem biogeochemical model for predicting plant productivity under future acid deposition scenarios.

Using single cell type proteomics to identify Al-induced proteomes in outer layer cells and interior tissues in the apical meristem/cell division regions of tomato root-tips.

Aluminum (Al) toxicity primarily targets the root tips, inhibiting root growth and function and leading to crop yield losses on acidic soils. Previously we reported using laser capture microdissection (LCM) proteomics to identify Al-induced proteins in the outer layer cells in the transitional zone of tomato root-tips. This study aims to further characterize Al-induced proteomic dynamics from the outer to interior tissues, thus providing a panoramic view reflecting Al resistance in the root tip as a whole in tomatoes. Three types of cells were isolated via LCM from the basal 350-400 µm (below cell elongation regions) of root tips using tomato (Solanum lycopersicum) 'Micro-Tom' plants. Type I and Type II were from Al-treated plants. Type I included cells of the outer three layers, i.e., the epidermis and cortex initials and the quiescent center (QC) in root apical meristem (RAM), and Type II possessed the interior tissues of the same region. Type III contained cells from the non-Al-treated root tips collected in the same region as Type I. Two tandem mass tag (TMT) proteomics analyses with three biological replicates for each sample type were conducted. The TMTexp1 (comparing Type I and Type II) identified 6575 quantifiable proteins and 178 different abundance proteins (DAPs). The TMTexp2 (comparing Type I and Type III) identified 7197 quantifiable proteins and 162 DAPs. Among all quantified proteins (7685) from the two TMT experiments, 6088 (79%) proteins, including 313 DAPs (92% of the 340 total), were identified in all tissues. A model reflecting the tissue-specific Al-resistance mechanism was proposed, in which the level of the citrate transporter MATE protein, involved in Al exclusion, accumulated to the highest level in the outer-layer cells but decreased toward the interior of root-tips (which concurs with the tissue-specific importance in Al resistance). Proteins for biosynthesis of ethylene and jasmonic acid, proteolytic enzymes, stress-responsive proteins, and cell wall modeling were affected by Al treatment, some in a cell type-specific manner. The KEGG metabolite pathways enriched with these DAPs changed depending on the cell types. This study demonstrated the advantage of using the tissue/cell-specific analysis for identifying proteins and their dynamic changes directly associated with Al resistance in the root-tip region. The proteomics datasets have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository (https://www.ebi.ac.uk/pride/) with the dataset identifier as PXD021994 under project title: Proteomics studies of outer and inner cellular layers of tomato root-tips for Al stress, Project DOI: 10.6019/ PXD021994; and PXD018234 under Project title: Al-induced root proteomics changes in stress-acclimated tomato plant, Project DOI: https://doi.org/10.6019/PXD018234. SIGNIFICANCE: This paper presents the method of using laser capture microdissection (LCM) to collect homogenous cell-type specific tissue samples from the outer layers and inner central regions of tomato root-tips. The tandem mass tag-proteomics analysis showed that the outer-layer cells expressed proteomes that were different from the inner tissues of Al-treated root-tips; proteins related to resistance/tolerance to Al toxicity were highly accumulated in the outer-layer cells. Furthermore, the Al-treated outer-layer cells expressed proteomes which were different from the non-Al treated counterpart cells. This study has provided the first dataset of proteins differentiating from the outer to inner layers of cells in Al-treated root-tips. It provided convincing experimental evidences demonstrating the single-cell type proteomics as a powerful analytical approach to identify Al tolerance mechanisms in plants. The analytical procedure of LCM-tandem mass tag-quantitative proteomics analysis has a broad application for proteomics analysis of spatially separated cells in complex tissues.