Root Exudates Research Articles

Carbon flow from roots to rhizobacterial networks: Grafting effects

Plants recruit microorganisms from bulk soil by secreting easily available organic carbon into the rhizosphere. Grafting often increases the disease resistance of agricultural plants by modifying this carbon flow from roots into rhizosphere and by recruiting active microorganisms that suppress pathogens. Here, we continuously labeled grafted and ungrafted watermelon plants in a 13CO2 atmosphere to identify the active microorganisms assimilating root exudates. Multi-omics associated technologies (amplicon sequencing, metagenomics and metabolomics) combined with 13C tracing were used to examine the carbon flows, microbial utilization and transformation in the rhizosphere. The number of potentially active bacterial species recruited in the rhizosphere of grafted plants and utilizing root exudates was four times more than in ungrafted plants. These potentially active species matched to metagenome-assembled-genomes (MAGs) mainly belonging to Sphingomonas in the rhizosphere of ungrafted plants, and to Sphingomonas, Chitinophaga, Dyadobacter and Pseudoxanthomonas in the rhizosphere of grafted plants. Sphingomonas possesses the functional potential to metabolize a plant self-toxic substance, namely 4-hydroxybenzoic acid. Furthermore, grafting shaped the complex metabolic interactions and changed the original metabolic dependence between the potentially active bacterial species. Grafting plants diversified belowground carbon flows, activating a greater number of beneficial microbes.

Locomotion of Bacillus subtilis SL-44 mediated by root exudate and carrier in Cr(OH)3-modified porous media

Plant growth promoting rhizobacteria (PGPR) have a remediation effect on Cr-contaminated soil; however, the remediation scope is only within a small area around the bacteria. Hence, the remediation effect depends on the migration ability of bacteria in the soil. Root exudates enhance the chemotaxis and locomotion of Bacillus subtilis SL-44 by reducing its adhesion coefficient katt and hydrodynamic dispersion coefficient D. The locomotion capacity was enhanced by 7.84%–20.00%. Among the root exudates, proline and sucrose remarkably improved the motility of SL44. Biochar and bentonite increased the katt and D of SL-44, inhibited bacterial locomotion, and improved the retention rate on the carrier surface. Bacterial locomotion was reduced by biochar and bentonite by 57.99% and 50.42%, respectively. These reductions were caused by macropore. SL-44 locomotion was positively correlated with the concentration of environmental root exudate (R2 = 0.88−0.92). The results of the simulated soil study were validated in actual agricultural Cr-contaminated soils through qPCR.

Rhizosphere flavonoids alleviates the inhibition of soybean nodulation caused by shading under maize-soybean strip intercropping

The flavonoids produced by legume roots are signal molecules that induce nod genes for symbiotic rhizobium. Nevertheless, the promoting effects of flavonoids in root exudates in intercropping system on soybean nodulation are still unknown. A two years of field experiments was carried with maize soybean strip intercropping, i.e., the interspecific row spacing of 30 cm (MS30), 45 cm (MS45), 60 cm (MS60), and sole soybean/maize:SS/MM, and root interaction, i.e., root no barrier (NB) and root polythene-plastic barrier (PB), to evaluate relationships between flavonoids in root exudates and nodulation. We found that root-root interaction between soybean and maize enhances the nodules number and fresh weight in intercropped soybean. This enhancement increase gradually with expansion of interspecific distance. Proportion of nodules with diameter greater than 0.4cm was higher in intercropped soybean with NB than with PB. The expressions of nodules-related genes (GmENOD40, GmNIN2b and GmEXPB2) were up-regulated. Furthermore, compared with monocropping, isoflavones secretion of soybean roots reduced, flavonoids and flavonols secretion of maize and soybean roots increased under intercropping. The secretion of differential metabolites of flavonoids in the rhizosphere of maize and soybean declined with root barrier. The expressions of GmCHS8 and GmIFS1 in soybean roots were up-regulated and GmICHG was down-regulated under root interaction. The most of the flavonoids and flavonol compounds were positively correlated with nodule diameter. The nodules number, the nodules fresh weight and the proportion of nodules with a diameter greater than 0.2 cm increased in different genotypes of soybean treated with maize root exudate, which promoted the improvement of nitrogen fixation capacity. Therefore, maize-soybean strip intercropping combined with reasonable spacing to enhance the positive effect of underground root interaction, and improve the nodulation and nitrogen fixation capacity of intercropping soybean.

Pepper root exudate alleviates cucumber root-knot nematode infection by recruiting a rhizobacterium

Root-knot nematodes (Meloidogyne spp.) have garnered significant attention from researchers due to their substantial damage to crops and worldwide distribution. However, controlling this nematode disease is challenging which results from limited chemical pesticides and biocontrol agents effective against them. Here, we demonstrate that pepper-rotation markedly reduces Meloidogyne incognita infection in cucumber and diminishes the presence of p-hydroxybenzoic acid in the soil, a compound known to exacerbate M. incognita infection. Pepper-rotation also structures the rhizobacterial community, leading to the colonization of two Pseudarthrobacter oxydans strains (RH60 and RH97) in the cucumber rhizosphere, facilitated by palmitic acid enrichment in pepper root exudates. Furthermore, both strains exhibit high nematocidal activity against M. incognita, and possess the ability to biosynthesize indoleacetic acid and biodegrade p-hydroxybenzoic acid. RH60 and RH97 additionally induce systemic resistance in cucumber plants and promote their growth. These data suggest that pepper root-exudate palmitic acid alleviates M. incognita infection by recruiting beneficial P. oxydans in the cucumber rhizosphere. Our analyses identify a novel chemical component in root exudates and uncover its pivotal role in crop rotation for disease attenuation, providing intriguing insights into the keystone function of root exudates in plant protection against root-knot nematode infection.

Plant and soil characteristics affected by the allelopathic pathways of Avena fatua and Lolium temulentum weeds

The potential of the most prevalent weeds should be characterized biologically and chemically in infected soil and crops for sustainable agriculture. Therefore, the allelopathic potential of Avena fatua L. and Lolium temulentum L. weeds were compared via leachates, root exudates, decayed residues in soil, and the decomposition in water pathways. Chemical measurements were taken on wheat (Triticum aestivum L.), and soil decomposed solution. Based on EC50, the allelopathic effect of leachates were higher in aboveground parts than in subterranean parts, influenced by plant parts and concentrations. The root exudates show EC50 by 655.9 μg. ml−1 for A. fatua and 625.66 μg. ml−1 for L. temulentumin the seedling biomass fresh weights of T. aestivum. The systematic inhibition by decayed residues was affected by plant types, concentration, and time and correlated with soil parameters and crop performance. The decomposition rate was higher under aerobic conditions than anaerobic conditions, with the inhibition pattern showing the reverse trend. These finding highlight the importance of environmental conditions in mediating allelopathic effects. The highest quantities of phenolic acids determined by LC-ES/MS in decomposed solutions were citric acid, with concentrations of 7.71 and 13.31 μg/ml in A. fatua under aerobic conditions, and coumaric acid, with concentrations of 9.21 and 16.99 μg/ml in L. temulentum under aerobic conditions. The allelopathic potentials of A. fatua and L. temulentum may play a crucial role in T. aestivum crop growth and soil parameters. In general weed residues can suppress crop growth and negatively affect soil parameters based on their quantity and type, therefore they should be managed carefully for sustainable crop production.

A root mucilage analogue from chia seeds reduces soil gas diffusivity

AbstractGas exchange in the soil is determined by the size and connectivity of air‐filled pores. Root mucilage reduces air‐filled pore connectivity and thus gas diffusivity. It is unclear to what extent mucilage affects soil pore connectivity and tortuosity. The aim of this study was to gain a better understanding of gas diffusion processes in the rhizosphere by explaining the geometric alterations of the soil pore space induced by mucilage. We quantified the effect of a root mucilage analogue collected from chia seeds without intrinsic respiratory activity on oxygen diffusion at different water contents during drying–rewetting cycles in a diffusion chamber experiment. Quantification of oxygen diffusion showed that mucilage decreased the gas diffusion coefficient in dry soil without affecting air‐filled porosity. Without mucilage, a hysteresis in gas diffusion coefficient during a drying–rewetting cycle was observed. The effect depended on particle size and diminished with increasing mucilage content. X‐ray computed tomography imaging indicated a hysteresis in the connectivity of the gas phase during a drying–rewetting cycle for samples without mucilage. This effect was attenuated with increasing mucilage content. Furthermore, electron microscopy showed that mucilage structures formed in drying soil increase with mucilage content, thereby progressively reducing the connectivity of the gas phase. In conclusion, the effect of mucilage on soil gas diffusion highly depends on soil texture and mucilage content. The diminishing hysteresis with the addition of mucilage suggests that plant roots secrete mucilage to balance oxygen availability and water content, even under fluctuating moisture conditions.

Hydraulic and mechanical response of loess to different chemical components in root exudates

Hydraulic and mechanical response of loess to different chemical components in root exudates

Coral mucus promotes the carbon metabolic potency of microorganisms in the coral reef ecosystem

AbstractCoral mucus, teeming with organic matter, releases nutrients and microorganisms, affecting element cycling and microbial communities in coral reefs. While terrestrial ecosystems exhibit well‐studied priming effects from root exudates, the influence of mucus in marine environments, particularly in coral reefs, remains underexplored. We hypothesize that coral mucus, through its nutrients and microbes, stimulates the surrounding microorganisms, regulating carbon metabolism and thus contributing to high coral reef productivity. Initially, natural samples (Acropora pruinosa mucus, seawater, and sediment) were collected for metagenomic assessment of microbial communities and functions. Results showed significant differences in microbial diversity, community structures, co‐occurrence modes, and unique functions among mucus, seawater, and sediment. Subsequent laboratory experiments demonstrated mucus's substantial influence on surrounding microorganisms. Analyses, including 16S rRNA sequencing and Eco‐plate results, revealed that mucus regulates microbial composition and activities. Quantitative gene‐chip analysis showed significant increase in the functional genes related to carbon fixation (e.g., the 3‐hydroxypropionate cycle) and degradation (e.g., pectin and hemicellulose hydrolysis) by 55.81% and 65.48%, respectively. Partial least squares path modeling identified mucus nutrients and microbial community composition as key contributors to carbon metabolic potential. This research confirms that mucus acts as a trigger, reshaping microbial profiles around corals and enhancing carbon utilization efficiency, highlighting its essential role in carbon metabolism and the maintenance of high productivity in coral reef ecosystems.

Unveiling the Role of Root Exudates in Plant Adaptation to Drought and Heat Stress

Unveiling the Role of Root Exudates in Plant Adaptation to Drought and Heat Stress

OsCYP706C2 diverts rice strigolactone biosynthesis to a noncanonical pathway branch.

Strigolactones exhibit dual functionality as regulators of plant architecture and signaling molecules in the rhizosphere. The important model crop rice exudes a blend of different strigolactones from its roots. Here, we identify the inaugural noncanonical strigolactone, 4-oxo-methyl carlactonoate (4-oxo-MeCLA), in rice root exudate. Comprehensive, cross-species coexpression analysis allowed us to identify a cytochrome P450, OsCYP706C2, and two methyl transferases as candidate enzymes for this noncanonical rice strigolactone biosynthetic pathway. Heterologous expression in yeast and Nicotiana benthamiana indeed demonstrated the role of these enzymes in the biosynthesis of 4-oxo-MeCLA, which, expectedly, is derived from carlactone as substrate. The oscyp706c2 mutants do not exhibit a tillering phenotype but do have delayed mycorrhizal colonization and altered root phenotype. This work sheds light onto the intricate complexity of strigolactone biosynthesis in rice and delineates its role in symbiosis and development.

Kinetics of uptake, translocation, and metabolism of organophosphate esters in japonica rice (Oryza sativa L.): Hydroponic experiment combined with model

Hydroponics combined with fugacity model was employed to investigate the kinetics of uptake, accumulation, and metabolism of organophosphate esters (OPEs) by japonica rice. The time-dependent process for uptake and accumulation of 5 OPEs and their diester-metabolites in both rice root and shoot fitted well with the pseudo-first-order kinetic model. The peak OPE accumulations in rice root and shoot were significantly positively or negatively correlated with their octanol-water partition coefficient (logKow) respectively, but not for their apparent accumulation rates. Root concentration factors (RCFs) and root-to-shoot translocation factors (TFs) of OPEs were found to be positively and negatively correlated with their logKow, respectively. Triphenyl phosphate with benzene ring substituents showed the highest RCF, but the lowest TF, because of its high potential for root adsorption due to the π electron-rich structures. Sterilized root exudates can hinder the root adsorption and absorption of OPEs from solution probably through competitive adsorption of OPEs with root surface. The first-hand transport and metabolism rates were also obtained by generating these rates to fit the dynamic fugacity model with the measurement values. The simulation indicated that the kinetics of OPE accumulation in rice plants may be controlled by multiple processes and physicochemical properties besides Kow.

Understanding the differential impact of PFOS and F-53B pollution on reed-mediated soil organic matter decomposition: Insights from rhizosphere priming effect

Plants affect soil microorganisms through the release of root exudates under pollution stress. This process may affect rhizosphere priming effect (RPE) and alter the rate of soil organic matter decomposition. However, the influence of plants on the decomposition of organic matter in soil subjected to pollution stress remains unclear. We studied the effects of exposure to perfluorooctanesulfonic (PFOS) and its alternative, chlorinated polyfluoroalkyl ether sulfonic (F-53B), at concentrations of 0.1 mg/kg and 50 mg/kg on the RPE of reed. We conducted our experiments in an artificial climate chamber and used the natural 13C tracer method to determine RPE. In the PFOS-exposed groups, the RPE was negative, with values of −11.45 mg C kg−1 soil d−1 in the low PFOS group and −8.04 mg C kg−1 soil d−1 in the high PFOS group. In contrast, in the F-53B-exposed groups, the RPE was positive, with values of 8.26 mg C kg−1 soil d−1 in the low F-53B group and 12.18 mg C kg−1 soil d−1 in the high F-53B group. Exposure of reeds to PFOS/F-53B stress resulted in differential effects on extracellular enzyme activities. The observed positive and negative RPE phenomena could be attributed to variations in extracellular enzyme activities. In conclusion, RPE responded differently under PFOS/F-53B exposure.

Long-Term Straw Returning Enhances Phosphorus Uptake by Zea mays L. through Mediating Microbial Biomass Phosphorus Turnover and Root Functional Traits.

The intensive use of chemical fertilizers in China to maintain high crop yields has led to significant environmental degradation and destabilized crop production. Returning straw to soil presents a potential alternative to reduce chemical fertilizer requirements and enhance soil fertility. This study investigates the effects of different nitrogen (N) input levels and straw additions on crop phosphorus (P) uptake and soil P availability based on a long-term N-fertilizer trial. The treatments included no fertilizer input (CK), conventional (NPK), reduced NPK (0.75NPK), and straw-amended (SNPK) treatments. Results indicate that SNPK significantly enhances shoot P uptake and crop yields by 43.7-61.9% and 29.3-39.6%, respectively. The SNPK treatment improved rhizosphere P availability and increased the phosphorus activation coefficient (PAC) by 1.72-fold compared to NPK alone. The enhanced soil P availability under SNPK was primarily attributed to an abundance of functional microbes, leading to higher P storage in the microbial biomass P pool and its turnover. Additionally, SNPK promoted root exudate and phosphate-mobilizing microbes, enhancing P mobilization and uptake. Nitrogen fertilization primarily influenced root functional traits related to P acquisition. These findings provide valuable insights for developing effective fertilizer management strategies in maize-oilseed rape rotation systems, emphasizing the benefits of integrating straw with chemical fertilizers.

Untargeted metabolomic analysis reveals a potential role of saponins in the partial resistance of pea (Pisum sativum) against a root rot pathogen, Aphanomyces euteiches.

In soil-borne diseases, the plant-pathogen interaction begins as soon as the seed germinates and develops into a seedling. Aphanomyces euteiches, an oomycete, stays dormant in soil and gets activated by sensing the host through chemical signals present in the root exudates. The composition of plant exudates may, thus, play an important role during the early phase of infection. To better understand the role of root exudates in plant resistance, we investigated the interaction between partially resistant lines (PI660736 and PI557500) and susceptible pea cultivars (CDC Meadow and AAC Chrome) against Aphanomyces euteiches during the pre-invasion phase. The root exudates of two sets of cultivars clearly distinguished from each other in inducing oospore germination. PI557500 root exudate not only had diminished induction but also inhibited the oospore germination. The contrast between the root exudates of resistance and susceptible cultivars was reflected in their metabolic profiles. Data from fractionation and oospore germination inhibitory experiments identified a group of saponins that accumulated differentially in susceptible and resistant cultivars. We detected 56 saponins and quantified 44 of them in pea root and 30 from root exudate; the majority of them, especially Soyasaponin I and dehydrosoyasaponin I with potent in vitro inhibitory activities, were present in significantly higher amounts in both roots and root exudates of PI660736 and PI557500 as compared to Meadow and Chrome. Our results provide evidence for saponins as deterrents against Aphanomyces euteiches, which might have contributed to the resistance against root rot in the studied pea cultivars.

Postfire extracellular enzyme activity in a temperate montane forest

AbstractWildfire is a disturbance expected to increase in frequency and severity, changes that may impact carbon (C) dynamics in the soil ecosystem. Fire changes the types of C sources available to soil microbes, increasing pyrogenic C and coarse downed wood, and if there is substantial tree mortality, decreasing C from root exudates and leaf litter. To investigate the impact of this shift in the composition of C resources on microbial processes driving C cycling, we examined microbial activity in soil sampled from an Oregon burn 1 year after fire from sites spanning a range in soil burn severity from unburned to highly burned. We found evidence that postfire rhizosphere priming loss may reduce soil C loss after fire. We measured the potential activity of C‐acquiring and nitrogen (N)‐acquiring extracellular enzymes and contextualized the microbial resource demand using measurements of mineralizable C and N. Subsurface mineralizable C and N were unaltered by fire and negatively correlated with hydrolytic extracellular enzyme activity (EEA) in unburned, but not burned sites. EEA was lower in burned sites by up to 46%, but only at depths below 5 cm, and with greater decreases in sites with high soil burn severity. These results are consistent with a subsurface mechanism driven by tree mortality. We infer that in sites with high tree mortality, subsurface EEAs decreased due to loss of rhizosphere priming and that inputs of dead roots contributed to mineralizable C stabilization.

Unlocking the roles of wheat root exudates in regulating laccase-catalyzed estrogen humification

While laccase humification has an efficient capacity to convert estrogenic pollutants, the roles of wheat (Triticum aestivum L.) root exudates (W-REs) in the enzymatic humification remain poorly understood. Herein, we presented the research into the effects of W-REs on 17β-estradiol (E2) and bisphenol A (BPA) conversion in vitro laccase humification. W-REs inhibited E2 removal but promoted BPA conversion in the enzymatic humification, and the first-order kinetic constants for E2 and BPA were 0.27–0.69 and 0.28–0.55 h−1, respectively. Specialized small phenols and amino acids in W-REs were susceptible to laccase humification, resulting in increased copolymerization of estrogen and W-REs. In greenhouse hydroponics, the accumulated amounts of E2 (BPA) in the roots and shoots were estimated to be 0.87 (2.15) and 0.43 (0.51) nmol·plant−1 at day 3, respectively. By forming low- and eventually non-toxic copolymeric precipitates between estrogen and W-REs, laccase humification lowered the phytotoxicity and bioavailability of estrogen in the rhizosphere solution, consequently relieving its uptake, accumulation, and distribution in the wheat cells. This work sheds light on the roles of W-REs in regulating laccase-catalyzed estrogen humification, and gives an insight into the path of addressing organic contamination in the rhizosphere and ensuring food safety.

Characterization of the Bioavailability of Per- and Polyfluoroalkyl Substances in Farmland Soils and the Factors Impacting Their Translocation to Edible Plant Tissues.

In this study, various crops and farmland soils were collected from the Fen-Wei Plain, China, to investigate the bioavailability of perfluoroalkyl substances (PFAS), their accumulation in edible plant tissues, and the factors impacting their accumulation. PFAS were frequently detected in all of the crops, with total concentrations ranging from 0.61 to 35.8 ng/g. The results of sequential extractions with water, basic methanol, and acidic methanol indicate that water extraction enables to characterize the bioavailability of PFAS in soil to edible plant tissues more accurately, especially for the shorter-chain homologues. The bioavailability of PFAS was remarkably enhanced in the rhizosphere (RS) soil, with the strongest effect observed for leafy vegetables. The water-extracted Σ16PFAS in RS soil was strongly correlated with the content of dissolved organic carbon in the soil. Tannins and lignin, identified as the main components of plant root exudates by Fourier transform-ion cyclotron resonance mass spectrometry, were found to enhance the bioavailability of PFAS significantly. Redundancy analysis provided strong evidence that the lipid and protein contents in edible plant tissues play important roles in the accumulation of short- and long-chain PFAS, respectively. Additionally, the high water demand of these tissues during the growth stage greatly facilitated the translocation of PFAS, particularly for the short-chain homologues and perfluorooctanoic acid.

Dual column chromatography combined with high-resolution mass spectrometry improves coverage of non-targeted analysis of plant root exudates

BackgroundWithin the plant kingdom, there is an exceptional amount of chemical diversity that has yet to be annotated. It is for this reason that non-targeted analysis is of interest for those working in novel natural products. To increase the number and diversity of compounds observable in root exudate extracts, several workflows which differ at three key stages were compared: 1) sample extraction, 2) chromatography, and 3) data preprocessing. ResultsPlants were grown in Hoagland's solution for two weeks, and exudates were initially extracted with water, followed by a 24-h regeneration period with subsequent extraction using methanol. Utilizing the second extraction showed improved results with less ion suppression and reduced retention time shifting compared to the first extraction. A single column method, utilizing a pentafluorophenyl column, paired with high-resolution mass spectrometry ionized and correctly identified 34 mock root exudate compounds, while the dual column method, incorporating a pentafluorophenyl column and a porous graphitic carbon column, retained and identified 43 compounds. In a pooled quality control sample of exudate extracts, the single column method detected 1,444 compounds. While the dual method detected fewer compounds overall (1,050), it revealed a larger number of small polar compounds. Three preprocessing methods (targeted, proprietary, and open source) successfully identified 43, 31, and 38 mock root exudate compounds to confidence level 1, respectively. SignificanceEnhancing signal strength and analytical method stability involves removing the high ionic strength nutrient solution before sampling root exudate extracts. Despite signal intensity loss, a dual column method enhances compound coverage, particularly for small polar metabolites. Open-source software proves a viable alternative for non-targeted analysis, even surpassing proprietary software in peak picking.

Root-secreted (-)-loliolide mediates chemical defense in rice and wheat against pests.

Plant chemical defense can be elicited by signaling chemicals. As yet, the elicitation is mainly known from volatile aboveground signals. Root-secreted belowground signals and their underlying mechanisms are largely unknown. This study examined a root-secreted signaling (-)-loliolide to trigger chemical defense in rice and wheat against pests by means of cocultivation and incubation experiments. Wild-type Arabidopsis (WT) and its root exudates with (-)-loliolide induced the production of defensive metabolites of rice and wheat and reduced the performance of weeds, pathogens and herbivores, while a carotenoid-deficient mutant (szl1-1) and its root exudates without (-)-loliolide had no similar effects. However, the induction and reduction occurred in the szl1-1 root exudates by (-)-loliolide supplementation with the level equal to that of WT. RNA-sequencing analysis revealed a significant change in the transcript level of defense-related genes in rice exposure to (-)-loliolide. Furthermore, (-)-loliolide enhanced rice resistance against Rhizoctonia solani through changing reactive oxygen species (ROS) system, and mediating jasmonic acid, salicylic acid and abscisic acid biosynthesis. Root-secreted signaling (-)-loliolide can trigger chemical defense in rice and wheat against their pests. Such perception-dependent chemical defenses provide an intriguing possibility for ecological pest management to increase crop productivity and sustainability. © 2024 Society of Chemical Industry.

Different patterns of fine-root exudation rates along soil depth of Pinus densiflora, Chamaecyparis obtusa, and Cryptomeria japonica in coniferous forests

Tree fine-root exudation and organic carbon compounds in the soil affect physiological functions through degradation by microorganisms around the roots and the enhancement of allelopathic effects on invasion attempts by other plant species. However, the underlying mechanism of fine-root action in vertically heterogeneous resource-distribution patterns along the soil profile is little known. Here, we quantified root exudation and its morphological and chemical traits in the soil layers down to 100-cm depth for three conifer species, Pinus densiflora (ectomycorrhiza), Chamaecyparis obtusa, and Cryptomeria japonica (arbuscular mycorrhiza), in a cool temperate Japanese forest. By using glass filter-trap method, root exudation rates in all three species varied with soil depth in a species dependent pattern. Specifically, the root exudation rate of P. densiflora was significantly higher in the middle soil layer, and lower in the topsoil layer, whereas the exudation rates significantly decreased in C. japonica and did not vary in C. obtusa along soil depth. Exudation strongly correlated with variation in root traits in C. obtusa and C. japonica but not in P. densiflora. These findings suggest that differences in exudation patterns among tree species are likely compensated for by structural modifications in fine roots in response to soil depth. In addition, the traits with mycorrhizal types might be related to adequate root exudation and enhanced soil-nutrient acquisition. Distinct differences among species in exudation, associated with vertical root distribution, will increase our understanding of root survival strategies for nutrient acquisition and carbon sequestration in forest tree species.