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The phenomenon of nitrogen deposition resulting in species loss in terrestrial ecosystems has been demonstrated in several experiments. Nitrogen (N) and phosphorus (P), as major nutrients required for plant growth, exhibit ecological stoichiometric coupling in many ecosystems. The increased availability of nitrogen can exacerbate the ecological effects of phosphorus. To reveal the ecological effects of phosphorus under nitrogen-limiting and non-limiting conditions, we conducted a controlled N-P interaction experiment over 5 years in the Hulunbuir meadow steppe, where two nitrogen addition levels were implemented: 0 g N·m-2·a-1 (nitrogen-limiting condition) and 10 g N·m-2·a-1 (nitrogen-non-limiting condition), together with six levels of phosphorus addition (0, 2, 4, 6, 8, and 10 g P·m-2·a-1). The results showed that nitrogen addition (under nitrogen-non-limiting conditions) significantly decreased species diversity in the steppe community, which was exacerbated under phosphorus addition. Under nitrogen-limiting conditions, phosphorus addition had no marked impact on species diversity compared to the control; however, there were substantial differences between different levels of phosphorus addition, exhibiting a unimodal change. Under both experimental nitrogen conditions, the addition of 6 g P·m-2·a-1 was the threshold for affecting the community species diversity. Nitrogen addition reduced the relative biomass of legumes, bunch grasses, and forbs, but substantially increased the relative biomass of rhizomatous grasses. In contrast, phosphorus addition only markedly affected the relative biomass of forbs and rhizomatous grasses, with the former showing a unimodal pattern of first increasing and then decreasing with increasing phosphorus addition level, and the latter exhibiting the opposite pattern. The different responses of rhizomatous grasses and other functional groups to nitrogen and phosphorus addition were observed to have a regulatory effect on the changes in grassland community structure. Phosphorus addition may increase the risk of nitrogen deposition-induced species loss. Both nitrogen and phosphorus addition lead to soil acidification and an increase in the dominance of the already-dominant species, and the consequent species loss in the forb functional group represents the main mechanism for the reduction in community species diversity.

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Increasing nitrogen (N) deposition alters the availability of soil nutrients and is likely to intensify phosphorus (P) limitations, especially in P-limited tropical and subtropical forests. Soil microorganisms play vital roles in carbon (C) and nutrient cycling, but it is unclear whether and how much N and P imbalances affect the soil's microbial metabolism and mechanisms of nutrient limitations. In this study, a 3-year field experiment of N and P addition (control (CK), 100 kg N ha-1 yr-1 (N), 50 kg P ha-1 yr-1 (P), and NP) was set up to analyze the extracellular enzyme activities and stoichiometry characteristics of the top mineral soils in Chinese fir plantations with different stand ages (7, 20, and 33 years old). The results showed that the enzyme activities associated with the acquisition of C (ß-1,4-glucosidase (BG) and ß-d-cellobiohydrolase (CBH)) and P (acid phosphatases (APs)) in the N treatment were significantly higher than those in the CK treatment. Moreover, vector analysis revealed that both the vector's length and angle increased in stands of all ages, which indicated that N addition aggravated microbial C and P limitations. The P and NP treatments both significantly decreased the activity of AP and the enzymes' N:P ratio, thereby alleviating microbial P limitations, as revealed by the reduction in the vector's angle. Stand age was found to promote all enzymatic activities but had no obvious effects on the limitation of microbial metabolism with or without added nutrients in the soils under Chinese fir. Available N, Olsen-P, and pH were the main drivers of microbial metabolic limitations related to C nutrients. These results provide useful data for understanding the change in soil microbial activity in response to environmental changes, and suggest that P fertilization should be considered for management to improve productivity and C sequestration in Chinese fir plantation in the context of increased deposition of N.

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Human activities have increased nitrogen (N) and phosphorus (P) inputs to the Yellow River Delta and the supply level of N and P affects plant growth as well as ecosystem structure and function directly. However, the root growth, stoichiometry, and antioxidant system of plants in response to N and P additions, especially for herbaceous halophyte in the Yellow River Delta (YRD), remain unknown. A field experiment with N addition (0, 5, 15, and 45 g N m-2 yr-1, respectively) as the main plot, and P addition (0 and 1 g N m-2 yr-1, respectively) as the subplot, was carried out with a split-plot design to investigate the effects on the root morphology, stoichiometry, and antioxidant system of Suaeda salsa. The results showed that N addition significantly increased the above-ground and root biomass as well as shoot-root ratio of S. salsa, which had a significant interaction with P addition. The highest biomass was found in the treatment with 45 g N m-2 yr-1 combined with P addition. N addition significantly increased TN content and decreased C:N ratio of root, while P addition significantly increased TP content and decreased C:P ratio. The main root length (MRL), total root length (TRL), specific root length (SRL), and root tissue density (RTD) of S. salsa root were significantly affected by N addition and P addition, as well as their interaction. The treatments with or without P addition at the 45 g N m-2 yr-1 of N addition significantly increased the superoxide dismutase (SOD), peroxidase (POD), catalase (CAT) activities and soluble protein content of roots, decreased malondialdehyde (MDA) content. And there was a significant interaction between the N and P addition on SOD activity. Therefore, N and P additions could improve the growth of S. salsa by altering the root morphology, increasing the root nutrient content, and stimulating antioxidant system.

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Introduction: Understanding the response of cross-domain co-occurrence networks of soil microorganisms to phosphorus stability and the resulting impacts is critical in ecosystems, but the underlying mechanism is unclear in artificial grassland ecosystems. Methods: In this study, the effects of four phosphorus concentrations, P0 (0 kg P ha-1), P1 (15.3 kg P ha-1), P2 (30.6 kg P ha-1), and P3 (45.9 kg P ha-1), on the cross-domain co-occurrence network of bacteria and fungi were investigated in an artificial Leymus chinensis grassland in an arid region. Results and discussion: The results of the present study showed that phosphorus addition significantly altered the stem number, biomass and plant height of the Leymus chinensis but had no significant effect on the soil bacterial or fungal alpha (ACE) diversity or beta diversity. The phosphorus treatments all increased the cross-domain co-occurrence network edge, node, proportion of positively correlated edges, edge density, average degree, proximity to centrality, and robustness and increased the complexity and stability of the bacterial-fungal cross-domain co-occurrence network after 3 years of continuous phosphorus addition. Among them, fungi (Ascomycota, Basidiomycota, Mortierellomycota and Glomeromycota) play important roles as keystone species in the co-occurrence network, and they are significantly associated with soil AN, AK and EC. Finally, the growth of Leymus chinensis was mainly due to the influence of the soil phosphorus content and AN. This study revealed the factors affecting the growth of Leymus chinense in artificial grasslands in arid areas and provided a theoretical basis for the construction of artificial grasslands.

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Soil microbial community composition and diversity are often affected by nutrient enrichment, which may influence soil microbes to affect nutrient cycling and plant community structure. However, the response of soil bacteria to nitrogen (N) and phosphorus (P) addition and whether it is influenced by plants remains unclear. By 16S rRNA sequencing, we investigated the response of the rhizosphere and bulk soil bacterial communities of different halophytes (salt-rejecting, salt-absorbing, and salt-secreting plant) in the Yellow River Delta to short-term N and P addition. The response of rhizosphere bacterial diversity to N and P addition was opposite in Phragmites communis and Suaeda salsa. N addition increased the rhizosphere soil bacterial α-diversity of S. salsa and Aeluropus sinensis, while P addition decreased the rhizosphere bacterial α-diversity bacteria of S. salsa. The N and P addition had a weak effect on the rhizosphere bacterial community composition and a significant effect on the bulk soil bacterial community composition of halophytes. The S. salsa and P. communis bulk soil bacterial community were mainly influenced by P addition, while it was influenced by N addition in A. sinensis. N and P addition reduced the difference in bacterial community composition between the two types of soil. N and P addition increased the eutrophic taxa (Proteobacteria and Bacteroidetes) and decreased the oligotrophic taxa (Acidobacteria). Redundancy analysis showed that soil organic matter, salt, and total N content had significant effects on the bacterial community composition. The results clarify that the response of soil bacterial communities to N and P additions is inconsistent across the three halophyte soils, and the effect of plant species on the bacterial community was stronger than short-term N and P addition. IMPORTANCE: The bulk soil bacterial community was more affected by nutrient addition. Nitrogen (N) and phosphorus (P) have different effects on bacterial community. Soil organic matter is a key factor influencing the response of bacterial community to nutrient addition. N and P influence on bacterial community changes with plants.

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Introduction: Phosphorus (P), which plays a vital role in plant growth, is continually added to soil to maximize biomass production, leading to excessive P accumulation and water eutrophication. Results: In this study, a pot experiment using a subtropical tobacco-growing soil fertilized with four P levels-no P, low P, medium P, and high P-was conducted and rhizosphere and bulk soils were analyzed. Results: P addition significantly increased tobacco biomass production (except under low P input) and total soil P and available P content (P<0.05), whereas total nitrogen content decreased in the rhizosphere soils, although this was only significant with medium P application. P fertilization also significantly altered the bacterial communities of rhizosphere soils (P<0.05), but those of bulk soils were unchanged (P>0.05). Moreover, a significant difference was found between rhizosphere soils with low (LR) and high (HR) P inputs (P<0.05). Additionally, compared with rhizosphere soils with no P (CKR), Shannon diversity showed a declining trend, which was significant with LR and HR (P<0.05), whereas an increasing tendency was observed for Chao1 diversity except in LR (P>0.05). Functional prediction revealed that P application significantly decreased the total P and N metabolism of microorganisms in rhizosphere soils (P<0.05). Discussion: Collectively, our results indicate that maintaining sustainable agricultural ecosystems under surplus P conditions requires more attention to be directed toward motivating the potential of soil functional microbes in P cycling, rather than just through continual P input.

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The escalation of global nitrogen deposition levels has heightened the inhibitory impact of phosphorus limitation on plant growth in subtropical forests. Plant roots area particularly sensitive tissue to nitrogen and phosphorus elements. Changes in the morphological characteristics of plant roots signify alterations in adaptive strategies. However, our understanding of resource-use strategies of roots in this environment remains limited. In this study, we conducted a 10-month experiment at the Castanopsis kawakamii Nature Reserve to evaluate the response of traits of seedling roots (such as specific root length, average diameter, nitrogen content, and phosphorus content) to nitrogen and phosphorus addition. The aim was to reveal the adaptation strategies of roots in different nitrogen and phosphorus addition concentrations. The results showed that: (1) The single phosphorus and nitrogen-phosphorus interaction addition increased the specific root length, surface area, and root phosphorus content. In addition, single nitrogen addition promotes an increase in the average root diameter. (2) Non-nitrogen phosphorus addition and single nitrogen addition tended to adopt a conservative resource-use strategy to maintain growth under low phosphorus conditions. (3) Under the single phosphorus addition and interactive addition of phosphorus and nitrogen, the roots adopted an acquisitive resource-use strategy to obtain more available phosphorus resources. Accordingly, the adaptation strategy of seedling roots can be regulated by adding appropriate concentrations of nitrogen or phosphorus, thereby promoting the natural regeneration of subtropical forests.

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Rice-fish coculture (RFC), as a traditional agricultural strategy in China, can optimally utilize the scarce resource, especially in subtropical regions where phosphorus (P) deficiency limits agricultural production. However, ammonia-oxidizing archaea (AOA) and bacteria (AOB) are involved in the ammonia oxidation, but it remains uncertain whether their community compositions are related to the RFC combined with and without P addition that improves soil nitrogen (N) use efficiency. Here, a microcosm experiment was conducted to assess the impacts of RFC combined with and without inorganic P (0 and 50 mg P kg-1 as KH2PO4) addition on AOA and AOB community diversities, enzyme activities and N availability. The results showed that RFC significantly increased available N content without P addition compared with P addition. Moreover, RFC significantly increased urease activity and AOA shannon diversity, and reduced NAG activity and AOB shannon diversity without P addition, respectively. Higher diversity of AOA compared with that of AOB causes greater competition for resources and energy within their habitats, thereby resulting in lower network complexity. Our findings indicated that the abundances of AOA and AOB are influenced through the introduction of fish and/or P availability, of which AOB is linked to N availability. Overall, RFC could improve paddy soil N availability without P addition in subtropical region, which provides a scientific reference for promoting the practices that reduce N fertilizer application in RFC.

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Understanding how nutrient addition affects the tree growth is critical for assessing forest ecosystem function and processes, especially in the context of increased nitrogen (N) and phosphorus (P) deposition. Subtropical forests are often considered N-rich and P-poor ecosystems, but few existing studies follow the traditional "P limitation" paradigm, possibly due to differences in nutrient requirements among trees of different size classes. We conducted a three-year fertilization experiment with four treatments (Control, N-treatment, P-treatment, and NP-treatment). We measured soil nutrient availability, leaf stoichiometry, and relative growth rate (RGR) of trees across three size classes (small, medium and large) in 64 plots. We found that N and NP-treatments increased the RGR of large trees. P-treatment increased the RGR of small trees. RGR was mainly affected by N addition, the total effect of P addition was only 10 % of that of N addition. The effect of nutrient addition on RGR was mainly regulated by leaf stoichiometry. This study reveals that nutrient limitation is size dependent, indicating that continuous unbalanced N and P deposition will inhibit the growth of small trees and increase the instability of subtropical forest stand structure, but may improve the carbon sink function of large trees.

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Anthropogenic eutrophication is known to impair the stability of aboveground net primary productivity (ANPP), but its effects on the stability of belowground (BNPP) and total (TNPP) net primary productivity remain poorly understood. Based on a nitrogen and phosphorus addition experiment in a Tibetan alpine grassland, we show that nitrogen addition had little impact on the temporal stability of ANPP, BNPP, and TNPP, whereas phosphorus addition reduced the temporal stability of BNPP and TNPP, but not ANPP. Significant interactive effects of nitrogen and phosphorus addition were observed on the stability of ANPP because of the opposite phosphorus effects under ambient and enriched nitrogen conditions. We found that the stability of TNPP was primarily driven by that of BNPP rather than that of ANPP. The responses of BNPP stability cannot be predicted by those of ANPP stability, as the variations in responses of ANPP and BNPP to enriched nutrient, with ANPP increased while BNPP remained unaffected, resulted in asymmetric responses in their stability. The dynamics of grasses, the most abundant plant functional group, instead of community species diversity, largely contributed to the ANPP stability. Under the enriched nutrient condition, the synchronization of grasses reduced the grass stability, while the latter had a significant but weak negative impact on the BNPP stability. These findings challenge the prevalent view that species diversity regulates the responses of ecosystem stability to nutrient enrichment. Our findings also suggest that the ecological consequences of nutrient enrichment on ecosystem stability cannot be accurately predicted from the responses of aboveground components and highlight the need for a better understanding of the belowground ecosystem dynamics.

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Root-associated fungi play a vital role in maintaining nutrient absorption and health of host plants. To compare the responses of root-associated fungal community structures to nitrogen (N) and/or phosphorus (P) additions across differential mycorrhizal types, we collected roots of nine plant species belonging to three mycorrhizal types (arbuscular mycorrhiza, ectomycorrhiza, and ericoid mycorrhiza) under control and N and/or P addition treatments from a subtropical forest, and detected the diversity and community composition of fungi inhabiting roots through the high-throughput sequencing technique. The results showed that root-associated fungal communities of all nine plant species were mainly composed of Basidiomycota and Ascomycota. The relative abundance of Ascomycota and Basidiomycota was significantly lower and higher under the P addition than that under control, respectively. The relative abundance of Ascomycota of ericoid mycorrhizal trees was significantly higher than those of arbuscular mycorrhizal and ectomycorrhizal trees, while the relative abundance of Basidiomycota was significantly lower than the other two mycorrhizal types. Compared with the control, P addition significantly reduced the α-diversity and changed community composition of root-associated fungi across different mycorrhizal plant types, while no effect of N addition or mycorrhizal type was observed. Compared with the control and N addition treatments, NP addition caused root-associated fungal communities of all plants becoming integrally divergent. In addition, the fungal communities of ectomycorrhizal mycorrhizal trees became apparently convergent in comparison with those of arbuscular and ericoid mycorrhizal trees under the NP addition. Collectively, our results highlighted that P was a critical factor influencing community structures of tree root-associated fungi in subtropical forest soils. This study would enhance our understanding of the responses and maintenance mechanisms of plant root-associated fungal diversity under global environmental changes in the subtropical region.

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Nitrogen (N) and phosphorus (P) addition, either individually or in combination, has been demonstrated to enhance plant productivity in grassland ecosystems. Soil bacterial community, which is the driver of litter decomposition and nutrient cycling, is assumed to control responses of terrestrial ecosystem structure and function to N and P addition. Using a high-throughput Illumina MiSeq sequencing platform, we conducted a 9-year field experiment of N (0, 5, 10, and 20 g N m-2 yr-1) and P (0 and 10 g P m-2 yr-1) additions in the Inner Mongolian steppes to elucidate long-term effects of N and P addition on soil bacterial richness, diversity and composition. We found that N addition reduced the relative abundance of Acidobacteria, Chloroflexi, and Nitrospirae, while increased that of Bacteroides. The results showed that the bacterial biomarker was enriched in P addition treatments, either individually or combined with N addition. Both N and P addition altered the bacterial community structure, while only N addition greatly decreased bacterial richness and diversity. More importantly, we showed that all of these effects were most significant in N3P treatment (20 g N m-2 yr-1 and 10 g P m-2 yr-1), implying that P coupled with a high-level N addition exerted a great influence on soil bacterial community. Structural equation models revealed that N and P addition had a great direct effect on soil bacterial community and an indirect effect on it mainly by changing the litter biomass. Our findings highlighted that severe niche differentiation was induced by P along with a high-level N, further emphasizing the importance of simultaneously evaluating response of soil bacterial community to N and P addition, especially in the context of increasing anthropogenic nutrient additions.

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Sugar, as a nutrient exchange substance between arbuscular mycorrhizal (AM) fungi and host plants, plays an important role in the abiotic stress response of mycorrhizal plants. This experiment aimed to study the effects of AM fungi and phosphorus (P) addition on the sugar metabolism and 14-3-3 gene expression of Populus cathayana under drought stress. The results showed that drought affects the process of sugar metabolism by increasing the activities of amylase and invertase, resulting in the decrease of starch content in leaves and roots and the accumulation of soluble sugars (including reducing sugar and sucrose) in roots. Under drought stress, the activity or content of sucrose synthetase, sucrose phosphate synthase, acid invertase, alkaline invertase, reducing sugar, soluble sugar, sucrose, and starch in the root showed the best mycorrhizal effect at the 100 mg P level. The expression levels of the 14-3-3 genes (PcGRF10 and PcGRF11) were significantly increased by mycorrhizal induction under drought stress. These levels were positively correlated with SS, SPS, sucrose, and starch phosphorylase in leaves, as well as with almost all sugar metabolism indicators in roots. However, they were negatively correlated with starch content in both leaves and roots. Sugar metabolism and 14-3-3 protein gene expression were induced by AM fungi and P addition in response to drought stress. The 14-3-3 genes induced by AM fungi may be involved in participating in osmotic regulation during drought stress. This study provides a new idea for the mechanism of sugar metabolism of mycorrhizal plants in arid regions.

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Plant invasion has severely damaged ecosystem stability and species diversity worldwide. The cooperation between arbuscular mycorrhizal fungi (AMF) and plant roots is often affected by changes in the external environment. Exogenous phosphorus (P) addition can alter the root absorption of soil resources, thus regulating the root growth and development of exotic and native plants. However, it remains unclear how exogenous P addition regulates the root growth and development of exotic and native plants mediated by AMF, affecting the exotic plant invasion. In this experiment, the invasive plant Eupatorium adenophorum and native plant Eupatorium lindleyanum were selected and cultured under intraspecific (Intra-) competition and interspecific (Inter-) competition conditions, involving inoculation with (M+) and without AMF (M-) and three different levels of P addition including no addition (P0), addition with 15 mg P kg-1 soil (P15), and addition with 25 mg P kg-1 soil (P25) for the two species. Root traits of the two species were analyzed to study the response of the two species' roots to AMF inoculation and P addition. The results showed that AMF significantly promoted the root biomass, length, surface area, volume, tips, branching points, and carbon (C), nitrogen (N), and P accumulation of the two species. Under M+ treatment, the Inter- competition decreased the root growth and nutrient accumulation of invasive E. adenophorum but increased the root growth and nutrient accumulation of native E. lindleyanum relative to the Intra- competition. Meanwhile, the exotic and native plants responded differently to P addition, exhibiting root growth and nutrient accumulation of invasive E. adenophorum increased with P addition, whereas native E. lindleyanum reduced with P addition. Further, the root growth and nutrition accumulation of native E. lindleyanum were higher than invasive E. adenophorum under Inter- competition. In conclusion, exogenous P addition promoted the invasive plant but reduced the native plant in root growth and nutrient accumulation regulated by AMF, although the native plant outcompeted the invasive plant when the two species competed. The findings provide a critical perspective that the anthropogenic P fertilizer addition might potentially contribute to the successful invasion of exotic plants.

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Methane (CH4) is a critical greenhouse gas, and wetlands are the largest natural emitters of CH4. Owing to global climate change and the intensification of anthropogenic activities, the input of exogenous nutrients such as nitrogen (N) and phosphorus (P) into wetland ecosystems has increased, which may significantly affect nutrient cycling and CH4 fluxes from wetlands. However, the environmental and microbial effects of the addition of N and P on CH4 emissions from alpine wetlands have not been thoroughly examined. We conducted a two-year field experiment with N and P addition to examine its impact on CH4 emissions from wetlands on the Qinghai-Tibet Plateau (QTP). The treatments comprised a blank control (CK), N addition (15 kg N ha-1 yr-1, N15), P addition (15 kg P ha-1 yr-1, P15), and NP co-addition (15 kg NP ha-1 yr-1, N15P15). We measured CH4 flux, soil environmental factors, and microbial community structure for each treatment plot. The results showed that the CH4 emissions of N and P addition were higher than CK. Specifically, the CH4 fluxes of N15, P15, and N15P15 treatments were 0.46 mg CH4 m-2 h-1, 4.83 mg CH4 m-2 h-1, and 0.95 mg CH4 m-2 h-1 higher than the CK. Additionally, the CH4 fluxes of N15P15 treatments was 3.88 mg CH4 m-2 h-1 lower than the P15 and 0.49 mg CH4 m-2 h-1 higher than the N15. This finding indicated that the CH4 flux in the alpine wetland soil was more sensitive to the addition of P. N and P addition increased not only soil organic carbon content (P < 0.05) but also the relative abundance of Chloroflexi and Actinobacteria in the soil, which may be the main reason for the promotion of CH4 emissions. Therefore, our results indicate that N and P addition can change the microbial abundance and community structure of wetland soil, and the distribution of soil carbon, promote CH4 emissions, and ultimately affect the carbon sink function of wetland ecosystems.

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Imbalanced nitrogen (N) and phosphorus (P) depositions are profoundly shifting terrestrial ecosystem biogeochemical processes. However, how P addition and its interaction with N addition influence the release of litter carbon (C), N, P, and especially metal nutrients in subtropical forests remains unclear. Herein, a two-year field litterbag experiment was conducted in a natural subtropical evergreen broadleaved forest of southwestern China using a factorial design with three levels of N addition (0, 10, and 20 g N m-2 y-1) and P addition (0, 5, 15 g P m-2 y-1). During two years of decomposition, N- and P-only addition treatments decreased the accumulated mass loss and release rates of litter C, N, P, K, Na, and Mn (p < 0.05); N and P coaddition treatments increased the accumulated mass loss and release rates of litter C, N, K, Na, Mn, and Cu (p < 0.05) and decreased the accumulated release rates of litter P and Mg (p < 0.05); the C/P and N/P ratios of the residual litter increased under the N-only addition treatments (p < 0.05) and decreased under the P-only addition and N and P coaddition treatments (p < 0.05). Overall, the results suggest that combined N and P supply can increase biological activities and thus accelerate the release of litter C, N, and most metal nutrients, as expected within the framework of ecological stoichiometry and growth rate hypothesis. Our study also highlights that the effect of N addition on litter C and nutrients release depends on P availability.

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Plants can alter soil microbial biomass and extracellular enzyme activities related with carbon (C), nitrogen (N), and phosphorus (P), through litter and root exudates, with consequences on soil carbon, nitrogen and phosphorus (P) cycling. However, it is not well known how the changes in soil phosphorus availability affect the relationships between plants and soil microorganisms. In this study, a factorial experiment was conducted to investigate the effects of Chinese fir (Cunninghamia lanceolata) planting and different levels of P addition (0, 1.95, 3.9, 7.8 and 15.6 g P·m-2·a-1) on soil microbial biomass and extracellular enzyme activities. The results showed that planting Chinese fir planting significantly altered soil microbial biomass and C- and N- and P-related extracellular enzyme activities, but the effects were dependent on P addition levels. Without P addition, Chinese fir planting significantly reduced soil nutrient availability and pH, which led to the aggravation of P limitation and lower soil microbial biomass. P addition relieved P limitation, and reduced soil acid phosphatase (ACP) activities by 30.0%, 30.5%, 35.3% and 47.1% with the increasing P addition level (1.95, 3.9, 7.8 and 15.6 g P·m-2·a-1). Under three P addition levels (1.95, 3.9 and 7.8 g P·m-2·a-1), the negative effects of Chinese fir planting on soil microbial growth were alleviated. Under the high P addition level (15.6 g P·m-2·a-1), the negative effects of Chinese fir planting on soil microbial growth occurred again due to soil N limitation. Taken together, Chinese fir planting and soil P availability generally affected soil microbial biomass and extracellular enzyme activities, and changed P limitation.

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Plant-soil feedback (PSF) is conventionally characterized by plant biomass growth, yet it remains unclear how PSF affects plant nutrient acquisition strategies (e.g., nutrient absorption and nutrient resorption) associated with plant growth, particularly under changing soil environments. A greenhouse experiment was performed with seedlings of Pinus elliottii Englem and conditioned soils of monoculture plantations (P. elliottii and Cunninghamia lanceolata Hook). Soil sterilization was designed to test plant phosphorus (P) acquisition strategy with and without native soil fungal communities. Soils from P. elliottii and C. lanceolata plantations were used to explore the specific soil legacy effects on two different P acquisition pathways (absorption and resorption). Phosphorus addition was also applied to examine the separate and combined effects of soil abiotic factors and soil fungal factors on P acquisition pathways. Due to diminished mycorrhizal symbiosis, PSF prompted plants to increasingly rely on P resorption under soil sterilization. In contrast, P absorption was employed preferentially in the heterospecific soil, where species-specific pathogenic fungi could not affect P absorption. Higher soil P availability diluted the effects of soil fungal factors on the trade-off between the two P acquisition pathways in terms of the absolute PSF. Moreover, P addition plays a limited role in terms of the relative PSF and does not affect the direction and strength of relative PSF. Our results reveal the role of PSF in regulating plant P acquisition pathways and highlight the interaction between mycorrhizal and pathogenic fungi as the underlying mechanism of PSF.

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The decoupling of microbial functional and taxonomic components refers to the phenomenon that a drastic change in microbial taxonomic composition leads to no or only a gentle change in functional composition. Although many studies have identified this phenomenon, the mechanisms underlying it are still unclear. Here we demonstrate, using metagenomics data from a steppe grassland soil under different grazing and phosphorus addition treatments, that there is no "decoupling" in the variation of taxonomic and metabolic functional composition of the microbial community within functional groups at species level. In contrast, the high consistency and complementarity between the abundance and functional gene diversity of two dominant species made metabolic functions unaffected by grazing and phosphorus addition. This complementarity between the two dominant species shapes a bistability pattern that differs from functional redundancy in that only two species cannot form observable redundancy in a large microbial community. In other words, the "monopoly" of metabolic functions by the two most abundant species leads to the disappearance of functional redundancy. Our findings imply that for soil microbial communities, the impact of species identity on metabolic functions is much greater than that of species diversity, and it is more important to monitor the dynamics of key dominant microorganisms for accurately predicting the changes in the metabolic functions of the ecosystems.

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BACKGROUND AND AIMS: Growing evidence has suggested that plant responses to model soil microorganisms are context dependent; however, few studies have investigated the effects of whole soil microbial communities on plant performance in different abiotic and biotic conditions. To address this, we examined how soil phosphorus (P) availability and different planting patterns regulate soil microbial effects on the growth of two native plant species in a semiarid steppe. METHODS: We carried out a glasshouse experiment to explore the effects of the whole indigenous soil microbiota on the growth and performance of Leymus chinensis and Cleistogenes squarrosa using soil sterilization with different soil P availabilities and planting patterns (monoculture and mixture). Transcriptome sequencing (RNA-seq) was used to explain the potential molecular mechanisms of the soil microbial effects on C. squarrosa. KEY RESULTS: The soil sterilization treatment significantly increased the biomass of L. chinensis and C. squarrosa in both monoculture and mixture conditions, which indicated that the soil microbiota had negative growth effects on both plants. The addition of P neutralized the negative microbial effects for both L. chinensis and C. squarrosa, whereas the mixture treatment amplified the negative microbial effects on L. chinensis but alleviated them on C. squarrosa. Transcriptomic analysis from C. squarrosa roots underscored that the negative soil microbial effects were induced by the upregulation of defence genes. The P addition treatment resulted in significant decreases in the number of differentially expressed genes attributable to the soil microbiota, and some defence genes were downregulated. CONCLUSIONS: Our results underline that indigenous soil microbiota have negative effects on the growth of two dominant plant species from a semiarid steppe, but their effects are highly dependent on the soil P availability and planting patterns. They also indicate that defence genes might play a key role in controlling plant growth responses to the soil microbiota.

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