Plant- and microbial-derived residues constitute the primary sources of soil organic carbon (SOC) in grassland ecosystems. However, their differential responses to chronic nitrogen (N) enrichment and the depth-dependent mechanisms governing their accumulation remain poorly characterized, particularly for water-limited grassland systems. Based on a 13-year field experiment in a semiarid grassland, we quantified the effects of long-term N addition on the accumulation of plant- (lignin phenols) and microbial-derived (amino sugars) residues. We found that N addition significantly increased lignin phenol content and its contribution to SOC in the topsoil, whereas lignin phenols exhibited a hump-shaped response peaking under moderate N levels in the subsoil. Amino sugar concentrations and their relative contribution to SOC increased in both soil layers under N addition but declined at the highest N input. The dominant factors regulating residue accumulation varied with soil depth: in the topsoil, microbial K-/r-traits and community composition primarily explained lignin phenol and amino sugar dynamics, while in the subsoil, mineral-associated protection and microbial composition were the key drivers. These findings underscore the depth-dependent nature of SOC formation pathways and highlight the importance of incorporating both plant- and microbial-derived residues into Earth System Models to improve projections of carbon-climate feedback under changing nitrogen regimes.Read the free for this article on the Journal blog.

Increased atmospheric nitrogen (N) deposition alters the structure and function of soil microbial communities in terrestrial ecosystems, consequently exerting a profound influence on ecosystem processes. However, the effects of N deposition on soil microbial network complexity and its regulation of soil carbon (C) processes in semiarid grassland ecosystems are poorly understood. In this study, based on a 13-year multilevel field N addition experiment in a semiarid grassland on the Loess Plateau, together with metagenomic sequencing and cooccurrence network analysis methods, we observed that the complexity of microbial co-occurrence network, characterized by the number of nodes and edges and the average path length, increased first and then decreased in a nonlinear response to N addition, with thresholds between 4.60 g N m- 2 yr- 1 and 9.20 g N m- 2 yr- 1 in both the topsoil and subsoil. Meanwhile, soil microbial network complexity was significantly positively correlated with plant root traits (e.g., root biomass), soil microbial properties (e.g., fungal community composition and bacterial Shannon diversity and community composition) and most physicochemical properties (e.g., soil water content, NH4+-N, and Fep). Structural equation model analysis (SEM) revealed that the major determinants of the soil microbial network complexity shifted from soil physicochemical properties to bacterial community composition along the N addition gradient. Further analysis revealed that N-induced alterations in microbial network complexity could modulate soil organic C (SOC) formation, preservation, and decomposition by affecting the functional potential of microbial communities. For instance, the microbial network complexity, abundance of functional genes involved in starch and hemicellulose degradation, and microbial C use efficiency decreased significantly under high levels of N addition. These results provide empirical evidence for the close linkages between soil microbial network complexity and soil C processes and highlight the need to disentangle the mechanisms underlying the nonlinear response of soil microbial interactions to atmospheric N deposition to improve soil C projections.

Under phosphate (Pi) limiting conditions, lipid remodeling serves as a critical mechanism for enhancing phosphorus (P) use efficiency in plants. This process also affects the photosynthetic process simultaneously, thereby influencing the accumulation of biomass. Our previous studies have proved that Zygophyllum xanthoxylum had a remarkable P remobilization capacity, and could maintain a high biomass under Pi deficiency environment. However, the specific patterns of membrane lipid remodeling and their regulatory effects on photosynthetic performance remain to be elucidated. In this study, the changes of photosynthetic parameters, chlorophyll fluorescence parameters, leaves lipid compositions, Pi content and ATPase activity of chloroplast were determined after 1D, 10D and 40D of Pi sufficient and Pi deficient treatments. We found that Pi deficiency did not cause a significant decrease in photosynthetic indices (except 40D treatment) and did not weaken the photosynthetic electron transport process. Under Pi deficiency treatment, the glyceroglycolipid content in leaves showed significant increase at 10D and 40D treatments, but the phospholipid content remained stable. The concentration of Pi and the activity of ATPase in chloroplasts at 1D and 10D treatments were significantly increased, but there was no significant difference between 40D treatment and that of the CP. The results showed that under Pi deficiency environment, Z. xanthoxylum provided structural and functional protection for electron transport process by maintaining the content stability of phospholipids and increasing the glyceroglycolipid content. In addition, more Pi was allocated to chloroplasts, enhancing ATPase activity and providing continuous and stable assimilatory power for the photosynthetic process.

Nitrogen (N) fertilization is known to impact the capacity of ecosystems to support multiple ecosystem services such as carbon sequestration and nutrient cycling, particularly in nutrient‐limited environments. Yet, little is known about how N fertilization may result in trade‐offs across contrasting soil ecosystem services. Moreover, the contribution of soil microbial networks as mediators of the impacts of fertilization on soil ecosystem services is poorly understood. Here we collected topsoil (0–10 cm) and subsoil (10–20 cm) samples from a 13‐year N addition experiment in a semiarid grassland to investigate how long‐term N additions affect soil multiservices. We found that soil multiservice predominantly exhibited a hump‐shaped response to the increasing levels of N addition across two soil depths. More importantly, changes in the complexity of soil microbial networks were positively correlated with ecosystem multiservices across the two soil depths. This relationship was especially important in explaining topsoil multiservice responses, while in subsoils, multiservices were more strongly associated with abiotic properties than network complexity. This distinction may be attributed to the lower microbial activity and reduced nutrient utilization capacity in subsoils, which allows abiotic factors to play a more dominant role on multiservices. Synthesis . Our results highlight that soil microbial network complexity is highly correlated with multiple ecosystem services in the context of global atmospheric N deposition.

Alpine grassland degradation is a major threat to global carbon cycles, yet the microbial mechanisms driving soil organic carbon (SOC) loss remain poorly understood. Ecological stoichiometry theory provides a framework for understanding how resource imbalances constrain microbial activity and metabolism. Here, we investigated how grassland degradation altered the stoichiometric imbalances between soil microbes and their resources and how microbes coped with such imbalances, as well as the implications of their responses for SOC stock. We established a degradation gradient (non-, light, moderate, and heavy) in both an alpine meadow and an alpine steppe on the Qinghai-Tibet Plateau, China, with analyzing vegetation nutrient storage, soil physicochemical properties, microbial biomass, dissolved organic nutrients, extracellular enzyme activities, and nutrient mineralization rates. Our results showed that C:N stoichiometric imbalance exhibited a hump-shaped response to grassland degradation with a maximum around moderate degradation, while C:P and N:P stoichiometric imbalances significantly decreased with increasing grassland degradation levels in both ecosystems. However, microbial responses were ecosystem-specific: meadow microbes showed strong C:N:P homeostasis, while steppe microbes showed weaker C:N and C:P homeostasis, indicating higher stoichiometric plasticity. Mechanistically, microbes coped with these shifting imbalances by adjusting extracellular enzyme stoichiometry, net N mineralization, and soil microbial respiration. For instance, C:P and N:P stoichiometric imbalances were strongly linked to the relative production of P-acquiring enzymes across both ecosystems, with slightly stronger correlations in meadows. These response mechanisms were significantly correlated with SOC stock, suggesting that microbial metabolic adjustments are a key pathway regulating the 14.8–71.5% decline in SOC stock decline observed during degradation. Our findings provide a mechanistic link between grassland degradation, microbial stoichiometric response, and carbon cycling, highlighting that ecosystem-specific microbial strategies are critical determinants of SOC vulnerability in these sensitive high-altitude ecosystems.

Soil net nitrogen mineralization (Nmin), a microbial-mediated conversion of organic to inorganic N, is critical for grassland productivity and biogeochemical cycling. Enhanced atmospheric N deposition has been shown to substantially increase both plant and soil N content, leading to a major change in Nmin. However, the mechanisms underlying microbial properties, particularly microbial functional genes, which drive the response of Nmin to elevated N deposition are still being discussed. Besides, it is still uncertain whether the relative importance of plant carbon (C) input, microbial properties, and mineral protection in regulating Nmin under continuous N addition would vary with the soil depth. Here, based on a 13-year multi-level field N addition experiment conducted in a typical grassland on the Loess Plateau, we elucidated how N-induced changes in plant C input, soil physicochemical properties, mineral properties, soil microbial community, and the soil Nmin rate (Rmin)-related functional genes drove the responses of Rmin to N addition in the topsoil and subsoil. The results showed that Rmin increased significantly in both topsoil and subsoil with increasing rates of N addition. Such a response was mainly dominated by the rate of soil nitrification. Structural equation modeling (SEM) revealed that a combination of microbial properties (functional genes and diversity) and mineral properties regulated the response of Rmin to N addition at both soil depths, thus leading to changes in the soil N availability. More importantly, the regulatory impacts of microbial and mineral properties on Rmin were depth-dependent: the influences of microbial properties weakened with soil depth, whereas the effects of mineral protection enhanced with soil depth. Collectively, these results highlight the need to incorporate the effects of differential microbial and mineral properties on Rmin at different soil depths into the Earth system models to better predict soil N cycling under further scenarios of N deposition.

Soil microbes are subject to stoichiometric imbalances, which are the dissimilarities in elemental stoichiometry between microbial biomass and resources. Shifts in dominant plant species co-occur with unparallel changes in the stoichiometry of soil microbial biomass and resources, leading to stoichiometric imbalances. However, how soil microbes deal with stoichiometric imbalances induced by changes in dominant plant species, and what the implications are for soil carbon cycling, remain unknown. Here, we compared the stoichiometric imbalances of five plant patch types with the dominant plant community (Kobresia pygmaea) in a Tibetan alpine grassland to examine how soil microbes respond physiologically to stoichiometric imbalances, thereby affecting soil microbial respiration (SMR). We found that C:N and C:P imbalances varied between plant patches and differed significantly from those in the soil of K. pygmaea. We also found that the regulation of extracellular enzyme production, SMR, potential microbial carbon use efficiency (CUE), net N mineralization, and net ammonification were essential mechanisms for soil microbes to deal with C:N imbalances. Simultaneously, soil microbes dealt with fluctuating C:P imbalances by regulating their net P mineralization and CUE. Further, structural equation modeling revealed that stoichiometric imbalances induced by changes in dominant plant species could indirectly affect SMR by regulating extracellular enzyme stoichiometry and net nutrient mineralization. These results highlight the importance of the stoichiometry of soil microbe/resource interactions in regulating metabolic activities and modifying terrestrial carbon flows in shifting plant communities.

Soil net nitrogen mineralization (Nmin), a microbial-mediated conversion of organic to inorganic N, is critical for grassland productivity and biogeochemical cycling. Enhanced atmospheric N deposition has been shown to substantially increase both plant and soil N content, leading to a major change in Nmin. However, the mechanisms underlying microbial properties, particularly microbial functional genes, which drive the response of Nmin to elevated N deposition are still being discussed. Besides, it is still uncertain whether the relative importance of plant carbon (C) input, microbial properties, and mineral protection in Nmin regulating under continuous N addition would vary with the soil depth. Here, based on a 13-year multi-level field N addition experiment conducted in a typical grassland on the Loess Plateau, we elucidated how N-induced changes in plant C input, soil physicochemical properties, mineral properties, soil microbial community, and soil net N mineralization (Rmin)-related functional genes drove the responses of Rmin to N addition in the topsoil and subsoil. The results showed that Rmin increased significantly in both topsoil and subsoil with increasing rates of N addition. Such a response was mainly dominated by the rate of soil nitrification. Structural equation modeling (SEM) revealed that a combination of microbial properties (functional genes and diversity) and mineral properties regulated the response of Rmin to N addition at both soil depths, thus leading to changes in the soil N availability. More importantly, the regulatory impacts of microbial and mineral properties on Rmin were depth-dependent: the influences of microbial properties weakened with soil depth, whereas the effects of mineral protection enhanced with soil depth. Collectively, these results highlight the need to incorporate the effects of differential microbial and mineral properties on Rmin at different soil depths into the Earth system models to better project soil N cycling under further scenarios of N deposition.

Elevated atmospheric N deposition can profoundly alter soil carbon (C) mineralization (Cmin) and nitrogen (N) mineralization (Nmin), which could severely impact long-term productivity of grassland ecosystem. However, little is known about how N addition, season, and their interaction affect soil Cmin and Nmin rates and their relationships by regulating soil abiotic and biotic factors. Here we investigated the seasonal variations in soil Cmin and Nmin rates and their relationship in response to multi-level N additions in a semiarid grassland in 2014-2015, and further identified direct and indirect pathways by which soil abiotic and biotic factors regulated these variations using structural equation modeling. We documented the statistically significant impacts of N addition and its interaction with season on soil Cmin rates. In contrast, only a significant seasonal effect on the soil Nmin rate was observed. Random forest analysis revealed that across all seasons, dissolved organic carbon (DOC), soil water content (SWC), catalase, urease, sucrase, microbial biomass carbon (MBC), soil organic carbon (SOC) to total nitrogen (TN) ratio, and TN were the most pivotal predictors of the soil Cmin rate. Comparatively, catalase, MBC, DOC, NO3--N, urease, TN, NH4+-N, SWC, and the MBC to microbial biomass nitrogen (MBN) ratio were the most dominant drivers of the soil Nmin rate. SEM results indicated that the identified potential drivers that regulated the soil Cmin and Nmin rates in response to N addition varied seasonally. Additionally, N addition decoupled the soil Cmin and Nmin rates, which was a consistent relationship among most seasons. In summary, our results show that, in this semiarid grassland, current N additions can enhance soil N immobilization across all seasons; however, its impacts on soil C sequestration were seasonally variable. These findings provide evidence that season and its interactions with elevated atmospheric N deposition have important implications for the grassland biogeochemical cycling.

Litter decomposition is a key component of global biogeochemical cycles that affects the availability of soil nutrients for plant productivity. Significant variations in litter decomposability between plant taxa are attributed to diverse functional traits including litter quality, plant nutrient production, and nutrient resorption efficiency which should influence the ecological fitness of plants in the community. However, no reports to date have explored the relationships between litter decomposition and plant community dynamics, that is, plant succession and interspecific competition. We conducted a litter decomposition experiment that focused on 21 plant species in an alpine meadow. The litter decomposition rates of these species were compared with their initial litter quality and nutrient use efficiencies to examine whether the plant litter indicators of the community structures were altered following grazing exclusion. We found that among these 21 plant species, those with higher nutrient use efficiencies had a lower litter decomposition rate. Meanwhile, lower decomposition rates were correlated with higher plant importance values, and this correlation became stronger over time in plant communities following grazing exclusion, except for dominant species. Our results suggested that litter decomposability can be used to predict the changing trajectories of plant communities following grazing exclusion, except for dominant species.

The maintenance and stability of soil structures are critical for the stability of alpine grassland ecosystems. To elucidate how soil structures are altered by the degradation of alpine grasslands, this study investigated the various characteristics of soil aggregate particle sizes, and associated interactions with soil variables and soil stability in lightly, moderately, and severely degraded alpine steppes and meadows in Tianjun County, Qinghai Province, China. The results revealed that grassland degradation culminated in the modification of soil particle sizes from macro-aggregates (2-0.25 mm) to micro aggregates (0.25-0.053 mm), to silt + clay fractions (<0.053 mm), and the stability of soil aggregates decreased. Moreover, the finer particle size grade might more clearly reflect soil disaggregation processes. Soil organic carbon (SOC), soil total nitrogen (STN), microbial biomass carbon (MBC), microbial biomass nitrogen (MBN) contents of both alpine steppes and alpine meadows significantly decreased with the worsening of grassland degradation. However, the microbial entropy carbon (qMBC) hardly changed, whereas the microbial entropy nitrogen (qMBN) of the severely degraded alpine steppes and alpine meadows were significantly higher than their counterparts in lightly and moderately degraded grasslands. Redundancy analysis (RDA) revealed that SOC, STN, MBC and MBN were positively correlated with the 2-1 mm fraction and the mean weight diameter (MWD) in both the alpine steppes and alpine meadows, but negatively correlated with the 0.25-0.1 mm fraction in the alpine steppes, and the <0.053 mm fraction in the alpine meadows. Structural equation model (SEM) results revealed that SOC and STN were the critical factors affecting the composition and stability of soil aggregates.

以白沙蒿为试验材料,在干旱(D)、UV-B辐射(U)和干旱与UV-B辐射复合(D+U)胁迫下,从幼苗生长、膜脂氧化、次生物质类黄酮、脂肪酸代谢及其基因表达等方面研究了干旱和UV-B辐射胁迫及其互作对白沙蒿抗性生理的影响.结果显示,D和U处理下,白沙蒿幼苗叶、茎、根生物量及总生物量积累减少,株高、叶面积和相对含水量(RWC)降低.D+U处理缓解了D和U处理造成的白沙蒿生物量的下降.D和U处理下,叶相对电导率(REC)显著升高;D+U处理的REC显著下降.D处理的丙二醛(MDA)含量和脂氧合酶(LOX)活性分别为对照(CK)的1.65和3.69倍,而U处理MDA含量和LOX活性无显著变化;D+U处理MDA含量和LOX活性分别为D处理的66.69％和44.00％.D和U处理类黄酮含量分别为CK的1.25和1.37倍;D+U处理类黄酮含量为D处理的1.57倍.D处理未引起不饱和脂肪酸指数(IUFA)显著变化,U处理造成IUFA显著降低,为CK的91.96％;D+U处理IUFA为U处理的1.08倍.结果表明,干旱和UV-B辐射胁迫引起的膜损伤是造成白沙蒿生物量下降的主要原因;干旱和UV-B辐射复合胁迫通过增加类黄酮含量、抑制LOX活性和提高脂肪酸不饱和度的效应产生叠加作用,缓解了彼此对白沙蒿造成的膜损伤.

Aims Grasslands are experiencing severe degradation globally, impacting aboveground vegetation and soil properties. The influences of grassland degradation on bacterial communities in soil are not well-understood. Methods The normalized difference vegetation index (NDVI) was calculated to represent grassland status and indicate grassland degradation (decreasing NDVI). Soil ph, bacterial communities, as well as nutrient and organic carbon concentrations were measured. Results Bacterial alpha diversity had negative relationships with soil moisture, soil organic carbon (SOC), total nitrogen (TN), and total phosphorus (TP). Bacterial community structure was significantly associated with NDVI, the change rate of NDVI, moisture, ph, SOC, TN, as well as soil C:N and C:P ratios. Bacterial phyla were differentially related with these environmental variables. Moreover, network analysis showed that the network of soil bacteria had strong cooperation relationships (positive correlations between taxa) and was grouped into three modules. According to modularity, 71 keystone taxa were detected as network connectors and module hubs. All the modules had close relationships with environmental variables, and many keystone taxa were negatively associated with soil moisture and SOC. Conclusions These results suggest that grassland degradation might be responsible for shifts in soil bacterial communities in terms of alpha diversity and community structure through changes in soil moisture, SOC, nutrients, and C:nutrient ratios. Moreover, in the network analyses, the strong co-occurrence relationships between taxa as well as close relationships between environmental variables and module structures and keystone taxa suggest low stability and high vulnerability of bacterial communities to the influences of grassland degradation.

Soil phosphate (Pi) deficiency is a global issue and a major constraint on plant growth. Plants typically acclimatize to low Pi by enhancing their P utilization and/or P acquisition efficiencies; however, different species have variable preferred strategies. RNA sequencing analysis was performed on the shoots and roots of Zygophyllum xanthoxylum, under 1 day and 10 days of Pi stress, to investigate their adaptation strategies to P deprivation. A total of 364,614 unigenes and 9,270 differentially expressed genes (DEGs) were obtained via transcriptome sequencing. An analysis of the DEGs revealed that under the 10D treatment, anthocyanin synthesis genes were upregulated under Pi stress, whereas gibberellin, ethylene, and cytokinins synthesis genes were upregulated, and abscisic acid synthesis genes were downregulated. Genes related to organic acid synthesis, encoding for purple acid phosphatases (APase) and nucleases (RNase) were upregulated under the 1D and 10D treatments, respectively. Furthermore, genes associated with Pi transport were induced by Pi stress. Zygophyllum xanthoxylum has special P adaptation strategies, the variation trends of genes involved in external P mobilization and acquisition, which were different from that of most other species; however, the expression levels of organophosphorus mobilization related genes, such as APases and RNases, were significantly increased. Meanwhile, PHT2s and TPTs, which distributed Pi to effective sites (e.g., chloroplast), played critical roles in the maintenance of photosynthesis. We speculated that these were economic and energy saving strategies, and there are critical adaptive mechanisms that Z. xanthoxylum employs to cope with deficits in Pi.

Background The allometric relationships of plants and their changes assist with elucidating the adaptive responses of plants to the environment. However, it remains unclear whether different species of the life-form ‘shrubs’ have consistent or similar allometric relationships between modular characteristics (including morphological characteristics and biomass allocations). Here, we selected eight xerophytic shrubs as samples to investigate the morphological characteristics, biomass allocations and their allometric relationships. Results The results showed that there were common allometric scaling exponents ( α RMA ) between modular characteristics with the exception of crown area ( C )-belowground biomass ( BGB ) and C -plant height ( H ). Moreover, The BGB- total biomass ( TGB ) of the eight species accorded with the significant isometric relationships, and the isometric or allometric relationships of different species in aboveground biomass ( AGB )- BGB and AGB - TGB were similar, which meant that the belowground biomass mainly determined the total biomass for xerophytic shrubs. Conclusions Our results suggested that there were similar trends of collaborative changes between modular characteristics of eight xerophytic shrubs with the exception of C - BGB and C - H , which reflected the convergent adaptation of different species in the life-form ‘shrubs’ to arid environments.

AbstractFlavonoids are a group of phenolic secondary metabolites in plants that have important physiological, ecological and economic value. In this study, using the desert plant Artemisia sphaerocephala Krasch. as the sample material, the content and components of the total flavonoids in its seeds at seven different developmental stages were determined. In addition, the genes involved in flavonoid metabolism were identified by full-length transcriptome sequencing (third-generation sequencing technology based on PacBio RS II). Their expression levels were analysed by RNA-seq short reading sequencing, to reveal the patterns and regulation mechanisms of flavonoid accumulation during seed development. The key results were as follows: the content of total flavonoids in mature seeds was 15.05 mg g−1, including five subclasses: flavonols, chalcones, flavones, flavanones and proanthocyanidins, among which flavonols accounted for 45.78%. The period of rapid accumulation of flavonoids was 40–70 d following anthesis. The high expression of phenylalanine ammonia-lyase (PAL), 4-coumarate-CoA ligase (4CL) and UDP-glucose:flavonoids 3-o-glucosyltransferase (UF3GT) promoted the accumulation of total flavonoids, while the high expression of flavonoids 3′-hydroxylase (F3′H) and flavonols synthase (FLS) made flavanols the main component. Transcription factors such as the MYB-bHLH-WDR (MBW) complex and Selenium-binding protein (SBP) directly regulated the structural genes of flavonoid metabolism, while C2H2-type zinc finger (C2H2), Zinc-finger transcription factor (GATA), Dehydration-responsive element binding (DREB), Global Transcription factor Group E protein (GTE), Trihelix DNA-binding factors (Trihelix) and Phytochrome-interacting factor (PIF) indirectly promoted the synthesis of flavonoids through hormones such as brassinoidsteroids (BRs) and abscisic acid (ABA). These results provided valuable resources for the application of related genes in genetics and breeding.

Artemisia sphaerocephala seeds are rich in polysaccharides and linoleic acid (C18:2), which have been widely used as traditional medicine and to improve food quality. The accumulation patterns and molecular regulatory mechanisms of polysaccharides during A. sphaerocephala seed development have been studied. However, the related research on seed oil and C18:2 remain unclear. For this study, A. sphaerocephala seeds at seven different development stages at 10, 20, 30, 40, 50, 60, and 70 days after flowering (designated as S1~S7), respectively, were employed as experimental samples, the accumulation patterns of oil and fatty acids (FA) and the underlying molecular regulatory mechanisms were analyzed. The results revealed that oil content increased from 10.1% to 20.0% in the early stages of seed development (S1~S2), and up to 32.0% in mature seeds, of which C18:2 accounted for 80.6% of the total FA. FA and triacylglycerol biosynthesis-related genes jointly involved in the rapid accumulation of oil in S1~S2. Weighted gene co-expression network analysis showed that transcription factors FUS3 and bHLH played a critical role in the seed oil biosynthesis. The perfect harmonization of the high expression of FAD2 with the extremely low expression of FAD3 regulated the accumulation of C18:2. This study uncovered the gene involved in oil biosynthesis and molecular regulatory mechanisms of high C18:2 accumulation in A. sphaerocephala seeds; thus, advancing research into unsaturated fatty acid metabolism in plants while generating valuable genetic resources for optimal C18:2 breeding.

Degradation of alpine meadows on the Qinghai-Tibetan Plateau is an important issue for ecological science, policy making, and the welfare of local herders. Destruction of alpine meadows results from degeneration of vegetation and soil systems and from the mechanical decoupling of the environment, grassland, livestock, and herders and, subsequently, discordance among these subsystems. In this study, systematic integration of restoration techniques based on the grassland agroecosystems coupling theory was developed for the management and restoration of degraded alpine meadows. To test the effectiveness of these integrated restoration techniques, we conducted restoration trials that included grazing management, enclosed, fertilization, overseeding, and sward ripping by evaluating the ecosystem coupling of soil, plant and livestock, and ecosystem functions. The results of this study suggest that comprehensive restoration practices include grazing and agronomy techniques (fertilizer, overseeding, and sward ripping) that result in the greatest level of ecosystem coupling, while the single restoration practice leads to poorly coupled ecosystems. Restoration practice changes in ecosystem functionality are positively related to changes in ecosystem coupling. Our results highlight the importance of diversified restoration practices for facilitating ecological coupling and functioning in the degraded alpine meadow. The restorative scheme also bridges the gap between restoration theory and practice by providing guidelines for herders and policy makers for the urgent task of restoring degraded alpine meadows.