Effects of Enterobacter cloacae insecticidal protein on the Duox-ROS system and midgut bacterial community and function of Galleria mellonella larvae.

BACKGROUND: Enterobacter cloacae insecticidal proteins have been reported to kill Galleria mellonella larvae through affecting their midgut microbiome. However, the mechanisms involved remain unclear. Here we aim to investigate how the insecticidal proteins act on the midgut Duox-ROS system and microbial community of G. mellonella larvae. METHODS: Reverse transcription qPCR and fluorescence probes were utilized to assess the Duox expression levels and to evaluate quantitative changes of the ROS levels. Sequencing of the 16S rRNA gene sequences of the midgut bacteria of G. mellonella larvae was conducted for further analyses of bacterial diversity, composition, and abundance. RESULTS: After the injection of the insecticidal proteins, the Duox expression levels first increased within 28 h, then dramatically peaked at 36 h, and slowly decreased thereafter. Simultaneously, the ROS levels increased significantly at 36 h, peaked at 48 h, and rapidly declined to the normal level at 60 h. Responsive to the change of the ROS levels, the structure of the midgut microbial community was altered substantially, compared to that of the untreated larvae. The relative abundance of Enterobacteriaceae and other specific pathogenic bacteria increased significantly, whereas that of Lactobacillus decreased sharply. Importantly, notable shifts were observed in the crucial midgut predicted metabolic functions, including membrane transportation, carbohydrate metabolism, and amino acid metabolism. CONCLUSION: Insecticidal proteins of E. cloacae kill G. mellonella larvae mainly through generation of high oxidative stress, alterations of the midgut microbial community and function, and damage to the physiological functions. These findings provide insights into the inhibition mechanism of E. cloacae insecticidal proteins to G. mellonella larvae.

Warming-induced shifts in alpine soil microbiome: An ecosystem-scale study with environmental context-dependent insights.

Climate warming is a pressing global issue with substantial impacts on soil health and function. However, the influence of environmental context on the responses of soil microorganisms to warming remains largely elusive, particularly in alpine ecosystems. This study examined the responses of the soil microbiome to in situ experimental warming across three elevations (3850 m, 4100 m, and 4250 m) in the meadow of Gongga Mountain, eastern Tibetan Plateau. Our findings demonstrate that soil microbial diversity is highly resilient to warming, with significant impacts observed only at specific elevations. Furthermore, the influence of warming on the composition of the soil microbial community is also elevation-dependent, underscoring the importance of local environmental context in shaping microbial evolution in alpine soils under climate warming. Notably, we identified soil moisture at 3850 m and carbon-to-nitrogen ratio at 4250 m as indirect predictors regulating the responses of microbial diversity to warming at specific elevations. These findings underscore the paramount importance of considering pre-existing environmental conditions in predicting the response of alpine soil microbiomes to climate warming. Our study provides novel insights into the intricate interactions between climate warming, soil microbiome, and environmental context in alpine ecosystems, illuminating the complex mechanisms governing soil microbial ecology in these fragile and sensitive environments.

Plant water source effects on plant-soil feedback for primary succession of terrestrial ecosystems in a glacier region in China.

Despite the extensive research conducted on plant-soil-water interactions, the understanding of the role of plant water sources in different plant successional stages remains limited. In this study, we employed a combination of water isotopes (δ2H and δ18O) and leaf δ13C to investigate water use patterns and leaf water use efficiency (WUE) during the growing season (May to September 2021) in Hailuogou glacier forefronts in China. Our findings revealed that surface soil water and soil nutrient gradually increased during primary succession. Dominant plant species exhibited a preference for upper soil water uptake during the peak leaf out period (June to August), while they relied more on lower soil water sources during the post-leaf out period (May) or senescence (September to October). Furthermore, plants in late successional stages showed higher rates of water uptake from uppermost soil layers. Notably, there was a significant positive correlation between the percentage of water uptake by plants and available soil water content in middle and late stages. Additionally, our results indicated a gradual decrease in WUE with progression through succession, with shallow soil moisture utilization negatively impacting overall WUE across all succession stages. Path analysis further highlighted that surface soil moisture (0- 20 cm) and middle layer nutrient availability (20- 50 cm) played crucial roles in determining WUE. Overall, this research emphasizes the critical influence of water source selection on plant succession dynamics while elucidating underlying mechanisms linking succession with plant water consumption.

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

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

Nitrogen dynamics of alpine swamp meadows are less responsive to climate warming than that of alpine meadows.

The freeze-thaw cycle mediates permafrost soil hydrothermal status, nitrogen (N) mineralization, and loss. Furthermore, it affects root development and competition among nitrophilic and other species, shaping the pattern of N distribution in alpine ecosystems. However, the specific N dynamics during the growing season and N loss during the non-growing season in response to climate warming under low- and high-moisture conditions are not well documented. Therefore, we added 15N tracers to trace the fate of N in warmed and ambient alpine meadows and alpine swamp meadows in the permafrost region of the Qinghai-Tibet Plateau. During the growing season, warming increased 15N recovery (15Nrec) in shoots of K. humilis, litters, 0-5 and 5-20 cm roots in the alpine meadow by 149.94 % ± 52.87 %, 114.58 % ± 24.43 %, 61.11 % ± 32.27 %, and 97.12 % ± 42.92 %, respectively, while increased 15Nrec of litters by 151.55 % ± 27.06 % in the alpine swamp meadow. During the non-growing season, warming reduced 15N stored in roots by 486.77 % ± 57.90 %, though increased the 15N recovery in 5-20 cm soil depth by 76.68 % ± 39.42 % in the alpine meadow, whereas it did not affect N loss during the non-growing season in the alpine swamp meadow. Overall, warming promoted N utilization by increasing the plant N pool during the growing season, and enhanced root N loss and downward migration during the non-growing season due to the freeze-thaw process, which may result in fine root turnover and cell destruction releasing N in the alpine meadow. Conversely, the N dynamics of alpine swamp meadows were less responsive to climate warming.

Soil development following glacier retreat shapes metagenomic and metabolomic functioning associated with asynchronous C and N accumulation.

Glacier retreat caused by global warming may result in the variation of soil organic carbon and nutrient cycling. Yet, the dynamic change of soil microbial functional profiles, especially C metabolism-related, with soil development following glacier retreat are still unclear. In the present study, we investigated the soil microbial communities, metagenomic functioning, and metabolomic profiles along the Hailuogou Glacier forefield representing a 120-year chronosequence. The alpha diversity indices of soil bacteria, protozoa and nifH genes showed an upward trend with increased soil ages, and the beta diversity of soil archaea, bacteria, fungi, protozoa, nifH and nirS genes were significantly correlated with soil ages, in which increasing soil C and P while decreased C/N and pH significantly contributed to the differences of soil microbial communities among the analyzed environmental variables. The metagenomic functional genes related to the metabolisms of Glycogen and Cellulosome, Iron Acquisition and Metabolism were significantly decreased with chronosequence, while the utilization of Xylose and Lactate, Potassium Metabolism, Sulfur Metabolism showing an upward trend with soil ages, in which soil C/N ratios and pH were the most influential factors. In addition, soil C and C/N ratios were also significantly correlated to metabolomic compositions, in which the complexity of the metabolite structure increased with soil ages. Our results indicate that glacier retreat may lead to the asynchronous C and N accumulation along the chronosequence, thereby affecting the metagenomic and metabolomic functioning of soil microbial communities related to C metabolisms during soil development following glacier retreat.

Suppression of the human malic enzyme 2 modifies energy metabolism and inhibits cellular respiration.

Human mitochondrial NAD(P)+-dependent malic enzyme (ME2) is well-known for its role in cell metabolism, which may be involved in cancer or epilepsy. We present potent ME2 inhibitors based on cyro-EM structures that target ME2 enzyme activity. Two structures of ME2-inhibitor complexes demonstrate that 5,5'-Methylenedisalicylic acid (MDSA) and embonic acid (EA) bind allosterically to ME2's fumarate-binding site. Mutagenesis studies demonstrate that Asn35 and the Gln64-Tyr562 network are required for both inhibitors' binding. ME2 overexpression increases pyruvate and NADH production while decreasing the cell's NAD+/NADH ratio; however, ME2 knockdown has the opposite effect. MDSA and EA inhibit pyruvate synthesis and thus increase the NAD+/NADH ratio, implying that these two inhibitors interfere with metabolic changes by inhibiting cellular ME2 activity. ME2 silence or inhibiting ME2 activity with MDSA or EA decreases cellular respiration and ATP synthesis. Our findings suggest that ME2 is crucial for mitochondrial pyruvate and energy metabolism, as well as cellular respiration, and that ME2 inhibitors could be useful in the treatment of cancer or other diseases that involve these processes.

Phyllosphere antibiotic resistome in a natural primary vegetation across a successional sequence after glacier retreat.

The spread of antibiotic-resistance genes (ARGs) has posed a significant threat to human health over the past decades. Despite the fact that the phyllosphere represents a crucial pool of microorganisms, little is known about the profile and drivers of ARGs in less human interference natural habitats. In order to minimize the influence of environmental factors, here we collected leaf samples from the early-, middle- and late-successional stages across a primary vegetation successional sequence within 2 km, to investigate how the phyllosphere ARGs develop in natural habitats. Phyllosphere ARGs were determined using high-throughput quantitative PCR. Bacterial community and leaf nutrient content were also measured to assess their contribution to the phyllosphere ARGs. A total of 151 unique ARGs were identified, covering almost all recognized major antibiotic classes. We further found that there was some stochastic and a core set of the phyllosphere ARGs during the plant community succession process, due to the fluctuant phyllosphere habitat and specific selection effect of plant individuals. The ARG abundance significantly decreased due to the reduction of the phyllosphere bacterial diversity, community complexity, and leaf nutrient content during the plant community succession process. While the closer links between soil and fallen leaf resulted in a higher ARG abundance in leaf litter than in fresh leaf. In summary, our study reveals that the phyllosphere harbors a broad spectrum of ARGs in the natural environment. These phyllosphere ARGs are driven by various environmental factors, including the plant community composition, host leaf properties, and the phyllosphere microbiome.

Niche-mediated bacterial community composition in continental glacier alluvial valleys under cold and arid environments.

Introduction: Bacteria are an essential component of glacier-fed ecosystems and play a dominant role in driving elemental cycling in the hydrosphere and pedosphere. However, studies of bacterial community composition mechanisms and their potential ecological functions from the alluvial valley of mountain glaciers are extremely scarce under cold and arid environments. Methods: Here, we analyzed the effects of major physicochemical parameters related to soil on the bacterial community compositions in an alluvial valley of the Laohugou Glacier No. 12 from the perspective of core, other, and unique taxa and explored their functional composition characteristics. Results and discussion: The different characteristics of core, other, and unique taxa highlighted the conservation and difference in bacterial community composition. The bacterial community structure of the glacial alluvial valley was mainly affected by the above sea level, soil organic carbon, and water holding capacity. In addition, the most common and active carbon metabolic pathways and their spatial distribution patterns along the glacial alluvial valley were revealed by FAPTOTAX. Collectively, this study provides new insights into the comprehensive assessment of glacier-fed ecosystems in glacial meltwater ceasing or glacier disappearance.

Livestock grazing-exclusion under global warming scenario decreases phosphorus mineralization by changing soil food web structure in a Tibetan alpine meadow.

The exclusion of grazing has been used extensively in alpine meadows on the Tibetan Plateau. Studies, however, have shown reported recent trends of decreasing concentrations of soil nutrients because of grazing exclusion and climate change. The effects of excluding grazing on the soil biogeochemical process of phosphorus cycling in alpine meadows are unclear, especially under climatic warming. We conducted a 5-year grazing-exclusion and warming-manipulation experiment to examine the effects of excluding grazing on fractions of soil phosphorus, microbial and nematode communities and enzymatic activities in treatments of low grazing intensity, grazing exclusion, and combined grazing exclusion and warming. Our results indicated that excluding grazing significantly decreased bacterivore and omnivore-predator densities, phoD gene abundance and alkaline phosphomonoesterase activity (in the 0-5 cm layer by -34, -41, -38 and -42 %) at altitudes of 3850 m, 4000 m, 4150 m and 4250 m, respectively. Structural equation modeling indicated that bacterivores positively affected phoD gene abundance, alkaline phosphomonoesterase activity and inorganic­phosphorus fractions. Combined grazing exclusion and warming significantly decreased bacterivore and omnivore-predator densities but significantly increased fungivore density (in the 0-5 cm layer by 238, 172, 119 and 65 %) at altitudes of 3850, 4000, 4150 and 4250 m, respectively. Structural equation modeling also indicated that the combined grazing-exclusion and warming treatment increased the soil fungi and fungivores, but the higher abundances of fungi and fungivores did not significantly affect acid phosphomonoesterase activity or inorganic­phosphorus fractions. Alternatively, the combined grazing-exclusion and warming treatment significantly increased the concentrations of amorphous and free aluminum, which were positively correlated with the maximum adsorption of phosphorus. The combined grazing-exclusion and warming treatment thus significantly decreased the availability of resin phosphorus (-63, -51, -81 and -67 %) in the 0-5 cm layer at altitudes of 3850, 4000, 4150 and 4250 m, respectively. Our results suggested that light grazing (0.5 yak ha-1 year-1) could increase phosphorus mineralization and the activity of soil enzymes in alpine meadows under global warming. An adequate load of livestock pressure at each altitude can be an effective management technique, mainly under warming, to maintain an adequate, sustainable and equilibrated phosphorus cycle in the plant-soil system.

Effects of revegetation and precipitation gradient on soil carbon and nitrogen variations in deep profiles on the Loess Plateau of China.

Precipitation is one of the most important factors affecting the variations in soil carbon (C) and nitrogen (N) following revegetation. However, the effects of revegetation and precipitation gradients on soil organic carbon (SOC), total nitrogen (TN) and C-N interactions in deep profiles over large scales are poorly understood. This study measured the SOC and TN stocks to depth of 300 cm in three revegetation types (grassland, shrubland and forestland) and paired cropland stands at seven sites along a precipitation gradient with mean annual precipitation (MAP) from 280 to 540 mm yr-1 in the Loess Plateau of China. The results showed that the SOC and TN stocks in the 0-300 cm profile increased along the precipitation gradient. Revegetation did not always result in accumulation of SOC and TN stocks, which depended on the precipitation condition and varied among different vegetation types. Grassland restoration resulted in more SOC and TN accumulation than shrubland and forestland in areas with MAP < 510 mm, whereas there were losses in SOC and TN following grass plantation in sites with MAP > 510 mm. The changes in SOC and TN stocks following revegetation (∆SOC and ∆TN) were significantly correlated with MAP in only the 0-20 cm layer, whereas the changes in the C/N ratio of each depth were significantly and negatively correlated with MAP. The correlations between ∆SOC and ∆TN were stronger in the 0-60 cm layer than that in the 60-300 cm layer, and an accumulation of 1 g TN was associated with approximately 7.9 g increase of SOC in the 0-300 cm profile following revegetation. This study indicated that the changes in soil C and N stocks following revegetation had different patterns along precipitation gradient and among depths, and grassland restoration and N fertilizer input benefitted soil C and N sequestration in drier areas.

Forest biomass carbon stocks and variation in Tibet's carbon-dense forests from 2001 to 2050.

Tibet's forests, in contrast to China's other forests, are characterized by primary forests, high carbon (C) density and less anthropogenic disturbance, and they function as an important carbon pool in China. Using the biomass C density data from 413 forest inventory sites and a spatial forest age map, we developed an allometric equation for the forest biomass C density and forest age to assess the spatial biomass C stocks and variation in Tibet's forests from 2001 to 2050. The results indicated that the forest biomass C stock would increase from 831.1 Tg C in 2001 to 969.4 Tg C in 2050, with a net C gain of 3.6 Tg C yr-1 between 2001 and 2010 and a decrease of 1.9 Tg C yr-1 between 2040 and 2050. Carbon tends to allocate more in the roots of fir forests and less in the roots of spruce and pine forests with increasing stand age. The increase of the biomass carbon pool does not promote significant augmentation of the soil carbon pool. Our findings suggest that Tibet's mature forests will remain a persistent C sink until 2050. However, afforestation or reforestation, especially with the larger carbon sink potential forest types, such as fir and spruce, should be carried out to maintain the high C sink capacity.

Control factors and scale analysis of annual river water, sediments and carbon transport in China.

Under the context of dramatic human disturbances on river system, the processes that control the transport of water, sediment, and carbon from river basins to coastal seas are not completely understood. Here we performed a quantitative synthesis for 121 sites across China to find control factors of annual river exports (Rc: runoff coefficient; TSSC: total suspended sediment concentration; TSSL: total suspended sediment loads; TOCL: total organic carbon loads) at different spatial scales. The results indicated that human activities such as dam construction and vegetation restoration might have a greater influence than climate on the transport of river sediment and carbon, although climate was a major driver of Rc. Multiple spatial scale analyses indicated that Rc increased from the small to medium scale by 20% and then decreased at the sizable scale by 20%. TSSC decreased from the small to sizeable scale but increase from the sizeable to large scales; however, TSSL significantly decreased from small (768 g·m(-2)·a(-1)) to medium spatial scale basins (258 g·m(-2)·a(-1)), and TOCL decreased from the medium to large scale. Our results will improve the understanding of water, sediment and carbon transport processes and contribute better water and land resources management strategies from different spatial scales.

Soil carbon and nitrogen changes following afforestation of marginal cropland across a precipitation gradient in Loess Plateau of China.

Cropland afforestation has been widely found to increase soil organic carbon (SOC) and soil total nitrogen (STN); however, the magnitudes of SOC and STN accumulation and regulating factors are less studied in dry, marginal lands, and therein the interaction between soil carbon and nitrogen is not well understood. We examined the changes in SOC and STN in younger (5-9-year-old) and older (25-30-year-old) black locust (Robinia pseudoacacia L., an N-fixing species) plantations that were established on former cropland along a precipitation gradient (380 to 650 mm) in the semi-arid Loess Plateau of China. The SOC and STN stocks of cropland and plantations increased linearly with precipitation increase, respectively, accompanying an increase in the plantation net primary productivity and the soil clay content along the increasing precipitation gradient. The SOC stock of cropland decreased in younger plantations and increased in older plantations after afforestation, and the amount of the initial loss of SOC during the younger plantations' establishment increased with precipitation increasing. By contrast, the STN stock of cropland showed no decrease in the initial afforestation while tending to increase with plantation age, and the changes in STN were not related to precipitation. The changes in STN and SOC showed correlated and were precipitation-dependent following afforestation, displaying a higher relative gain of SOC to STN as precipitation decreased. Our results suggest that the afforestation of marginal cropland in Loess Plateau can have a significant effect on the accumulation of SOC and STN, and that precipitation has a significant effect on SOC accumulation but little effect on STN retention. The limitation effect of soil nitrogen on soil carbon accumulation is more limited in the drier area rather than in the wetter sites.

A policy-driven large scale ecological restoration: quantifying ecosystem services changes in the Loess Plateau of China.

As one of the key tools for regulating human-ecosystem relations, environmental conservation policies can promote ecological rehabilitation across a variety of spatiotemporal scales. However, quantifying the ecological effects of such policies at the regional level is difficult. A case study was conducted at the regional level in the ecologically vulnerable region of the Loess Plateau, China, through the use of several methods including the Universal Soil Loss Equation (USLE), hydrological modeling and multivariate analysis. An assessment of the changes over the period of 2000-2008 in four key ecosystem services was undertaken to determine the effects of the Chinese government's ecological rehabilitation initiatives implemented in 1999. These ecosystem services included water regulation, soil conservation, carbon sequestration and grain production. Significant conversions of farmland to woodland and grassland were found to have resulted in enhanced soil conservation and carbon sequestration, but decreased regional water yield under a warming and drying climate trend. The total grain production increased in spite of a significant decline in farmland acreage. These trends have been attributed to the strong socioeconomic incentives embedded in the ecological rehabilitation policy. Although some positive policy results have been achieved over the last decade, large uncertainty remains regarding long-term policy effects on the sustainability of ecological rehabilitation performance and ecosystem service enhancement. To reduce such uncertainty, this study calls for an adaptive management approach to regional ecological rehabilitation policy to be adopted, with a focus on the dynamic interactions between people and their environments in a changing world.

Soil carbon sequestration potential for "Grain for Green" project in Loess Plateau, China.

Conversion of cropland into perennial vegetation land can increase soil organic carbon (SOC) accumulation, which might be an important mitigation measure to sequester carbon dioxide from the atmosphere. The "Grain for Green" project, one of the most ambitious ecological programmes launched in modern China, aims at transforming the low-yield slope cropland into grassland and woodland. The Loess Plateau in China is the most important target of this project due to its serious soil erosion. The objectives of this study are to answer three questions: (1) what is the rate of the SOC accumulation for this "Grain for Green" project in Loess Plateau? (2) Is there a difference in SOC sequestration among different restoration types, including grassland, shrub and forest? (3) Is the effect of restoration types on SOC accumulation different among northern, middle and southern regions of the Loess Plateau? Based on analysis of the data collected from the literature conducted in the Loess Plateau, we found that SOC increased at a rate of 0.712 TgC/year in the top 20 cm soil layer for 60 years under this project across the entire Loess Plateau. This was a relatively reliable estimation based on current data, although there were some uncertainties. Compared to grassland, forest had a significantly greater effect on SOC accumulation in middle and southern Loess Plateau but had a weaker effect in the northern Loess Plateau. There were no differences found in SOC sequestration between shrub and grassland across the entire Loess Plateau. Grassland had a stronger effect on SOC sequestration in the northern Loess Plateau than in the middle and southern regions. In contrast, forest could increase more SOC in the middle and southern Loess Plateau than in the northern Loess Plateau, whereas shrub had a similar effect on SOC sequestration across the Loess Plateau. Our results suggest that the "Grain for Green" project can significantly increase the SOC storage in Loess Plateau, and it is recommended to expand grassland and shrub areas in the northern Loess Plateau and forest in the middle and southern Loess Plateau to enhance the SOC sequestration in this area.

[Variation of below-ground carbon sequestration under long term cultivation and grazing in the typical steppe of Nei Monggol in North China].

Human activities have changed the earth surface mostly, which caused many environment issues now. We aimed to detect the process that human activities exert on ecosystem by investigating and analyzing the changes of plant community, especially underground soil and root carbon sequestration under long-term cultivation and grazing in typical steppe area of Nei Monggol, North China. The carbon sequestration on the root and soil in three plots of typical steppe area: 26-year exclosure grassland (E26), continuous grazing grassland (LG) and 35-year cultivated tillage (LC) were measured. The carbon storage in the layer of 0-40 cm showed a trend that E26 (7 307.59 g x m(-2) and 950.32 g x m(-2)) approximately LG (7834.01 g x m(-2) and 843.43 g x m(-2)) > LC (4537.04 g x m(-2) and 277.35 g x m(-2)), occupied 88.49%, 90.28% and 94.24% of total soil-root carbon respectively. The original composition structure of plant-soil system was completely destroyed by human cultivation, and it also led to sever soil erosion as well. The sand content in soil of LC at layers 0-10 cm, 10-20 cm were increasing by 81% and 39% compared to E26. On the other hand, the root biomass of LG at 0-40 cm decreased by 71%. Cultivation resulted in significant decrease of the carbon storages in soil and root. Therefore, the present cultivation should be ceased and the optimum measurements should be taken to make the tillage restore to natural grassland condition. Continuous grazing led to the significantly changes of the above-ground vegetation characteristics such as community height, coverage and biomass. While the changes of soil organic content and root biomass was not remarkable under grazing. However, the bulk density of surface soil (0-10 cm) exhibited significant increase in LG compared to E26, which indicated that the present grazing pressure have been reaching the threshold of grassland capacity. Therefore, the present grazing pressure should be decreased properly in order to avoid more serious degradation.