Enumeration of Methanotrophic Bacteria in the Cover Soil of an Aged Municipal Landfill

The enumeration of methanotrophic bacteria in the cover soil of an aged municipal landfill was carried out using (1) fluorescent in situ hybridization (FISH) with horseradish peroxidase-labeled oligonucleotide probes and tyramide signal amplification, also known as catalyzed reporter deposition-FISH (CARD-FISH), and (2) most probable number (MPN) method. The number of methanotrophs was determined in cover soil samples collected during April–November 2003 from a point with low CH₄ emission. The number of types I and II methanotrophs obtained by CARD-FISH varied from 15 ± 2 to 56 ± 7 × 10⁸ cells g⁻¹ absolute dry mass (adm) of soil and methanotrophs of type I dominated over type II. The average number of methanotrophs throughout the cover soil profile was highest during May–September when the cover soil temperature was above 13°C. Methanotrophs accounted for about 50% of the total bacterial population in the deepest cover soil layer owing to higher availability of substrate (CH₄). A lower number of methanotrophs (7 × 10² to 17 × 10⁵ cells g⁻¹ adm of soil) was determined by the MPN method compared to the CARD-FISH counts, thus confirming previous results that the MPN method is limited to the estimation of the culturable species that can be grown under the incubation conditions used. The number of culturable methanotrophs correlated with the methane-oxidizing activity measured in laboratory assays. In comparison to the incubation-based measurements, the number of methanotrophs determined by CARD-FISH better reflected the actual characteristics of the environment, such as release and uptake of CH₄, temperature, and moisture, and availability of substrates.     

Revealing the community and metabolic potential of active methanotrophs by targeted metagenomics in the Zoige wetland of the Tibetan Plateau

The Zoige wetland of the Tibetan Plateau is one of the largest alpine wetlands in the world and a major emission source of methane. Methane oxidation by methanotrophs can counteract the global warming effect of methane released in the wetlands. Understanding methanotroph activity, diversity and metabolism at the molecular level can guide the isolation of the uncultured microorganisms and inform strategy-making decisions and policies to counteract global warming in this unique ecosystem. Here we applied DNA stable isotope probing using ¹³C-labelled methane to label the genomes of active methanotrophs, examine the methane oxidation potential and recover metagenome-assembled genomes (MAGs) of active methanotrophs. We found that gammaproteobacteria of type I methanotrophs are responsible for methane oxidation in the wetland. We recovered two phylogenetically novel methanotroph MAGs distantly related to extant Methylobacter and Methylovulum. They belong to type I methanotrophs of gammaproteobacteria, contain both mxaF and xoxF types of methanol dehydrogenase coding genes, and participate in methane oxidation via H₄MPT and RuMP pathways. Overall, the community structure of active methanotrophs and their methanotrophic pathways revealed by DNA-SIP metagenomics and retrieved methanotroph MAGs highlight the importance of methanotrophs in suppressing methane emission in the wetland under the scenario of global warming.     

Grazing intensifies degradation of a Tibetan Plateau alpine meadow through plant–pest interaction

Understanding the plant–pest interaction under warming with grazing conditions is critical to predict the response of alpine meadow to future climate change. We investigated the effects of experimental warming and grazing on the interaction between plants and the grassland caterpillar Gynaephora menyuanensis in an alpine meadow on the Tibetan Plateau in 2010 and 2011. Our results showed that grazing significantly increased nitrogen concentration in graminoids and sward openness with a lower sward height, sward coverage, and plant litter mass in the community. Grazing significantly increased G. menyuanensis body size and potential fecundity in 2010. The increases in female body size were about twofold greater than in males. In addition, grazing significantly increased G. menyuanensis density and its negative effects on aboveground biomass and graminoid coverage in 2011. We found that G. menyuanensis body size was significantly positively correlated with nitrogen concentration in graminoids but negatively correlated with plant litter mass. Even though warming did not significantly increased G. menyuanensis performance and the negative effects of G. menyuanensis on alpine meadow, the increases in G. menyuanensis growth rate and its negative effect on aboveground biomass under the warming with grazing treatment were significantly higher than those under the no warming with grazing treatment. The positive effects of grazing on G. menyuanensis performance and its damage were exacerbated by the warming treatment. Our results suggest that the fitness of G. menyuanensis would increase under future warming with grazing conditions, thereby posing a greater risk to alpine meadow and livestock production.     

青藏高原不同草地利用方式对土壤粒径分形特征的影响

研究青藏高原草地土壤粒径结构分形特征,为该地区土壤质量评价和生态恢复提供科学依据。以青藏高原4种高寒草地(放牧、围栏禁牧、围栏禁牧+补植、未干扰)为对象,采用分形理论,研究不同利用方式对高寒草地土壤颗粒组成及分形特征的影响,明确土壤粒径分形特征的影响因素。结果表明：与放牧和围栏禁牧+补植相比,围栏禁牧草地中黏粒和粉粒体积分数分别增加了60%—91.1%、43.5%—80.1%,禁牧能够促进土壤砂粒向黏粒和粉粒转变。不同草地利用方式对分形维数有显著影响,单重分形维数D值依次为放牧草地<围栏禁牧+补植草地<未干扰草地=围栏禁牧草地,多重分形维数,包括信息维数D₁、信息维数/容量维数比值D₁/D₀和关联维数D₂依次为放牧草地<围栏禁牧+补植草地<围栏禁牧草地<未干扰草地。单重分形维数D与土壤黏粒、粉粒呈极显著正相关(P<0.01);砂粒、黏粒、粉粒、有机碳和全氮是多重分形维数的限制因素。信息维数D₁、信息维数/容量维数比值D₁...     

Moderate grazing increased alpine meadow soils bacterial abundance and diversity index on the Tibetan Plateau

The response of grassland soil bacterial community characteristics to different grazing intensities is central ecological topics. However, the underlying mechanisms between bacterial abundance, diversity index, and grazing intensity remain unclear. We measured alpine meadow soil bacterial gene richness and diversity index under four grazing intensities using 16S rDNA sequence analysis on the Tibetan Plateau. The results suggest that extreme grazing significantly decreased alpine meadow both bacterial gene abundance and diversity index (p < .05). The lowest operational taxonomic unit numbers were 3,012 ± 447 copies under heavy grazing in the growing season. It was significantly lower than heavy grazing with approximately 3,958 ± 119 copies (p < .05). The Shannon index for medium and high grazing grassland bacterial diversity was slightly higher than for light grazing in the growing season. Furthermore, the lowest index was approximately 9.20 ± 0.50 for extreme grazing of grassland in the growing season. The average bacterial gene abundance and diversity index in the dormancy period were slightly higher than that in the growing season. Soil bulk density, pH, ammonium, and nitrate nitrogen were the main positive factors driving grazed grassland bacterial communities. Our study provides insight into the response of alpine meadows to grazing intensity, demonstrating that moderate grazing increases bacterial community diversity in grazed grasslands.     

The microbially mediated soil organic carbon loss under degenerative succession in an alpine meadow

Land‐cover change has long been recognized as having marked effect on the amount of soil organic carbon (SOC). However, the microbially mediated processes and mechanisms on SOC are still unclear. In this study, the soil samples in a degenerative succession from alpine meadow to alpine steppe meadow in the Qinghai–Tibetan Plateau were analysed using high‐throughput technologies, including Illumina sequencing and geochip functional gene arrays. The soil microbial community structure and diversity were significantly (p < .05) different between alpine meadow and alpine steppe meadow; the microbial ɑ‐diversity in alpine steppe meadow was significantly (p < .01) higher than in alpine meadow. Molecular ecological network analysis indicated that the microbial community structure in alpine steppe meadow was more complex and tighter than in the alpine meadow. The relative abundance of soil microbial labile carbon degradation genes (e.g., pectin and hemicellulose) was significantly higher in alpine steppe meadow than in alpine meadow, but the relative abundance of soil recalcitrant carbon degradation genes (e.g., chitin and lignin) showed the opposite tendency. The Biolog Ecoplate experiment showed that microbially mediated soil carbon utilization was more active in alpine steppe meadow than in alpine meadow. Consequently, more soil labile carbon might be decomposed in alpine steppe meadow than in alpine meadow. Therefore, the degenerative succession of alpine meadow because of climate change or anthropogenic activities would most likely decrease SOC and nutrients medicated by changing soil microbial community structure and their functional potentials for carbon decomposition.     

生长季放牧对高寒草甸传粉网络的影响

植物-传粉者相互作用面临人类活动的威胁。在青藏高原地区,放牧是一项主要的人类活动干扰,过度放牧导致高寒草甸植被严重退化。然而在该区域放牧如何影响植物-传粉者相互作用网络还不得而知。在青藏高原东部的高寒草甸选取了两个研究样点,每个样点包括一块禁牧样地（生长季禁牧）和放牧样地（全年放牧）。在2016年至2018年每年的生长季（7月和8月）进行了连续3年观测,共构建16个传粉网络。结果发现,在研究区域的高寒草甸生态系统中,传粉者群落的物种组成以双翅目昆虫为主。放牧后双翅目、鳞翅目以及鞘翅目传粉者的种类数减少,但膜翅目传粉者的种类数未受到放牧的影响。放牧干扰显著降低了群落中植物、传粉者以及它们之间相互作用的多样性,但对传粉网络的嵌套性和特化程度（H₂''）没有显著影响,说明网络的稳定性和恢复力没有受到放牧的影响。探讨了放牧对传粉网络的影响,发现区域放牧强度过大,降低了传粉昆虫和传粉网络的多样性。未来需要进一步深入研究高寒草甸生态系统中放牧强度对传粉网络的影响模式,以期为合理的放牧制度模式提供理论依据。     

Considerable methane uptake by alpine grasslands despite the cold climate: in situ measurements on the central Tibetan Plateau, 2008–2013

The uptake of CH₄ by aerate soil plays a secondary role in the removal of tropospheric CH₄, but it is still highly uncertain in terms of its magnitude, spatial, and temporal variation. In an attempt to quantify the sink of the vast alpine grasslands (1 400 000 km²) of the Tibetan Plateau, we conducted in situ measurements in an alpine steppe (4730 m) and alpine meadow (4900 m) using the static chamber and gas chromatograph method. For the alpine steppe, measurements (2008–2013) suggested that there is large interannual variability in CH₄ uptake, ranging from ?48.8 to ?95.8 μg CH₄ m^?2 h^?1 (averaged of ?71.5 ± 2.5 μg CH₄ m^?2 h^?1), due to the variability in precipitation seasonality. The seasonal pattern of CH₄ uptakes in the form of stronger uptake in the early growing season and weaker uptake in the rainy season closely matched the precipitation seasonality and subsequent soil moisture variation. The relationships between alpine steppe CH₄ uptake and soil moisture/temperature are best depicted by a quadratic function and an exponential function (Q₁₀ = 1.67) respectively. Our measurements also showed that the alpine meadow soil (average of ?59.2 ± 3.7 μg CH₄ m^?2 h^?1) uptake less CH₄ than the alpine steppe and produces a similar seasonal pattern, which is negatively regulated by soil moisture. Our measurements quantified – at values far higher than those estimated by process‐based models – that both the alpine steppe and alpine meadow are considerable CH₄ sinks, despite the cold weather of this high‐altitude area. The consecutive measurements gathered in this study also highlight that precipitation seasonality tends to drive the interannual variation in CH₄ uptake, indicating that future study should be done to better characterize how CH₄ cycling might feedback to the more extreme climate.     

The combined effects of warming and drying suppress CO2 and N2O emission rates in an alpine meadow of the eastern Tibetan Plateau

The eastern Tibetan Plateau has become increasingly warmer and drier since the 1990s. Such warming and drying has a great impact on ecosystem processes on the eastern Tibetan Plateau. To determine their combined effects on CO₂ and N₂O emission rates, we conducted a field manipulative experiment in an alpine meadow of the eastern Tibetan Plateau during the growing season of 2009. The experiment showed that warming manipulation increased soil temperature by 1?°C, and drying manipulation decreased soil water content by 6.8?%. We found that by counteracting the effect of low temperature in the area, experimental warming significantly increased soil microbial biomass, the number of bacteria, fungi, actinomycetes, ammonifying bacteria, nitrobacteria and denitrifying bacteria, and facilitated the emission rates of CO₂ and N₂O by 33.4 and 31.5?%, respectively. However, decreased precipitation further aggravated soil water stress and inhibited the numbers of these organisms, and reduced the emission rates of CO₂ and N₂O by 47.4 and 37.9?%, respectively. So decreased soil water content tended to offset the positive effect of warming. Compared to the positive effects of warming, decreased soil water content was shown in our study to have even greater impact on the eastern Tibetan Plateau during the growing season. Therefore, inhibition of CO₂ and N₂O emission rates (32.3 and 29.3?%, respectively) by warming and drying will intensify if the combined effects of these climatic trends persist in the region.     

Different responses of ecosystem carbon exchange to warming in three types of alpine grassland on the central Qinghai–Tibetan Plateau

Climate is a driver of terrestrial ecosystem carbon exchange, which is an important product of ecosystem function. The Qinghai–Tibetan Plateau has recently been subjected to a marked increase in temperature as a consequence of global warming. To explore the effects of warming on carbon exchange in grassland ecosystems, we conducted a whole‐year warming experiment between 2012 and 2014 using open‐top chambers placed in an alpine meadow, an alpine steppe, and a cultivated grassland on the central Qinghai–Tibetan Plateau. We measured the gross primary productivity, net ecosystem CO₂ exchange (NEE), ecosystem respiration, and soil respiration using a chamber‐based method during the growing season. The results show that after 3 years of warming, there was significant stimulation of carbon assimilation and emission in the alpine meadow, but both these processes declined in the alpine steppe and the cultivated grassland. Under warming conditions, the soil water content was more important in stimulating ecosystem carbon exchange in the meadow and cultivated grassland than was soil temperature. In the steppe, the soil temperature was negatively correlated with ecosystem carbon exchange. We found that the ambient soil water content was significantly correlated with the magnitude of warming‐induced change in NEE. Under high soil moisture condition, warming has a significant positive effect on NEE, while it has a negative effect under low soil moisture condition. Our results highlight that the NEE in steppe and cultivated grassland have negative responses to warming; after reclamation, the natural meadow would subject to loose more C in warmer condition. Therefore, under future warmer condition, the overextension of cultivated grassland should be avoided and scientific planning of cultivated grassland should be achieved.     

Warming decreased and grazing increased plant uptake of amino acids in an alpine meadow

Organic nitrogen (N) uptake by plants has been recognized as a significant component of terrestrial N cycle. Several studies indicated that plants have the ability to switch their preference between inorganic and organic forms of N in diverse environments; however, research on plant community response in organic nitrogen uptake to warming and grazing is scarce. Here, we demonstrated that organic N uptake by an alpine plant community decreased under warming with ¹³C–¹⁵N‐enriched glycine addition method. After 6 years of treatment, warming decreased plant organic N uptake by 37% as compared to control treatment. Under the condition of grazing, warming reduced plant organic N uptake by 44%. Grazing alone significantly increased organic N absorption by 15%, whereas under warming condition grazing did not affect organic N uptake by the Kobresia humilis community on Tibetan Plateau. Besides, soil NO₃–N content explained more than 70% of the variability observed in glycine uptake, and C:N ratio in soil dissolved organic matter remarkably increased under warming treatment. These results suggested warming promoted soil microbial activity and dissolved organic N mineralization. Grazing stimulated organic N uptake by plants, which counteracted the effect of warming.     

Rice roots select for type I methanotrophs in rice field soil

Methanotrophs are an important regulator for reducing methane (CH₄) emissions from rice field soils. The type I group of the proteobacterial methanotrophs are generally favored at low CH₄ concentration and high O₂ availability, while the type II group lives better under high CH₄ and limiting O₂ conditions. Such physiological differences are possibly reflected in their ecological preferences. In the present study, methanotrophic compositions were compared between rice-planted soil and non-planted soil and between the rhizosphere and rice roots by using terminal restriction fragment length polymorphism (T-RFLP) analysis of particulate methane monooxygenase (pmoA) genes. In addition, the effects of rice variety and nitrogen fertilizer were evaluated. The results showed that the terminal restriction fragments (T-RFs), which were characteristic for type I methanotrophs, substantially increased in the rhizosphere and on the roots compared with non-planted soils. Furthermore, the relative abundances of the type I methanotroph T-RFs were greater on roots than in the rhizosphere. Of type I methanotrophs, the 79 bp T-RF, which was characteristic for an unknown group or Methylococcus/Methylocaldum, markedly increased in field samples, while the 437 bp, which possibly represented Methylomonas, dominated in microcosm samples. These results suggested that type I methanotrophs were enriched or selected for by rice roots compared to type II methanotrophs. However, the members of type I methanotrophs are dynamic and sensitive to environmental change. Rice planting appeared to increase the copy number of pmoA genes relative to the non-planted soils. However, neither the rice variety nor the N fertilizer significantly influenced the dynamics of the methanotrophic community.     

川西北高寒草甸不同放牧模式对土壤呼吸的影响

采用土壤二氧化碳(CO_2)通量自动测量系统,对不同放牧模式(全年禁牧、夏季放牧、冬季放牧和自由放牧)下川西北高寒草甸的土壤呼吸进行监测,比较了不同放牧模式下土壤呼吸的季节动态和温度敏感性。研究发现:1)放牧模式可以改变高寒草甸土壤呼吸的季节动态变化。禁牧、夏季放牧以及自由放牧样地的土壤呼吸在季节上的变化趋势基本相似,而冬季放牧样地的土壤呼吸最大值与前者相比明显向后推迟;2)放牧模式并不改变高寒草甸年平均土壤呼吸速率,但对不同季节土壤呼吸速率的影响不同;3)不同放牧模式可以改变土壤呼吸对温度的敏感性(Q_(10))。不同放牧模式下土壤呼吸Q_(10)值大小依次为:禁牧1a(8.13)冬季放牧(7.49)禁牧3a(5.46)夏季放牧(5.20)自由放牧(4.53)。该地区土壤呼吸的Q_(10)值均明显高于热带和其它温带草地土壤呼吸的Q_(10)值。结果表明,放牧模式是影响高寒草甸土壤碳排放的一个重要因素。此外,在未来全球气候变暖背景下,在生长季节无放牧干扰的高寒草甸可能比放牧干扰的高寒草甸释放出更多的CO_2到大气中。     

Diversity and activity of methanotrophs in landfill cover soils with and without landfill gas recovery systems

Aerobic CH₄ oxidation plays an important role in mitigating CH₄ release from landfills to the atmosphere. Therefore, in this study, oxidation activity and community of methanotrophs were investigated in a subtropical landfill. Among the three sites investigated, the highest CH₄ concentration was detected in the landfill cover soil of the site (A) without a landfill gas (LFG) recovery system, although the refuse in the site had been deposited for a longer time (∼14–15 years) compared to the other two sites (∼6–11 years) where a LFG recovery system was applied. In April and September, the higher CH₄ flux was detected in site A with 72.4 and 51.7 g m⁻² d⁻¹, respectively, compared to the other sites. The abundance of methanotrophs assessed by quantification of pmoA varied with location and season. A linear relationship was observed between the abundance of methanotrophs and CH₄ concentrations in the landfill cover soils (R = 0.827, P < 0.001). The key factors influencing the methanotrophic diversity in the landfill cover soils were pH, the water content and the CH₄ concentration in the soil, of which pH was the most important factor. Type I methanotrophs, including Methylococcus, Methylosarcina, Methylomicrobium and Methylobacter, and type II methanotrophs (Methylocystis) were all detected in the landfill cover soils, with Methylocystis and Methylosarcina being the dominant genera. Methylocystis was abundant in the slightly acidic landfill cover soil, especially in September, and represented more than 89% of the total terminal-restriction fragment abundance. These findings indicated that the LFG recovery system, as well as physical and chemical parameters, affected the diversity and activity of methanotrophs in landfill cover soils.     

Methane emission by plant communities in an alpine meadow on the Qinghai-Tibetan Plateau: a new experimental study of alpine meadows and oat pasture

Recently, plant-derived methane (CH₄) emission has been questioned because limited evidence of the chemical mechanism has been identified to account for the process. We conducted an experiment with four treatments (i.e. winter-grazed, natural alpine meadow; naturally restored alpine meadow eight years after cultivation; oat pasture and bare soil without roots) during the growing seasons of 2007 and 2008 to examine the question of CH₄ emission by plant communities in the alpine meadow. Each treatment consumed CH₄ in closed, opaque chambers in the field, but two types of alpine meadow vegetation reduced CH₄ consumption compared with bare soil, whereas oat pasture increased consumption. This result could imply that meadow vegetation produces CH₄. However, measurements of soil temperature and water content showed significant differences between vegetated and bare soil and appeared to explain differences in CH₄ production between treatments. Our study strongly suggests that the apparent CH₄ production by vegetation, when compared with bare soil in some previous studies, might represent differences in soil temperature and water-filled pore space and not the true vegetation sources of CH₄.     

增温对青藏高原高寒草甸呼吸作用的影响

生态系统呼吸(ER)和土壤呼吸(SR)是草地生态系统碳排放的关键环节,其对气候变化极为敏感。高寒草甸是青藏高原典型的草地生态系统,其呼吸作用对气候变化的响应对区域碳排放具有重要的影响。以高寒草甸生态系统为对象,于2012—2016年采用模拟增温的方法研究呼吸作用对增温的响应。结果表明:增温对高寒草甸ER的影响存在年际差异,2013年和2014年增温对ER无显著影响,其他年份显著增加ER(P0.05),综合5年结果,平均增幅达22.3%。增温显著促进了高寒草甸SR(P0.05),较对照处理5年平均增幅高达67.1%;增温总体上提高了SR在ER中的比例(P0.05),最高增幅达到59.9%。ER和SR与土壤温度有显著的正相关关系(P0.05),与土壤水分没有显著的相关关系(P0.05)。对照样地中,土壤温度分别能解释33.0%和18.5%的ER和SR变化。在增温条件下,土壤温度可以解释20.5%和13.0%的ER和SR变化。在增温条件下,SR的温度敏感性显著增加,而ER的温度敏感性变化较小,导致SR的比重进一步增加。因此,在未来气候变暖条件下,青藏高原高寒草甸生态系统碳排放,尤其是土壤碳排放有可能进一步增加,土壤碳流失风险增加。     

Field application of nitrogen and phenylacetylene to mitigate greenhouse gas emissions from landfill cover soils: effects on microbial community structure

Landfills are large sources of CH₄, but a considerable amount of CH₄ can be removed in situ by methanotrophs if their activity can be stimulated through the addition of nitrogen. Nitrogen can, however, lead to increased N₂O production. To examine the effects of nitrogen and a selective inhibitor on CH₄ oxidation and N₂O production in situ, 0.5 M of NH₄Cl and 0.25 M of KNO₃, with and without 0.01% (w/v) phenylacetylene, were applied to test plots at a landfill in Kalamazoo, MI from 2007 November to 2009 July. Nitrogen amendments stimulated N₂O production but had no effect on CH₄ oxidation. The addition of phenylacetylene stimulated CH₄ oxidation while reducing N₂O production. Methanotrophs possessing particulate methane monooxygenase and archaeal ammonia-oxidizers (AOAs) were abundant. The addition of nitrogen reduced methanotrophic diversity, particularly for type I methanotrophs. The simultaneous addition of phenylacetylene increased methanotrophic diversity and the presence of type I methanotrophs. Clone libraries of the archaeal amoA gene showed that the addition of nitrogen increased AOAs affiliated with Crenarchaeal group 1.1b, while they decreased with the simultaneous addition of phenylacetylene. These results suggest that the addition of phenylacetylene with nitrogen reduces N₂O production by selectively inhibiting AOAs and/or type II methanotrophs.     

高寒草地植物群落关键种对不同放牧家畜组合放牧的响应

如何通过合理的利用方式提高高寒草地管理水平,实现其可持续利用一直是草地生态学领域的研究热点。为明确不同放牧家畜组合下高寒草地植物群落关键种的演替规律及其驱动因素,基于中等放牧强度设置了不同放牧家畜组合放牧样地（牦牛单牧、藏羊单牧、牦牛藏羊1 ： 2混牧、牦牛藏羊1 ： 4混牧、牦牛藏羊1 ： 6混牧）以及围封样地,并在连续放牧处理7年后系统分析了不同放牧家畜组合下植物群落特征与土壤理化性质变化,以期确定最优放牧组合。结果表明：（1）不同放牧家畜组合均会显著降低高寒草地植物盖度,但对其物种丰富度与多样性指数的影响并不显著。（2）牦牛藏羊1 ： 2混牧下植物群落特征与禁牧处理下植物群落特征较为相似,且牦牛藏羊1 ： 2混牧下的植物群落稳定性最强、组织水平最高。（3）牦牛单牧与1 ： 2混牧下关键种为矮生嵩草;藏羊单牧下关键种为天山针茅;1 ： 4混牧下关键种为星毛委陵菜;1 ： 6混牧与围封下关键种为赖草。（4）围封能够显著降低土壤容重,藏羊单牧则会显著增加土壤容重与土壤速效氮、磷含量。（5）土壤速效氮、容重与含水量是驱动不同放牧家畜组合下植物群落关键种演替的重要理化因子。综上所述,中等放牧强度下,牦牛藏羊1 ： 2混牧是青藏高原高寒草地较为理想的利用方式。此外,相较于单纯增加草地物种丰富度或多样性,建立植物群落物种之间的有效关联是提高青藏高原高寒草地管理水平的另一关键途径。