The effects of organic fertilizer application on the soil microbial community in grassland systems have been extensively studied. However, the effects of organic fertilizers on the structure of rhizosphere microbial communities are still limited. In this study, the diversity and composition of rhizosphere microbial communities of a gramineous grass Elymus nutans under organic fertilizer treatment were studied in an artificial pasture on Qinghai–Tibet Plateau. After a growing season, the application of organic fertilizer not only increased the height and biomass of Elymus nutans, but also changed the rhizosphere microbial compositions. In particular, organic fertilizer increased the diversity of rhizosphere bacterial community and inhibited the growth of pathogenic bacteria such as Acinetobacter, but the opposite trend was observed for the diversity of fungal community. The assembly process of fungal community was changed from a stochastic process to a deterministic process, indicating that selection was strengthened. Additionally, both the infection rate of arbuscular mycorrhizal fungi (AMF) toward host plants and the development of AMF-related structures were significantly increased after the application of organic fertilizer. Our study demonstrated that the addition of organic fertilizer to artificial pasture could improve the growth of grass through the alteration of the rhizosphere microbial communities. Organic fertilizer had a greater selectivity for the bacterial and the fungal communities that enhanced the niche filtration in this community, further benefiting the yield of forages.

Chemical fertilizers are gradually being replaced with new biological fertilizers, which can improve the soil and soil microorganisms. In this experiment, leguminous forage (Medicago sativa cv. Beilin 201) was used as the research object. By measuring alfalfa root systems and soil properties and using high-throughput sequencing technology, we investigated the effect of biological (rhizobial) fertilizer at different concentrations on soil fertility and alfalfa rhizosphere microbiota in alpine grasslands. The results demonstrated that the treatment with biofertilizer significantly reduced total nitrogen (TN) and total organic carbon (TOC) content in soils, increased root densities, and significantly increased the number of root nodules in alfalfa. There were differences in the response of rhizosphere microorganisms to different concentrations of biofertilizer, and the treatment with biofertilizer led to pronounced changes in the microbial community structure. The abundance of beneficial bacteria such as Rhizobium, Arthrobacter, and Pseudomonas was significantly increased. The Pearson correlation analysis showed that soil moisture and soil conductivity were significantly positively correlated with the observed richness of rhizosphere microbiota (p < 0.05). Meanwhile, Actinobacteria showed a significantly positive correlation with nitrate, TOC, and TN (p < 0.01). These results indicated that biofertilizers enhanced soil fertility and altered the rhizosphere microbiota of alfalfa in alpine grassland.

The Qinghai–Tibet Plateau, a critical ecological barrier and major livestock region, faces deteriorating grasslands and rising forage demand under its harsh alpine climate. Oat (Avena sativa L.), valued for its cold tolerance, rapid biomass accumulation, and ability to thrive in nutrient-poor soils, can expand winter feed reserves and partly alleviate grazing pressure on native rangelands. However, genetic improvement alone has not been sufficient to address the environmental challenges. This issue is particularly severe in the Qaidam Basin, where soil salinization, characterized by high pH, poor soil structure, and low nutrient availability, significantly limits crop performance. Rhizosphere growth-promoting bacteria (PGPR) are environmentally friendly biofertilizers known to enhance crop growth, yield, and soil quality, but their application in the saline soil of the Qaidam Basin remains limited. We evaluated two PGPR application rates (B1 = 75 kg hm⁻² and B2 = 150 kg hm⁻²) on ‘Qingtian No. 1’ oat, assessing plant growth, soil physicochemical properties, and rhizosphere microbial communities. The results indicated that both treatments significantly increased oat productivity, raised the comprehensive growth index, augmented soil organic matter, and lowered soil pH; B1 chiefly enhanced above-ground biomass and fungal community stability, whereas B2 more strongly promoted root development and bacterial community stability. Structural equation modeling showed that PGPR exerted direct effects on the comprehensive growth index and indirect effects through soil and microbial pathways, with soil properties contributing slightly more than microbial factors. Notably, rhizosphere organic matter, fungal β-diversity, and overall microbial community stability emerged as positive key drivers of the comprehensive growth index. These findings provide a theoretical basis for optimizing PGPR dosage in alpine forage systems and support the sustainable deployment of microbial fertilizers under saline soil conditions in the Qaidam Basin.

It has been widely recognized that organic fertilizer (OF) application under monoculture and continuous cropping can change the microbial community and increase forage biomass in the Qinghai–Tibet Plateau. However, as a commonly used grassland planting pattern, the way in which grass–legume mixtures respond to OF application remains unclear. To clarify application effects of organic fertilizer in the grass–legume mixtures, we conducted a field experiment at the Qinghai–Tibet Plateau and collected the rhizospheric and bulk soils to reveal their microbial community by using high-throughput sequencing and molecular ecological networks. It was found that OF application changed the microbial community and increased the forage biomass under monoculture. However, in grass–legume mixtures, we found that OF application did not promote the increase of forage (Gramineae) biomass (Student t-test: p > 0.05). By analyzing both prokaryote and fungal communities, it was found that OF application had a greater impact on bulk soil microorganisms than on those of the rhizosphere in grass–legume mixtures. Co-occurrence network analysis showed that the rhizosphere and bulk soil networks of grass–legume mixtures were significantly more vulnerable under OF treatment (vulnerability of prokaryotes in grass: 0.1222; vulnerability of prokaryotes in legumes: 0.1730; fungal vulnerability in grass: 0.0116; fungal vulnerability in legumes: 0.0223) than non-OF treatment (vulnerability of prokaryotes in grass: 0.1015; vulnerability of prokaryotes in legumes: 0.1337; fungal vulnerability in grass: 0.0046; fungal vulnerability in legumes: 0.0126), which indicated that OF application did not provide favorable conditions for microbial interactions in grass–legume mixtures. In addition, structural equation modeling showed that OF application had some significant negative impacts on soil physicochemical properties and the robustness of the prokaryote community. The robustness of fungi had a significant negative (p < 0.001) impact on forage biomass, but OF application had no significant (p > 0.05) direct impact on the forage biomass, which indicated that the OF did not promote forage biomass in grass–legume mixtures. These results suggest that the application of organic fertilizer is unnecessary for grass–legume mixtures, because it does not promote the interactions between rhizospheric microbes and forage.

Soil salinization limits global agricultural sustainability, and extensive areas of saline–alkaline soils on the Qinghai–Tibet Plateau remain underutilized. Against this backdrop, this study evaluated the effects and ecological regulatory mechanisms of potassium fulvate (PF) application on oat (Avena sativa L.) growth, soil properties, and rhizosphere microbial communities in the saline–alkali soils of the Qaidam Basin. The results showed that PF significantly enhanced both aboveground and belowground biomass and improved root morphological traits, with the higher application rate (150 kg·hm⁻²) showing superior performance. PF also effectively improved soil nutrient conditions (organic matter, ammonium nitrogen, and potassium), reduced the integrated salinity–alkalinity index, significantly optimized the composition of rhizosphere soil cations (increased K⁺ and Ca²⁺; decreased Na⁺ and Mg²⁺), and induced a marked reshaping of the composition and structure of rhizosphere microbial communities. Notably, microbial β-diversity exhibited a significant regulatory effect on the comprehensive growth of oats. Structural equation modeling (SEM) revealed that PF primarily promoted oat growth indirectly by improving soil physicochemical properties (direct effect = 0.94), while the microbial community structure served as a synergistic ecological mediator. This study clarifies the regulatory mechanisms of PF in oat cultivation under alpine saline–alkali conditions, providing both theoretical and practical support for improving soil quality, enhancing forage productivity, and promoting sustainable agriculture in cold regions.

Introduction: This study aimed to investigate the seed germination characteristics and metabolic response mechanisms of Medicago ruthenica under saline-alkali stress, with the goal of elucidating its physiological and molecular mechanisms of salt-alkali tolerance.Methods: It was systematically analyzed the germination characteristics of four M. ruthenica germplasm resources (YSZ, XHZ, Shoulu, and Longzhong 1) from different origins with various concentrations of individual salts (NaCl, Na₂SO₄, NaHCO₃) and compound saline-alkali stress treatments. Additionally, the metabolite changes in the domesticated species under saline-alkali stress were examined using metabolomics technology.Results: The results indicated that low concentrations of NaCl stress did not significantly inhibit the germination of M. ruthenica seeds; rather, it promoted germination to some extent. In contrast, high concentrations of NaHCO₃ and compound saline-alkali stress significantly inhibited both seed germination and seedling growth. The germination characteristics of M. ruthenica germplasm resources from different sources exhibit variability under saline-alkali stress. Domesticated species demonstrated strong tolerance to salt-alkali conditions. Metabolomic analyses indicated that saline-alkali stress significantly impacts key metabolic pathways, including amino acid metabolism, sugar metabolism, and lipid metabolism in M. ruthenica sprouts, with a notable increase in the accumulation of amino acids and their derivatives. Correlation analyses revealed that amino acids such as L-arginine, histidine, and glutamine are significantly positively correlated with germination rate and root length, suggesting that these amino acids play a crucial role in M. ruthenica’s response to saline-alkali stress.Discussion: This study provided a new theoretical foundation for understanding the salt-alkali tolerance mechanisms of M. ruthenica and serves as an important reference for breeding salt-alkali tolerant forage varieties and for the ecological restoration of saline-alkali land.