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Non-rhizobial endophytes (NREs) are active colonizers inhabiting the root nodules. Though their active role in the lentil agroecosystem is not well defined, here we observed that these NREs might promote the growth of lentils, modulate rhizospheric community structure and could be used as promising organisms for optimal use of rice fallow soil. NREs from root nodules of lentils were isolated and examined for plant growth-promoting traits, exopolysaccharide (EPS) and biofilm production, root metabolites, and the presence of nifH and nifK elements. The greenhouse experiment with the chosen NREs, i.e., Serratia plymuthica 33GS and Serratia sp. R6 significantly increased the germination rate, vigour index, development of nodules (in non-sterile soil) and fresh weight of nodules (33GS 94%, R6 61% growth) and length of the shoot (33GS 86%, R6 51.16%) as well as chlorophyll levels when compared to the uninoculated control. Scanning Electron Microscopy (SEM) revealed that both isolates could successfully colonize the roots and elicit root hair growth. The inoculation of the NREs resulted in specific changes in root exudation patterns. The plants with 33GS and R6 treatment significantly stimulated the exudation of triterpenes, fatty acids, and their methyl esters in comparison to the uninoculated plants, altering the rhizospheric microbial community structure. Proteobacteria dominated the rhizospheric microbiota in all the treatments. Treatment with 33GS or R6 also enhanced the relative abundance of other favourable microbes, including Rhizobium, Mesorhizobium, and Bradyrhizobium. The correlation network analysis of relative abundances resulted in numerous bacterial taxa, which were in cooperation with each other, having a possible role in plant growth promotion. The results indicate the significant role of NREs as plant growth promoters, which also includes their role in root exudation patterns, enhancement of soil nutrient status and modulation of rhizospheric microbiota, suggesting their prospects in sustainable, and bio-based agriculture.

Aims Specific soil bacteria can sense and respond to the selective rhizosphere recruitment of root exudates using unique systems of chemotaxis that mediate plant-microbe and microbe-microbe interactions. This study investigates how the bacterial chemotaxis systems have been impacted by selection during the domestication of rice ( Oryza species). Methods Shotgun metagenomic sequencing and 16S rRNA gene amplicon sequencing were performed to investigate the bacterial chemotaxis systems and chemotactic bacteria in the rhizospheres of wild and cultivated rice. Metabolomics analysis was performed to examine the root metabolites of different accessions of rice. Results The bacterial chemotaxis genes exhibited a higher abundance in the rhizospheres of wild rice than cultivated rice, and that the compositional profile of chemotaxis genes was distinctly different between types of rice. Differential selection of chemotaxis systems was at least partially driven by changes in the metabolite profiles of rice roots that were affected by domestication. A core group of chemotactic bacteria was also identified, and specific chemotactic bacteria were found to function as hub taxa in the rhizosphere bacterial community. Conclusion The present study provides novel insights into the composition and function of the bacterial chemotaxis systems in the rhizospheres of wild and domesticated rice. It also provides a new perspective on the impact of rice domestication on the assembly of rhizomicrobiome.

The cultivation of Paeonia suffruticosa ‘Luoyang Hong’, a valuable ornamental crop, faces significant challenges due to replanting issues. However, the dynamics of its rhizosphere micro-ecosystem under continuous cropping remain poorly understood. This study systematically investigates the successional patterns of the rhizosphere micro-ecosystem over a 12- to 42-year chronosequence to identify the underlying drivers of these issues. Using an integrated multi-omics approach combining metagenomics and non-targeted metabolomics, we deciphered the rhizosphere mechanisms associated with replanting issues in Paeonia suffruticosa ‘Luoyang Hong’. Based on differential changes in metabolites within the soil and root systems, key substances such as succinic acid, trans-ferulic acid, vanillic acid, and Leu-Val-Arg-Lys were identified. The microbial succession demonstrated a distinct temporal progression. Initially, at the 12-year stage, the rhizosphere was enriched with beneficial bacterial genera. However, around the 20-year stage, the abundance of these beneficial genera significantly declined. Subsequently, at the 34-year stage, the community shifted to a dominance of genera associated with organic matter degradation. Finally, at the 42-year stage, a partial recovery of certain beneficial genera and their functions was observed. Despite this recovery, the overall system continued to exhibit signs of continuous degradation. Integrated multi-omics analysis further revealed significant positive correlations, such as that between N,N-dimethyldodecylamine N-oxide and several differential microbial genera, underscoring the complex interactions between metabolites and microbes. Our findings provide a systematic perspective on the micro-ecological dynamics in the rhizosphere of Paeonia suffruticosa ‘Luoyang Hong’, offering deeper insights into replanting issues and supporting future mitigation strategies.

Grazing disturbance can change the structure of plant rhizosphere microbial communities and thereby alter the feedback to promote plant growth or induce plant defenses. However, little is known about how such changes occur and vary under different grazing pressures or the roles of root metabolites in altering the composition of rhizosphere microbial communities. In this study, the effects of different grazing pressures on the composition of microbial communities were investigated, and the mechanisms by which different grazing pressures changed rhizosphere microbiomes were explored with metabolomics. Grazing changed composition, functions, and co-expression networks of microbial communities. Under light grazing (LG), some saprophytic fungi, such as Lentinus sp., Ramichloridium sp., Ascobolus sp. and Hyphoderma sp., were significantly enriched, whereas under heavy grazing (HG), potentially beneficial rhizobacteria, such as Stenotrophomonas sp., Microbacterium sp., and Lysobacter sp., were significantly enriched. The beneficial mycorrhizal fungus Schizothecium sp. was significantly enriched in both LG and HG. Moreover, all enriched beneficial microorganisms were positively correlated with root metabolites, including amino acids (AAs), short-chain organic acids (SCOAs), and alkaloids. This suggests that these significantly enriched rhizosphere microbial changes may be caused by these differential root metabolites. Under LG, it is inferred that root metabolites, especially AAs such as L-Histidine, may regulate specific saprophytic fungi to participate in material transformations and the energy cycle and promote plant growth. Furthermore, to help alleviate the stress of HG and improve plant defenses, it is inferred that the root system actively regulates the synthesis of these root metabolites such as AAs, SCOAs, and alkaloids under grazing interference, and then secretes them to promote the growth of some specific plant growth-promoting rhizobacteria and fungi. To summarize, grasses can regulate beneficial microorganisms by changing root metabolites composition, and the response strategies vary under different grazing pressure in typical grassland ecosystems.

Continuous cropping in soybean is increasingly practiced in Heilongjiang Province, leading to substantial yield reductions and quality degradation. Arbuscular mycorrhizal fungi (AMF) are soil microorganisms that form mutualistic interactions with plant roots and can restore the plant rhizosphere microenvironment. In this study, two soybean lines (HN48 and HN66) were chosen as experimental materials, which were planted in different years of continuous cropping soybean soils and were inoculated or not with Funneliformis mosseae in potted-experiments. Ultimately, analysis of root tissue metabolome and root exudates, soil physicochemical properties, plant biomass, as well as rhizosphere soil properties in different experimental treatments, inoculated or not with F. mosseae, was performed. Experimental results showed that: (a) The disease index of soybean root rot was significantly lower in the treatment group than in the control group, and there were differences in disease index and the resistance effect of F. mosseae between the two cultivars; (b) compared with the control, the root tissue metabolome and root exudates remained unchanged, but there were changes in the relative amounts in the treatment group, and the abundant metabolites differed by soybean cultivar; (c) soybean biomass was significantly higher in the treatment group than in the control group, and the effect of F. mosseae on biomass differed with respect to the soybean cultivar; and (d) there were differences in the physiochemical indexes of soybean rhizosphere soil between the treatment and control groups, and the repairing effect of F. mosseae differed between the two cultivars. Therefore, F. mosseae can increase the biomass of continuously cropped soybean, improve the physicochemical properties of the rhizosphere soil, regulate the root metabolite profiles, and alleviate barriers to continuous cropping in potted-experiments of soybean.

The development of cereal crops with high nitrogen use efficiency (NUE) is a priority for worldwide agriculture. In addition to conventional plant breeding and genetic engineering, the use of the plant microbiome offers another approach to improving crop NUE. To gain insight into the bacterial communities associated with sorghum lines that differ in NUE, a field experiment was designed comparing 24 diverse Sorghum bicolor lines under sufficient and deficient nitrogen (N). Amplicon sequencing and untargeted gas chromatography–mass spectrometry were used to characterize the bacterial communities and the root metabolome associated with sorghum genotypes varying in sensitivity to low N. We demonstrated that N stress and sorghum type (energy, sweet, and grain sorghum) significantly impacted the root-associated bacterial communities and root metabolite composition of sorghum. We found a positive correlation between sorghum NUE and bacterial richness and diversity in the rhizosphere. The greater alpha diversity in high NUE lines was associated with the decreased abundance of a dominant bacterial taxon, Pseudomonas . Multiple strong correlations were detected between root metabolites and rhizosphere bacterial communities in response to low N stress. This indicates that the shift in the sorghum microbiome due to low N is associated with the root metabolites of the host plant. Taken together, our findings suggest that host genetic regulation of root metabolites plays a role in defining the root-associated microbiome of sorghum genotypes differing in NUE and tolerance to low N stress. The development of crops that are more nitrogen use-efficient (NUE) is critical for the future of the enhanced sustainability of agriculture worldwide. This objective has been pursued mainly through plant breeding and plant molecular engineering, but these approaches have had only limited success. Therefore, a different strategy that leverages soil microbes needs to be fully explored because it is known that soil microbes improve plant growth through multiple mechanisms. To design approaches that use the soil microbiome to increase NUE, it will first be essential to understand the relationship among soil microbes, root metabolites, and crop productivity. Using this approach, we demonstrated that certain key metabolites and specific microbes are associated with high and low sorghum NUE in a field study. This important information provides a new path forward for developing crop genotypes that have increased NUE through the positive contribution of soil microbes.

Intercropping can enable more efficient resource use and increase yield. Most current studies focus on the correlation between soil nutrients and crop yield under intercropping conditions. However, the mechanisms related to root exudates and soil nutrients remain unclear. Therefore, this study explored the correlation between rhizosphere soil nutrients and root exudates in sugarcane/peanut intercropping. Root extracts, root exudates, rhizosphere soil enzyme activities, and soil nutrients were analyzed and compared in monocultured and intercropped peanut and sugarcane at different growth stages. The root metabolites were annotated using the Kyoto Encyclopedia of Genes and Genomes pathways to further identify the connection between soil nutrients and root exudates. The effects of intercropping differed in peanut and sugarcane at different growth stages, and the difference between podding and pod-filling stages was significant. Intercropping generally had a great effect on peanut; it not only significantly increased the organic acid, soluble sugars, and phenolic acids in root exudates and extracts from peanuts, but also significantly increased rhizosphere soil enzyme activities and soil nutrient levels. Intercropping peanuts promoted fumaric acid secretion from roots and significantly affected the metabolic pathways of alanine, aspartate, and glutamate. Sugarcane/peanut intercropping can increase root exudates and effectively improve soil nutrients. The changes in soil nutrients are closely related to the effects of fumaric acid on alanine, aspartate, and glutamate metabolism.

The root traits and response strategies of plants play crucial roles in mediating interactions between plant root systems. Current research on the role of root exudates as underground chemical signals mediating these interactions has focused mainly on crops, with less attention given to desert plants in arid regions. In this study, we focused on the typical desert plant Haloxylon ammodendron and conducted a pot experiment using three root isolation methods (plastic film separation, nylon mesh separation, and no separation). We found that (1) as the degree of isolation increased, plant biomass significantly increased (p < 0.05), while root organic carbon content exhibited the opposite trend; (2) soil electrical conductivity (EC), soil total nitrogen (STN), soil total phosphorus (STP), and soil organic carbon (SOC) were significantly greater in the plastic film and nylon mesh separation treatments than in the no separation treatment (p < 0.05), and the abundance of Proteobacteria and Actinobacteriota was significantly greater in the plastic film separation treatment than in the no separation treatment (p < 0.05); (3) both plastic film and nylon mesh separations increased the secretion of alkaloids derived from tryptophan and phenylalanine in the plant root system compared with that in the no separation treatment; and (4) Pseudomonas, Proteobacteria, sesquiterpenes, triterpenes, and coumarins showed positive correlations, while both pseudomonas and proteobacteria were significantly positively correlated with soil EC, STN, STP, and SOC (p < 0.05). Aurachin D was negatively correlated with Gemmatimonadota and Proteobacteria, and both were significantly correlated with soil pH, EC, STN, STP, and SOC. The present study revealed strong negative interactions between the root systems of H. ammodendron seedlings, in which sesquiterpenoids, triterpenoids, coumarins, and alkaloids released by the roots played an important role in the subterranean competitive relationship. This study provides a deeper understanding of intraspecific interactions in the desert plant H. ammodendron and offers some guidance for future cultivation of this species in the northwestern region of China.

Two years of field testing were conducted in naturally Plasmopara halstedii -infested soils to assess the effectiveness of Trichoderma harzianum isolate (TRIC8) against downy mildew ( P. halstedii ) on sunflower ( Helianthus annuus ). Seed treatment with this antagonist offered protection of 73.64% in the first year. The effects of this treatment on oil content of the seeds and metabolite profiles of roots, along with disease severity, were evaluated in the second year by comparing to the fungicide treatment (Metalaxyl M). Oil content in seeds and metabolite profile in roots were studied using Nuclear Magnetic Resonance (NMR) and gas chromatography/mass spectrometry (GC/MS), respectively. TRIC8 and fungicide treatments were found to cause significantly lower disease severity when compared to that of the control, although weather conditions were more favorable for the disease in the second year than in the first year. The oil content in seeds with TRIC8 treatment was superior to that in seeds with fungicide treatment or control. The metabolites in roots from treated plants and disease severity were subjected to principal component analysis (PCA). TRIC8 treatment increased the accumulation of some alcohols [2-ethyl-2-methyltridecan-1-ol, 2-(propylamino)ethanol], aldehydes [2,5-bis(trimethylsilyloxy)benzaldehyde, dodecanal], alkanes (dotriacontane, eicosane, heneicosane, nonacosane, nonylcyclopropane, tetracontane, tetracosane), ketones [(1-hydroxycyclohexyl)-phenylmethanone, (4-methylphenyl)-phenylmethanone], an ester (6-ethyloctan-3-yl 6-bromohexanoate) and a terpene (falcarinol), which were among the metabolites that contributed over 0.70 in PC1 and were negatively correlated with disease severity. We suggest these metabolites might be involved in defense response of sunflower against P. halstedii in the field.

With continued anthropogenic inputs of nitrogen (N) into the environment, non-point source N pollutants produced in winter cannot be ignored. As the water-soil interface zones, riparian wetlands play important roles in intercepting and buffering N pollutants. However, winter has the antagonistic effect on the N removal. Substrate improvement has been suggested as a strategy to optimize wetland performance and there remain many uncertainties about the inner mechanism. This study explores the effects of substrate improvement on N removal in winter and rhizospheric crosstalk between reed ( Phragmites australis ) and microbes in subtropical riparian reed wetlands. The rates of wetland N removal in winter, root metabolite profiles, and rhizosphere soil microbial community compositions were determined following the addition of different substrates (gravel, gravel + biochar, ceramsite + biochar, and modified ceramsite + biochar) to natural riparian soil. The results showed that the addition of different substrates to initial soil enhanced N removal from the microcosms in winter. Gravel addition increased NH 4 + -N removal by 8.3% ( P  < 0.05). Gravel + biochar addition increased both TN and NH 4 + -N removals by 8.9% ( P  < 0.05). The root metabolite characteristics and microbial community compositions showed some variations under different substrate additions compared to the initial soil. The three treatments involving biochar addition decreased lipid metabolites and enhanced the contents and variety of carbon sources in rhizosphere soil, while modified ceramsite + biochar addition treatment had a greater impact on the microbial community structure. There was evidence for a complex crosstalk between plants and microbes in the rhizosphere, and some rhizosphere metabolites were seen to be significantly correlated with the bacterial composition of the rhizospheric microbial community. These results highlighted the importance of rhizospheric crosstalk in regulating winter N removal in riparian reed wetland, provided a scientific reference for the protection and restoration of riparian reed areas and the prevention and control of non-point source pollution.