The rhizosphere hosts a complex web of prokaryotes interacting with one another that may modulate crucial functions related to plant growth and health. Identifying the key factors structuring the prokaryotic community of the plant rhizosphere is a necessary step toward the enhancement of plant production and crop yield with beneficial associative microorganisms. We used a long-term field experiment conducted at three locations in the Canadian prairies to verify that: (1) the level of cropping system diversity influences the α- and β-diversity of the prokaryotic community of canola (Brassica napus) rhizosphere; (2) the canola rhizosphere community has a stable prokaryotic core; and (3) some highly connected taxa of this community fit the description of hub-taxa. We sampled the rhizosphere of canola grown in monoculture, in a 2-phase rotation (canola-wheat), in a 3-phase rotation (pea-barley-canola), and in a highly diversified 6-phase rotation, five and eight years after cropping system establishment. We detected only one core bacterial Amplicon Sequence Variant (ASV) in the prokaryotic component of the microbiota of canola rhizosphere, a hub taxon identified as cf. Pseudarthrobacter sp. This ASV was also the only hub taxon found in the networks of interactions present in both years and at all three sites. We highlight a cohort of bacteria and archaea that were always connected with the core taxon in the network analyses.

The temporal distribution patterns of bacterial communities, as an important group in mountain soil, are affected by various environmental factors. To improve knowledge regarding the successional seasonal dynamics of the mountain soil bacterial communities, the rhizospheric soil of a 30-year-old natural secondary Pinus tabulaeformis forest, located in the high-altitude (1900 m a.s.l.) of the temperate Qinling Mountains, was sampled and studied during four different seasons. The bacterial community composition and structure in the rhizospheric soil were studied using an Illumina MiSeq Sequencing platform. Furthermore, the edaphic properties and soil enzymatic activities (urease, phosphatase, and catalase) were measured in order to identify the main impact factors on the soil bacterial community. According to the results, all of the edaphic properties and soil enzymatic activities were significantly affected by the seasonal changes, except for the C/N ratio. Although the biomasses of soil bacterial communities increased during the summer and autumn (warm seasons), their Shannon diversity and Pielou's evenness were decreased. Proteobacteria, Acidobacteria, Actinobacteria, Planctomycetes, and Bacteroidetes were the predominant bacterial groups in all of the soil samples, and the genera of Ktedonobacter, Sphingobium as well as an unclassified member of the Ktedonobacteria were the keystone taxa. The composition and structure of soil bacterial communities were strongly impacted by the edaphic properties, especially the temperature, moisture, ammoniacal nitrogen, available phosphorus and total phosphorus which were the crucial factors to drive the temporal distribution of the soil bacterial community and diversity. In conclusion, the soil temperature, moisture and the nutrients N and P were the crucial edaphic factors for shaping the rhizospheric soil bacterial communities as season and climate change in a P. tabulaeformis forest of Qinling Mountains.

Agriculturally-driven land transformation is increasing globally. Improving phosphorus (P) use efficiency to sustain optimum productivity in diverse ecosystems, based on knowledge of soil P dynamics, is also globally important in light of potential shortages of rock phosphate to manufacture P fertilizer. We investigated P chemical speciation and P cycling with solution 31P nuclear magnetic resonance, P K-edge X-ray absorption near-edge structure spectroscopy, phosphatase activity assays, and shotgun metagenomics in soil samples from long-term agricultural fields containing four different land-use types (native and tame grasslands, annual croplands, and roadside ditches). Across these land use types, native and tame grasslands showed high accumulation of organic P, principally orthophosphate monoesters, and high acid phosphomonoesterase activity but the lowest abundance of P cycling genes. The proportion of inositol hexaphosphates (IHP), especially the neo-IHP stereoisomer that likely originates from microbes rather than plants, was significantly increased in native grasslands than croplands. Annual croplands had the largest variances of soil P composition, and the highest potential capacity for P cycling processes based on the abundance of genes coding for P cycling processes. In contrast, roadside soils had the highest soil Olsen-P concentrations, lowest organic P, and highest tricalcium phosphate concentrations, which were likely facilitated by the neutral pH and high exchangeable Ca of these soils. Redundancy analysis demonstrated that IHP by NMR, potential phosphatase activity, Olsen-P, and pH were important P chemistry predictors of the P cycling bacterial community and functional gene composition. Combining chemical and metagenomics results provides important insights into soil P processes and dynamics in different land-use ecosystems.

Canola is one of the most economically important crops in Canada, and the root and rhizosphere microbiomes of a canola plant likely impact its growth and nutrient uptake. The aim of this study was to determine whether canola has a core root microbiome (i.e., set of microbes that are consistently selected in the root environment), and whether this is distinct from the core microbiomes of other crops that are commonly grown in the Canadian Prairies, pea, and wheat. We also assessed whether selected agronomic treatments can modify the canola microbiome, and whether this was associated to enhanced yield. We used a field experiment with a randomized complete block design, which was repeated at three locations across the canola-growing zone of Canada. Roots and rhizosphere soil were harvested at the flowering stage of canola. We separately isolated total extractable DNA from plant roots and from adjacent rhizosphere soil, and constructed MiSeq amplicon libraries for each of 60 samples, targeting bacterial, and archaeal 16S rRNA genes and the fungal ITS region. We determined that the microbiome of the roots and rhizosphere of canola was consistently different from those of wheat and pea. These microbiomes comprise several putative plant-growth-promoting rhizobacteria, including Amycolatopsis sp., Serratia proteamaculans, Pedobacter sp., Arthrobacter sp., Stenotrophomonas sp., Fusarium merismoides, and Fusicolla sp., which correlated positively with canola yield. Crop species had a significant influence on bacterial and fungal assemblages, especially within the roots, while higher nutrient input or seeding density did not significantly alter the global composition of bacterial, fungal, or archaeal assemblages associated with canola roots. However, the relative abundance of Olpidium brassicae, a known pathogen of members of the Brassicaceae, was significantly reduced in the roots of canola planted at higher seeding density. Our results suggest that seeding density and plant nutrition management modified the abundance of other bacterial and fungal taxa forming the core microbiomes of canola that are expected to impact crop growth. This work helps us to understand the microbial assemblages associated with canola grown under common agronomic practices and indicates microorganisms that can potentially benefit or reduce the yield of canola.

It is possible that arbuscular mycorrhizal fungi play a pivotal role in root development and Pb phytostabilization in plants grown in Pb-contaminated soil. In this study, a pot experiment was conducted over 4 months to evaluate the effects of Funneliformis mosseae strain BGCXJ01A on root characteristics of black locust (Robinia pseudoacacia L.) seedlings in Pb-contaminated soil. Four Pb treatments (0, 90, 900, and 3,000 mg kg-1) were applied to soil in the presence and absence of F. mosseae. Inoculation with F. mosseae prominently improved root length, surface area, volume, and tip number in the plants across all Pb treatments. The F. mosseae inoculation also increased root diameter and fork number, especially under high Pb treatments. The presence of F. mosseae significantly increased the root activity and root tolerance index. However, there was little difference in specific root length between inoculated and non-inoculated plants. The biomass of roots, stems, and leaves all increased following inoculation with F. mosseae. Inoculated plants had greater accumulation and translocation capacities for Pb in the roots and stems, but lower capacities were found in the leaves when compared with those in non-inoculated plants. These results highlight that F. mosseae can alleviate the toxic effects of Pb on root development and can immobilize Pb in the roots and stems of R. pseudoacacia grown in Pb-contaminated soil. This study provides a model system for phytoremediation of Pb-contaminated soil via reciprocal symbiosis between arbuscular mycorrhizal fungi and woody legumes.

Melatonin is a new kind of plant growth regulator. The aim of this study was to figure out the effect of melatonin on arbuscular mycorrhizal (AM) symbiosis and heavy metal tolerance. A three-factor experiment was conducted to determine the effect of melatonin application on the growth, AM symbiosis, and stress tolerance of Medicago truncatula. A two-factor (AM inoculation and Pb stress) experiment was conducted to determine the effect of AM fungus on melatonin accumulation under Pb stress. AM plants under Pb stress had a higher melatonin accumulation than non-mycorrhizal (NM) plants under Pb stress. Acetylserotonin methyltransferase (ASMT) is the enzymatic reaction of the last step in melatonin synthesis. The accumulation of melatonin may be related to the expression of MtASMT. Melatonin application increased the relative expression of MtPT4 and AM colonization in AM plants. Melatonin application decreased Pb uptake with and without AM inoculation. Both melatonin application and AM inoculation improved M. truncatula growth and increased antioxidant response with Pb stress. These results indicated that melatonin application has positive effects on AM symbiosis and Pb stress tolerance under Pb stress. AM inoculation improve melatonin synthesis capacity under Pb stress. Melatonin application may improve AM plant growth by enhancing AM symbiosis, stimulating antioxidant response, and inhibiting Pb uptake.

Root microbiota have a significant effect on plant health. However, the role of root microbiota in the resistance of Rhododendron against root rot is not known. In this study, we employed amplicon 16S and ITS sequencing to investigate the bacterial and fungal communities associated with four distinct niches (bulk soil, rhizosphere, rhizoplane, and endosphere) of both healthy and diseased Rhododendron plants in the Baili Rhododendron nature reserve in China. The amplicon data analysis identified 182 bacterial genera and 141 fungal genera that were impacted by root rot across all niches. Specifically, the rhizoplane appeared to exert a selective gating effect, resulting in a reduction in the complexity of bacterial communities, but not fungal communities, in wild Rhododendron delavayi Franch roots. Nevertheless, the stress induced by root rot led to alterations in the root microbiota and compromised the gating function of the rhizoplane, thereby significantly increasing the complexity of the bacterial community within the plant root. In the root tissue following root rot outbreak, the relative abundance of the pathogenic species Pezicula brunnea and Diaporthe helianthi was enriched by as much as 6.13% and 1.71%, respectively. These findings provide novel insights into the contribution of enrichment of root-associated microbiota to wild plant hosts under the disease stress of root rot. The root rot-causing pathogenic fungi may interact with beneficial bacteria and induce plants to send out "cry for help" signals, which may encourage the specific assembly of microbiota. In the Rhododendron delavayi Franch root microbiota, we found 23 potentially beneficial microbes. Notably, certain beneficial bacteria, such as Sporolactobacillus and Stenotrophomonas, were found to accumulate in the rhizoplane and endosphere under root rot disease stress. Overall, our results lend support to our hypothesis that Rhododendron recruits protective microbes as a strategy to suppress root rot outbreaks. Future endeavors in isolating beneficial microbes capable of mitigating root rot have the potential to enhance plant resilience against root diseases.

The association of plants and microbial communities is crucial for crop production, and host plants influence the composition of rhizosphere microbiomes. Pulse crops play an important role in the development of sustainable cropping systems, and producers in the Canadian prairies often increase the frequency of pulses in their cropping systems. In this study, we determined the shifts in the fungal community of pea (Pisum sativum L.) rhizosphere, as influenced by the frequency of pulses in rotation, using high throughput sequencing. Six cropping systems containing pea (P), lentil (Lens culinaris Medik., L), hybrid canola (Brassica napus L., C), wheat (Triticum aestivum L., W), and oat (Avena sativa L., O) in different intensities were tested. The fungal communities were assessed at the flowering stage in the fourth and fifth year of the 4-year rotations. Cropping system had a significant impact on the composition of the rhizosphere fungal community, and the effect of crop rotation sequence was greater and explained more of the variation than the effect of previous crops. The rotation with consecutive pulses (WPLP) decreased fungal evenness and increased the proportion of pathotrophs. Fusarium was a dominant and ubiquitous pathotrophic genus. Olpidium virulentus, Botrytis cinerea, Fusarium solani, F. graminearum, and Alternaria eichhorniae were generally more abundant in pulse intensive rotations (WPLP, WLOP, and WPOP), the exception being F. solani which was not promoted by lentil. Reads of O. virulentus and B. cinerea were most abundant in pea preceded by lentil followed by the reads of Mortierella elongata in pea preceded by wheat. Pea consistently had higher grain yield when grown in diversified rotations including wheat, canola/lentil, and oat than rotations with two repeated crops (canola or pea). Cropping system affected the soil physicochemical properties, and soil pH was the main driver of fungal community shift. No evidence of beneficial microorganisms involvement in plant productivity was observed, but the high abundance of pathotrophs in pulse intensified rotations suggests the possibility of pathogen buildup in the soil with increasing pulse frequency. Diversifying rotation sequences minimized disease risk and increased pea production, in this study. Careful selection of plant species appears as a strategy for the management of rhizosphere fungal communities and the maintenance of crop production system's health.

Plants and other organisms in the ecosystem compete for the limited nitrogen (N) in the soil. Formation of a symbiotic relationship with arbuscular mycorrhizal fungi (AMF) may influence plant competitiveness for N. However, the effects of AMF on plant nitrate (NO3 -) uptake capacity remain unknown. In this study, a pot experiment was conducted to investigate the effects of N application and Rhizophagus irregularis inoculation on the root absorbing area, uptake kinetics of NO3 -, and the expression of NO3 - transporter (NRT) genes in Populus × canadensis 'Neva'. The results showed that R. irregularis colonized more than 70% of the roots of the poplar and increased root active absorbing area/total absorbing area. The uptake kinetics of NO3 - by poplar fitted the Michaelis-Menten equation. Mycorrhizal plants had a higher maximum uptake rate (V max) value than non-mycorrhizal plants, indicating that R. irregularis enhanced the NO3 - uptake capacity of poplar. The expression of NRTs in roots, namely, NRT1;2, NRT2;4B, NRT2;4C, NRT3;1A, NRT3;1B, and NRT3;1C, was decreased by R. irregularis under conditions of 0 and 1 mM NH4NO3. This study demonstrated that the improved NO3 - uptake capacity by R. irregularis was not achieved by up-regulating the expression of NRTs in roots. The mycorrhizal pathway might repress root direct pathway in the NO3 - uptake by mycorrhizal plants.

In terrestrial ecosystems, mycorrhizal roots play a key role in the cycling of soil carbon (C) and other nutrients. The impact of environmental factors on the mycorrhizal fungal community has been well studied; however, the seasonal variations in the root-associated fungal microbiota affected by environmental changes are less clear. To improve the understanding of how environmental factors shape the fungal microbiota in mycorrhizal roots, seasonal changes in Pinus tabuliformis root-associated fungi were investigated. In the present study, the seasonal dynamics of edaphic properties, soil enzymatic activities, root fungal colonization rates, and root-associated fungal microbiota in P. tabuliformis forests were studied across four seasons during a whole year to reveal their correlations with environmental changes. The results indicate that the soil functions, such as the enzymatic activities related to nitrogen (N) and phosphorus (P) degradation, were varied with the seasonal changes in microclimate factors, resulting in a significant fluctuation of edaphic properties. In addition, the ectomycorrhizal fungal colonization rate in the host pine tree roots increased during warm seasons (summer and autumn), while the fungal colonization rate of dark septate endophyte was declined. Moreover, the present study indicates that the fungal biomass increased in both the pine roots and rhizospheric soils during warm seasons, while the fungal species richness and diversity decreased. While the Basidiomycota and Ascomycota were the two dominant phyla in both root and soil fungal communities, the higher relative abundance of Basidiomycota taxa presented in warm seasons. In addition, the fungal microbial network complexity declined under the higher temperature and humidity conditions. The present study illustrates that the varieties in connectivity between the microbial networks and in functional taxa of root-associated fungal microbiota significantly influence the soil ecosystem functions, especially the N and P cycling.

Pine wilt disease (PWD) is a deadly disease to pines (Pinus spp.) worldwide. The occurrence of PWD can reduce the relative abundance of root ectomycorrhizal fungi (ECMF) and dark septate endophytes (DSE). However, the effects of exogenous ECMF/DSE inoculation on the rhizosphere microbial community structure of Pinus tabulaeformis infected by pine wood nematode (PWN) is little known. Here, we tested how ECMF/DSE may improve resistance to PWD by quantifying microbial carbon biomass and soil enzymatic activity among different treatments at 6 and 9 months after PWN infection. Denaturing gradient gel electrophoresis (DGGE) was used to study the microbial community structure at 3, 6, and 9 months after PWN infection in the rhizosphere of P. tabulaeformis seedlings inoculated with ECMF/DSE. The results showed that exogenous ECMF/DSE inoculation reduced the disease severity caused by PWN infection. After PWN infection, the rhizosphere microbial carbon of seedlings inoculated with Amanita vaginata, Suillus bovinus, Gaeumannomyces cylindrosporus, and Paraphoma chrysanthemicola was 38.16, 49.67, 42.11, and 96.05% higher than that of the control group, respectively. Inoculation of ECMF/DSE inhibited the decrease of rhizosphere microbial biomass caused by PWN infection. The richness and diversity of P. tabulaeformis rhizosphere fungi at 9 months were reduced by PWN infection but partially recovered by the exogenous fungi (ECMF/DSE) inoculation except for P. chrysanthemicola, which indicates a role of ECMF/DSE in maintaining stability of the microbial community. Inoculation with ECMF/DSE increased the beneficial bacterial (Thauera sp., Mesorhizobium sp., etc.) and fungal groups (Tomentella ellisii, Wilcoxina mikolae, etc.) of in the rhizosphere. In summary, exogenous ECMF/DSE inoculation could increase P. tabulaeformis resistance to PWD probably by improving the rhizosphere microenvironment.