Rhizosphere Microbiome Research Articles

Multigenerational Adaptation Can Enhance the Pathogen Resistance of Plants via Changes in Rhizosphere Microbial Community Assembly.

Plants withstand pathogen attacks by recruiting beneficial bacteria to the rhizosphere and passing their legacy on to the next generation. However, the underlying mechanisms involved in this process remain unclear. In our study, we combined microbiomic and transcriptomic analyses to reveal how the rhizosphere microbiome assembled through multiple generations and defense-related genes expressed in Arabidopsis thaliana under pathogen attack stress. Our results showed that continuous exposure to the pathogen Pseudomonas syringae pv tomato DC3000 led to improved growth and increased disease resistance in a third generation of rps2 mutant Arabidopsis thaliana. It could be attributed to the enrichment of specific rhizosphere bacteria, such as Bacillus and Bacteroides. Pathways associated with plant immunity and growth in A. thaliana, such as MAPK signaling pathways, phytohormone signal transduction, ABC transporter proteins, and flavonoid biosynthesis, were activated under the influence of rhizosphere bacterial communities. Our findings provide a scientific basis for explaining the relationship between beneficial microbes and defense-related gene expression. Understanding microbial communities and the mechanisms involved in plant responses to disease can contribute to better plant management and reduction of pesticide use.

Plant-guided microbiome selection produces transient effects on insect performance and rhizosphere community assembly

Microbial communities are essential for plant health, but using these relationships to enhance growth and pest protection is challenging. Leveraging the natural mechanisms plants employ to manage relationships with microbes is one promising means to selectively engineer whole microbial communities with beneficial properties. This approach, known as host-guided selection, has been successful in some model species targeting performance traits (e.g., biomass). However, few studies use crop plants or focus on defensive traits (e.g., pest resistance). Our goal was to naturally engineer tomato root-associated microbiomes that increase resistance to insect pests. First, we used an iterative soil microbial inoculation process to engineer insect-suppressive rhizosphere soil microbiomes that reduce damage from aphid feeding (Macrosiphum euphorbiae) or caterpillar defoliation (Manduca sexta, Spodoptera exigua). We then characterized the bacterial and fungal microbial communities of soils associated with differences in insect performance using metabarcoding approaches. Overall, soil microbiome selection produced transient differences in aphid performance, but caterpillar growth was unaffected. In 4 of 9 generations, aphid population growth rate was significantly less on plants with rhizospheres selected for “Low” insect performance, where abundance was reduced by up to 20%. Correspondingly minor shifts in fungal and bacterial relative abundance occurred in insect-suppressive communities. However, network analysis indicated aphid feeding disrupted rhizosphere microbiome assembly, resulting in lower community complexity and connectivity, and fewer structurally important taxa compared to un-infested controls. Overall, our results highlight critical factors for successful engineering of beneficial microbiomes, particularly insect feeding guild and microbial community stability.

Climate change and plant rhizosphere microbiomes: an experiential course-embedded research project.

The current and ongoing challenges brought on by climate change will require future scientists who have hands-on experience using advanced molecular techniques, can work with large data sets, and can make correlations between metadata and microbial diversity. A course-embedded research project can prepare students to answer complex research questions that might help plants adapt to climate change. The project described herein uses plants as a host to study the impact of climate change-induced drought on host-microbe interactions through next-generation DNA sequencing and analysis using a command-line program. Specifically, the project studies the impact of simulated drought on the rhizosphere microbiome of Fast Plants rapid cycling Brassica rapa using inexpensive greenhouse supplies and 16S rRNA V3/V4 Illumina sequencing. Data analysis is performed with the freely accessible Python-based microbiome bioinformatics platform QIIME 2.

Intercropping enhances maize growth and nutrient uptake by driving the link between rhizosphere metabolites and microbiomes.

Intercropping leads to different plant roots directly influencing belowground processes and has gained interest for its promotion of increased crop yields and resource utilization. However, the precise mechanisms through which the interactions between rhizosphere metabolites and the microbiome contribute to plant production remain ambiguous, thus impeding the understanding of the yield-enhancing advantages of intercropping. This study conducted field experiments (initiated in 2013) and pot experiments, coupled with multi-omics analysis, to investigate plant-metabolite-microbiome interactions in the rhizosphere of maize. Field-based data revealed significant differences in metabolite and microbiome profiles between the rhizosphere soils of maize monoculture and intercropping. In particular, intercropping soils exhibited higher microbial diversity and metabolite chemodiversity. The chemodiversity and composition of rhizosphere metabolites were significantly related to the diversity, community composition, and network complexity of soil microbiomes, and this relationship further impacted plant nutrient uptake. Pot-based findings demonstrated that the exogenous application of a metabolic mixture comprising key components enriched by intercropping (soyasapogenol B, 6-hydroxynicotinic acid, lycorine, shikimic acid, and phosphocreatine) significantly enhanced root activity, nutrient content, and biomass of maize in natural soil, but not in sterilized soil. Overall, this study emphasized the significance of rhizosphere metabolite-microbe interactions in enhancing yields in intercropping systems. It can provide new insights into rhizosphere controls within intensive agroecosystems, aiming to enhance crop production and ecosystem services.

Structure, acquisition, assembly, and function of the root-associated microbiomes in Japonica rice and hybrid rice

Understanding the genetic and environmental factors that structure root-associated microbiomes is necessary to inform rice (Oryza sativa L.) breeding and fertilizer. Herein, we comprehensivly explored the microbiomes along four fine-scale root compartments, bulk soil, rhizosphere, rhizoplane and endosphere, between traditional Japonica rice and improved hybrid rice acorss three spatially distant locations. Bacterial diversity decreased monotonically from external bulk soil to internal endosphere. The acquisition of endosphere symbionts is governed conjointly by a two-step selection process and an amplification selection process, which depends on the bacterial functional group. The bacterial communities are primarly affected by root compartment and planting location, followed by relatively samll but signifcant rice genotype effect. The exterior bulk soil, rhizosphere and rhizoplane communities are govrned by variable selection. In contrast, the internal endosphere counterparts are shapped by homogenizing selection, which are undiscernible between the two rice genotypes. After ruling out planting location effect, we screened 12 rice genotype-discriminatory orders that contributed an overall 97.22 % accuracy in diagnosing rice genotype. Hybrid rice is inferior to Japonica rice in carbon and nitrogen fixation, nitrogen and phosphorus biomineralization from organic sources. Collectively, we proposed a conceptual model to infer rice genotype effect on the rhizosphere microbiomes, and the subsequent requirement of external fertilizer for sustaining yield. Our findings deepen our understanding of the complex rice genotype–root microbiomes–fertilizer interactions from the prespective of microbially-mediated nutrient cycles.

The rhizosphere microbiome of 51 potato cultivars with diverse plant growth characteristics.

Rhizosphere microbial communities play a substantial role in plant productivity. We studied the rhizosphere bacteria and fungi of 51 distinct potato cultivars grown under similar greenhouse conditions using a metabarcoding approach. As expected, individual cultivars were the most important determining factor of the rhizosphere microbial composition; however, differences were also obtained when grouping cultivars according to their growth characteristics. We showed that plant growth characteristics were related to deterministic and stochastic assembly processes of bacterial and fungal communities, respectively. The bacterial genera Arthrobacter and Massilia (known to produce indole acetic acid and siderophores) exhibited greater relative abundance in high- and medium-performing cultivars. Bacterial co-occurrence networks were larger in the rhizosphere of these cultivars and were characterized by a distinctive combination of plant beneficial Proteobacteria and Actinobacteria along with a module of diazotrophs namely Azospira, Azoarcus, and Azohydromonas. Conversely, the network within low-performing cultivars revealed the lowest nodes, hub taxa, edges density, robustness, and the highest average path length resulting in reduced microbial associations, which may potentially limit their effectiveness in promoting plant growth. Our findings established a clear pattern between plant productivity and the rhizosphere microbiome composition and structure for the investigated potato cultivars, offering insights for future management practices.

Rhizosphere metagenomics provides insights into the environmental effect on the secondary metabolism of Ligusticum chuanxiong

Rhizosphere microbiomes influence the secondary metabolism of plants, but the mechanism has not been analyzed. This study aimed to explore relationship between abiotic environments, microbiomes and secondary metabolites of Ligusticum chuanxiong. By combining metagenomic, metatranscriptomic, and high-performance liquid chromatography, the microbial community structure and key microbiomes in Ligusticum chuanxiong were identified. Furthermore, the relationship between root-associated microbiomes and the secondary metabolites in Ligusticum chuanxiong was revealed. The microbial community structures of L. chuanxiong differed in each individual. The key microbiome related to secondary metabolism were similar, while there were significant differences in relative abundance across all samples. Four families (Burkholderiaceae, Comamonadaceae, Pseudomonadaceae, and Streptomycetaceae) and four species (Gemmatirosa kalamazoonesis, Luteitalea pratensis, Candidatus Solibacter usitatus, and Sorangium cellulosum) were identified as conservative core microbes, and Chloroflexi was the main biomarker. Functional annotation results showed that the metabolic function strength of the microbial community was positively correlated with the secondary metabolite content. According to correlation results, rhizosphere microbiomes related with secondary metabolites in two aspects including community diversity and microbial abundance. In conclusion, environmental factors might affect secondary metabolites of Ligusticum chuanxiong Hort. by influencing microbial diversity and the abundance of key microbes. This study can provide references for improving the artificial cultivation of Ligusticum chuanxiong, further developing and utilizing this medicinal resource.

Altitude-dependent influence of Artemisia brevifolia on its rhizosphere microbiome in Ladakh region of the Western Himalayas

Altitude-induced variations significantly influence plant physiology, shaping the diversity and composition of prokaryotic and fungal communities in the rhizosphere soil. This study investigates the impact of altitude on the rhizosphere microbiome of Artemisia brevifolia in the Ladakh region of the Western Himalayas. Alpha diversity indices reveal a decline in both prokaryotic and fungal diversity in rhizosphere soil compared to bulk soil, with a more pronounced effect on prokaryotic communities at different altitudes. Altitude variation significantly impacts prokaryotic and fungal beta-diversity, leading to distinct microbial communities at different elevations. Notably, the rhizosphere communities at higher altitudes exhibit little overlap with bulk soil communities, indicating a unique microbial assemblage possibly in response to harsh environmental conditions. The dominant prokaryotic phyla include Actinobacteriota, Proteobacteria, Chloroflexi, and Acidobacteriota, while the fungal communities are dominated by Ascomycota and Basidiomycota. A significantly higher frequency of rhizobia was noted in the A. brevifolia rhizosphere at both locations, with the genus Rhizobium prevalent at lower altitudes and Mesorhizobium at the higher altitudes. Predicted functions of prokaryotic communities revealed a higher abundance of pathways associated with the biosynthesis of siderophores, secondary metabolites and aromatic-compounds, and denitrification in the rhizosphere. Additionally, a higher abundance of gene clusters associated with nitrogen fixation and phosphorus metabolisms was noted in rhizosphere particularly at higher elevations.In summary, this comprehensive study highlights the unique structure and function of microbial communities along increasing elevations, particularly in the rhizosphere, potentially aiding the host plant in thriving under the stressful conditions of high altitudes in the climate-sensitive Himalayas.

Evaluation of the Rhizosphere Resistome of Cultivated Soils Polluted with Antibiotics from Reclaimed Wastewater

The use of reclaimed wastewater to irrigate crops is a valuable option due to water scarcity. However, the presence of antibiotics residues that are not removed in the tertiary treatments reaches crop soils when irrigated and it poses a serious concern for human health. Crops rhizosphere is considered a hotspot of antibiotic resistant genes (ARG) being in addition a link to plant phyllosphere and human microbiome. Understanding the structure of the soil microbiota is crucial before applying any bioremediation or biostimulation strategy. The aim of this work was firstly to confirm the presence of antibiotics residues in soil and fruits in two greenhouses at the south of Spain irrigated with reclaimed water. Secondly, to characterize the rhizosphere microbiome of three crops (Capsicum annuum, Cucumis melo and Solanum melongena) cultured in those greenhouses. Finally, a predictive functional analysis was done using PICRUSt2 to figure out the rhizosphere resistome. The presence of residues of antibiotics was confirmed both soil and fruits. Antibiotics absorbed by plants correlated with those on soil. The most abundant resistance gene was the multidrug in all the three crops tested. Compatibility of basidiomycete fungi (i.e., Pleurotus) with soil bacteria could be indicative of their possible use for restoration of agricultural soils polluted with antibiotics.

Assembly patterns and key taxa of bacterial communities in the rhizosphere soil of moso bamboo (Phyllostachys pubescens) under different Cd and Pb pollution

Moso bamboo is excellent candidate for cadmium (Cd)/lead (Pb) phytoremediation, while rhizosphere microbiome has significant impact on phytoremediation efficiency of host plant. However, little is known about the rhizosphere bacterial communities of moso bamboo in Cd/Pb contaminated soils. Therefore, this study investigated the assembly patterns and key taxa of rhizosphere bacterial communities of moso bamboo in Cd/Pb polluted and unpolluted soils, by field sampling, chemical analysis, and 16S rRNA gene sequencing. The results indicated α-diversity between Cd/Pb polluted and unpolluted soils showed a similar pattern (p > 0.05), while β-diversity was significantly different (p < 0.05). The relative abundance analysis indicated α-proteobacteria (37%) and actinobacteria (31%) were dominant in Cd/Pb polluted soils, while γ-proteobacteria (40%) and α-proteobacteria (22%) were dominant in unpolluted soils. Co-occurrence network analysis indicated microbial networks were less complex and more negative in polluted soils than in unpolluted soils. Mantel analysis indicated soil available phosphorus, organic matter, and available Pb were the most important environmental factors affecting microbial community structure. Correlation analysis showed 11 bacterial genera were significantly positively related to Cd/Pb. Overall, this study identified the bacterial community composition of bamboo rhizosphere in responding to Cd/Pb contamination and provides a theoretical basis for microbe-assistant phytoremediation in the future.

Developing stable, simplified, functional consortia from Brachypodium rhizosphere for microbial application in sustainable agriculture

The rhizosphere microbiome plays a crucial role in supporting plant productivity and ecosystem functioning by regulating nutrient cycling, soil integrity, and carbon storage. However, deciphering the intricate interplay between microbial relationships within the rhizosphere is challenging due to the overwhelming taxonomic and functional diversity. Here we present our systematic design framework built on microbial colocalization and microbial interaction, toward successful assembly of multiple rhizosphere-derived Reduced Complexity Consortia (RCC). We enriched co-localized microbes from Brachypodium roots grown in field soil with carbon substrates mimicking Brachypodium root exudates, generating 768 enrichments. By transferring the enrichments every 3 or 7 days for 10 generations, we developed both fast and slow-growing reduced complexity microbial communities. Most carbon substrates led to highly stable RCC just after a few transfers. 16S rRNA gene amplicon analysis revealed distinct community compositions based on inoculum and carbon source, with complex carbon enriching slow growing yet functionally important soil taxa like Acidobacteria and Verrucomicrobia. Network analysis showed that microbial consortia, whether differentiated by growth rate (fast vs. slow) or by succession (across generations), had significantly different network centralities. Besides, the keystone taxa identified within these networks belong to genera with plant growth-promoting traits, underscoring their critical function in shaping rhizospheric microbiome networks. Furthermore, tested consortia demonstrated high stability and reproducibility, assuring successful revival from glycerol stocks for long-term viability and use. Our study represents a significant step toward developing a framework for assembling rhizosphere consortia based on microbial colocalization and interaction, with future implications for sustainable agriculture and environmental management.

Impact of Rhizospheric Microbiome on Rice Cultivation.

The rhizosphere niche is extremely important for the overall growth and development of plants. Evidently, it is necessary to understand the complete mechanism of plant microbe interactions of the rhizosphere for sustainable and low input productivity. To meet the increasing global food demand, rice (Oryza sativa L.) agriculture seeks optimal conditions. The unique oxic-anoxic interface of rice-growing soil has invited divergent microbes with dynamic biogeochemical cycles. This review provides the systematic analysis of microbes associated with the major biogeochemical cycles with the aim to generate better management strategies of rhizospheric microbiome in the field of rice agriculture. For instance, several methanogenic and methanotrophic bacteria in the rice rhizosphere make an equilibrium for methane concentration in the environment. The carbon sequestration in paddy soil is again done through many rhizospheric microorganisms that can directly assimilate CO2 with their photoautotrophic mode of nutrition. Also the phosphate solubilizing microbes remain to be the most important keys for the PGPR activity of the paddy ecosystem. In addition, rhizospheric microbiome remain crucial in degradation and solubilization of organo-sulfur and insoluble inorganic sulfides which can be taken by the plants. Further, this review elucidates on the advantages of using metagenomic and metaproteomic approaches as an alternative of traditional approaches to understand the overall metabolic pathways operational in paddy-field. These knowledges are expected to open new possibilities for designing the balanced microbiome used as inoculum for intensive farming and will eventually lead to exert positive impacts on rice cultivation.

Phages enhance both phytopathogen density control and rhizosphere microbiome suppressiveness.

Ralstonia solanacearum is a highly destructive plant-pathogenic bacterium with the ability to cause bacterial wilt in several crucial crop plants. Given the limitations of conventional chemical control methods, the use of bacterial viruses (phages) has been explored as an alternative biological control strategy. In this study, we show that increasing the phage application frequency can improve the density control of R. solanacearum, leading to a significant reduction in bacterial wilt disease. Furthermore, we found that repeated phage application increased the diversity of rhizosphere microbiota and specifically enriched Actinobacterial taxa that showed synergistic pathogen suppression when combined with phages due to resource and interference competition. Together, our study unravels an undiscovered benefit of phages, where phages trigger a second line of defense by the pathogen-suppressing bacteria present in resident microbial communities. Phage therapies could, hence, potentially be tailored according to host microbiota composition to unlock the pre-existing benefits provided by resident microbiota.

Effects of Slope Position on the Rhizosphere and Fine Root Microbiomes of Cupressus gigantea on the Tibet Plateau, China

Effects of Slope Position on the Rhizosphere and Fine Root Microbiomes of Cupressus gigantea on the Tibet Plateau, China

Climate–resilient strategy for shortening the crop cycle in rice and the host influenced rhizosphere microbiome

We present the field rapid generation advancement procedure for shortening the crop cycle in rice. The practical protocol developed by us using different maturity groups of rice showed promising early flower induction. We observed a shortening of crop cycle to about 35–40 days depending on the variety. The spacing between the plants was the major influencer for flower induction compared to other interventions tested viz., clipping, potassium di-hydrogen phosphate and paclobutrazol spray. Our raised bed direct seeding strategy completely avoided the transplantation. The work flow for flower induction can be a reference to increase generations of rice breeding. The rhizosphere bacterial dynamics using 16srRNA gene amplicon sequencing showed the Acinetobacter population abundance as a key mediator and marker for flowering time. The alpha diversity at flowered and unflowered stage showed the species richness as Control < Pusa Sugandh 5 < BPT-5204. The rice rhizosphere of this ecosystem had an abundance of Methylotrophs that utilize methane as a carbon source suggesting that the developed method is a green technique suitable for generation advancement that can be replaced for flooded condition assisted breeding in rice. Selective enrichment of functional abundance between varieties suggested the host - influenced microbiome for early flowering in rice. This protocol is expected to greatly accelerate the process of new variety breeding and the construction of mapping populations.

Rice rhizosphere microbiome is more diverse but less variable along environmental gradients compared to bulk soil

Rice rhizosphere microbiome is more diverse but less variable along environmental gradients compared to bulk soil

Genetic variation in Zea mays influences microbial nitrification and denitrification in conventional agroecosystems

Abstract Background and Aims Nitrogenous fertilizers provide a short-lived benefit to crops in agroecosystems, but stimulate nitrification and denitrification, processes that result in nitrate pollution, N2O production, and reduced soil fertility. Recent advances in plant microbiome science suggest that genetic variation in plants can modulate the composition and activity of rhizosphere N-cycling microorganisms. Here we attempted to determine whether genetic variation exists in Zea mays for the ability to influence the rhizosphere nitrifier and denitrifier microbiome under “real-world” conventional agricultural conditions. Methods To capture an extensive amount of genetic diversity within maize we grew and sampled the rhizosphere microbiome of a diversity panel of germplasm that included ex-PVP inbreds (Z. mays ssp. mays), ex-PVP hybrids (Z. mays ssp. mays), and teosinte (Z. mays ssp. mexicana and Z. mays ssp. parviglumis). From these samples, we characterized the microbiome, a suite of microbial genes involved in nitrification and denitrification and carried out N-cycling potential assays. Results Here we are showing that populations/genotypes of a single species can vary in their ecological interaction with denitrifers and nitrifers. Some hybrid and teosinte genotypes supported microbial communities with lower potential nitrification and potential denitrification activity in the rhizosphere, while inbred genotypes stimulated/did not inhibit these N-cycling activities. These potential differences translated to functional differences in N2O fluxes, with teosinte plots producing less GHG than maize plots. Conclusion Taken together, these results suggest that Zea genetic variation can lead to changes in N-cycling processes that result in N leaching and N2O production, and thereby are selectable targets for crop improvement. Understanding the underlying genetic variation contributing to belowground microbiome N-cycling into our conventional agricultural system could be useful for sustainability.

The rhizosphere microbiome and its influence on the accumulation of metabolites in Bletilla striata (Thunb.) Reichb. f

BackgroundBletilla striata (Thunb.) Reichb. f. (B. striata) is a perennial herbaceous plant in the Orchidaceae family known for its diverse pharmacological activities, such as promoting wound healing, hemostasis, anti-inflammatory effects, antioxidant properties, and immune regulation. Nevertheless, the microbe-plant-metabolite regulation patterns for B. striata remain largely undetermined, especially in the field of rhizosphere microbes. To elucidate the interrelationships between soil physics and chemistry and rhizosphere microbes and metabolites, a comprehensive approach combining metagenome analysis and targeted metabolomics was employed to investigate the rhizosphere soil and tubers from four provinces and eight production areas in China.ResultsOur study reveals that the core rhizosphere microbiome of B. striata is predominantly comprised of Paraburkholderia, Methylibium, Bradyrhizobium, Chitinophaga, and Mycobacterium. These microbial species are recognized as potentially beneficial for plants health. Comprehensive analysis revealed a significant association between the accumulation of metabolites, such as militarine and polysaccharides in B. striata and the composition of rhizosphere microbes at the genus level. Furthermore, we found that the soil environment indirectly influenced the metabolite profile of B. striata by affecting the composition of rhizosphere microbes. Notably, our research identifies soil organic carbon as a primary driving factor influencing metabolite accumulation in B. striata.ConclusionOur fndings contribute to an enhanced understanding of the comprehensive regulatory mechanism involving microbe-plant-metabolite interactions. This research provides a theoretical basis for the cultivation of high-quality traditional Chinese medicine B. striata.

Role of Bacillus subtilis exopolymeric genes in modulating rhizosphere microbiome assembly

BackgroundBacillus subtilis is well known for promoting plant growth and reducing abiotic and biotic stresses. Mutant gene-defective models can be created to understand important traits associated with rhizosphere fitness. This study aimed to analyze the role of exopolymeric genes in modulating tomato rhizosphere microbiome assembly under a gradient of soil microbiome diversities using the B. subtilis wild-type strain UD1022 and its corresponding mutant strain UD1022eps−TasA, which is defective in exopolysaccharide (EPS) and TasA protein production.ResultsqPCR revealed that the B. subtilis UD1022eps−TasA− strain has a diminished capacity to colonize tomato roots in soils with diluted microbial diversity. The analysis of bacterial β-diversity revealed significant differences in bacterial and fungal community structures following inoculation with either the wild-type or mutant B. subtilis strains. The Verrucomicrobiota, Patescibacteria, and Nitrospirota phyla were more enriched with the wild-type strain inoculation than with the mutant inoculation. Co-occurrence analysis revealed that when the mutant was inoculated in tomato, the rhizosphere microbial community exhibited a lower level of modularity, fewer nodes, and fewer communities compared to communities inoculated with wild-type B. subtilis.ConclusionThis study advances our understanding of the EPS and TasA genes, which are not only important for root colonization but also play a significant role in shaping rhizosphere microbiome assembly. Future research should concentrate on specific microbiome genetic traits and their implications for rhizosphere colonization, coupled with rhizosphere microbiome modulation. These efforts will be crucial for optimizing PGPR-based approaches in agriculture.

Bacillus velezensis B63 and chitosan control root rot, improve growth and alter the rhizosphere microbiome of geranium

The root rot complex of geranium plants caused by Rhizoctonia solani and Macrophomina phaseolina is a major threat, and control of these pathogens predominantly relies on chemicals. This study explored multifaceted applications of Bacillus velezensis (strain B63) and chitosan, assessing their biocontrol efficacy against root rot, and their subsequent effects on rhizosphere communities. Strain B63 was antagonistic to R. solani and M. phaseolina. Under field conditions, greatest efficacy was obtained with strain B63 (36% and 33% disease reductions in, respectively, two growing seasons), chitosan soaking + foliar spray 0.2% (CSF 0.2%) (33 and 27% reductions), and 0.1% chitosan soaking + foliar spray (CSF 0.1%) (33 and 26% reductions). These treatments also changed rhizosphere microbiota, as shown by numbers of colony-forming units (CFU) and 16S rRNA gene microbiome analyses. Concomitant with rhizosphere shifts, essential oil yields and composition were positively affected, as shown by gas chromatography analyses. Chitosan soaking + foliar spray 0.2% increased concentrations of citronellol (1.36-fold), geraniol (1.37-fold), citronellyl formate (1.54-fold), and geranyl formate (1.94-fold) in geranium essential oil, compared with the experimental controls. Strain B63 also increased these essential oils by 1.04- to 1.27-fold. B63 also enhanced eugenol levels by 1.35-fold. Treatments with B63 were more effective than chitosan in improving the geranium plant morphological parameters (plant height, numbers of branches, biomass). These results show that B. velezensis strain B63 treatments have potential for enhancing yields and product quality from geranium plant under root rot infection.