Plant-microbe Interactions Research Articles

Light availability and plant shade tolerance modify plant-microbial interactions and feedbacks in subtropical trees.

Plant-soil feedbacks (PSFs) are an important mechanism of species coexistence in forest communities. However, evidence remains limited for how light availability regulates PSFs in species with different shade tolerance via changes in plant-microbial interactions. Here we tested in a glasshouse experiment how PSFs changed as a function of light availability and tree shade tolerance. Soil bacterial and fungal communities were profiled using the 16S rRNA and ITS2 gene sequencing, respectively. Under low light, individual PSFs were positively related to shade tolerance, while the least shade-tolerant species produced the most positive PSFs under high light. Pairwise PSFs between species with contrasting shade tolerance were strongly positive under high light but negative under low light, thereby promoting the dominance of less shade-tolerant species in forest gaps and species coexistence under closed canopy, respectively. Under high light, PSFs were related to soil microbial composition and diversity, with the relative abundance of arbuscular mycorrhizal fungi being the primary driver of PSFs. Under low light, none of soil microbial properties were significantly related to PSFs. These findings indicate PSFs and plant shade tolerance interact to promote species coexistence and improve our understanding of how soil microbes contribute to variation in PSFs.

Transcriptome and metabolome analyses reveal that Bacillus subtilis BS-Z15 lipopeptides mycosubtilin homologue mediates plant defense responses.

Microbial-plant interactions protect plants from external stimuli, releasing various elicitor that activate the plants defense response and regulate its growth. Bacillus subtilis BS-Z15 was screened from cotton inter-rhizosphere soil, antagonized various plant pathogens, and protected cotton against Verticillium dahliae. This study showed that the BS-Z15 lipopeptide mycosubtilin homologue could act as an elicitor to induce systemic resistance (ISR) in plants. Mycosubtilin homologue induced ROS burst and deposition, callose deposition, MAPK cascade phosphorylation, and up-regulated PR1 and PDF1.2 gene expression in Arabidopsis seedlings, moreover enhanced resistance of Arabidopsis to Pseudomonas syringae pv. Tomato DC3000 (Pst DC3000) and V. dahliae. Transcriptome analysis was then used to evaluate the impact of mycosubtilin homologue on plant gene expression control. Mycosubtilin homologues activated Arabidopsis ISR on genes in metabolic pathways such as Arabidopsis plant-pathogen interactions, phenylpropanoid biosynthesis, MAPK signaling pathway, and phytohormone signaling. These analyses revealed that mycosubtilin homologues mediate the regulation of plant systemic resistance and growth and development by affecting related metabolites in glycolysis and gluconeogenesis, pentose phosphate pathway, tricarboxylic acid cycle, and amino acid metabolism in Arabidopsis. These findings confirmed that a mycosubtilin homologue could trigger the initiation of the Arabidopsis ISR by interacting with a variety of PTI components and transcriptional metabolic signaling pathways.

Transcriptome Landscapes of Salt-Susceptible Rice Cultivar IR29 Associated with a Plant Growth Promoting Endophytic Streptomyces

Plant growth-promoting endophytic (PGPE) actinomycetes have been known to enhance plant growth and mitigate plant from abiotic stresses via their PGP-traits. In this study, PGPE Streptomyces sp. GKU 895 promoted growth and alleviated salt tolerance of salt-susceptible rice cultivar IR29 by augmentation of plant weight and declined ROS after irrigation with 150 mM NaCl in a pot experiment. Transcriptome analysis of IR29 exposed to the combination of strain GKU 895 and salinity demonstrated up and downregulated differentially expressed genes (DEGs) classified by gene ontology and plant reactome. Streptomyces sp. GKU 895 induced changes in expression of rice genes including transcription factors under salt treatment which involved in growth and development, photosynthesis, plant hormones, ROS scavenging, ion transport and homeostasis, and plant–microbe interactions regarding pathogenesis- and symbiosis-related proteins. Taken together, these data demonstrate that PGPE Streptomyces sp. GKU 895 colonized and enhanced growth of rice IR29 and triggered salt tolerance phenotype. Our findings suggest that utilisation of beneficial endophytes in the saline fields could allow for the use of such marginal soils for growing rice and possibly other crops.

PGPR Promotes the Recovery of Submerged Macrophytes via Indigenous Microbiome Modulations under Combined Abiotic Stress

The restoration of submerged macrophytes is commonly limited by abiotic stress. Here, we isolated PGPR strains from the rhizosphere of submerged macrophytes using ACC deaminase selective medium, and evaluated their growth promoting effects on Vallisneria natans (V. natans) under low light intensity and (or) high sediment organic matter load, and also explored the indigenous microbiome response of V. natans seedlings to PGPR inoculants. Ten isolates were chosen from the 252 isolated strains based on the ACC deaminase activity and the production of IAA and siderophore. M1 (Pseudomonas vancouverensis) and E15 (Enterobacter ludwigii) had the best growth promoting effects under low light stress and under double stress of low light and high sediment organic matter load, and the shoot height increased by 36% and 46%, respectively. The results of indigenous microbiome analysis showed that PGPR inoculants could regulate the relative abundance of unclassified_f_Enterobacteriaceae and improve the α-diversity of the rhizosphere bacterial community. Under high sediment organic matter load, inoculation of PGPR obviously shifted the β-diversity of rhizosphere bacterial communities to promote the early growth of V. natans. This study expands the application of plant–microbe interaction in the field of freshwater ecological restoration.

Computational models for prediction of protein-protein interaction in rice and Magnaporthe grisea.

Plant-microbe interactions play a vital role in the development of strategies to manage pathogen-induced destructive diseases that cause enormous crop losses every year. Rice blast is one of the severe diseases to rice Oryza sativa (O. sativa) due to Magnaporthe grisea (M. grisea) fungus. Protein-protein interaction (PPI) between rice and fungus plays a key role in causing rice blast disease. In this paper, four genomic information-based models such as (i) the interolog, (ii) the domain, (iii) the gene ontology, and (iv) the phylogenetic-based model are developed for predicting the interaction between O. sativa and M. grisea in a whole-genome scale. A total of 59,430 interacting pairs between 1,801 rice proteins and 135 blast fungus proteins are obtained from the four models. Furthermore, a machine learning model is developed to assess the predicted interactions. Using composition-based amino acid composition (AAC) and conjoint triad (CT) features, an accuracy of 88% and 89% is achieved, respectively. When tested on the experimental dataset, the CT feature provides the highest accuracy of 95%. Furthermore, the specificity of the model is verified with other pathogen-host datasets where less accuracy is obtained, which confirmed that the model is specific to O. sativa and M. grisea. Understanding the molecular processes behind rice resistance to blast fungus begins with the identification of PPIs, and these predicted PPIs will be useful for drug design in the plant science community.

Natural rubber reduces herbivory and alters the microbiome below ground.

Laticifers are hypothesized to mediate both plant-herbivore and plant-microbe interactions. However, there is little evidence for this dual function. We investigated whether the major constituent of natural rubber, cis-1,4-polyisoprene, a phylogenetically widespread and economically important latex polymer, alters plant resistance and the root microbiome of the Russian dandelion (Taraxacum koksaghyz) under attack of a root herbivore, the larva of the May cockchafer (Melolontha melolontha). Rubber-depleted transgenic plants lost more shoot and root biomass upon herbivory than normal rubber content near-isogenic lines. Melolontha melolontha preferred to feed on artificial diet supplemented with rubber-depleted rather than normal rubber content latex. Likewise, adding purified cis-1,4-polyisoprene in ecologically relevant concentrations to diet deterred larval feeding and reduced larval weight gain. Metagenomics and metabarcoding revealed that abolishing biosynthesis of natural rubber alters the structure but not the diversity of the rhizosphere and root microbiota (ecto- and endophytes) and that these changes depended on M. melolontha damage. However, the assumption that rubber reduces microbial colonization or pathogen load is contradicted by four lines of evidence. Taken together, our data demonstrate that natural rubber biosynthesis reduces herbivory and alters the plant microbiota, which highlights the role of plant-specialized metabolites and secretory structures in shaping multitrophic interactions.

Apoplast-Localized β-Glucosidase Elevates Isoflavone Accumulation in the Soybean Rhizosphere.

Plant specialized metabolites (PSMs) are often stored as glycosides within cells and released from the roots with some chemical modifications. While isoflavones are known to function as symbiotic signals with rhizobia and to modulate the soybean rhizosphere microbiome, the underlying mechanisms of root-to-soil delivery are poorly understood. In addition to transporter-mediated secretion, the hydrolysis of isoflavone glycosides in the apoplast by an isoflavone conjugate-hydrolyzing β-glucosidase (ICHG) has been proposed but not yet verified. To clarify the role of ICHG in isoflavone supply to the rhizosphere, we have isolated two independent mutants defective in ICHG activity from a soybean high-density mutant library. In the root apoplastic fraction of ichg mutants, the isoflavone glycoside contents were significantly increased, while isoflavone aglycone contents were decreased, indicating that ICHG hydrolyzes isoflavone glycosides into aglycones in the root apoplast. When grown in a field, the lack of ICHG activity considerably reduced isoflavone aglycone contents in roots and the rhizosphere soil, although the transcriptomes showed no distinct differences between the ichg mutants and wild-types (WTs). Despite the change in isoflavone contents and composition of the root and rhizosphere of the mutants, root and rhizosphere bacterial communities were not distinctive from those of the WTs. Root bacterial communities and nodulation capacities of the ichg mutants did not differ from the WTs under nitrogen-deficient conditions either. Taken together, these results indicate that ICHG elevates the accumulation of isoflavones in the soybean rhizosphere but is not essential for isoflavone-mediated plant-microbe interactions.

Phytoremediation technologies and their mechanism for removal of heavy metal from contaminated soil: An approach for a sustainable environment.

The contamination of soils with heavy metals and its associated hazardous effects are a thrust area of today's research. Rapid industrialization, emissions from automobiles, agricultural inputs, improper disposal of waste, etc., are the major causes of soil contamination with heavy metals. These contaminants not only contaminate soil but also groundwater, reducing agricultural land and hence food quality. These contaminants enter the food chain and have a severe effect on human health. It is important to remove these contaminants from the soil. Various economic and ecological strategies are required to restore the soils contaminated with heavy metals. Phytoremediation is an emerging technology that is non-invasive, cost-effective, and aesthetically pleasing. Many metal-binding proteins (MBPs) of the plants are significantly involved in the phytoremediation of heavy metals; the MBPs include metallothioneins; phytochelatins; metalloenzymes; metal-activated enzymes; and many metal storage proteins, carrier proteins, and channel proteins. Plants are genetically modified to enhance their phytoremediation capacity. In Arabidopsis, the expression of the mercuric ion-binding protein in Bacillus megaterium improves the metal accumulation capacity. The phytoremediation efficiency of plants is also enhanced when assisted with microorganisms, biochar, and/or chemicals. Removing heavy metals from agricultural land without challenging food security is almost impossible. As a result, crop selections with the ability to sequester heavy metals and provide food security are in high demand. This paper summarizes the role of plant proteins and plant-microbe interaction in remediating soils contaminated with heavy metals. Biotechnological approaches or genetic engineering can also be used to tackle the problem of heavy metal contamination.

Plant–Microbe Interactions under the Action of Heavy Metals and under the Conditions of Flooding

Heavy metals and flooding are among the primary environmental factors affecting plants and microorganisms. This review separately considers the impact of heavy metal contamination of soils on microorganisms and plants, on plant and microbial biodiversity, and on plant–microorganism interactions. The use of beneficial microorganisms is considered one of the most promising methods of increasing stress tolerance since plant-associated microbes reduce metal accumulation, so the review focuses on plant–microorganism interactions and their practical application in phytoremediation. The impact of flooding as an adverse environmental factor is outlined. It has been shown that plants and bacteria under flooding conditions primarily suffer from a lack of oxygen and activation of anaerobic microflora. The combined effects of heavy metals and flooding on microorganisms and plants are also discussed. In conclusion, we summarize the combined effects of heavy metals and flooding on microorganisms and plants.

The Key Roles of ROS and RNS as a Signaling Molecule in Plant-Microbe Interactions.

Reactive oxygen species (ROS) and reactive nitrogen species (RNS) play a pivotal role in the dynamic cell signaling systems in plants, even under biotic and abiotic stress conditions. Over the past two decades, various studies have endorsed the notion that these molecules can act as intracellular and intercellular signaling molecules at a very low concentration to control plant growth and development, symbiotic association, and defense mechanisms in response to biotic and abiotic stress conditions. However, the upsurge of ROS and RNS under stressful conditions can lead to cell damage, retarded growth, and delayed development of plants. As signaling molecules, ROS and RNS have gained great attention from plant scientists and have been studied under different developmental stages of plants. However, the role of RNS and RNS signaling in plant-microbe interactions is still unknown. Different organelles of plant cells contain the enzymes necessary for the formation of ROS and RNS as well as their scavengers, and the spatial and temporal positions of these enzymes determine the signaling pathways. In the present review, we aimed to report the production of ROS and RNS, their role as signaling molecules during plant-microbe interactions, and the antioxidant system as a balancing system in the synthesis and elimination of these species.

Insect egg-induced innate immunity: Who benefits?

Plants perceive the presence of insect eggs deposited on leaves as a cue of imminent herbivore attack. Consequential plant signaling events include the accumulation of salicylic acid and reactive oxygen species, transcriptional reprogramming, and cell death. Interestingly, egg-induced innate immunity shows similarities with immune responses triggered upon recognition of microbial pathogens, and in recent years, it became apparent that egg perception affects plant-microbe interactions. Here, we highlight recent findings on insect egg-induced innate immunity and how egg-mediated signaling impacts plant-microbe interactions. Ecological considerations beg the question: Who benefits from egg perception in these complex interactions?

Interaction between bacterial endophytes and host plants.

Endophytic bacteria are mainly present in the plant's root systems. Endophytic bacteria improve plant health and are sometimes necessary to fight against adverse conditions. There is an increasing trend for the use of bacterial endophytes as bio-fertilizers. However, new challenges are also arising regarding the management of these newly discovered bacterial endophytes. Plant growth-promoting bacterial endophytes exist in a wide host range as part of their microbiome, and are proven to exhibit positive effects on plant growth. Endophytic bacterial communities within plant hosts are dynamic and affected by abiotic/biotic factors such as soil conditions, geographical distribution, climate, plant species, and plant-microbe interaction at a large scale. Therefore, there is a need to evaluate the mechanism of bacterial endophytes' interaction with plants under field conditions before their application. Bacterial endophytes have both beneficial and harmful impacts on plants but the exact mechanism of interaction is poorly understood. A basic approach to exploit the potential genetic elements involved in an endophytic lifestyle is to compare the genomes of rhizospheric plant growth-promoting bacteria with endophytic bacteria. In this mini-review, we will be focused to characterize the genetic diversity and dynamics of endophyte interaction in different host plants.

Changes in Phyllosphere Microbial Communities of Pinus tabuliformis after Infestation by Bursaphelenchus xylophilus

Phyllosphere microbial communities have an important role in plant growth and resistance to pathogen infection and are partially influenced by leaf characteristics. Pinewood nematode, Bursaphelenchus xylophilus, is one of the greatest threats to pine trees and is spreading all over the world. However, studies on the resistance of plant–microbe interactions to pathogens during the nematode’s pathogenesis and the relationships of leaf chemical characteristics caused by pinewood nematode and phyllosphere microbial communities are limited. In this study, different stages of Pinus tabuliformis that were healthy or infected with B. xylophilus-associated leaf characteristics and phyllosphere bacterial and fungal communities were compared. These results demonstrated that soluble sugar and starch contents decreased based on the extent of infection. Phyllosphere microbial community changes potentially caused by B. xylophilus infection of P. tabuliformis and the fungal community compositions of healthy P. tabuliformis trees (Ya) were clearly different from diseased P. tabuliformis trees at an early stage of B. xylophilus infection (Yb) and P. tabuliformis trees in the last stage of B. xylophilus infection (Yc), particularly along the first coordinate axis. According to a linear discriminant effect size (LEfSe) analysis, the biomarker species in the phyllosphere of Yb were Acidobacteria, Deinococcus-Thermus, and Patescibacteria, while those in the phyllosphere of Ya were Proteobacteria, Aureobasidium, Dictyosporium, Alternariaster, Knufia, Microstroma, and Naganishia. Particularly at the end of PWD (pine wilt disease) infection, the majority of microbial taxa tended to co-exclude rather than co-occur with PWD infection. The result of a canonical correlation analysis (CCA) showed that the chemical properties of leaves, such as carbon and nitrogen, have significant impacts on phyllosphere microbial communities. These results expanded the possible connections between the phyllosphere communities and plant health.

The putative transporter MtUMAMIT14 participates in nodule formation in Medicago truncatula

Transport systems are crucial in many plant processes, including plant–microbe interactions. Nodule formation and function in legumes involve the expression and regulation of multiple transport proteins, and many are still uncharacterized, particularly for nitrogen transport. Amino acids originating from the nitrogen-fixing process are an essential form of nitrogen for legumes. This work evaluates the role of MtN21 (henceforth MtUMAMIT14), a putative transport system from the MtN21/EamA-like/UMAMIT family, in nodule formation and nitrogen fixation in Medicago truncatula. To dissect this transporter’s role, we assessed the expression of MtUMAMIT14 using GUS staining, localized the corresponding protein in M. truncatula root and tobacco leaf cells, and investigated two independent MtUMAMIT14 mutant lines. Our results indicate that MtUMAMIT14 is localized in endosomal structures and is expressed in both the infection zone and interzone of nodules. Comparison of mutant and wild-type M. truncatula indicates MtUMAMIT14, the expression of which is dependent on the presence of NIN, DNF1, and DNF2, plays a role in nodule formation and nitrogen-fixation. While the function of the transporter is still unclear, our results connect root nodule nitrogen fixation in legumes with the UMAMIT family.

Overviewing Drought and Heat Stress Amelioration—From Plant Responses to Microbe-Mediated Mitigation

Microbes (e.g., plant-growth-promoting rhizobacteria, arbuscular mycorrhizal fungi and endophytes) are the natural inhabitants of the soil-plant-environment ecosystem having the potential to ameliorate the negative effects of environmental extremities. Plant-microbe interactions are integral events of agricultural ecosystems which must be studied in order to modulate the systemic mechanisms in field crops. Under changing climatic scenarios, drought and heat stresses tend to induce numerous physiological, morphological, metabolic and biochemical alterations in crop plants, while microbes hold the potential to mitigate these adverse impacts in a sustainable way. However, plant-microbe interaction mechanisms remain understudied owing to their complexities in the rhizosphere and within the cellular systems of plants. In this review, we have attempted to summarize microbes’ interactions with crop plants that tend to influence hormonal and nutrients balance, and the biosynthesis of metabolites and phytohormones, etc. In particular, focus has been kept on the underlying mechanisms related to plant-microbe interactions which confer abiotic stress tolerance. Moreover, various physiological, morphological, metabolic and biochemical responses of plants subjected to water scarcity and elevated temperatures have been synthesized objectively. Lastly, from the perspective of microbes’ application as biofertilizers, both challenges and future research needs to develop microbe-mediated tolerance as a biologically potent strategy have been strategically pointed out.

Advances in studies on the plant rhizosphere microorganisms in wetlands: A visualization analysis based on CiteSpace

Rhizosphere microorganisms and their interactions with plants in wetlands have recently attracted much attention due to their importance in enhancing plant environmental adaptation, removing wetland pollutants, and alleviating climate change. However, the fluctuating hydrological environment of wetlands leads to more complex dynamics in the rhizosphere environment. Research progress and hotspots concerning plant-rhizosphere microorganisms under special wetland environments are still kept unclear. To better understand the current research status, hotspots and trends of rhizosphere microorganisms in wetlands, we used CiteSpace bibliometric software to visualize and analyze 231 English-language publications from the Web of Science core collection database. Here, we reviewed the role played by various countries, institutions, and scholars in the studies of plant rhizosphere microorganisms in wetlands based on cooperation network analysis. We discussed the shift from bioremediation and nutrient removal to rhizosphere microbial community composition as a research hotspot for plant rhizosphere microorganisms in wetlands according to keyword co-occurrence and clustering analysis. Finally, we highlighted that more attention should be paid to the ecological functions of rhizosphere microorganisms in different wetland ecosystems, and the plant‒microbe microinterface processes and interaction patterns should be explored in depth to provide new indicators for the evaluation of wetland ecosystem functions.

Plant growth and microbial responses from urban agriculture soils amended with excavated local sediments and municipal composts

Abstract With increasing urbanization and critical issues of food insecurity emerging globally, urban agriculture is expanding as an agroecosystem with a distinct soil type. Growing food in cities is challenged by legacy contaminants in soils, which necessitates the use of imported, safe soils and composts. To promote the long-term sustainability of urban agriculture, we examined the agronomic potential of constructing safe, locally sourced soils to support food production. We collected composts from four municipal composting facilities in New York City: Big Reuse, Long Island City, Queens (BRL), New York Department of Sanitation, Fresh Kills, Staten Island (DNY), Lower Eastside Ecology Center (LES) and Queens Botanic Garden (QBG). We then created two types of constructed soils using each compost: 100% pure compost and a 50:50 blend of compost and clean excavated sediments from the New York City Clean Soil Bank. We then assessed the growth of tomato, pepper and kale in the constructed soils within a plant growth chamber facility. We found Clean Soil Bank sediments enhanced tomato aboveground biomass production by 98%, kale aboveground biomass production by 50% and pepper plant height by 52% when mixed with compost from BRL. At the same time, Clean Soil Bank Sediments decreased tomato plant height by 16% and aboveground biomass production by 29% in LES compost and tomato plant height by 18% in QBG compost, likely due to compost properties. The addition of Clean Soil Bank sediments showed no decline in the symbiosis of arbuscular mycorrhizal fungi across all composts, which is an important beneficial plant–microbe interaction in agroecosystems. A positive ecosystem service was found when Clean Soil Bank sediments were added to municipal composts, with up to a 74% decrease in greenhouse gas emissions of soil CO2 in BRL compost. The results indicate that urban agricultural soils can be constructed using clean, locally sourced materials, such as composted organic waste and excavated sediments from city development sites to support sustainable urban agriculture while enhancing ecosystem services.

Optical Sensing Technologies to Elucidate the Interplay between Plant and Microbes.

Plant-microbe interactions are critical for ecosystem functioning and driving rhizosphere processes. To fully understand the communication pathways between plants and rhizosphere microbes, it is crucial to measure the numerous processes that occur in the plant and the rhizosphere. The present review first provides an overview of how plants interact with their surrounding microbial communities, and in turn, are affected by them. Next, different optical biosensing technologies that elucidate the plant-microbe interactions and provide pathogenic detection are summarized. Currently, most of the biosensors used for detecting plant parameters or microbial communities in soil are centered around genetically encoded optical and electrochemical biosensors that are often not suitable for field applications. Such sensors require substantial effort and cost to develop and have their limitations. With a particular focus on the detection of root exudates and phytohormones under biotic and abiotic stress conditions, novel low-cost and in-situ biosensors must become available to plant scientists.

QTL for induced resistance against leaf rust in barley.

Leaf rust caused by Puccinia hordei is one of the major diseases of barley (Hordeum vulgare L.) leading to yield losses up to 60%. Even though, resistance genes Rph1 to Rph28 are known, most of these are already overcome. In this context, priming may promote enhanced resistance to P. hordei. Several bacterial communities such as the soil bacterium Ensifer (syn. Sinorhizobium) meliloti are reported to induce resistance by priming. During quorum sensing in populations of gram negative bacteria, they produce N-acyl homoserine-lactones (AHL), which induce resistance in plants in a species- and genotype-specific manner. Therefore, the present study aims to detect genotypic differences in the response of barley to AHL, followed by the identification of genomic regions involved in priming efficiency of barley. A diverse set of 198 spring barley accessions was treated with a repaired E. meliloti natural mutant strain expR+ch producing a substantial amount of AHL and a transformed E. meliloti strain carrying the lactonase gene attM from Agrobacterium tumefaciens. For P. hordei resistance the diseased leaf area and the infection type were scored 12 dpi (days post-inoculation), and the corresponding relative infection and priming efficiency were calculated. Results revealed significant effects (p<0.001) of the bacterial treatment indicating a positive effect of priming on resistance to P. hordei. In a genome-wide association study (GWAS), based on the observed phenotypic differences and 493,846 filtered SNPs derived from the Illumina 9k iSelect chip, genotyping by sequencing (GBS), and exome capture data, 11 quantitative trait loci (QTL) were identified with a hot spot on the short arm of the barley chromosome 6H, associated to improved resistance to P. hordei after priming with E. meliloti expR+ch. Genes in these QTL regions represent promising candidates for future research on the mechanisms of plant-microbe interactions.

New dual functional CYP450 gene involves in isoflavone biosynthesis in Glycine max L.

Glycine max L. accumulates a large amount of isoflavonoid compounds, which is beneficial for plant defense, plant-microbe symbiotic interactions, and human health. Several CYP450 subfamily genes are involved in the flavonoid biosynthetic pathway in plants. In the present study, we found 24 CYP82 subfamily genes were differentially expressed in various tissues of soybean, in Phytophthora sojae-infected soybean varieties and in soybean hairy roots treated with cell wall glucan elicitor. Six of them (GmCYP82A2, GmCYP82A3, GmCYP82A4, GmCYP82A23, GmCYP82C20 and GmCYP82D26) were co-expressed with other known isoflavonoid pathway genes in soybean. Their enzymatic activity in yeast feeding assays showed that only GmCYP82D26 was able to convert naringenin to daidzein with both aryl migration and dehydration function. When GmCYP82D26 was over-expressed in soybean hairy roots, the contents of the two major isoflavonoid aglycones in soybean (daidzein and genistein) were reduced, but total flavonoids were not affected. When GmCYP82D26 was suppressed by RNAi in the hairy roots, daidzein content was decreased but genistein content was increased, with unchanged total flavonoid content. GmCYP82D26 was found to be localized in the endoplasmic reticulum at subcellular level when transiently expressed in tobacco leaf epidermis. GmCYP82D26 gene was preferentially expressed in roots, with low expression level in other tissues in soybean. Homology modeling and molecular docking showed that GmCYP82D26 could form hydrogen bond with both HEM and naringenin at C5–OH and C4 carbonyl. All these results indicated that GmCYP82D26 possesses new and dual enzymatic activity, which bridges the two branches (daidzein and genistein branch) of isoflavonoid pathway in soybean.