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Arbuscular mycorrhizal (AM) symbiosis is the oldest and most widespread mutualistic association on Earth and involves plants and soil fungi belonging to Glomeromycotina. A complex molecular, cellular, and genetic developmental program enables partner recognition, fungal accommodation in plant tissues, and activation of symbiotic functions such as transfer of phosphorus in exchange for carbohydrates and lipids. AM fungi, as ancient obligate biotrophs, have evolved strategies to circumvent plant defense responses to guarantee an intimate and long-lasting mutualism. They are among those root-associated microorganisms able to boost plants' ability to cope with biotic stresses leading to mycorrhiza-induced resistance (MIR), which can be effective across diverse hosts and against different attackers. Here, we examine the molecular mechanisms underlying the modulation of plant immunity during colonization by AM fungi and at the onset and display of MIR against belowground and aboveground pests and pathogens. Understanding the MIR efficiency spectrum and its regulation is of great importance to optimizing the biotechnological application of these beneficial microbes for sustainable crop protection.

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Knowledge is scarce to what extent environmental drivers and native symbiotic fungi in soil induce abrupt (short-term), systemic (multiple traits), or specific (a subset of traits) shifts in C3 plants' ecophysiological/mycorrhizal responses. We cultivated an emblematic native C3 species (Capsicum annuum var. glabriusculum, "Chiltepín") to look at how the extreme heat of the Sonoran desert, sunlight regimes (low = 2, intermediate = 15, high = 46 mol m2 d-1) and density of native arbuscular mycorrhizal fungi in soil (low AMF = 1% v/v, high AMF = 100% v/v), drive shifts on mycorrhizal responses through multiple functional traits (106 traits). The warming thresholds were relentlessly harsh even under intensive shade (e.g. superheat maximum thresholds reached ranged between 47-63°C), and several pivotal traits were synergistically driven by AMF (e.g. photosynthetic capacity, biomass gain/allometry, and mycorrhizal colonization traits); whereas concurrently, sunlight regimes promoted most (76%) alterations in functional acclimation traits in the short-term and opposite directions (e.g. survival, phenology, photosynthetic, carbon/nitrogen economy). Multidimensional reduction analysis suggests that the AMF promotes a synergistic impact on plants' phenotypic integration and functional plasticity in response to sunlight regimes; however, complex relationships among traits suggest that phenotypic variation determines the robustness degree of ecophysiological/mycorrhizal phenotypes between/within environments. Photosynthetic canopy surface expansion, Rubisco activity, photosynthetic nitrogen allocation, carbon gain, and differential colonization traits could be central to plants' overall ecophysiological/mycorrhizal fitness strengthening. In conclusion, we found evidence that a strong combined effect among environmental factors in which AMF are key effectors could drive important trade-offs on plants' ecophysiological/mycorrhizal fitness in the short term.

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BACKGROUND: Potassium (K) is an essential nutrient for plant growth and development. Maize (Zea mays) is a widely planted crops in the world and requires a huge amount of K fertilizer. Arbuscular mycorrhizal fungi (AMF) are closely related to the K uptake of maize. Genetic improvement of maize K utilization efficiency will require elucidating the molecular mechanisms of maize K uptake through the mycorrhizal pathway. Here, we employed transcriptome and gene family analysis to elucidate the mechanism influencing the K uptake and utilization efficiency of mycorrhizal maize. METHODS AND RESULTS: The transcriptomes of maize were studied with and without AMF inoculation and under different K conditions. AM symbiosis increased the K concentration and dry weight of maize plants. RNA sequencing revealed that genes associated with the activity of the apoplast and nutrient reservoir were significantly enriched in mycorrhizal roots under low-K conditions but not under high-K conditions. Weighted gene correlation network analysis revealed that three modules were strongly correlated with K content. Twenty-one hub genes enriched in pathways associated with glycerophospholipid metabolism, glycerolipid metabolism, starch and sucrose metabolism, and anthocyanin biosynthesis were further identified. In general, these hub genes were upregulated in AMF-colonized roots under low-K conditions. Additionally, the members of 14 gene families associated with K obtain were identified (ARF: 38, ILK: 4, RBOH: 12, RUPO: 20, MAPKK: 89, CBL: 14, CIPK: 44, CPK: 40, PIN: 10, MYB: 174, NPF: 79, KT: 19, HAK/HKT/KUP: 38, and CPA: 8) from maize. The transcript levels of these genes showed that 92 genes (ARF:6, CBL:5, CIPK:13, CPK:2, HAK/HKT/KUP:7, PIN:2, MYB:26, NPF:16, RBOH:1, MAPKK:12 and RUPO:2) were upregulated with AM symbiosis under low-K conditions. CONCLUSIONS: This study indicated that AMF increase the resistance of maize to low-K stress by regulating K uptake at the gene transcription level. Our findings provide a genome-level resource for the functional assignment of genes regulated by K treatment and AM symbiosis in K uptake-related gene families in maize. This may contribute to elucidate the molecular mechanisms of maize response to low K stress with AMF inoculation, and provided a theoretical basis for AMF application in the crop field.

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Salinity stress negatively affects the growth and yield of crops worldwide. Onion (Allium cepa L.) is moderately sensitive to salinity. Beneficial microorganisms can potentially confer salinity tolerance. This study investigated the effects of endomycorrhizal fungi (M), Pseudomonas putida (Ps) and their combination (MPs) on onion growth under control (0 ppm), moderate (2000 ppm) and high (4000 ppm) NaCl salinity levels. A pot experiment was conducted with sandy loam soil and onion cultivar Giza 20. Results showed that salinity reduced growth attributes, leaf pigments, biomass and bulb yield while increasing oxidative stress markers. However, individual or combined inoculations significantly increased plant height, bulb diameter and biomass production compared to uninoculated plants under saline conditions. MPs treatment provided the highest stimulation, followed by Pseudomonas and mycorrhizae alone. Overall, dual microbial inoculation showed synergistic interaction, conferring maximum benefits for onion growth, bulbing through integrated physiological and biochemical processes under salinity. Bulb yield showed 3.5, 36 and 83% increase over control at 0, 2000 and 4000 ppm salinity, respectively. In conclusion, combined application of mycorrhizal-Pseudomonas inoculations (MPs) effectively mitigate salinity stress. This approach serves as a promising biotechnology for ensuring sustainable onion productivity under saline conditions.

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Mycorrhizal fungi, a category of fungi that form symbiotic relationships with plant roots, can participate in the induction of plant disease resistance by secreting phosphatase enzymes. While extensive research exists on the mechanisms by which mycorrhizal fungi induce resistance, the specific contributions of phosphatases to these processes require further elucidation. This article reviews the spectrum of mycorrhizal fungi-induced resistance mechanisms and synthesizes a current understanding of how phosphatases mediate these effects, such as the induction of defense structures in plants, the negative regulation of plant immune responses, and the limitation of pathogen invasion and spread. It explores the role of phosphatases in the resistance induced by mycorrhizal fungi and provides prospective future research directions in this field.

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Terrestrial plants form primarily mutualistic symbiosis with mycorrhizal fungi based on a compatible exchange of solutes between plant and fungal partners. A key attribute of this symbiosis is the acquisition of soil nutrients by the fungus for the benefit of the plant in exchange for a carbon supply to the fungus. The interaction can range from mutualistic to parasitic depending on environmental and physiological contexts. This review considers current knowledge of the functionality of ectomycorrhizal (EM) symbiosis in the mobilisation and acquisition of soil nitrogen (N) in northern hemisphere forest ecosystems, highlighting the functional diversity of the fungi and the variation of symbiotic benefits, including the dynamics of N transfer to the plant. It provides an overview of recent advances in understanding 'mycorrhizal decomposition' for N release from organic or mineral-organic forms. Additionally, it emphasises the taxon-specific traits of EM fungi in soil N uptake. While the effects of EM communities on tree N are likely consistent across different communities regardless of species composition, the sink activities of various fungal taxa for tree carbon and N resources drive the dynamic continuum of mutualistic interactions. We posit that ectomycorrhizas contribute in a species-specific but complementary manner to benefit tree N nutrition. Therefore, alterations in diversity may impact fungal-plant resource exchange and, ultimately, the role of ectomycorrhizas in tree N nutrition. Understanding the dynamics of EM functions along the mutualism-parasitism continuum in forest ecosystems is essential for the effective management of ecosystem restoration and resilience amidst climate change. KEY POINTS: • Mycorrhizal symbiosis spans a continuum from invested to appropriated benefits. • Ectomycorrhizal fungal communities exhibit a high functional diversity. • Tree nitrogen nutrition benefits from the diversity of ectomycorrhizal fungi.

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Interview with Caroline Gutjahr, who studies the molecular factors influencing the formation and function of mycorrhiza at the Max Planck Institute of Molecular Plant Physiology, where she is also the Director.

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Improving the cultivation mode and technology for traditional Chinese medicine has become important for its sustainable development. Monoculture enhances plant diseases, which decreases yield and quality. Intercropping is an effective measure to counterbalance that negative effect. In this study, we focused on Panax quinquefolium L. (ginseng) and four treatments were set up: the control without intercropping, P. quinquefolius + ryegrass (Lolium perenne L.), P. quinquefolius + red clover (Trifolium pratense L.), and P. quinquefolius + ryegrass + red clover. An LC-MS/MS system was used to detect the changes in the P. quinquefolius secondary metabolites, and high-throughput sequencing technology was used to determine the changes in the P. quinquefolius' rhizosphere soil microorganisms. Ginsenoside content, soil enzyme activities, and arbuscular mycorrhizal infection rate of P. quinquefolius were also measured using HPLC, ELISA kits, and microscopy, respectively. Co-intertia and Pearson's analysis were performed to explore the relationship between the metabolites and the P. quinquefolius microorganisms. Intercropping significantly increased the content of ginsenoside metabolites and recruited a large number of beneficial bacteria to the P. quinquefolius rhizosphere. The P. quinquefolius secondary metabolites were associated with the rhizosphere microbial community. For example, the dominant microorganisms, such as Acidobacteriota and Chloroflexi, played a key role in promoting the synthesis of ginsenoside Rd and (20R) ginsenoside Rg3 by P. quinquefolius. Intercropping led to changes in the P. quinquefolius secondary metabolites by driving and reshaping the rhizosphere microorganisms. These findings revealed the potential application of intercropping for improving the quality of P. quinquefolius.

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Arbuscular mycorrhizal (AM) fungi are well known for enhancing phosphorus uptake in plants; however, their regulating roles in cation transporting gene family, such as natural resistance-associated macrophage protein (NRAMP), are still limited. Here, we performed bioinformatics analysis and quantitative expression assays of tomato SlNRAMP 1 to 5 genes under nutrient deficiency and cadmium (Cd) stress in response to AM symbiosis. These five SlNRAMP members are mainly located in the plasma or vacuolar membrane and can be divided into two subfamilies. Cis-element analysis revealed several motifs involved in phytohormonal and abiotic regulation in their promoters. SlNRAMP2 was downregulated by iron deficiency, while SlNRAMP1, SlNRAMP3, SlNRAMP4, and SlNRAMP5 responded positively to copper-, zinc-, and manganese-deficient conditions. AM colonization reduced Cd accumulation and expression of SlNRAMP3 but enhanced SlNRAMP1, SlNRAMP2, and SlNRMAP4 in plants under Cd stress. These findings provide valuable genetic information for improving tomato resilience to nutrient deficiency and heavy metal stress by developing AM symbiosis.

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The cell and molecular bases of arbuscular mycorrhizal (AM) symbiosis, a crucial plant-fungal interaction for nutrient acquisition, have been extensively investigated by coupling traditional RNA sequencing techniques of roots sampled in bulk, with methods to capture subsets of cells such as laser microdissection. These approaches have revealed central regulators of this complex relationship, yet the requisite level of detail to effectively untangle the intricacies of temporal and spatial development remains elusive.The recent adoption of single-cell RNA sequencing (scRNA-seq) techniques in plant research is revolutionizing our ability to dissect the intricate transcriptional profiles of plant-microbe interactions, offering unparalleled insights into the diversity and dynamics of individual cells during symbiosis. The isolation of plant cells is particularly challenging due to the presence of cell walls, leading plant researchers to widely adopt nuclei isolation methods. Despite the increased resolution that single-cell analyses offer, it also comes at the cost of spatial perspective, hence, it is necessary the integration of these approaches with spatial transcriptomics to obtain a comprehensive overview.To date, few single-cell studies on plant-microbe interactions have been published, most of which provide high-resolution cell atlases that will become crucial for fully deciphering symbiotic interactions and addressing future questions. In AM symbiosis research, key processes such as the mutual recognition of partners during arbuscule development within cortical cells, or arbuscule senescence and degeneration, remain poorly understood, and these advancements are expected to shed light on these processes and contribute to a deeper understanding of this plant-fungal interaction.

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In many ecosystems, a large fraction of gross primary production is invested in mycorrhiza. Ectomycorrhizal (ECM) mycelium is involved in regulating soil carbon and nutrient cycling. However, little is known about how mycelial biomass, production and turnover differ depending on ECM fungal community composition and associated tree species. We quantified fine root biomass and length using soil cores, and mycelial traits (biomass, production, and turnover) using mesh-bags and ergosterol analysis, and identified ECM exploration types by Illumina MiSeq sequencing of four ECM-dominated tree species (Picea asperata, Larix gmelinii, Quercus aquifolioides and Betula albosinensis) in subalpine forest. The ECM fungal community composition separated between needle-leaved and broadleaved species, and between evergreen and deciduous species. The ratio of mycelial to fine root biomass was similar across the species regardless of genus-scale community composition and the relative abundance of exploration types. Compared to the other species, Q. aquifolioides displayed higher fine root biomass and mycelial biomass and production, dominated by contact-short exploration type. Mycelial turnover rate tended to be lowest in P. asperata, dominated by medium-long exploration type. Much higher production of mycelium and only slightly higher turnover rate in Q. aquifolioides suggests that its steady-state mycelial biomass would be higher than of the other species. Moreover, compared to the two deciduous species, with similar production but somewhat lower turnover rate, the standing crop of mycelium in P. asperata may stabilize at a higher value. Our findings, that exploration type may affect production and turnover, highlight the importance of characterizing ECM fungal communities by exploration types when estimating the contribution of mycelium biomass to forest carbon sink and storage.

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The excessive application of phosphorus (P) fertiliser and its poor utilisation efficiency have led to significant amounts of P being retained in agricultural soils in unavailable forms. The application of alkaline lignin to soil and its inoculation with arbuscular mycorrhizal fungi (AMF) have both been shown to improve plant P nutrition. However, their combined effects on soil P transformation remain unclear, particularly in cadmium (Cd)-contaminated soils. A potting experiment was conducted to examine the combined effects of AMF and alkaline lignin on soil P and Cd bioavailability and on the uptake of P and Cd by lettuce (Lactuca sativa L.) that were grown for 56 d in a growth chamber. Combined AMF and alkaline lignin treatment increased soil P availability and alkaline phosphatase activity. It furthermore increased bioavailable Cd concentrations of rhizosphere and bulk soils by 48 % and 72 %, respectively, and the Cd concentration in roots by 85 %, but the Cd concentration was not affected in the edible parts (shoots) of the lettuce. Moreover, the combined treatment increased shoot biomass by 26-70 % and root biomass by 99-164 %. Our findings suggested that the combined use of AMF and alkaline lignin mobilised both P and Cd in soil but did not increase the accumulation of Cd in the shoots of plants growing in Cd-contaminated soils, these results would provide guideline for increasing Cd tolerance of plants and their yield.

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AIMS: Arbuscular mycorrhizal fungi (AMF) can perform significant functions within sustainable agricultural ecosystems, including vineyards. Increased AMF diversity can be beneficial in promoting plant growth and increasing resilience to environmental changes. To effectively utilize AMF communities and their benefits in vineyard ecosystems, a better understanding of how management systems influence AMF community composition is needed. Moreover, it is unknown whether AMF communities in organically managed vineyards are distinct from those in conventionally managed vineyards. METHODS AND RESULTS: In this study, vineyards were surveyed across the Marlborough region, New Zealand to identify the AMF communities colonizing the roots of different rootstocks grafted with Sauvignon Blanc and Pinot Noir in both conventional and organic systems. The AMF communities were identified based on spores isolated from trap cultures established with the collected grapevine roots, and by next-generation sequencing technologies (Illumina MiSeq). The identified AMF species/genera belonged to Glomeraceae, Entrophosporaceae, and Diversisporaceae. The results revealed a significant difference in AMF community composition between rootstocks and in their interaction with management systems. CONCLUSIONS: These outcomes indicated that vineyard management systems influence AMF recruitment by rootstocks and some rootstocks may therefore be more suited to organic systems due to the AMF communities they support. This could provide an increased benefit to organic systems by supporting higher biodiversity.

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The genomes of ectomycorrhizal (ECM) fungi have a reduced number of genes encoding Carbohydrate-Active EnZymes (CAZymes), expansions in transposable elements (TEs) and small secreted proteins (SSPs) compared with saprotrophs. Fewer genes for specific peptidases and lipases in ECM fungi are also reported. It is unclear whether these changes occur at the shift to the ECM habit or are more gradual throughout the evolution of ECM lineages. We generated a genomic dataset of 20 species in the ECM lineage Inocybaceae and compared them with six saprotrophic species. Inocybaceae genomes have fewer CAZymes, peptidases, lipases, secondary metabolite clusters and SSPs and higher TE content than their saprotrophic relatives. There was an increase in the rate of gene family evolution along the branch with the transition to the ECM lifestyle. This branch had very high rate of evolution in CAZymes and had the largest number of contractions. Other significant changes along this branch included expansions in transporters, transposons-related genes and communication genes such as fungal kinases. There is a high concentration of changes in proximity to the transition to the ECM lifestyle, which correspond to the identified key changes for the gain of this lifestyle.

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Arbuscular mycorrhizal fungi (AMF) have been shown to effectively mitigate the detrimental effects of heavy metal stress on their plant hosts. Nevertheless, the biological activities of AMF were concurrently compromised. Biochar (BC), as an abiotic factor, had the potential compensate for this limitation. To elucidate the synergistic effects of biotic and abiotic factors, a pot experiment was conducted to assess the impact of biochar and AMF on the growth, physiological traits, and genetic expression in rice plants subjected to Cd stress. The results demonstrated that biochar significantly increased the mycorrhizal colonization rate by 22.19 %, while the combined application of biochar and AMF led to a remarkable enhancement of rice root biomass by 42.2 %. This resulted in a shift in spatial growth patterns that preferentially promoted enhanced underground development. Biochar effectively mitigated the stomatal limitations imposed by Cd on photosynthetic processes. The decrease in IBRv2 (Integrated Biomarker Response version 2) values suggested that the antioxidant system was experiencing a state of remission. An increase of Cd content within the rice root systems was observed, ranging from 33.71 % to 48.71 %, accompanied by a reduction in Cd bioavailability and mobility curtailed its translocation to the aboveground tissues. Under conditions of low soil Cd concentration (Cd ≤ 1 mg·kg-1), the Cd content in rice seeds from the group subjected to the combined treatment remained below the national standard (Cd ≤ 0.2 mg·kg-1). Furthermore, the combined treatment modulated the uptake of Fe and Zn by rice, while simultaneously suppressing the expression of genes associated with Cd transport. Collectively, the integration of biological and abiotic factors provided a novel perspective and methodological framework for safe in-situ utilization of soils with low Cd contamination.

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Arbuscular mycorrhizal (AM) fungi improve plant growth, nutrition, fitness and stress tolerance while AM fungi obtain carbohydrates and lipids from the host. This whole process of mutual benefit requires substantial alterations in the structural and functional aspects of the host root cells. These modifications ultimately culminate in the formation of arbuscules, which are specialized intraradical and highly branched fungal structures. Arbuscule-containing cells undergo massive reprogramming to hosting arbuscule and members of the GRAS transcription factor family have been characterized as AM inducible genes which play a pivotal role in these process. Here, we show a functional analysis for the GRAS transcription factor SCL3/SlGRAS18 in tomato. SlGRAS18 interacts with SlDELLA, a central regulator of AM formation. Silencing of SlGRAS18 positively impacts arbuscule development and the improvement in symbiotic status, favouring flowering and therefore progress in the formation and development of fruits in SlGRAS18 silenced plants which parallel to a discernible pattern of mineral nutrient redistribution in leaves. Our results advance the knowledge of GRAS transcription factors involved in the formation and establishment of AM symbiosis and provide experimental evidence for how specific genetic alterations can lead to more effective AM symbiosis.

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Strigolactones (SLs) constitute essential phytohormones that control pathogen defense, resilience to phosphate deficiency and abiotic stresses. Furthermore, SLs are released into the soil by roots, especially in conditions in which there is inadequate phosphate or nitrogen available. SLs have the aptitude to stimulate the root parasite plants and symbiotic cooperation with arbuscular mycorrhizal (AM) fungi in rhizosphere. The use of mineral resources, especially phosphorus (P), by host plants is accelerated by AMF, which also improves plant growth and resilience to a series of biotic and abiotic stresses. Thus, these SL treatments that promote rhizobial symbiosis are substitutes for artificial fertilizers and other chemicals, supporting ecologically friendly farming practices. Moreover, SLs have become a fascinating target for abiotic stress adaptation in plants, with an array of uses in sustainable agriculture. In this review, the biological activity has been summarized that SLs as a signaling hormone for AMF symbiosis, nutrient acquisition, and abiotic stress tolerance through interaction with other hormones. Furthermore, the processes behind the alterations in the microbial population caused by SL are clarified, emphasizing the interplay with other signaling mechanisms. This review covers the latest developments in SL studies as well as the properties of SLs on microbial populations, plant hormone transductions, interactions and abiotic stress tolerance.

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Castanea sativa Miller, a high-valuable crop for Mediterranean countries, is facing frequent and prolonged periods of heat and drought, severely affecting chestnut production. Aiming to tackle this problem, this study unraveled the influence of mycorrhizal association with the fungi Paxillus involutus (Batsch) on young chestnut plants' responses to combined heat (42 °C; 4 h/day) and drought (no irrigation until soil moisture reached 25%) over 21 days of stress exposure. Heat stress had no harmful effects on growth, photosynthesis, nor induced oxidative stress in either mycorrhizal (MR) or non-mycorrhizal (NMR) chestnut plants. However, drought (alone or combined) reduced the growth of NMR plants, affecting water content, leaf production, and foliar area, while also hampering net CO2 assimilation and carbon relations. The mycorrhizal association, however, mitigated the detrimental effects of both stresses, resulting in less susceptibility and fewer growth limitations in MR chestnut plants, which were capable of ensuring a proper carbon flow. Evaluation of the oxidative metabolism revealed increased lipid peroxidation and hydrogen peroxide levels in NMR plants under water scarcity, supporting their higher susceptibility to stress. Conversely, MR plants activated defense mechanisms by accumulating antioxidant metabolites (ascorbate, proline and glutathione), preventing oxidative damage, especially under the combined stress. Overall, drought was the most detrimental condition for chestnut growth, with heat exacerbating stress susceptibility. Moreover, mycorrhizal association with P. involutus substantially alleviated these effects by improving growth, water relations, photosynthesis, and activating defense mechanisms. Thus, this research highlights mycorrhization's potential to enhance C. sativa resilience against climate change, especially at early developmental stages.

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Arbuscular mycorrhizal fungi (AMF) and Chitooligosaccharide (COS) can increase the resistance of plants to disease. COS can also promote the symbiosis between AMF and plants. However, the effects of AMF & COS combined application on the rhizosphere soil microbial community of tobacco and the improvement of tobacco's resistance to black shank disease are poorly understood.·We treated tobacco with AMF, COS, and combined application of AMF & COS (AC), respectively. Then studied the incidence, physio-biochemical changes, root exudates, and soil microbial diversity of tobacco seedling that was inoculated with Phytophthora nicotianae. The antioxidant enzyme activity and root vigor of tobacco showed a regular of AC > AMF > COS > CK, while the severity of tobacco disease showed the opposite regular. AMF and COS enhance the resistance to black shank disease by enhancing root vigor, and antioxidant capacity, and inducing changes in the rhizosphere microecology of tobacco. We have identified key root exudates and critical soil microorganisms that can inhibit the growth of P. nicotianae. The presence of caprylic acid in root exudates and Bacillus (WdhR-2) in rhizosphere soil microorganisms is the key factor that inhibits P. nicotianae growth. AC can significantly increase the content of caprylic acid in tobacco root exudates compared to AMF and COS. Both AMF and COS can significantly increase the abundance of Bacillus in tobacco rhizosphere soil, but the abundance of Bacillus in AC is significantly higher than that in AMF and COS. This indicates that the combined application of AMF and COS is more effective than their individual use. These findings suggest that exogenous stimuli can induce changes in plant root exudates, regulate plant rhizosphere microbial community, and then inhibit the growth of pathogens, thereby improving plant resistance to diseases.

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