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Background: Nutrient limitation is a universal phenomenon in terrestrial ecosystems. Root and mycorrhizal are critical to plant nutrient absorption in nutrient-limited ecosystems. However, how they are modified by N and P limitations with advancing vegetation successions in karst forests remains poorly understood. The present study compared the diversity indices, composition, and co-occurrence network of arbuscular mycorrhizal fungi (AMF) between grassland, shrubland, shrub-tree forest, and tree forest in subtropical karst forests, as well as soil nutrients and fine root functional traits (e.g., specific root length (SRL), specific root area (SRA), diameter, biomass, and N and P contents)., Results: The fine roots diameter, biomass, and N and P contents increased with advancing succession, whereas SRL and SRA decreased. Network complexity and Richness and Chao1 indices of AMF increased from grassland to shrub-tree forest but decreased in tree forest. The fine roots N and P contents were positively related to their diameter and biomass, soil nutrients, and AMF composition but were negatively correlated with SRL and SRA. Moreover, these two parameters increased with the increase of soil nutrients. The variations in fine roots N and P contents were mainly explained by soil nutrients and fine root functional traits in grassland and by the interactions of soil nutrients, fine root functional traits, and AMF in the other three stages. Additionally, the interactive explanation with AMF increased from shrubland to shrub-tree forest but decreased in tree forest., Conclusions: Our results indicated that mycorrhizal strategy might be the main nutrient acquisition strategy under N and P co-limitation. In contrast, the root strategy is the main one when an individual is subject to limitations in N or P in karst ecosystems. Root and mycorrhizal nutrient acquisition strategies are generally mutualistic, mycorrhizal strategy enhances plant nutrient acquisition under N and P co-limitation., Competing Interests: Declarations. Ethics approval and consent to participate: Not applicable. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

To investigate the effects of row ratio configurations on intercropping advantages and related rhizosphere microbial communities, a field experiment involving five treatments of different rows of broomcorn millet, i.e., P1M1 (1 row of broomcorn millet intercropped with 1 row of alfalfa), P2M3, P1M2, P1M3 and broomcorn millet alone (SP), was conducted on the Loess Plateau of China. We analyzed the yield, nutritional content of broomcorn millet, the soil nutrient availability and the diversity and community composition of AMF (arbuscular mycorrhizal fungi) and diazotrophs in the rhizosphere of broomcorn millet. The results showed that compared with monocultures, alfalfa-millet intercropping system under different row ratio configurations significantly increased the yield of broomcorn millet and the absorption of PTP and PTK (total phosphorus and potassium of broomcorn millet). In addition, the broomcorn millet-alfalfa intercropping system also improved soil nutrition, with the decrease of the row ratio of broomcorn millet, the changes of TN, NH 4 + -N and microbial biomass in the rhizosphere of broomcorn millet were consistent, which was opposite to NO 3 - -N. Moreover, co-occurrence network and PLS-PM (partial least squares path modelling) analysis showed alfalfa-broomcorn millet intercropping system changed the community diversity and composition of soil microorganisms, increased the improvement of soil nutrition (TN, NH 4 + -N and microbial biomass), and promoted the absorption of different nutrients by plants (N, P and K) mainly through the negative regulation of AMF and the synergistic effect of AMF on diazotrophs, and finally increased crop yield. This shows that broomcorn millet-alfalfa intercropping can increase plant nutrient content by adjusting soil nutrients and soil microbial activities, thereby increasing yield. Furthermore, we found that 1P2M was the best ratio of alfalfa-millet intercropping system, which may provide reliable suggestions and selection basis for future agricultural production practices., Competing Interests: Declarations. Ethics approval and consent to participate: All methods were carried out in compliance with local and national regulations. Clinical trial number: not applicable. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Background: Adventitious root (AR) formation is the key step for successful cutting propagation of tea plants (Camellia sinensis L.). Studies showed that arbuscular mycorrhizal fungus (AMF) can promote the rooting ability, and auxin pathway in basal stem of cuttings was involved in this process. However, auxin and abscisic acid (another important regulator on AR formation) in the other parts of cuttings at different rooting stages responding to AMF inoculation are not well studied. Therefore, in this paper, contents, enzymes and genes related to these two plant hormones were comprehensively determined aiming to unveil how endogenous indole-3-acetic acid (IAA) and abscisic acid (ABA) involve in the AMF regulating AR development of tea cuttings., Results: Inoculating with AMF significantly increased the proportion of cuttings at S2 stage (AR formation), which was more than twice as much as the control. And the total rooting rate in mycorrhizal treatment was also higher than that in the control with an increase of 8.66%. Enzyme activity assays showed that except for decreased polyphenol oxidase (PPO) activity at the S3 stage and peroxidase (POD) activity in middle stem of S3 stage, AMF inoculation increased activities of POD, PPO, superoxide dismutase (SOD) and catalase (CAT) to varying degrees in leaf, middle stem and basal stem of tea cuttings. After inoculation with AMF, the indoleacetic acid oxidase (IAAO) activity decreased to a certain extent in the first three stages of tea cuttings, which showed a trend of 'low-high-low' in the basal stem of all treatments. Besides, there was a significantly positive correlation between SOD activity and AR formation, especially for the proportion of cuttings at S2 and S3 stages. Higher IAA level and IAA/ABA ratio was found in basal stem of cuttings at S1 stage induced by AMF, which promoting the AR formation as revealed by correlation analysis. At the same time, AMF significantly elevated the level of IAA in leaf at S1 stage. By screening differentially expressed genes (DEGs) related to IAA and ABA pathways, together with redundant analysis, it was indicated that auxin biosynthesis and transport, as well as ABA transport and signal transduction, were involved in AMF regulating the rooting of tea cuttings., Conclusions: Overall, both endogenous IAA and ABA played roles in the regulation of AR formation of tea cuttings by AMF inoculating, enriching the theoretical basis of AMF regulating rooting of cuttings and providing foundations for cutting propagation of tea plants., Competing Interests: Declarations. Ethics approval and consent to participate: The authors all declare that any human participants, human data, or human tissue are not involved in the study of this manuscript. Therefore, it is not applicable. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Tuta absoluta (Lepidoptera: Gelechiidae) is one of the most significant invasive and destructive pests worldwide, causing serious economic losses to the tomato industry. Rhizosphere microorganism, such as arbuscular mycorrhizal fungi (AMF) and Pseudomonas bacteria, can interact with plants individually or collectively to improve plant growth and resistance to pests and disease. However, the effects of AMF, Pseudomonas, and their interactions on plant responses to insect herbivores remain unclear. A pot experiment was conducted to investigate the effects of single/dual inoculation with AMF (Funneliformis mosseae, M) and Pseudomonas putida (P) on the growth and defense of tomato variety Dafen (Solanum lycopersicum L.) in response to infestation by T. absoluta, as well as the growth, development, and enzyme activity of insect. The results showed that M, P, and MP promoted tomato growth by increasing nutrient concentrations, with the growth-promoting effect of dual-inoculation significantly surpassing that of single inoculation. M, P, and MP still improved tomato growth in T. absoluta infestation, with biomass increases of 57.34%, 54.46%, and 255.49%. M, P, and MP significantly increased the defense ability of tomato, with jasmonic acid concentrations increasing by 42.15%, 60.87% and 90.02%, and phenylalanine ammonia-lyase activity increasing by 47.40%, 47.68%, and 59.97%. The inoculation treatments inhibited the growth and development of T. absoluta, reduced its feeding, prolonged its growth and development, decreased egg weight, and increased the activity of protective and detoxifying enzymes. Overall, our results indicated that AMF and bacteria can stimulate each other, positively influence tomato growth and enhance resistance to T. absoluta. These findings indicate the feasibility of AMF and bacteria in combinations as potential biocontrol agents for the management of T. absoluta., Competing Interests: Declarations. Ethics approval and consent to participate: Not applicable. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Background: Arbuscular mycorrhizal fungi (AMF) can regulate metal(loid) tolerance in plants and their capacity for phytoremediation. These effects can vary depending on the host plant and the AMF species. The impact of different AMF species on the ability of safflower (Carthamus tinctorius L.) for arsenic (As) phytoremediation is still largely unknown. Therefore, this study aimed to assess the effect of two AMF species, Rhizophagus irregularis, and Funneliformis mosseae, on the tolerance and accumulation of As in safflower in soils spiked with varying arsenate concentrations (0, 25, 50, and 100 mg kg -1 )., Results: The results indicated that both AMF species established effective symbiotic relationships with safflower. However, plants inoculated with R. irregularis exhibited higher mycorrhizal dependency and root colonization, especially under 100 mg kg -1 As. Both AMF species significantly improved plant growth parameters, chlorophyll content, and phosphorus (P) nutrition, which resulted in increased P/As ratio and enhanced tolerance index in safflower plants. In addition, AMF inoculation reduced As-induced lipid peroxidation by enhancing catalase and peroxidase activity in leaves and roots. While the mycorrhizal symbiosis didn't affect As availability in soils, it significantly reduced shoot As concentration and the translocation factor under all As levels. Furthermore, mycorrhizal inoculation, especially with R. irregularis, increased As concentration and modified-bioconcentration factor in the roots and enhanced total As uptake per plant., Conclusions: Based on the results and multivariate analyses, both AMF species, particularly R. irregularis, enhanced safflower's As tolerance by retaining As in roots, improving phosphorus nutrition, and increasing antioxidant enzyme activity, showcasing their potential to enhance phytostabilization in safflower plants., Competing Interests: Declarations. Ethics approval and consent to participate: Not applicable. This manuscript does not involve researching about humans or animals. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

This experiment was conducted to study the effect of inoculation with arbuscular mycorrhizal fungi (AMF) as an ecofriendly strategy on physiological, biochemical and yield characters of wheat plants. Our results showed a significant decrease in chlorophyll a and b as well as the maximum quantum efficiency of PSII (Fv/Fm) in wheat plants under drought conditions compared to control in the two seasons. Drought stress significantly reduced relative water content (RWC%) in the stressed plants compared with the control. Additionally, 1000 grain weight (g) and biological yield (t h - 1 ) were reduced significantly under drought stress. Furthermore, hydrogen peroxide (H 2 O 2 ), Superoxide (O 2 - ), electrolyte leakage (EL%), lipid peroxidation (MDA) and total phenolic compounds were increased significantly in the stressed plants under drought conditions. However, wheat plants treated with AMF displayed a significant increase in chlorophyll concentrations and the maximum quantum efficiency of PSII as well as RWC% in the stressed wheat plants when compared with the stressed untreated plants. Our findings also indicated that application of AMF led to regulate the antioxidant enzymes activity, proline content and decrease hydrogen peroxide (H 2 O 2 ), Superoxide (O 2 - ), electrolyte leakage (EL%) and lipid peroxidation (MDA) levels in the drought stressed wheat plants. Eventually, application of AMF as ecofriendly approach can improve wheat growth and grain yield of wheat plants by mitigating the adverse effects of drought stress. These results provide evidence for the important role of AMF in agricultural production and maximizing wheat grain yield., Competing Interests: Declarations. Ethics approval and consent to participate: Not applicable. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Background and Aims: Continuous vegetable production under plastic tunnels faces challenges like soil degradation, increased soil-borne pathogens, and diminished eggplant yield. These factors collectively threaten the long-term sustainability of food security by diminishing the productivity and resilience of agricultural soils. This research examined the use of raw garlic stalk (RGS) waste and arbuscular mycorrhizal fungi (AMF) as a sustainable solution for these issues in eggplant monoculture. We hypothesized that the combined application of RGS waste and AMF would improve soil physicochemical properties compared to untreated soil in eggplant monoculture. The combined use of RGS and AMF was expected to suppress soil-borne pathogens, increase the abundance of soil beneficial microorganisms and alter fungal community structure. The combined application of RGS and AMF will significantly enhance eggplant yield compared to untreated plots. This study aimed to determine whether AMF and RGS, individually or in combination, can ameliorate the adverse effects of monoculture on eggplant soil. We also investigated whether these treatments could enhance eggplant yield., Methods: The experiment was arranged in a completely randomized design with four treatments: AMF, RGS, and a combined treatment of AMF + RGS (ARGS), along with a control. Each treatment was replicated three times, Eggplant seedlings inoculated with AMF and treated with RGS amendments, both individually and combined. The effects on root traits, soil physicochemical properties, soil enzyme activity, and fungal community structure were investigated., Results: RGS amendments and AMF inoculation improved root length, volume, and mycorrhizal colonization. The combined treatment showed the most significant improvement. RGS and AMF application increased soil nutrient availability (N, P, K) and organic matter content. Enzyme activities also increased with RGS and AMF treatments, with the combined application showing the highest activity. Soil electrical conductivity (EC) increased, while soil pH decreased with RGS and AMF amendments. Sequencing revealed a shift in the fungal community structure. Ascomycota abundance decreased, while Basidiomycota abundance increased with RGS and AMF application. The combined treatment reduced the abundance of pathogenic genera (Fusarium) and enriched beneficial taxa (Chaetomium, Coprinellus, Aspergillus). Pearson correlations supported the hypothesis that soil physicochemical properties influence fungal community composition., Conclusions: This study demonstrates the potential of co-applying RGS and AMF in continuous cropping systems. It enhances soil physicochemical properties, reduces soil-borne pathogens, and promotes beneficial microbial communities and eggplant yield. This combined approach offers a sustainable strategy to address the challenges associated with eggplant monoculture under plastic tunnels., Competing Interests: Declarations Ethics declarations We declare that the manuscript reporting the research does not involve human subjects, data, or tissue. Therefore, it is not applicable. Consent for publication Not applicable. Competing interests The authors declare no competing interests., (© 2024. The Author(s).)

The plant kingdom exhibits a diversity of nutritional strategies, extending beyond complete autotrophy. In addition to full mycoheterotrophs and holoparasites, it is now recognized that a greater number of green plants than previously assumed use partly of fungal carbon. These are termed partial mycoheterotrophs or mixotrophs. Notably, some species exhibit a dependency on fungi exclusively during early ontogenetic stages, referred to as initial mycoheterotrophy. Japonolirion osense, a rare plant thriving in serpentinite soils, emerges as a potential candidate for initial mycoheterotrophy or mixotrophy. Several factors support this hypothesis, including its diminutive sizes of shoot and and seeds, the establishment of Paris-type arbuscular mycorrhizal associations, its placement within the Petrosaviales-largely composed of fully mycoheterotrophic species-and its ability to face the challenging conditions of its environment. To explore these possibilities, our study adopts a multidisciplinary approach, encompassing stable isotope abundance analyses, in vitro experiments, anatomical analyses, and comparative plastome analyses. Our study aims to (1) determine whether J. osense relies on fungal carbon during germination, indicating initial mycoheterotrophy, (2) determine if it employs a dual carbon acquisition strategy as an adult, and (3) investigate potential genomic reductions in photosynthetic capabilities. Contrary to expectations, our comprehensive findings strongly indicate that J. osense maintains complete autotrophy throughout its life cycle. This underscores the contrasting nutritional strategies evolved by species within the Petrosaviales., (© 2024. The Author(s).)

Colonization by Ectomycorrhizal (EcM) fungi is key for the health and performance of plants under different stress scenarios, such as those faced by trees in urban environments. Because urban environments can be lacking EcM fungi, we here assessed the benefits of inoculating Tilia tomentosa seedlings in a pre-transplantation nursery context with the EcM fungi Lactarius deliciosus and Paxillus involutus, using substrates of different pH and facing water-stress. P. involutus had a more evident positive effect in T. tomentosa seedlings and had a good performance in both acidic and alkaline substrate. In acidic substrate the fungus increased the plant height by 0.91-fold, increased the mycorrhization rate by 3.23-fold, expansion rate by 5.03-fold and formation of secondary roots by 0.46-fold, compared to the non-inoculated control. This species also improved the phosphorus content of leaves, which revealed a promotion of nutrient uptake. In alkaline substrate P. involutus increased root dry weight by 3.92-fold and the mycorrhization parameters. In contrast, L. deliciosus only had a positive effect in the improvement of mycorrhization and expansion rates and phosphorus content in the root, effects visible only in alkaline substrate. When exposed to water-stress the increase of proline content was visible in acidic substrate for both fungi, L. deliciosus and P. involutus, and in alkaline substrate for the fungus P. involutus, a response indicative of the enhancement of defenses in stressing scenarios such as water scarcity. We conclude that fungal inoculation improves the vigour and resilience of Tilia seedlings and that it is of utmost importance to select a suitable EcM fungus and to consider the soil pH of the transplanting site. The inoculation approach can be a valuable tool to produce robust seedlings which may have a better performance when transplanted to the challenging urban environment., (© 2024. The Author(s).)

Arbuscular mycorrhizal fungi (AMF) have profound effects on plant growth and nitrogen (N) and phosphorus (P) nutrition. However, a comprehensive evaluation of how plant N and P respond to AMF inoculation is still unavailable. Here, we complied data from 187 original researches and carried out a meta-analysis to assess the effects of AMF inoculation on plant growth and N and P nutrition. We observe overall positive effects of AMF inoculation on plant performance. The mean increases of plant biomass, N concentration, P concentration, N and P uptake of whole plant are 47%, 16%, 27%, 67%, and 105%, respectively. AMF inoculation induces more increases in plant concentrations and storage of P than N. Plant responses to AMF inoculation are substantially higher with single AMF species than with mixed AMF species, in laboratory experiments than in field experiments, and in legumes than in non-legumes. The response ratios of plant N and P nutrition are positively correlated with AMF colonization rate, N addition, P addition, and water condition, while unvaried with experiment duration. The biggest and smallest effect sizes of AMF inoculation on plant performance are observed in the application of nitrate and ammonium, respectively. Accordingly, this meta-analysis study clearly suggests that AMF inoculation improves both plant N and P nutrients and systematically clarifies the variation patterns in AMF effects with various biotic and abiotic factors. These findings highlight the important role of AMF inoculation in enhancing plant N and P resource acquisitions and provide useful references for evaluating the AMF functions under the future global changes., (© 2024. The Author(s).)

Bio-transformations refer to the chemical modifications made by an organism on a chemical compound that often involves the interaction of plants with microbes to alter the chemical composition of soil or plant. Integrating bio-transformations and entomopathogenic fungi into litchi cultivation can enhance symbiotic relationships, microbial enzymatic activity in rhizosphere, disease suppression and promote overall plant health. The integration of biological formulations and entomopathogenic fungi can significantly influence growth, nutrient dynamics, physiology, and rhizosphere microbiome of air-layered litchi (Litchi chinensis Sonn.) saplings. Biological modifications included, K-mobilizers, AM fungi, Pseudomonas florescence and Azotobacter chroococcum along with Metarhizium, entomopathogenic fungi have been used. The treatments included, T 1 -Litchi orchard soil + sand (1:1); T 2 -Sand + AM fungi + Azotobacter chroococcum (1:2:1); T 3 -Sand + Pseudomonas florecence + K-mobilizer (1:1:1); T 4 - AM fungi + K-mobilizers (1:1); T 5 , P. Florecence + A. chroococcum + K-mobilizer (1:1:1); T 6 -Sand + P. florecence (1:2) and T 7 -Uninoculated control for field performance. Treatments T 4 -T 6 were further uniformly amended with drenching of Metarrhizium in rhizosphere. T 2 application significantly increased resident microbe survival, total chlorophyll content and root soil ratio in seedlings. A. chroococcum, Pseudomonas, K-mobilizers and AM fungi increased in microbial biomass of 2.59, 3.39, 2.42 and 2.77 times, respectively. Acidic phosphatases, dehydrogenases and alkaline phosphatases were increased in rhizosphere. Leaf nutrients reflected through DOP were considerably altered by T 2 treatment. Based on Eigen value, PCA-induced changes at biological modifications showed maximum total variance. The study inferred that the bio-transformations through microbial inoculants and entomopathogenic fungi could be an encouraging strategy to enhance the growth of plants, health and productivity. Such practices align well with the goals of sustainable agriculture through biological means by reducing dependency on chemical inputs. By delving into these aspects, the research gaps including microbial processes, competitive and symbiotic relationships, resistance in microbes and how complex interactions among bio-transformations, entomopathogenic fungi and microbes can significantly impact the health and productivity of litchi. Understanding and harnessing these interactions can lead to more effective and sustainable farming practices., (© 2024. The Author(s).)

Background: Gadolinium (Gd) is an increasingly found lanthanide element in soil; thus, understanding its impact on plant physiology, biochemistry, and molecular responses is crucial. Here, we aimed to provide a comprehensive understanding of Gd (150 mg kg - 1 ) impacts on guar (Cyamopsis tetragonoloba L.) plant yield and metabolism and whether the symbiotic relationship with arbuscular mycorrhizal fungi (AMF) can mitigate Gd toxicity of soil contamination., Results: AMF treatment improved mineral nutrient uptake and seed yield by 38-41% under Gd stress compared to non-inoculated stressed plants. Metabolic analysis unveiled the defense mechanisms adopted by AMF-treated plants, revealing carbon and nitrogen metabolism adaptations to withstand Gd contamination. This included an increase in the synthesis of primary metabolites, such as total sugar (+ 39% compared to control), soluble sugars (+ 29%), starch (+ 30%), and some main amino acids like proline (+ 57%) and phenylalanine (+ 87%) in the seeds of AMF-treated plants grown under Gd contamination. Furthermore, fatty acid and organic acid profile changes were accompanied by the production of secondary metabolites, including tocopherols, polyamines, phenolic acids, flavones, and anthocyanins., Conclusions: Overall, the coordinated synthesis of these compounds underscores the intricate regulatory mechanisms underlying plant-AMF interactions and highlights the potential of AMF to modulate plant secondary metabolism for enhanced Gd stress tolerance., (© 2024. The Author(s).)

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., (© 2024. The Author(s).)

Backround: The utilization of high-quality water in agriculture is increasingly constrained by climate change, affecting availability, quality, and distribution due to altered precipitation patterns, increased evaporation, extreme weather events, and rising salinity levels. Salinity significantly challenges salt-sensitive vegetables like lettuce, particularly in a greenhouse. Hydroponics water quality ensures nutrient solution stability, enhances nutrient uptake, prevents contamination, regulates pH and electrical conductivity, and maintains system components. This study aimed to mitigate salt-induced damage in lettuce grown via the floating culture method under 50 mM NaCl salinity by applying biostimulants., Results: We examined lettuce's physiological, biochemical, and agronomical responses to salt stress after applying biostimulants such as amino acids, arbuscular mycorrhizal fungi, plant growth-promoting rhizobacteria (PGPR), fulvic acid, and chitosan. The experiment was conducted in a greenhouse with a randomized complete block design, and each treatment was replicated four times. Biostimulant applications alleviated salt's detrimental effects on plant weight, height, leaf number, and leaf area. Yield increases under 50 mM NaCl were 75%, 51%, 31%, 34%, and 33% using vermicompost, PGPR, fulvic acid, amino acid, and chitosan, respectively. Biostimulants improved stomatal conductance (58-189%), chlorophyll content (4-10%), nutrient uptake (15-109%), and water status (9-107%). They also reduced MDA content by 26-42%. PGPR (1.0 ml L ‒1 ), vermicompost (2 ml L ‒1 ), and fulvic acid (40 mg L ‒1 ) were particularly effective, enhancing growth, yield, phenol, and mineral content while reducing nitrate levels under saline conditions., Conclusions: Biostimulants activated antioxidative defense systems, offering a sustainable, cost-effective solution for mitigating salt stress in hydroponic lettuce cultivation., (© 2024. The Author(s).)

Background: The symbiosis among plants, rhizobia, and arbuscular mycorrhizal fungi (AMF) is one of the most well-known symbiotic relationships in nature. However, it is still unclear how bilateral/tripartite symbiosis works under resource-limited conditions and the diverse genetic backgrounds of the host., Results: Using a full factorial design, we manipulated mungbean accessions/subspecies, rhizobia, and AMF to test their effects on each other. Rhizobia functions as a typical facilitator by increasing plant nitrogen content, plant weight, chlorophyll content, and AMF colonization. In contrast, AMF resulted in a tradeoff in plants (reducing biomass for phosphorus acquisition) and behaved as a competitor in reducing rhizobia fitness (nodule weight). Plant genotype did not have a significant effect on AMF fitness, but different mungbean accessions had distinct rhizobia affinities. In contrast to previous studies, the positive relationship between plant and rhizobia fitness was attenuated in the presence of AMF, with wild mungbean being more responsive to the beneficial effect of rhizobia and attenuation by AMF., Conclusions: We showed that this complex tripartite relationship does not unconditionally benefit all parties. Moreover, rhizobia species and host genetic background affect the symbiotic relationship significantly. This study provides a new opportunity to re-evaluate the relationships between legume plants and their symbiotic partners., (© 2024. The Author(s).)

Background: Legumes utilize a long-distance signaling feedback pathway, termed Autoregulation of Nodulation (AON), to regulate the establishment and maintenance of their symbiosis with rhizobia. Several proteins key to this pathway have been discovered, but the AON pathway is not completely understood., Results: We report a new hypernodulating mutant, defective in autoregulation, with disruption of a gene, DAR (Medtr2g450550/MtrunA17_Chr2g0304631), previously unknown to play a role in AON. The dar-1 mutant produces ten-fold more nodules than wild type, similar to AON mutants with disrupted SUNN gene function. As in sunn mutants, suppression of nodulation by CLE peptides MtCLE12 and MtCLE13 is abolished in dar. Furthermore, dar-1 also shows increased root length colonization by an arbuscular mycorrhizal fungus, suggesting a role for DAR in autoregulation of mycorrhizal symbiosis (AOM). However, unlike SUNN which functions in the shoot to control nodulation, DAR functions in the root., Conclusions: DAR encodes a membrane protein that is a member of a small protein family in M. truncatula. Our results suggest that DAR could be involved in the subcellular transport of signals involved in symbiosis regulation, but it is not upregulated during symbiosis. DAR gene family members are also present in Arabidopsis, lycophytes, mosses, and microalgae, suggesting the AON and AOM may use pathway components common to other plants, even those that do not undergo either symbiosis., (© 2024. The Author(s).)

Background: Soybean is the main oil crop in Northeast China. Continuous monocropping is more commonly used for soybean production due to rising market demand and arable land constraints. However, autotoxic substances, such as phenolic acids, produced by continuously cropped soybean can reduce yield and quality. The mycorrhiza formed of Arbuscular mycorrhizal fungi (AMF) and plant roots regulate the metabolic activities of the host plant and increase its disease resistance. The main purpose of this study was to inhibit the production of phenolic acids and determine the adverse effects on the growth of continuous monocropping soybean by inoculating Funneliformis mosseae (F. mosseae)., Results: Transcriptomics results showed that the production of phenolic acids in continuous monocropping soybean roots was mainly regulated by the expression of the CHS6, PCL1, SAMT, SRG1, and ACO1 genes, and the expression of these genes was significantly downregulated after inoculation with F. mosseae. Metabolomics results showed that continuous monocropping soybean roots inoculated with F. mosseae inhibited phenolic acid production through the phenylpropane biosynthetic, α-linoleic acid, linoleic acid, and other metabolic pathways. Phenolic acids in the phenylpropane metabolic pathway, such as 4-hydroxybenzoic acid, phthalic acid, and vanillic acid, decreased significantly after inoculation with F. mosseae. The combined analysis of the two showed that genes such as YLS9 and ARF3 were positively correlated with 4-hydroxybenzoic acid and so on, while genes such as CHS6 and SRG1 were negatively correlated with butyric acid and so on., Conclusion: F. mosseae regulated the expression of functional genes and related phenolic acid metabolic pathways produced by continuous monocropping soybean roots, inhibiting the production of phenolic acid autotoxic substances in continuous cropped soybean, and slowing down the disturbance of continuous monocropping. This study provides a new solution for continuous monocropping of plants to overcome the autotoxicity barrier and provides a new basis for the development and utilization of AMF as a biological agent., (© 2024. The Author(s).)

Background: Opuntia ficus-indica (L.) Miller is dominantly growing on degraded soils in arid and semi-arid areas. The plants might establish a strong association with arbuscular mycorrhizal fungi (AMF) to adapt to nutrient, drought, and herbivore insect stress. The objective of this study was to investigate the effects of AMF inoculations and variable soil water levels (SWA) on the biomass, nutrient concentration, nutritional composition, and nutrient digestibility of the spiny and spineless O. ficus-indica by inducing resistance to cochineal stress. One mother Opuntia ficus-indica cladode was planted in a single pot in each field with 24 kg mixed soil. AMF inoculums were cultured in sorghum plants in a greenhouse and were inoculated in the planted cladodes. The planted cladodes were arranged using a completely randomized design (CRD) with three factors: AMF (present and absent); O. ficus-indica type (spiny and spineless) and four water treatments with 0-25% of plant available soil water (SWA), 25-50% of SWA, 50-75% of SWA, and 75-100% of SWA., Results: Drought stress reduced the below and above-ground biomass, cladode nutrient content, nutritional composition, and in vitro dry matter digestibility (IVDMD) and in vitro organic matter digestibility (IVOMD). AMF colonization significantly increased biomass production with significant changes in the macro and micro-nutrient concentrations of O. ficus-indica. AMF inoculation significantly increased the IVDMD and IVOMD of both O. ficus-indica types by improving the biomass, organic matter (OM), crude protein (CP), and reduced fiber and ash contents. AMF-inoculated cladodes improved the nutrient concentrations of the cladodes. AMF caused an increase in biomass production, increased tolerance to cochineal stress, and improved nutrient concentration, nutritional composition, and nutrient digestibility performance of O. ficus-indica plants., Conclusions: AMF improved the performance of the O. ficus-indica plant to resist drought and cochineal stress and increased the biomass, nutrient concentration, nutritional composition, and nutrient digestibility. The potential of O. ficus-indica to adapt to cochineal stress is controlled by the macro and micro-nutrient concentration brought by the AMF association., (© 2024. The Author(s).)

Recent studies have exhibited a very promising role of copper nanoparticles (CuNPs) in mitigation of abiotic stresses in plants. Arbuscular mycorrhizae fungi (AMF) assisted plants to trigger their defense mechanism against abiotic stresses. Arsenic (As) is a non-essential and injurious heavy-metal contaminant. Current research work was designed to elucidate role of CuNPs (100, 200 and 300 mM) and a commercial inoculum of Glomus species (Clonex ® Root Maximizer) either alone or in combination (CuNPs + Clonex) on physiology, growth, and stress alleviation mechanisms of E. sibiricus growing in As spiked soils (0, 50, and 100 mg Kg - 1 soil). Arsenic induced oxidative stress, enhanced biosynthesis of hydrogen peroxide, lipid peroxidation and methylglyoxal (MG) in E. sibiricus. Moreover, As-phytotoxicity reduced photosynthetic activities and growth of plants. Results showed that individual and combined treatments, CuNPs (100 mM) as well as soil inoculation of AMF significantly enhanced root growth and shoot growth by declining As content in root tissues and shoot tissues in As polluted soils. E. sibiricus plants treated with CuNPs (100 mM) and/or AMF alleviated As induced phytotoxicity through upregulating the activity of antioxidative enzymes such as catalase (CAT) and superoxide dismutase (SOD) besides the biosynthesis of non-enzymatic antioxidants including phytochelatin (PC) and glutathione (GSH). In brief, supplementation of CuNPs (100 mM) alone or in combination with AMF reduced As uptake and alleviated the As-phytotoxicity in E. sibiricus by inducing stress tolerance mechanism resulting in the improvement of the plant growth parameters., (© 2024. The Author(s).)

Background: Long-term continuous cropping has resulted in the frequent occurrence of fusarium wilt of watermelon (Citrullus lanatus). AMF inoculation can alleviate the continuous cropping barrier and reduce the incidence of fusarium wilt of watermelon. Our previous study found that the root exudates of mycorrhizal watermelon can enhance watermelon resistance to this disorder. It is necessary to further isolate and identify the specific compounds in root exudates of mycorrhizal watermelon and explore their control effects on fusarium wilt of continuous cropping watermelon., Result: The results of this study showed that the root system of watermelon seedlings inoculated with AMF (Funneliformis mosseae or Glomus versiforme) secreted diisooctyl phthalate (A) and dibutyl phthalate (B). Compared with water treatment, treatment with 0.1 ml/L (A1, B1), 0.5 ml/L (A2, B2) and 1 ml/L (A3, B3) of A or B significantly increased soil enzyme activities, the numbers of bacteria and actinomycetes, and the bacteria/fungi ratio in the rhizosphere. Furthermore, the Disease indexes (DI) of A1 and B3 were 25% and 20%, respectively, while the prevention and control effects (PCE) were 68.8% and 75%, respectively. In addition, diisooctyl phthalate or dibutyl phthalate increased the proportions of Gemmatimonadetes, Chloroflexi, and Acidobacteria in the rhizosphere of continuous cropping watermelon, and decreased the proportions of Proteobacteria and Firmicutes, with Novosphingobium, Kaistobacter, Bacillus, and Acinetobacter as the predominant bacteria. Compared with the water treatment, the abundance of Neosphingosaceae, Kateybacterium and Bacillus in the A1 group was increased by 7.33, 2.14 and 2.18 times, respectively, while that in the B2 group was increased by 60.05%, 80.24% and 1 time, respectively. In addition, exogenous diisooctyl phthalate and dibutyl phthalate were shown to promote growth parameters (vine length, stem diameter, fresh weight and dry weight) and antioxidant enzyme system activities (SOD, POD and CAT) of continuous cropping watermelon., Conclusion: Lower watermelon fusarium wilt incidence in mycorrhizal watermelons was associated with phthalate secretion in watermelons after AMF inoculation. Exogenous diisooctyl phthalate and dibutyl phthalate could alleviate the continuous cropping disorder of watermelon, reduce the incidence of fusarium wilt, and promote the growth of watermelon by increasing the enzyme activities and the proportion of beneficial bacteria in rhizosphere soil. In addition, the low concentration of phthalate diisooctyl and high concentration of phthalic acid dibutyl works best. Therefore, a certain concentration of phthalates in the soil can help alleviate continuous cropping obstacles., (© 2024. The Author(s).)

Background: This study aimed to investigate the alterations in biochemical and physiological responses of oat plants exposed to antimony (Sb) contamination in soil. Specifically, we evaluated the effectiveness of an arbuscular mycorrhizal fungus (AMF) and olive mill waste (OMW) in mitigating the effects of Sb contamination. The soil was treated with a commercial strain of AMF (Rhizophagus irregularis) and OMW (4% w/w) under two different levels of Sb (0 and 1500 mg kg -1 soil)., Results: The combined treatment (OMW + AMF) enhanced the photosynthetic rate (+ 40%) and chlorophyll a (+ 91%) and chlorophyll b (+ 50%) content under Sb condition, which in turn induced more biomass production (+ 67-78%) compared to the contaminated control plants. More photosynthesis in OMW + AMF-treated plants gives a route for phenylalanine amino acid synthesis (+ 69%), which is used as a precursor for the biosynthesis of secondary metabolites, including flavonoids (+ 110%), polyphenols (+ 26%), and anthocyanins (+ 63%) compared to control plants. More activation of phenylalanine ammonia-lyase (+ 38%) and chalcone synthase (+ 26%) enzymes in OMW + AMF-treated plants under Sb stress indicated the activation of phenylpropanoid pathways in antioxidant metabolites biosynthesis. There was also improved shifting of antioxidant enzyme activities in the ASC/GSH and catalytic pathways in plants in response to OMW + AMF and Sb contamination, remarkably reducing oxidative damage markers., Conclusions: While individual applications of OMW and AMF also demonstrated some degree of plant tolerance induction, the combined presence of AMF with OMW supplementation significantly enhanced plant biomass production and adaptability to oxidative stress induced by soil Sb contamination., (© 2024. The Author(s).)

Background: Selenium (Se) fertilizer and arbuscular mycorrhizal fungi (AMF) are known to modulate cadmium (Cd) toxicity in plants. However, the effects of their co-application on wheat growth and soil microbial communities in Cd-contaminated soil are unclear., Results: A pot experiment inoculation with two types of AMF and the application of Se fertilizer under Cd stress in wheat showed that inoculation AMF alone or combined with Se fertilizer significantly increased wheat biomass. Se and AMF alone or in combination significantly reduced available Cd concentration in wheat and soil, especially in the Se combined with Ri treatment. High throughput sequencing of soil samples indicated that Se and AMF application had stronger influence on bacterial community compared to fungal community and the bacterial network seemed to have more complex interconnections than the fungal network, and finally shaped the formation of specific microflora to affect Cd availability., Conclusion: These results indicate that the application of Se and AMF, particularly in combination, could successfully decrease soil Cd availability and relieve the harm of Cd in wheat by modifying rhizosphere soil microbial communities., (© 2024. The Author(s).)

Background: The establishment of mycorrhizal relationships between a fungus and a plant typically enhances nutrient and water uptake for the latter while securing a carbon source for the fungus. However, under a particular set of environmental conditions, such as low availability of light and abundant nutrients in the soil, the resources invested in the maintenance of the fungi surpass the benefits obtained by the host. In those cases, facultative mycorrhizal plants are capable of surviving without symbiosis. Facultative mycorrhization in ferns has been overlooked until now. The present study measured the response of Struthiopteris spicant L. Weiss, and its root-associated fungi to different levels of light and nutrient availability in terms of growth, mycorrhizal presence, and leaf nutrient content. This fern species exhibits a great tolerance to variable light, nutrient, and pH conditions, and it has been found with and without mycorrhizae. We conducted a greenhouse experiment with 80 specimens of S. spicant and three factors (Light, Phosphorus, and Nitrogen) resulting in eight treatments., Results: We found a significant influence of the factor light on fungal community composition, plant biomass, and nutrient accumulation. Departing from a lack of colonization at the initial stage, plants showed a remarkable increment of more than 80% in the arbuscular mycorrhizal fungi (AMF) richness and abundance in their roots when grown under high light conditions, compared with the ones in low light. We also observed an upward trend of C:P and C:N ratios and the above- and belowground biomass production when AMF abundance increased. Furthermore, the compositional analysis of the whole fungal communities associated with S. spicant roots revealed clear differences among low-light and high-light treatments., Conclusions: This study is the first to investigate the importance of light and nutrient availability in determining fern-AMF relationships. We confirmed that Struthiopteris spicant is a facultative mycorrhizal plant. The composition and diversity of AMF found in the roots of this fern are strongly influenced by light and less by nutrient conditions. Our study shows that ferns respond very sensitively to changes in environmental factors, leading to shifts in the associated mycorrhizal communities., (© 2024. The Author(s).)

Chromium (Cr) toxicity significantly threatens sunflower growth and productivity by interfering with enzymatic activity and generating reactive oxygen species (ROS). Zinc quantum dot biochar (ZQDB) and arbuscular mycorrhizal fungi (AMF) have become popular to resolve this issue. AMF can facilitate root growth, while biochar tends to minimize Cr mobility in soil. The current study aimed to explore AMF and ZQDB combined effects on sunflower plants in response to Cr toxicity. Four treatments were applied, i.e. NoAMF + NoZQDB, AMF + 0.40%ZQDB, AMF + 0.80%ZQDB, and AMF + 1.20%ZQDB, under different stress levels of Cr, i.e. no Cr (control), 150 and 200 mg Cr/kg soil. Results showed that AMF + 1.20%ZQDB was the treatment that caused the greatest improvement in plant height, stem diameter, head diameter, number of leaves per plant, achenes per head, 1000 achenes weight, achene yield, biological yield, transpiration rate, stomatal conductance, chlorophyll content and oleic acid, relative to the condition NoAMF + No ZQDB at 200 mg Cr/kg soil. A significant decline in peroxidase (POD), superoxide dismutase (SOD), and catalase (CAT) while improvement in ascorbate peroxidase (APx), oil content, and protein content further supported the effectiveness of AMF + 1.20%ZQDB against Cr toxicity. Our results suggest that the treatment AMF + 1.20%ZQDB can efficiently alleviate Cr stress in sunflowers., (© 2023. The Author(s).)

Background: Plant respiratory burst oxidase homolog (Rboh) gene family produces reactive oxygen species (ROS), and it plays key roles in plant-microbe interaction. Most Rboh gene family-related studies mainly focused on dicotyledonous plants; however, little is known about the roles of Rboh genes in gramineae., Results: A total of 106 Rboh genes were identified in seven gramineae species, including Zea mays, Sorghum bicolor, Brachypodium distachyon, Oryza sativa, Setaria italica, Hordeum vulgare, and Triticum aestivum. The Rboh protein sequences showed high similarities, suggesting that they may have conserved functions across different species. Duplication mode analysis detected whole-genome/segmental duplication (WGD)/(SD) and dispersed in the seven species. Interestingly, two local duplication (LD, including tandem and proximal duplication) modes were found in Z. mays, S. italica and H. vulgare, while four LD were detected in T. aestivum, indicating that these genes may have similar functions. Collinearity analysis indicated that Rboh genes are at a stable evolution state in all the seven species. Besides, Rboh genes from Z. mays were closely related to those from S. bicolor, consistent with the current understanding of plant evolutionary history. Phylogenetic analysis showed that the genes in the subgroups I and II may participate in plant-AM fungus symbiosis. Cis-element analysis showed that different numbers of elements are related to fungal induction in the promoter region. Expression profiles of Rboh genes in Z. mays suggested that Rboh genes had distinct spatial expression patterns. By inoculation with AM fungi, our transcriptome analysis showed that the expression of Rboh genes varies upon AM fungal inoculation. In particularly, ZmRbohF was significantly upregulated after inoculation with AM fungi. pZmRbohF::GUS expression analyses indicated that ZmRbohF was induced by arbuscular mycorrhizal fungi in maize. By comparing WT and ZmRbohF mutant, we found ZmRbohF had limited impact on the establishment of maize-AM fungi symbiosis, but play critical roles in regulating the proper development of arbuscules., Conclusions: This study provides a comprehensive analysis of the evolution relationship of Rboh genes in seven gramineae species. Results showed that several Rboh genes regulate maize-AM fungal symbiosis process. This study provides valuable information for further studies of Rboh genes in gramineae., (© 2023. The Author(s).)

Salinity stress is one of the major hurdles in agriculture which adversely affects crop production. It can cause osmotic imbalance, ion toxicity that disrupts essential nutrient balance, impaired nutrient uptake, stunted growth, increased oxidative stress, altered metabolism, and diminished crop yield and quality. However, foliar application of osmoprotectant is becoming popular to resolve this issue in crops. These osmoprotectants regulate the cellular osmotic balance and protect plants from the detrimental effects of high salt concentrations. Furthermore, the role of arbuscular mycorrhizae (AMF) is also established in this regard. These AMF effectively reduce the salinity negative effects by improving the essential nutrient balance via the promotion of root growth. That's why keeping in mind the effectiveness of osmoprotectants current study was conducted on cotton. Total of six levels of γ-Aminobutyric acid (GABA = 0 mM, 0. 5 mM, and 1 mM) and ectoine (ECT = 0 mM, 0.25 mM, and 0.5 mM) were applied as treatments in 3 replications. Results showed that 0.5 mM γ-Aminobutyric acid and ectoine performed significantly best for the improvement in cotton growth attributes. It also caused significant enhancement in K and Ca contents of the leaf, stem, bur, and seeds compared to the control. Furthermore, 0.5 mM γ-Aminobutyric acid and ectoine also caused a significant decline in Cl and Na contents of leaf, stem, bur, and seeds of cotton compared to control under salinity stress. A significant enhancement in chlorophyll contents, gas exchange attributes, and decline in electrolyte leakage validated the effectiveness of 0.5 mM γ-Aminobutyric acid and ectoine over control. In conclusion, 0.5 mM γ-Aminobutyric acid and ectoine have the potential to mitigate the salinity stress in cotton., (© 2023. BioMed Central Ltd., part of Springer Nature.)

Background: Heavy metals (HMs) phytoremediation is a well-recognized protocol to remove toxic elements from the soil. As known, arbuscular mycorrhizal fungi (AMF) enhance the plants' growth responses. The idea of the present study was to assay the response of lavender plants to HMs stress under AMF inoculation. We hypothesized that mycorrhiza will enhance the phytoremediation and simultaneously reduce the harmful effects of heavy HMs. So, lavender (Lavandula angustifolia L.) plants were inoculated with AMF (0 and 5 g Kg -1 soil) under Pb [150 and 225 mg kg -1 soil from Pb (NO 3 ) 2 ] and Ni [220 and 330 mg kg -1 soil from Ni (NO 3 ) 2 ] pollution, in the greenhouse conditions. The control treatment was plants not treated with AMF and HMs. Doing this, the root colonization, HMs uptake, enzymatic and non-enzymatic antioxidants pool, MDA, proline, total phenolics (TPC), flavonoids (TFC), anthocyanins, and essential oil (EO) components were evaluated., Results: According to the findings, the AMF inoculation enhanced shoot and root Pb and Ni content, antioxidant enzymes activity, the total antioxidant activity by DPPH and FRAP methods, TPC, TFC, anthocyanins, and H 2 O 2 content in the lavender plants subjected to Pb and Ni stress. Moreover, the highest (28.91%) and the least (15.81%) percentages of borneol were identified in the lavender plants subjected to AMF under 150 mg kg -1 of Pb and the control plants without AMF application, respectively. Furthermore, the top 1,8-cineole (12.75%) content was recorded in AMF-inoculated plants., Conclusions: The overall results verify that AMF inoculation can be a reliable methodology to enhance the phytoremediation of Pb and Ni by lavender plants while maintaining reliable growth potential. The treatments improved the main EO constituents content, especially under moderate HMs stress conditions. With more detailed studies, the results will be advisable for the extension section for the phytoremediation of polluted soils., (© 2023. The Author(s).)

Background: Arbuscular Mycorrhizal Fungi (AMF) are beneficial microorganisms in soil-plant interactions; however, the underlying mechanisms regarding their roles in legumes environmental stress remain elusive. Present trials were undertaken to study the effect of AMF on the ameliorating of salt, drought, and cold stress in peanut (Arachis hypogaea L.) plants. A new product of AMF combined with Rhizophagus irregularis SA, Rhizophagus clarus BEG142, Glomus lamellosum ON393, and Funneliformis mosseae BEG95 (1: 1: 1: 1, w/w/w/w) was inoculated with peanut and the physiological and metabolomic responses of the AMF-inoculated and non-inoculated peanut plants to salt, drought, and cold stress were comprehensively characterized, respectively., Results: AMF-inoculated plants exhibited higher plant growth, leaf relative water content (RWC), net photosynthetic rate, maximal photochemical efficiency of photosystem II (PSII) (Fv/Fm), activities of antioxidant enzymes, and K + : Na + ratio while lower leaf relative electrolyte conductivity (REC), concentration of malondialdehyde (MDA), and the accumulation of reactive oxygen species (ROS) under stressful conditions. Moreover, the structures of chloroplast thylakoids and mitochondria in AMF-inoculated plants were less damaged by these stresses. Non-targeted metabolomics indicated that AMF altered numerous pathways associated with organic acids and amino acid metabolisms in peanut roots under both normal-growth and stressful conditions, which were further improved by the osmolytes accumulation data., Conclusion: This study provides a promising AMF product and demonstrates that this AMF combination could enhance peanut salt, drought, and cold stress tolerance through improving plant growth, protecting photosystem, enhancing antioxidant system, and regulating osmotic adjustment., (© 2023. The Author(s).)

Background: The increase in the human consumption of fish results in the production of organic fish wastes (FW). For enhanced soil fertility and plant growth at a lower cost and without the negative impacts of chemical fertilizers, these wastes could be employed as a valuable organic fertilizer. To determine the synergistic bio-efficacy of Trichoderma sp. and arbuscular mycorrhizal (AM) fungi in stimulating the morphological and physiological characteristics of FW-fertilized Alium cepa, as well as to investigate their involvement in boosting soil fertility, the current study was carried out. Overall, eight treatments were applied as follows: AM, Trichoderma sp., AM + Trichoderma sp., FW, AM + FW, Trichoderma sp. + FW, AM + Trichoderma sp. + FW, and control. Growth and physiological assessments of onion plants were taken after 8 weeks from FW application., Results: Our results showed that FW application combined with AM fungi and Trichoderma sp. inoculations increased aggregate stability of the soil (glomalin content) and soil chitinase activity. Moreover, using the bio-inoculations along with FW amendments significantly (p < 0.05) improved the photosynthetic pigments, protein, carbohydrates, and nutrients content of onion plants. It's interesting to note that the triple interaction of AM + Trichoderma sp. + FW led to the greatest increase in plant height, root length, number of leaves, and leaf area as well as total fresh and dry weights of shoots and roots. Besides, AM fungal colonization was at its highest percentage with Trichoderma sp. inoculation, although this percentage decreased with FW addition., Conclusion: We concluded that the combined treatments of AM fungi and Trichoderma sp. along with FW application to the soil can be proposed as a successful strategy for plant performance in nutrient-deficient soils as both fungal inoculants are capable of degrading these wastes and converting them into manure suitable for farming so plants can uptake the minerals effortlessly., (© 2022. The Author(s).)

Background: While it is known that arbuscular mycorrhizal fungi (AMF) can improve nutrient acquisition and herbivore resistance in crops, the mechanisms by which AMF influence plant defense remain unknown. Plants respond to herbivory with a cascade of gene expression and phytochemical biosynthesis. Given that the production of defensive phytochemicals requires nutrients, a commonly invoked hypothesis is that the improvement to plant defense when grown with AMF is simply due to an increased availability of nutrients. An alternative hypothesis is that the AMF effect on herbivory is due to changes in plant defense gene expression that are not simply due to nutrient availability. In this study, we tested whether changes in plant defenses are regulated by nutritional provisioning alone or the response of plant to AMF associations. Maize plants grown with or without AMF and with one of three fertilizer treatments (standard, 2 × nitrogen, or 2 × phosphorous) were infested with fall armyworm (Spodoptera frugiperda; FAW) for 72 h. We measured general plant characteristics (e.g. height, number of leaves), relative gene expression (rtPCR) of three defensive genes (lox3, mpi, and pr5), total plant N and P nutrient content, and change in FAW mass per plant., Results: We found that AMF drove the defense response of maize by increasing the expression of mpi and pr5. Furthermore, while AMF increased the total phosphorous content of maize it had no impact on maize nitrogen. Fertilization alone did not alter upregulation of any of the 3 induced defense genes tested, suggesting the mechanism through which AMF upregulate defenses is not solely via increased N or P plant nutrition., Conclusion: This work supports that maize defense may be optimized by AMF associations alone, reducing the need for artificial inputs when managing FAW., (© 2022. The Author(s).)

Background: Apple (Malus domestica Borkh.) is an important economic crop. The pathological effects of Fusarium solani, a species complex of soilborne pathogens, on the root systems of apple plants was unknown. It was unclear how mycorrhizal apple seedlings resist infection by F. solani. The transcriptional profiles of mycorrhizal and non-mycorrhizal plants infected by F. solani were compared using RNA-Seq., Results: Infection with F. solani significantly reduced the dry weight of apple roots, and the roots of mycorrhizal apple plants were less damaged when the plants were infected with F. solani. They also had enhanced activity of antioxidant enzymes and a reduction in the oxidation of membrane lipids. A total of 1839 differentially expressed genes (DEGs) were obtained after mycorrhizal and non-mycorrhizal apple plants were infected with F. solani. A gene ontogeny (GO) analysis showed that most of the DEGs were involved in the binding of ADP and calcium ions. In addition, based on a MapMan analysis, a large number of DEGs were found to be involved in the response of mycorrhizal plants to stress. Among them, the overexpressed transcription factor MdWRKY40 significantly improved the resistance of the apple 'Orin' callus to F. solani and the expression of the resistance gene MdGLU by binding the promoter of MdGLU., Conclusion: This paper outlines how the inoculation of apple seedlings roots by arbuscular mycorrhizal fungi responded to infection with F. solani at the transcriptional level. In addition, MdWRKY40 played an important role in the resistance of mycorrhizal apple seedlings to infection with F. solani., (© 2022. The Author(s).)

Background: Eucalyptus spp. are candidates for phytoremediation in heavy metal (HM)-polluted soils as they can adapt to harsh environments, grow rapidly, and have good economic value. Arbuscular mycorrhizal fungi (AMF) are the most widely distributed plant symbiotic fungi in nature, and they play an important role in promoting the phytoremediation of HM-polluted soils. However, few studies have evaluated the HM detoxification mechanism of E. spp. in symbiosis with AMF, and thus, the molecular mechanism remains unclear., Results: The gene transcription and metabolic pathways of E. grandis were studied with and without inoculation with AMF and at different zinc (Zn) concentrations. Here, we focused on the transcript level of six HM-related gene families (ZNT, COPT/Ctr, YSL, ZIFL and CE). Under high-Zn conditions, thirteen genes (ZNT:2, COPT/Ctr:5, YSL:3, ZIFL:1, CE:2) were upregulated, whereas ten genes (ZNT:3, COPT/Ctr:2, YSL:3, ZIFL:1, CE:1) were downregulated. With AMF symbiosis under high-Zn conditions, ten genes (ZNT:4, COPT/Ctr:2, YSL:3, CE:1) were upregulated, whereas nineteen genes (ZNT:9, COPT/Ctr:2, YSL:3, ZIFL:4, CE:1) were downregulated. Under high-Zn conditions, genes of three potassium-related transporters, six phosphate transporters (PHTs), and two nitrate transporters (NRTs) were upregulated, whereas genes of four potassium-related transporters,four PHTs, and four nitrogen-related transporters were downregulated. With AMF symbiosis under high-Zn conditions, genes of two potassium-related transporters, six ammonium transporters (AMTs) and five PHTs were upregulated, whereas genes of six potassium-related transporters, two AMTs and five PHTs were downregulated., Conclusions: Our results indicates that AMF increases the resistance of E. grandis to high-Zn stress by improving nutrients uptake and regulating Zn uptake at the gene transcription level. Meanwhile, our findings provide a genome-level resource for the functional assignments of key genes regulated by Zn treatment and AM symbiosis in six HM-associated gene families and macromineral nutrient-related gene families of E. grandis. This may contribute to the elucidation of the molecular mechanisms of the response to Zn stress in E. grandis with AM symbiosis at the aspect of the interaction between HM tolerance and nutrient acquisition., (© 2022. The Author(s).)

Background: Cremastra appendiculata is a rare terrestrial orchid with a high market value as an ornamental and medicinal plant. However, the species depends entirely on fungi for seed germination under natural conditions. In a previous study, we have successfully isolated and identified the mycorrhizal fungus Coprinellus disseminatus which was able to induce the germination of C. appendiculata seeds. We then speculated that C. disseminatus may do so by breaking the testa imposed dormancy of the seeds. In this study, biochemical and transcriptomic analyses were used to characterize the germination of C. appendiculata seeds, collected at different stages of germination, as affected by C. disseminatus., Results: The lignocellulose in the seeds coat of C. appendiculata was degraded by the mycorrhizal fungus resulting in facilitated absorption of water. The rate of decline in lignin content was 67 and 73% at 6 and 12 days after sowing, respectively. The water content increased from 13 to 90% during symbiosis. A total of 15,382 genes showing significantly different levels of expression (log 2 FPKM≥2.0, Qvalue≤0.05) were successfully identified among all libraries, where the highest number of DEGs was shared between 6 days versus 0 day after symbiotic germination. Gene annotation results suggested that 15 key genes related water-status, such as DHN gene family and Xero 1 were down-regulated. The genes zeaxanthin epoxidase ZEP, 9-cis-epoxycarotenoid dioxygenase NCED3 and β-carotene hydroxylase involved in the biosynthesis of abscisic acid (ABA) were significantly down-regulated in 6 days as compared to 0 day after symbiotic germination., Conclusions: This work demonstrates that mycorrhizal fungus C. disseminatus can stimulate C. appendiculata seeds germination through a mechanism of breaking the testa imposed dormancy and inducing water absorption of the embryo., (© 2022. The Author(s).)

Background: Inoculation of arbuscular mycorrhizal (AM) fungi has the potential to alleviate salt stress in host plants through the mitigation of ionic imbalance. However, inoculation effects vary, and the underlying mechanisms remain unclear. Two maize genotypes (JD52, salt-tolerant with large root system, and FSY1, salt-sensitive with small root system) inoculated with or without AM fungus Funneliformis mosseae were grown in pots containing soil amended with 0 or 100 mM NaCl (incrementally added 32 days after sowing, DAS) in a greenhouse. Plants were assessed 59 DAS for plant growth, tissue Na + and K + contents, the expression of plant transporter genes responsible for Na + and/or K + uptake, translocation or compartmentation, and chloroplast ultrastructure alterations., Results: Under 100 mM NaCl, AM plants of both genotypes grew better with denser root systems than non-AM plants. Relative to non-AM plants, the accumulation of Na + and K + was decreased in AM plant shoots but increased in AM roots with a decrease in the shoot: root Na + ratio particularly in FSY1, accompanied by differential regulation of ion transporter genes (i.e., ZmSOS1, ZmHKT1, and ZmNHX). This induced a relatively higher Na + efflux (recirculating) rate than K + in AM shoots while the converse outcoming (higher Na + influx rate than K + ) in AM roots. The higher K + : Na + ratio in AM shoots contributed to the maintenance of structural and functional integrity of chloroplasts in mesophyll cells., Conclusion: AM symbiosis improved maize salt tolerance by accelerating Na + shoot-to-root translocation rate and mediating Na + /K + distribution between shoots and roots., (© 2021. The Author(s).)

Blueberry (Vaccinium ssp.) is a perennial shrub belonging to the family Ericaceae, which is highly tolerant of acid soils and heavy metal pollution. In the present study, blueberry was subjected to cadmium (Cd) stress in simulated pot culture. The transcriptomics and rhizosphere fungal diversity of blueberry were analyzed, and the iron (Fe), manganese (Mn), copper (Cu), zinc (Zn) and cadmium (Cd) content of blueberry tissues, soil and DGT was determined. A correlation analysis was also performed. A total of 84 374 annotated genes were identified in the root, stem, leaf and fruit tissue of blueberry, of which 3370 were DEGs, and in stem tissue, of which 2521 were DEGs. The annotation data showed that these DEGs were mainly concentrated in a series of metabolic pathways related to signal transduction, defense and the plant-pathogen response. Blueberry transferred excess Cd from the root to the stem for storage, and the highest levels of Cd were found in stem tissue, consistent with the results of transcriptome analysis, while the lowest Cd concentration occurred in the fruit, Cd also inhibited the absorption of other metal elements by blueberry. A series of genes related to Cd regulation were screened by analyzing the correlation between heavy metal content and transcriptome results. The roots of blueberry rely on mycorrhiza to absorb nutrients from the soil. The presence of Cd has a significant effect on the microbial community composition of the blueberry rhizosphere. The fungal family Coniochaetaceae, which is extremely extremelytolerant, has gradually become the dominant population. The results of this study increase our understanding of the plant regulation mechanism for heavy metals, and suggest potential methods of soil remediation using blueberry., (© 2021. The Author(s).)

Background: PROTEIN PHOSPHATASE 2A (PP2A) expression is crucial for the symbiotic association between plants and various microbes, and knowledge on these symbiotic processes is important for sustainable agriculture. Here we tested the hypothesis that PP2A regulatory subunits, especially B'φ and B'θ, are involved in signalling between plants and mycorrhizal fungi or plant-growth promoting bacteria., Results: Treatment of tomato plants (Solanum lycopersicum) with the plant growth-promoting rhizobacteria (PGPR) Azospirillum brasilense and Pseudomonas simiae indicated a role for the PP2A B'θ subunit in responses to PGPR. Arbuscular mycorrhizal fungi influenced B'θ transcript levels in soil-grown plants with canonical arbuscular mycorrhizae. In plant roots, transcripts of B'φ were scarce under all conditions tested and at a lower level than all other PP2A subunit transcripts. In transformed tomato plants with 10-fold enhanced B'φ expression, mycorrhization frequency was decreased in vermiculite-grown plants. Furthermore, the high B'φ expression was related to abscisic acid and gibberellic acid responses known to be involved in plant growth and mycorrhization. B'φ overexpressor plants showed less vigorous growth, and although fruits were normal size, the number of seeds per fruit was reduced by 60% compared to the original cultivar., Conclusions: Expression of the B'θ gene in tomato roots is strongly influenced by beneficial microbes. Analysis of B'φ overexpressor tomato plants and established tomato cultivars substantiated a function of B'φ in growth and development in addition to a role in mycorrhization.

Background: A better understanding of non-structural carbohydrate (NSC) dynamics in trees under drought stress is critical to elucidate the mechanisms underlying forest decline and tree mortality from extended periods of drought. This study aimed to assess the contribution of ectomycorrhizal (ECM) fungus (Suillus variegatus) to hydraulic function and NSC in roots, stems, and leaves of Pinus tabulaeformis subjected to different water deficit intensity. We performed a continuous controlled drought pot experiment from July 10 to September 10, 2019 using P. tabulaeformis seedlings under 80, 40, and 20% of the field moisture capacity that represented the absence of non-drought, moderate drought, and severe drought stress, respectively., Results: Results indicated that S. variegatus decreased the mortality rate and increased height, root biomass, and leaf biomass of P. tabulaeformis seedlings under moderate and severe drought stress. Meanwhile, the photosynthetic rates, stomatal conductance, and transpiration rates of P. tabulaeformis were significantly increased after S. variegatus inoculation. Moreover, the inoculation of S. variegatus also significantly increased the NSC concentrations of all seedling tissues, enhanced the soluble sugars content, and increased the ratios of soluble sugars to starch on all tissues under severe drought. Overall, the inoculation of S. variegatus has great potential for improving the hydraulic function, increasing the NSC storage, and improving the growth of P. tabulaeformis under severe drought., Conclusions: Therefore, the S. variegatus can be used as a potential application strain for ecological restoration on arid regions of the Loess Plateau, especially in the P. tabulaeformis woodlands.

Background: To decipher the root and microbial interaction, secondary metabolite accumulation in roots and the microbial community's succession model during the plant's growth period demands an in-depth investigation. However, till now, no comprehensive study is available on the succession of endophytic fungi and arbuscular mycorrhizal fungi (AMF) with roots of medicinal licorice plants and the effects of endophytic fungi and AMF on the secondary metabolite accumulation in licorice plant's root., Results: In the current study, interaction between root and microbes in 1-3 years old medicinal licorice plant's root and rhizospheric soil was investigated. Secondary metabolites content in licorice root was determined using high-performance liquid chromatography (HPLC). The composition and diversity of endophytic and AMF in the root and soil were deciphered using high-throughput sequencing technology. During the plant's growth period, as compared to AMF, time and species significantly affected the diversity and richness of endophytic fungi, such as Ascomycota, Basidiomycota, Fusarium, Cladosporium, Sarocladium. The growth period also influenced the AMF diversity, evident by the significant increase in the relative abundance of Glomus and the significant decrease in the relative abundance of Diversispora. It indicated a different succession pattern between the endophytic fungal and AMF communities. Meanwhile, distance-based redundancy analysis and Mantel tests revealed root's water content and secondary metabolites (glycyrrhizic acid, liquiritin, and total flavonoids), which conferred endophytic fungi and AMF diversity. Additionally, plant growth significantly altered soil's physicochemical properties, which influenced the distribution of endophytic fungal and AMF communities., Conclusions: This study indicated a different succession pattern between the endophytic fungal and AMF communities. During the plant's growth period, the contents of three secondary metabolites in roots increased per year, which contributed to the overall differences in composition and distribution of endophytic fungal and AMF communities. The endophytic fungal communities were more sensitive to secondary metabolites than AMF communities. The current study provides novel insights into the interaction between rhizospheric microbes and root exudates.

Background: Despite significant limitations of growth medium reuse, a large amount of organic substrate is reused in soilless cultivation of horticultural crops in China. Arbuscular mycorrhizal fungi (AMF) can promote nutrient absorption and improve plant tolerance to biotic and abiotic stresses. However, the mechanisms governing the effects of AMF on crop growth in organic continuous cropping substrates have not been elucidated., Results: In this study, we showed that the inoculation of AMF in continuous cropping substrates promoted growth and root development, and increased the root and NADP-malic enzyme (NADP-ME) activity of tomato seedlings. Root transcriptome analysis demonstrated that the plant hormone signal transduction pathway was highly enriched, and 109 genes that positively correlated with the AMF-inoculated plant phenotype were obtained by gene set enrichment analysis (GSEA), which identified 9 genes related to indole acetic acid (IAA). Importantly, the levels of endogenous IAA in tomato seedlings significantly increased after AMF inoculation. Furthermore, the application of AMF significantly increased the expression levels of NADP-ME1 and NADP-ME2, as well as the activity of NADP-ME, and enhanced the root activity of tomato seedlings in comparison to that observed without inoculation of AMF. However, these effects were blocked in plants treated with 2,3,5-triiodobenzoic acid (TIBA), a polar transport inhibitor of IAA., Conclusions: These results suggest that IAA mediates the AMF-promoted tomato growth and expression of NADP-MEs in continuous cropping substrates. The study provides convincing evidence for the reuse of continuous cropping substrates by adding AMF as an amendment.

Background: Pinus massoniana Lamb. is an important afforestation tree species with high economic, ecological and medicinal values. Aluminum (Al) toxicity driven by soil acidification causes dieback of P. massoniana plantations. Previous studies showed that ectomycorrhizal fungi alleviate Al stress damages in Pinus, but the underlying molecular mechanisms and key genes induced by ectomycorrhizal fungi inoculation under Al stress in Pinus have not been explored. Herein, we applied Al stress for 60 days to P. massoniana seedlings inoculated with Suillus luteus (SL) and those non-inoculated. Then, we compared their growth parameters and transcriptome in order to detect candidate genes induced by SL conferring Al tolerance in P. massoniana., Result: Our results showed that SL inoculation confers Al stress tolerance in P. massoniana through improved growth performance, strong antioxidant enzyme activities and reduced malondialdehyde accumulation as compared to non-inoculated seedlings. Transcriptome sequencing further supported these findings as very few genes (51 genes) were transcriptionally altered by Al in SL inoculated plants as compared to non-inoculated plants (2140 genes). We identified three core genes (cox1, cox3 and Nd1) that were strongly up-regulated by Al in the SL inoculated plants but were down-regulated in the non-inoculated plants. We also identified 42 genes specifically regulated by SL inoculated plants under Al stress, which are involved in a wide range of biological processes such as antioxidative response, transporters, hormone signaling and plant pathogen infection responses., Conclusions: Altogether, our data suggest that SL inoculation induces priming of key stress response pathways and triggers specific genes that efficiently alleviate Al stress effects in P. massoniana. The candidate genes resources generated in this study are of utmost importance for functional characterization and molecular studies aiming at improving Al tolerance in plants.

Background: Phosphate is an essential plant macronutrient required to achieve maximum crop yield. Roots are able to uptake soil phosphate from the immediate root area, thus creating a nutrient depletion zone. Many plants are able to exploit phosphate from beyond this root nutrient depletion zone through symbiotic association with Arbuscular Mycorrhizal Fungi (AMF). Here we characterise the relationship between root architecture, AMF association and low phosphate tolerance in strawberries. The contrasting root architecture in the parental strawberry cultivars 'Redgauntlet' and 'Hapil' was studied through a mapping population of 168 progeny. Low phosphate tolerance and AMF association was quantified for each genotype to allow assessment of the phenotypic and genotypic relationships between traits., Results: A "phosphate scavenging" root phenotype where individuals exhibit a high proportion of surface lateral roots was associated with a reduction in root system size across genotypes. A genetic correlation between "root system size" traits was observed with a network of pleiotropic QTL found to represent five "root system size" traits. By contrast, average root diameter and the distribution of roots appeared to be under two discrete methods of genetic control. A total of 18 QTL were associated with plant traits, 4 of which were associated with solidity that explained 46% of the observed variation. Investigations into the relationship between AMF association and root architecture found that a higher root density was associated with greater AMF colonisation across genotypes. However, no phenotypic correlation or genotypic association was found between low phosphate tolerance and the propensity for AMF association, nor root architectural traits when plants are grown under optimal nutrient conditions., Conclusions: Understanding the genetic relationships underpinning phosphate capture can inform the breeding of strawberry varieties with better nutrient use efficiency. Solid root systems were associated with greater AMF colonisation. However, low P-tolerance was not phenotypically or genotypically associated with root architecture traits in strawberry plants. Furthermore, a trade-off was observed between root system size and root architecture type, highlighting the energetic costs associated with a "phosphate scavenging" root architecture.

Background: Increasing zinc (Zn) concentrations in crops is important for alleviation of human Zn deficiency. Arbuscular mycorrhizal fungi (AMF) contribute to plant Zn uptake, but their contribution to Zn in the edible portion of crops has not yet been investigated. This study aimed to quantify the mycorrhizal pathway of Zn uptake into grain of wheat and barley under varying soil Zn availabilities. Bread wheat (Triticum aestivum) and barley (Hordeum vulgare) were grown in pots with a hyphal compartment containing 65 Zn. Plants were inoculated with Rhizophagus irregularis and grown at three soil Zn concentrations. Radioactive Zn in grain and straw was measured and the contribution of AMF to Zn uptake was calculated., Results: The mycorrhizal pathway of Zn uptake contributed up to 24.3% of total above-ground Zn in wheat, and up to 12.7% of that Zn in barley. The greatest contribution by the mycorrhizal pathway was observed in barley at the lowest Zn addition, and in wheat at the highest one. In addition, grain yield of bread wheat was increased by AMF., Conclusions: These results suggest that AMF have a substantial role in uptake of Zn into cereals, and the proportional contribution by the MPU is dependent on plant species, as well as available soil Zn.

Background: Below-ground linkage between plant and fungal communities is one of the major drivers of terrestrial ecosystem dynamics. However, we still have limited knowledge of how such plant-fungus associations vary in their community-scale properties depending on fungal functional groups and geographic locations., Methods: By compiling a high-throughput sequencing dataset of root-associated fungi in eight forests along the Japanese Archipelago, we performed a comparative analysis of arbuscular mycorrhizal, ectomycorrhizal, and saprotrophic/endophytic associations across a latitudinal gradient from cool-temperate to subtropical regions., Results: In most of the plant-fungus networks analyzed, host-symbiont associations were significantly specialized but lacked "nested" architecture, which has been commonly reported in plant-pollinator and plant-seed disperser networks. In particular, the entire networks involving all functional groups of plants and fungi and partial networks consisting of ectomycorrhizal plant and fungal species/taxa displayed "anti-nested" architecture (i.e., negative nestedness scores) in many of the forests examined. Our data also suggested that geographic factors affected the organization of plant-fungus network structure. For example, the southernmost subtropical site analyzed in this study displayed lower network-level specificity of host-symbiont associations and higher (but still low) nestedness than northern localities., Conclusions: Our comparative analyses suggest that arbuscular mycorrhizal, ectomycorrhizal, and saprotrophic/endophytic plant-fungus associations often lack nested network architecture, while those associations can vary, to some extent, in their community-scale properties along a latitudinal gradient. Overall, this study provides a basis for future studies that will examine how different types of plant-fungus associations collectively structure terrestrial ecosystems.

Background: The interaction between arbuscular mycorrhizal fungi (AMF) and AMF spore associated bacteria (SAB) were previously found to improve mycorrhizal symbiotic efficiency under saline stress, however, the information about the molecular basis of this interaction remain unknown. Therefore, the present study aimed to investigate the response of maize plants to co-inoculation of AMF and SAB under salinity stress., Results: The co-inoculation of AMF and SAB significantly improved plant dry weight, nutrient content of shoot and root tissues under 25 or 50 mM NaCl. Importantly, co-inoculation significantly reduced the accumulation of proline in shoots and Na + in roots. Co-inoculated maize plants also exhibited high K + /Na + ratios in roots at 25 mM NaCl concentration. Mycorrhizal colonization significantly positively altered the expression of ZmAKT2, ZmSOS1, and ZmSKOR genes, to maintain K + and Na + ion homeostasis. Confocal laser scanning microscope (CLSM) view showed that SAB were able to move and localize into inter- and intracellular spaces of maize roots and were closely associated with the spore outer hyaline layer., Conclusion: These new findings indicate that co-inoculation of AMF and SAB effectively alleviates the detrimental effects of salinity through regulation of SOS pathway gene expression and K + /Na + homeostasis to improve maize plant growth.

Background: Strigolactones (SLs) are considered to be a novel class of phytohormone involved in plant defense responses. Currently, their relationships with other plant hormones, such as abscisic acid (ABA), during responses to salinity stress are largely unknown., Results: In this study, the relationship between SL and ABA during the induction of H 2 O 2 - mediated tolerance to salt stress were studied in arbuscular mycorrhizal (AM) Sesbania cannabina seedlings. The SL levels increased after ABA treatments and decreased when ABA biosynthesis was inhibited in AM plants. Additionally, the expression levels of SL-biosynthesis genes in AM plants increased following treatments with exogenous ABA and H 2 O 2 . Furthermore, ABA-induced SL production was blocked by a pre-treatment with dimethylthiourea, which scavenges H 2 O 2 . In contrast, ABA production was unaffected by dimethylthiourea. Abscisic acid induced only partial and transient increases in the salt tolerance of TIS108 (a SL synthesis inhibitor) treated AM plants, whereas SL induced considerable and prolonged increases in salt tolerance after a pre-treatment with tungstate., Conclusions: These results strongly suggest that ABA is regulating the induction of salt tolerance by SL in AM S. cannabina seedlings.

Background: In nature, orchid plants depend completely on symbiotic fungi for their nutrition at the germination and the subsequent seedling (protocorm) stages. However, only limited quantitative methods for evaluating the orchid-fungus interactions at the protocorm stage are currently available, which greatly constrains our understanding of the symbiosis. Here, we aimed to improve and integrate quantitative evaluations of the growth and fungal colonization in the protocorms of a terrestrial orchid, Blettila striata, growing on a plate medium., Results: We achieved both symbiotic and asymbiotic germinations for the terrestrial orchid B. striata. The protocorms produced by the two germination methods grew almost synchronously for the first three weeks. At week four, however, the length was significantly lower in the symbiotic protocorms. Interestingly, the dry weight of symbiotic protocorms did not significantly change during the growth period, which implies that there was only limited transfer of carbon compounds from the fungus to the protocorms in this relationship. Next, to evaluate the orchid-fungus interactions, we developed an ink-staining method to observe the hyphal coils in protocorms without preparing thin sections. Crushing the protocorm under the coverglass enables us to observe all hyphal coils in the protocorms with high resolution. For this observation, we established a criterion to categorize the stages of hyphal coils, depending on development and degradation. By counting the symbiotic cells within each stage, it was possible to quantitatively evaluate the orchid-fungus symbiosis., Conclusions: We describe a method for quantitative evaluation of orchid-fungus symbiosis by integrating the measurements of plant growth and fungal colonization. The current study revealed that although fungal colonization was observed in the symbiotic protocorms, the weight of the protocorm did not significantly increase, which is probably due to the incompatibility of the fungus in this symbiosis. These results suggest that fungal colonization and nutrition transfer can be differentially regulated in the symbiosis. The evaluation methods developed in this study can be used to study various quantitative aspects of the orchid-fungus symbiosis.

Background: Genes involved in arbuscular mycorrhizal (AM) symbiosis have been identified primarily by mutant screens, followed by identification of the mutated genes (forward genetics). In addition, a number of AM-related genes has been identified by their AM-related expression patterns, and their function has subsequently been elucidated by knock-down or knock-out approaches (reverse genetics). However, genes that are members of functionally redundant gene families, or genes that have a vital function and therefore result in lethal mutant phenotypes, are difficult to identify. If such genes are constitutively expressed and therefore escape differential expression analyses, they remain elusive. The goal of this study was to systematically search for AM-related genes with a bioinformatics strategy that is insensitive to these problems. The central element of our approach is based on the fact that many AM-related genes are conserved only among AM-competent species., Results: Our approach involves genome-wide comparisons at the proteome level of AM-competent host species with non-mycorrhizal species. Using a clustering method we first established orthologous/paralogous relationships and subsequently identified protein clusters that contain members only of the AM-competent species. Proteins of these clusters were then analyzed in an extended set of 16 plant species and ranked based on their relatedness among AM-competent monocot and dicot species, relative to non-mycorrhizal species. In addition, we combined the information on the protein-coding sequence with gene expression data and with promoter analysis. As a result we present a list of yet uncharacterized proteins that show a strongly AM-related pattern of sequence conservation, indicating that the respective genes may have been under selection for a function in AM. Among the top candidates are three genes that encode a small family of similar receptor-like kinases that are related to the S-locus receptor kinases involved in sporophytic self-incompatibility., Conclusions: We present a new systematic strategy of gene discovery based on conservation of the protein-coding sequence that complements classical forward and reverse genetics. This strategy can be applied to diverse other biological phenomena if species with established genome sequences fall into distinguished groups that differ in a defined functional trait of interest.

Background: Induced aboveground plant defenses against pathogens can have negative effects on belowground microbial symbionts. While a considerable number of studies have utilized chemical elicitors to experimentally induce such defenses, there is surprisingly little evidence that actual aboveground pathogens affect root-associated microbes. We report here that an aboveground fungal pathogen of common bean (Phaseolus vulgaris) induces a defense response that inhibits both the belowground formation of root nodules elicited by rhizobia and the colonization with arbuscular mycorrhizal fungi (AMF)., Results: Foliage of plants inoculated with either rhizobia or AMF was treated with both live Colletotrichum gloeosporioides-a generalist hemibiotrophic plant pathogen-and C. gloeosporioides fragments. Polyphenol oxidase (PPO), chitinase and β-1,3-glucanase activity in leaves and roots, as well as the number of rhizobia nodules and the extent of AMF colonization, were measured after pathogen treatments. Both the live pathogen and pathogen fragments significantly increased PPO, chitinase and β-1,3-glucanase activity in the leaves, but only PPO activity was increased in roots. The number of rhizobia nodules and the extent of AMF colonization was significantly reduced in treatment plants when compared to controls., Conclusion: We demonstrate that aboveground fungal pathogens can affect belowground mutualism with two very different types of microbial symbionts-rhizobia and AMF. Our results suggest that systemically induced PPO activity is functionally involved in this above-belowground interaction. We predict that the top-down effects we show here can drastically impact plant performance in soils with limited nutrients and water; abiotic stress conditions usually mitigated by microbial belowground mutualists.

Background: Phosphorus (P) deficiency is one of the major nutrient stresses limiting plant growth. The uptake of P by plants is well considered to be mediated by a number of high-affinity phosphate (Pi) transporters belonging to the Pht1 family. Although the Pht1 genes have been extensively identified in several plant species, there is a lack of systematic analysis of the Pht1 gene family in any solanaceous species thus far., Results: Here, we report the genome-wide analysis, phylogenetic evolution and expression patterns of the Pht1 genes in tomato (Solanum lycopersicum). A total of eight putative Pht1 genes (LePT1 to 8), distributed on three chromosomes (3, 6 and 9), were identified through extensive searches of the released tomato genome sequence database. Chromosomal organization and phylogenetic tree analysis suggested that the six Pht1 paralogues, LePT1/3, LePT2/6 and LePT4/5, which were assigned into three pairs with very close physical distance, were produced from recent tandem duplication events that occurred after Solanaceae splitting with other dicot families. Expression analysis of these Pht1 members revealed that except LePT8, of which the transcript was undetectable in all tissues, the other seven paralogues showed differential but partial-overlapping expression patterns. LePT1 and LePT7 were ubiquitously expressed in all tissues examined, and their transcripts were induced abundantly in response to Pi starvation; LePT2 and LePT6, the two paralogues harboring identical coding sequence, were predominantly expressed in Pi-deficient roots; LePT3, LePT4 and LePT5 were strongly activated in the roots colonized by arbuscular mycorrhizal fungi under low-P, but not high-P condition. Histochemical analysis revealed that a 1250-bp LePT3 promoter fragment and a 471-bp LePT5 promoter fragment containing the two elements, MYCS and P1BS, were sufficient to direct the GUS reporter expression in mycorrhizal roots and were limited to distinct cells harboring AM fungal structures. Additionally, the four paralogues, LePT1, LePT2, LePT6 and LePT7, were very significantly down-regulated in the mycorrhizal roots under low Pi supply condition., Conclusions: The results obtained from this study provide new insights into the evolutionary expansion, functional divergence and genetic redundancy of the Pht1 genes in response to Pi deficiency and mycorrhizal symbiosis in tomato.

Background: Repeated oscillations in intracellular calcium (Ca2+) concentration, known as Ca2+ spiking signals, have been described in plants for a limited number of cellular responses to biotic or abiotic stimuli and most notably the common symbiotic signaling pathway (CSSP) which mediates the recognition by their plant hosts of two endosymbiotic microbes, arbuscular mycorrhizal (AM) fungi and nitrogen fixing rhizobia. The detailed analysis of the complexity and variability of the Ca2+ spiking patterns which have been revealed in recent studies requires both extensive datasets and sophisticated statistical tools., Results: As a contribution, we have developed automated Ca2+ spiking analysis (CaSA) software that performs i) automated peak detection, ii) statistical analyses based on the detected peaks, iii) autocorrelation analysis of peak-to-peak intervals to highlight major traits in the spiking pattern.We have evaluated CaSA in two experimental studies. In the first, CaSA highlighted unpredicted differences in the spiking patterns induced in Medicago truncatula root epidermal cells by exudates of the AM fungus Gigaspora margarita as a function of the phosphate concentration in the growth medium of both host and fungus. In the second study we compared the spiking patterns triggered by either AM fungal or rhizobial symbiotic signals. CaSA revealed the existence of different patterns in signal periodicity, which are thought to contribute to the so-called Ca2+ signature., Conclusions: We therefore propose CaSA as a useful tool for characterizing oscillatory biological phenomena such as Ca2+ spiking.