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The study aimed to explore the genetic and functional potential of mycorrhizal helper bacteria (MHB) strains isolated from polluted soil, focusing on their ability to enhance plant growth and ameliorate the adverse effects of polycyclic aromatic hydrocarbons (PAHs). We sequenced the genomes of three MHB strains isolated from soil contaminated with PAHs and phenol. Moreover, experiments were carried out to check if these bacteria have ability to stimulate the growth of arbuscular mycorrhizal fungi (AMF) and promote plant development. Phylogenomic analysis identified the strains as belonging to the Streptomyces, Pantoea, and Bacillus genera, all exhibiting high tolerance to hydrocarbons. Genome mining revealed genes encoding enzymes for the degradation of aromatic compounds, alongside biosynthetic gene clusters for secondary metabolites such as siderophores and antibiotics. Laboratory experiments confirmed that the studied MHB strains enhance AMF development and spore production while exhibiting plant growth-promoting mechanisms such as siderophore and ammonia production, phosphate solubilization, and cellulolytic enzyme synthesis. These findings highlight the potential application of MHB in microbial-assisted remediation of hydrocarbon-contaminated soils through the tripartite plant-MHB-AMF system. [ABSTRACT FROM AUTHOR]

Bacterial-fungal interaction (BFI) is found ubiquitously and plays important roles in various environmental settings, thus being responsible for numerous biophysical and chemical processes in nature. In terms of BFI, the capacity of the bacterium to enhance the growth of fungal mycelia is an indication of the roles of the bacterium in mutualistic interaction, since increasing mycelial growth results in higher changes for fungal establishment. In this review, the interaction between mycelial growth-promoting bacterium (MGPB) and its fungal counterpart in agricultural settings and the promotion of mycelial growth as an outcome of mutual interactions in various environmental niches were evaluated. The beneficial relationships included endohyphal interaction, association of bacteria with mushrooms, bacteria-mycorrhizae symbiosis, and geomicrobiology. Furthermore, the mode of interaction between MGPB and their fungal counterparts was also explained. There are two fundamental modes of interaction involved, namely physical interaction and chemical interaction. The first involved endosymbiosis and bacterial attachment, while the latter comprised quorum sensing, volatile metabolites, enzymatic activity, and chemotaxis. Particularly, the growth stimulants secreted by the bacteria, which promote the growth of hyphae, are discussed thoroughly. Moreover, the chance of trade-off metabolites between fungi and their MGPBs as a consequence of mutualistic interaction will also be observed. Finally, the agricultural relevance of BFI, particularly the relation between fungi and MGPBs, will also be provided, including key technologies and future bioprospects for optimum application. • Mycelial growth-promoting bacterium (MGPB) enhance fungal establishment. • The modes of interaction between MGPBs and fungi include physical and chemical. • The bacterial secondary metabolites involved in the mutualistic interaction. • There is a possibility of trade-off metabolites between fungi and MGPBs. • The relevance of interaction can be found in various agricultural sectors. [ABSTRACT FROM AUTHOR]

Arbuscular mycorrhizal fungi are major components of soil microbiota and mainly interact with other microorganisms in the rhizosphere. Mycorrhiza establishment impacts the plant physiology and some nutritional and physical properties of the rhizospheric soil. These effects alter the development of the root or mycorrhizas resulting from the activity of soil microorganisms. The rhizosphere of mycorrhizal plants (mycorrhizosphere), is inhabited by large microbial activities responsible for several key ecosystem processes. This review is focused on the microbial interactions between mycorrhizal fungi and components of rhizosphere microbiota and highlight the agronomic potentialities of the Mycorrhiza Helper Bacteria on mycorrhiza formation. The main conclusion is that this MHB effect in the rhizosphere of mycorrhizal plants, enhance plant fitness and soil quality and are of great interest to ensure sustainable agricultural development and ecosystem functioning. [ABSTRACT FROM AUTHOR]

By dint of the development of agroecological practices and organic farming, stakeholders are becoming more and more aware of the importance of soil life and banning a growing number of pesticide molecules, promoting the use of plant bio-stimulants. To justify and promote the use of microbes in agroecological practices and sustainable agriculture, a number of functions or services often are invoked: (i) soil health, (ii) plant growth promotion, (iii) biocontrol, (iv) nutrient acquiring, (v) soil carbon storage, etc. In this paper, a review and a hierarchical classification of plant fungal partners according to their ecosystemic potential with regard to the available technologies aiming at field uses will be discussed with a particular focus on interactive microbial associations and functions such as Mycorrhiza Helper Bacteria (MHB) and nurse plants. [ABSTRACT FROM AUTHOR]

The desert truffle Terfezia boudieri is an ascomycete fungus that forms ect-endomycorrhiza in the roots of plants belonging to Cistaceae. The fungus forms hypogeous edible fruit bodies, appreciated as gourmet food. Truffles and host plants are colonized by various microbes, which may contribute to their development. However, the diversity and composition of the bacterial community under field conditions in the Negev desert are still unknown. The overall goal of this research was to identify the rhizosphere microbial community supporting the establishment of a symbiotic association between T. boudieri and Helianthemum sessiliflorum. The bacterial community was characterized by fruiting bodies, mycorrhized roots, and rhizosphere soil. Based on next-generation sequencing meta-analyses of the 16S rRNA gene, we discovered diverse bacterial communities of fruit bodies that differed from those found in the roots and rhizosphere. Families of Proteobacteria, Planctomycetes, and Actinobacteria were present in all four samples. Alpha diversity analysis revealed that the rhizosphere and roots contain significantly higher bacterial species numbers compared to the fruit. Additionally, ANOSIM and PCoA provided a comparative analysis of the bacterial taxa associated with fruiting bodies, roots, and rhizosphere. The core microbiome described consists of groups whose biological role triggers important traits supporting plant growth and fruit body development. [ABSTRACT FROM AUTHOR]

Arbuscular mycorrhizal fungi are major components of soil microbiota and mainly interact with other microorganisms in the rhizosphere. Mycorrhiza establishment impacts the plant physiology and some nutritional and physical properties of the rhizospheric soil. These effects alter the development of the root or mycorrhizas resulting from the activity of soil microorganisms. The rhizosphere of mycorrhizal plants (mycorrhizosphere), is inhabited by large microbial activities responsible for several key ecosystem processes. This review is focused on the microbial interactions between mycorrhizal fungi and components of rhizosphere microbiota and highlight the agronomic potentialities of the Mycorrhiza Helper Bacteria on mycorrhiza formation. The main conclusion is that this MHB effect in the rhizosphere of mycorrhizal plants, enhance plant fitness and soil quality and are of great interest to ensure sustainable agricultural development and ecosystem functioning.

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Rhizosphere microbiome is one of the main sources of plant protection against drought. Beneficial symbiotic microorganisms, such as ectomycorrhizal fungi (ECMF) and mycorrhiza helper bacteria (MHB), interact with each other for increasing or maintaining host plant fitness. This mutual support benefits all three partners and comprises a natural system for drought acclimation in plants. Cork oak (Quercus suber L.) tolerance to drought scenarios is widely known, but adaptation to climate changes has been a challenge for forest sustainability protection. In this work, ECMF and MHB communities from cork oak forests were cross-linked and correlated with climates. Cenococcum, Russula and Tuber were the most abundant ECMF capable of interacting with MHB (ECMF~MHB) genera in cork oak stands, while Bacillus, Burkholderia and Streptomyces were the most conspicuous MHB. Integrating all microbial data, two consortia Lactarius/Bacillaceae and Russula/Burkholderaceae have singled out but revealed a negative interaction with each other. Russula/Burkholderaceae might have an important role for cork oak forest sustainability in arid environments, which will be complemented by the lower drought adaptation of competitive Lactarius/Bacillaceae. These microbial consortia could play an essential role on cork oak forest resilience to upcoming climatic changes. [ABSTRACT FROM AUTHOR]

Abstract Background Plants, fungi, and bacteria form complex, mutually-beneficial communities within the soil environment. In return for photosynthetically derived sugars in the form of exudates from plant roots, the microbial symbionts in these rhizosphere communities provide their host plants access to otherwise inaccessible nutrients in soils and help defend the plant against biotic and abiotic stresses. One role that bacteria may play in these communities is that of Mycorrhizal Helper Bacteria (MHB). MHB are bacteria that facilitate the interactions between plant roots and symbiotic mycorrhizal fungi and, while the effects of MHB on the formation of plant-fungal symbiosis and on plant health have been well documented, the specific molecular mechanisms by which MHB drive gene regulation in plant roots leading to these benefits remain largely uncharacterized. Results Here, we investigate the effects of the bacterium Pseudomonas fluorescens SBW25 (SBW25) on aspen root transcriptome using a tripartite laboratory community comprised of Populus tremuloides (aspen) seedlings and the ectomycorrhizal fungus Laccaria bicolor (Laccaria). We show that SBW25 has MHB activity and promotes mycorrhization of aspen roots by Laccaria. Using transcriptomic analysis of aspen roots under multiple community compositions, we identify clusters of co-regulated genes associated with mycorrhization, the presence of SBW25, and MHB-associated functions, and we generate a combinatorial logic network that links causal relationships in observed patterns of gene expression in aspen seedling roots in a single Boolean circuit diagram. The predicted regulatory circuit is used to infer regulatory mechanisms associated with MHB activity. Conclusions In our laboratory conditions, SBW25 increases the ability of Laccaria to form ectomycorrhizal interactions with aspen seedling roots through the suppression of aspen root antifungal defense responses. Analysis of transcriptomic data identifies that potential molecular mechanisms in aspen roots that respond to MHB activity are proteins with homology to pollen recognition sensors. Pollen recognition sensors integrate multiple environmental signals to down-regulate pollenization-associated gene clusters, making proteins with homology to this system an excellent fit for a predicted mechanism that integrates information from the rhizosphere to down-regulate antifungal defense response genes in the root. These results provide a deeper understanding of aspen gene regulation in response to MHB and suggest additional, hypothesis-driven biological experiments to validate putative molecular mechanisms of MHB activity in the aspen-Laccaria ectomycorrhizal symbiosis.

Copyright of Journal of Shenyang Agricultural University is the property of Journal of Shenyang Agricultural University Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Chryseobacterium sp. MHB01, Rhodococcus qingshengii MHB02, and Agrobacterium tumefaciens MHB03 were isolated from superabsorbent polymer granules cultured with an arbuscular mycorrhizal fungus. Whole-genome sequencing of these three strains revealed genome sizes of 4.57 Mb, 7.13 Mb, and 5.49 Mb with G + C contents of 36.9%, 62.5%, and 58.2%, respectively., Competing Interests: The authors declare no conflict of interest.

The ecological restoration of nickel mining-degraded areas in New Caledonia is strongly limited by low availability of soil mineral nutrients, metal toxicity, and slow growth rates of native plant species. In order to improve plant growth for restoration programs, special attention was paid to interactions between plant and soil microorganisms. In this study, we evaluated the influence of inoculation with Curtobacterium citreum BE isolated from a New Caledonian ultramafic soil on arbuscular mycorrhizal symbiosis and growth of Tetraria comosa, an endemic sedge used in restoration programs. A greenhouse experiment on ultramafic substrate was conducted with an inoculum comprising two arbuscular mycorrhizal fungi (AMF) species isolated from New Caledonian ultramafic soils: Rhizophagus neocaledonicus and Claroideoglomus etunicatum. The effects on plant growth of the AMF and C. citreum BE inoculated separately were not significant, but their co-inoculation significantly enhanced the dry weight of T. comosa compared with the non-inoculated control. These differences were positively correlated with mycorrhizal colonization which was improved by C. citreum BE. Compared with the control, co-inoculated plants were characterized by better mineral nutrition, a higher Ca/Mg ratio, and lower metal translocation. However, for Ca/Mg ratio and metal translocation, there were no significant differences between the effects of AMF inoculation and co-inoculation. [ABSTRACT FROM AUTHOR]

Arbuscular mycorrhizal (AM) fungi are ecologically important for the growth and survival of most vascular plants. These fungi are known as obligate biotrophs that acquire carbon solely from host plants. A 13C-labeling experiment revealed the ability of axenically grown Rhizophagus irregularis DAOM 197198 to derive carbon from axenic culture on a relatively novel medium containing two sources of palmitic acid developed by Ishii (designated IH medium). In a separate experiment, this model fungus grew larger mycelia and produced more daughter spores on the IH medium in the presence of two Variovorax paradoxus strains than in axenic culture. In contrast, a strain of Mycobacterium sp. did not influence the growth of the AM fungus. Rhizophagus irregularis produced branched absorbing structures on the IH medium and, in monoxenic culture with V. paradoxus, sometimes formed densely packed hyphal coils. In this study, we report for the first time the formation of coarse terminal pelotons and of terminal and intercalary very fine (≈ 1 μm diameter) hyphal elongations, which could form daughter spores in the presence of V. paradoxus. This study shows the value of IH medium and certain rhizobacteria in the culture of R. irregularis DAOM 197198 in vitro. [ABSTRACT FROM AUTHOR]

Boletus edulis (Be) and mycorrhiza helper bacteria (MHB) in the shiroes soil of Pinus thunbergii were isolated and their positive effects on the growth of P.thunbergii were investigated. The potential MHB strains isolated from rhizosphere soil were evaluated, based on the positive effect their extracellular metabolites had on the growth of Be. The MHB strain was screened based on seedling height, ground diameter of the stem, stem root ratio, and root infection rate of P. thunbergii seedlings. The MHB strain was then identified after morphological, biochemical, and physiological analyses, in conjunction with 16S rDNA gene sequencing. The results showed that metabolites exuded by the isolated strain H12 had a significant growth-promoting effect on Be. Co-inoculation of P. thunbergii seedlings with Be+H12 resulted in a 37.9% decrease in stem root ratio. However, the seedling height, ground diameter, and root infection rate increased by 23.5%, 50.5%, and 472.9%, respectively. The strain H12 was identified as Paenibacillus alvei, which exerts a considerably positive effect on mycorrhizal growth. Therefore, it might be developed as a biological fertilizer in rhizosphere soil. [ABSTRACT FROM AUTHOR]

The inoculation of forest seedlings with mycorrhizal fungi and rhizobacteria can improve the morphology and physiology of the seedlings and benefit the reforestation of Mediterranean areas and the reintroduction of mycorrhizal fungal inocula into these areas. Pinus nigra subsp. salzmannii,a forest component of the Mediterranean natural ecosystems, is currently used in the reforestation of Mediterranean regions. Its roots are able to form an ectomycorrhizal symbiosis with the Ascomycetes fungus Tuber melanosporum Vitt., the black truffle. The ecological, economic and social values of this ectomycorrhizal fungus is well known. Previously, we demonstrated that the inoculation of Pinus halepensis seedlings with Pseudomonas fluorescens CECT 844 rhizobacteria and the black truffle T. melanosporum improved the plant growth and N absorption of the seedlings. Furthermore, the addition of P. fluorescens CECT 844 doubled the rate of mycorrhization of T. melanosporum. In the present work, P. nigra seedlings were produced in a nursery under well-watered conditions. We studied the morphophysiological response of these seedlings to a combined T. melanosporum and/or a rhizobacteria P. fluorescens CECT 844 inoculation. Five months after inoculation, the growth parameters (seedling height, basal diameter, and shoot and root dry weight), mycorrhizal colonization, water parameters (osmotic potential at both full and zero turgor and modulus of elasticity), and the total contents and concentrations of N, P, and K in the seedlings roots and shoots were measured. The root growth potentials were subsequently estimated. The addition of P. fluorescens CECT 844 did not significantly improve the mycorrhizal colonization by T. melanosporum on P. nigra seedlings. Additionally, the P. fluorescens inoculation caused few significant improvements in the growth and water parameters. Moreover, apparently opposing effects were observed between the two inoculations regarding the seedlings P absorption. We discuss whether P. fluorescens CECT 844 could act as a Mycorrhizal Helper Bacterium (MHB) through different mechanisms depending on the environmental conditions.

Exploration of the effect of soil bacteria on growth and metabolism of beneficial root endophytic fungi is relevant to promote favorable associations between microorganisms of the plant rhizosphere. Hence, the interaction between the plant-growth-promoting fungus Piriformospora indica and different soil bacteria was investigated. The parameters studied were fungal growth and its amino acid composition during the interaction. Fungus and bacteria were confronted in dual cultures in Petri dishes, either through agar or separated by a Perspex wall that only allowed the bacterial volatiles to be effective. Fungal growth was stimulated by Azotobacter chroococcum, whereas Streptomyces anulatus AcH 1003 inhibited it and Streptomyces sp. Nov AcH 505 had no effect. To analyze amino acid concentration data, targeted metabolomics was implemented under supervised analysis according to fungal-bacteria interaction and time. Orthogonal partial least squares-discriminant analysis (OPLS-DA) model clearly discriminated P. indica−A. chroococcum and P. indica−S. anulatus interactions, according to the respective score plot in comparison to the control. The most observable responses were in the glutamine and alanine size groups: While Streptomyces AcH 1003 increased the amount of glutamine, A. chroococcum decreased it. The fungal growth and the increase of alanine content might be associated with the assimilation of nitrogen in the presence of glucose as a carbon source. The N-fixing bacterium A. chroococcum should stimulate fungal amino acid metabolism via glutamine synthetase-glutamate synthase (GS-GOGAT). The data pointed to a stimulated glycolytic activity in the fungus observed by the accumulation of alanine, possibly via alanine aminotransferase. The responses toward the growth-inhibiting Streptomyces AcH 1003 suggest an (oxidative) stress response of the fungus.

Microorganisms can be versatile in their interactions with each other, being variously beneficial, neutral or antagonistic in their effect. Although this versatility has been observed among many microorganisms and in many environments, little is known regarding the mechanisms leading to these changes in behavior. In the present work, we analyzed the mechanism by which the soil bacterium Pseudomonas fluorescens BBc6R8 shifts from stimulating the growth of the ectomycorrhizal fungus Laccaria bicolor S238N to killing the fungus. We show that among the three secondary metabolites produced by the bacterial strain--the siderophores enantio-pyochelin and pyoverdine, and the biosurfactant viscosin--the siderophores are mainly responsible for the antagonistic activity of the bacterium under iron-limited conditions. While the bacterial strain continues to produce beneficial factors, their effects are overridden by the action of their siderophores. This antagonistic activity of the strain P. fluorescens BBC6R8 in iron-depleted environments is not restricted to its influence on L. bicolor, since it was also seen to inhibit the growth of the actinomycete Streptomyces ambofaciens ATCC23877. We show that the strain P. fluorescens BBc6R8 uses different strategies to acquire iron, depending on certain biotic and abiotic factors. [ABSTRACT FROM AUTHOR]

Mycorrhiza helper bacteria (MHB) are known to increase host root colonization by mycorrhizal fungi but the molecular mechanisms and potential tripartite interactions are poorly understood. Through an effort to study Populus microbiome, we isolated 21 Pseudomonas strains from native Populus deltoides roots. These bacterial isolates were characterized and screened for MHB effectiveness on the Populus-Laccaria system. Two additional Pseudomonas strains (i.e., Pf-5 and BBc6R8) from existing collections were included for comparative purposes. We analyzed the effect of co-cultivation of these 23 individual Pseudomonas strains on Laccaria bicolor ‘S238N’ growth rate, mycelial architecture and transcriptional changes. Nineteen of the 23 Pseudomonas strains tested had positive effects on L. bicolor S238N growth, as well as on mycelial architecture, with strains GM41 and GM18 having the most significant effect. Four of seven L. bicolor reporter genes, Tra1, Tectonin2, Gcn5 and Cipc1, thought to be regulated during the interaction with MHB strain BBc6R8, were induced or repressed, while interacting with Pseudomonas strains GM17, GM33, GM41, GM48, Pf-5 and BBc6R8. Strain GM41 promoted the highest roots colonization across three Populus species but most notably in P. deltoides, which is otherwise poorly colonized by L. bicolor. Here we report novel MHB strains isolated from native Populus that improve L. bicolor root colonization on Populus. This tripartite relationship could be exploited for Populus species/genotypes nursery production as a means of improving establishment and survival in marginal lands.

Mycorrhiza helper bacteria (MHB) are known to increase host root colonization by mycorrhizal fungi but the molecular mechanisms and potential tripartite interactions are poorly understood. Through an effort to study Populus microbiome, we isolated 21 Pseudomonas strains from native Populus deltoides roots. These bacterial isolates were characterized and screened for MHB effectiveness on the Populus-Laccaria system. Two additional Pseudomonas strains (i.e., Pf-5 and BBc6R8) from existing collections were included for comparative purposes. We analyzed the effect of co-cultivation of these 23 individual Pseudomonas strains on Laccaria bicolor "S238N" growth rate, mycelial architecture and transcriptional changes. Nineteen of the 23 Pseudomonas strains tested had positive effects on L. bicolor S238N growth, as well as on mycelial architecture, with strains GM41 and GM18 having the most significant effect. Four of seven L. bicolor reporter genes, Tra1, Tectonin2, Gcn5, and Cipc1, thought to be regulated during the interaction with MHB strain BBc6R8, were induced or repressed, while interacting with Pseudomonas strains GM17, GM33, GM41, GM48, Pf-5, and BBc6R8. Strain GM41 promoted the highest roots colonization across three Populus species but most notably in P. deltoides, which is otherwise poorly colonized by L. bicolor. Here we report novel MHB strains isolated from native Populus that improve L. bicolor root colonization on Populus. This tripartite relationship could be exploited for Populus species/genotypes nursery production as a means of improving establishment and survival in marginal lands. [ABSTRACT FROM AUTHOR]

Paenibacillus sp. EJP73 has been previously demonstrated as a mycorrhization helper bacterium (MHB) for the Lactarius rufus- Pinus sylvestris symbiosis in both laboratory and glasshouse experiments. In the present study, the effect of Paenibacillus sp. EJP73 metabolites on L. rufus EO3 pre-symbiotic growth was tested in two agar plate-based systems. Specifically, volatile metabolites were investigated using a dual plate system, in which the presence of strain EJP73 resulted in a significant negative effect on L. rufus EO3 hyphal radial growth but enhanced hyphal branching and reduced internode distance. Soluble metabolites produced by strain EJP73 were tested on L. rufus EO3 growth in single-agar plate assays by incorporating bacterial cell-free whole or molecular weight fraction spent broth into the agar. Whole spent broth had a negative effect on hyphal growth, whereas a low molecular weight fraction (100-1,000) promoted colony radial growth. Headspace and spent broth analysis of strain EJP73 cultures revealed 2,5-diisopropylpyrazine to be the most significant component. Synthesised 2,5-diisopropylpyrazine and elevated CO (2,000 ppm) were tested as specific volatile metabolites in the dual plate system, but neither produced the response shown when strain EJP73 was present. Increased pre-symbiotic hyphal branching leading to increased likelihood of plant infection may be an important MHB mechanism for strain EJP73. Although the precise signal molecules could not be identified, the work suggests a number of metabolites may work synergistically to increase L. rufus root colonisation. [ABSTRACT FROM AUTHOR]

The present greenhouse study was undertaken to evaluate the effects of co-inoculating the ectomycorrhizal (ECM) fungus Boletus edulis with the mycorrhiza helper bacterium Bacillus cereus HB12 or HB59 on the growth and nutrient uptake of Pinus thunbergii. The inoculation with mycorrhiza helper bacterium significantly ( P ≤ 0.05) increased the ectomycorrhizal colonization. Treatments with dual inoculum (the mycorrhiza helper bacterium plus mycorrhiza) significantly ( P ≤ 0.05) increased the P. thunbergii growth. Bacteria-mycorrhizae interactions resulted in a great utilization of phosphate and potassium. The single inoculation resulted in a higher root activity than the control while the co-inoculation led to the highest root activity. The 6-CFDA staining assay showed that B. cereus enhanced fungal activity in ectomycorrhizal symbiosis. The results conclusively suggest that B. cereus isolated from the rhizosphere of P. thunbergii can potentially be used as individual inoculant or co-inoculated with ECM fungi to increase the production in sustainable ecological systems. These results support the potential use of B. cereus (HB12 or HB59) and B. edulis as mixed inoculants stimulating growth of P. thunbergii. [ABSTRACT FROM AUTHOR]

Abstract: The bacterium Wautersia [Ralstonia] basilensis has been shown to enhance the mycorrhizal symbiosis between Suillus granulatus and Pinus thunbergii (Japanese black pine). However, no information is available about this bacterium under field conditions. The objectives of this study were to detect W. basilensis in bulk and mycorhizosphere soils in a Japanese pine plantation in the Tottori Sand Dunes, determine the density of W. basilensis in soil, and determine the optimal cell density of W. basilensis for mycorrhizal formation in pine seedlings. We designed and validated 16S rRNA gene-targeted specific primers for detection and quantification of W. basilensis. SYBR Green I real-time PCR assay was used. A standard curve relating cultured W. basilensis cell density (103–108 cells ml−1) to amplification of DNA showed a strong linear relationship (R = 0.9968). The specificity of the reaction was confirmed by analyzing DNA melting curves and sequencing of the amplicon. The average cell density of W. basilensis was >4.8 × 107 cells g−1 of soil in the mycorrhizosphere and 7.0 × 106 cells g−1 in the bulk soil. We evaluated the W. basilensis cell density required for mycorrhizal formation using an in vitro microcosm with various inoculum densities ranging from 102 to 107 cells g−1 soil (104–109 cells ml−1). Cell densities of W. basilensis of >106 cells g−1 of soil were required to stimulate mycorrhizal formation. In vivo and in vitro experiments showed that W. basilensis was sufficiently abundant to enhance mycorrhizal formation in the mycorrhizosphere of Japanese black pine sampled from the Tottori Sand Dunes. [Copyright &y& Elsevier]

• The mycorrhiza helper Pseudomonas fluorescens BBc6R8 promotes the presymbiotic survival and growth of the ectomycorrhizal fungus Laccaria bicolor S238N in the soil. • An in vitro fungal–bacterial confrontation bioassay mimicking the promoting effects of the bacteria on fungal growth was set up to analyse the fungal morphological and transcriptional changes induced by the helper bacteria at three successive stages of the interaction. The specificity of the P. fluorescens BBc6R8 effect was assessed in comparison with six other rhizobacterial strains possessing mycorrhiza helper or pathogen antagonistic abilities. • The helper BBc6R8 strain was the only strain to induce increases in the radial growth of the colony, hyphal apex density and branching angle. These morphological modifications were coupled with pleiotropic alterations of the fungal transcriptome, which varied throughout the interaction. Early stage-responsive genes were presumably involved in recognition processes and transcription regulation, while late stage-responsive genes encoded proteins of primary metabolism. Some of the responsive genes were partly specific to the interaction with P. fluorescens BBc6R8, whereas others were mutually regulated by different rhizobacteria. • The results highlight the fact that the helper BBc6R8 strain has a specific priming effect on growth, morphology and gene expression of its fungal associate L. bicolor S238N. [ABSTRACT FROM AUTHOR]

• The interaction between the mycorrhiza helper bacteria Streptomyces nov. sp. 505 (AcH 505) and Streptomyces annulatus 1003 (AcH 1003) with fly agaric ( Amanita muscaria) and spruce ( Picea abies) was investigated. • The effects of both bacteria on the mycelial growth of different ectomycorrhizal fungi, on ectomycorrhiza formation, and on fungal gene expression in dual culture with AcH 505 were determined. • The fungus specificities of the streptomycetes were similar. Both bacterial species showed the strongest effect on the growth of mycelia at 9 wk of dual culture. The effect of AcH 505 on gene expression of A. muscaria was examined using the suppressive subtractive hybridization approach. The responsive fungal genes included those involved in signalling pathways, metabolism, cell structure, and the cell growth response. • These results suggest that AcH 505 and AcH 1003 enhance mycorrhiza formation mainly as a result of promotion of fungal growth, leading to changes in fungal gene expression. Differential A. muscaria transcript accumulation in dual culture may result from a direct response to bacterial substances. [ABSTRACT FROM AUTHOR]

Comments on a diverse set of reviews on evolutionary genomics published in "New Phytologist" which illustrates how challenging the genome will be to understand. Discussion on the duplicitous nuclear genome; Observation on a review article that explores epigenetics; Details of a molecular and evolutionary genetic work on understanding how plants prevent self-fertilization.

Abstract: This article reviews recent developments in in situ bioremediation of trace metal contaminated soils, with particular reference to the microbial dynamics in the rhizospheres of plants growing on such soils and their significance in phytoremediation. In non-agricultural conditions, the natural role of plant growth promoting rhizobacteria (PGPR), P-solubilizing bacteria, mycorrhizal-helping bacteria (MHB) and arbuscular mycorrhizal fungi (AMF) in maintaining soil fertility is more important than in conventional agriculture, horticulture, and forestry where higher use of agrochemicals minimize their significance. These microbes initiate a concerted action when a particular population density is achieved, i.e. quorum sensing. AMF also recognize their host by signals released by host roots, allowing a functional symbiosis. AM fungi produce an insoluble glycoprotein, glomalin, which sequester trace elements and it should be considered for biostabilization leading to remediation of contaminated soils. Conclusions drawn from studies of metal uptake kinetics in solution cultures may not be valid for more complex field conditions and use of some combination of glasshouse and field experiments with organisms that occur within the same plant community is suggested. Phytoextraction strategies, such as inoculation of plants to be used for phytoremediation with appropriate heavy metal adapted rhizobial microflora, co-cropping system involving a non-mycorrhizal hyperaccumulator plant and a non-accumulator but mycorrhizal with appropriate AMF, or pre-cropping with mycotrophic crop systems to optimize phytoremediation processes, merit further field level investigations. There is also a need to improve our understanding of the mechanisms involved in transfer and mobilization of trace elements by rhizosphere microbiota and to conduct research on selection of microbial isolates from rhizosphere of plants growing on heavy metal contaminated soils for specific restoration programmes. This is necessary if we are to improve the chances of successful phytoremediation. [Copyright &y& Elsevier]

Mycorrhizal helper bacteria (MHB) play an important role in mediating mycorrhizal symbiosis, which improves the growth and nutrient uptake of plants. This study examined the growth-promoting effects and mechanisms of pine growth after inoculation with the MHB Bacillus pumilus HR10 and/or Hymenochaete sp. Rl. The effect of B. pumilus HR10 on Hymenochaete sp. Rl growth, enzyme activity and gene expression related to mycorrhiza formation were determined. The growth, root activity, nitrogen, phosphorus, and potassium content and chlorophyll fluorescence activity of Pinus thunbergii and the mycorrhizal colonization intensity of Hymenochaete sp. Rl-inoculated pine seedlings after inoculation with B. pumilus HR10 were also evaluated. The results showed that B. pumilus HR10 promoted growth, regulated the expression of mycorrhizal-related genes and affected the β-1,3-glucanase activity of Hymenochaete sp. Rl. The mycorrhizal colonization intensity of pine seedlings co-inoculated with B. pumilus HR10 and Hymenochaete sp. Rl was 1.58-fold higher than seedlings inoculated with only Hymenochaete sp. Rl. Inoculation with B. pumilus HR10 and/or Hymenochaete sp. Rl increased lateral root number and root activity of pine seedlings and chlorophyll fluorescence activity of pine needles compared with the control. Bacillus pumilus HR10 facilitated nutrient uptake by enhancing the mycorrhizal proliferation of pine and induced greater photosynthesis and root activity of pine seedlings, which confirms its role as an outstanding plant-growth-promoting rhizobacterium. These findings improve our understanding of the mechanism of B. pumilus HR10 promotion of mycorrhizal symbiosis., (© The Author(s) 2021. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Exploration of the effect of soil bacteria on growth and metabolism of beneficial root endophytic fungi is relevant to promote favorable associations between microorganisms of the plant rhizosphere. Hence, the interaction between the plant-growth-promoting fungus Piriformospora indica and different soil bacteria was investigated. The parameters studied were fungal growth and its amino acid composition during the interaction. Fungus and bacteria were confronted in dual cultures in Petri dishes, either through agar or separated by a Perspex wall that only allowed the bacterial volatiles to be effective. Fungal growth was stimulated by Azotobacter chroococcum, whereas Streptomyces anulatus AcH 1003 inhibited it and Streptomyces sp. Nov AcH 505 had no effect. To analyze amino acid concentration data, targeted metabolomics was implemented under supervised analysis according to fungal-bacteria interaction and time. Orthogonal partial least squares-discriminant analysis (OPLS-DA) model clearly discriminated P. indica–A. chroococcum and P. indica–S. anulatus interactions, according to the respective score plot in comparison to the control. The most observable responses were in the glutamine and alanine size groups: While Streptomyces AcH 1003 increased the amount of glutamine, A. chroococcum decreased it. The fungal growth and the increase of alanine content might be associated with the assimilation of nitrogen in the presence of glucose as a carbon source. The N-fixing bacterium A. chroococcum should stimulate fungal amino acid metabolism via glutamine synthetase-glutamate synthase (GS-GOGAT). The data pointed to a stimulated glycolytic activity in the fungus observed by the accumulation of alanine, possibly via alanine aminotransferase. The responses toward the growth-inhibiting Streptomyces AcH 1003 suggest an (oxidative) stress response of the fungus. [ABSTRACT FROM AUTHOR]