Intransitivity in plant-soil feedbacks is rare but is associated with multispecies coexistence.

Although plant-soil feedback (PSF) is being recognized as an important driver of plant recruitment, our understanding of its role in species coexistence in natural communities remains limited by the scarcity of experimental studies on multispecies assemblages. Here, we experimentally estimated PSFs affecting seedling recruitment in 10 co-occurring Mediterranean woody species. We estimated weak but significant species-specific feedback. Pairwise PSFs impose similarly strong fitness differences and stabilizing-destabilizing forces, most often impeding species coexistence. Moreover, a model of community dynamics driven exclusively by PSFs suggests that few species would coexist stably, the largest assemblage with no more than six species. Thus, PSFs alone do not suffice to explain coexistence in the studied community. A topological analysis of all subcommunities in the interaction network shows that full intransitivity (with all species involved in an intransitive loop) would be rare but it would lead to species coexistence through either stable or cyclic dynamics.

Arbuscular mycorrhizal fungi from acidic soils favors production of tomatoes and lycopene concentration.

BACKGROUND: Tomato is widely consumed throughout the world for its flavor and nutritional value. This functional food largely depends on the implementation of new strategies to maintain the nutraceutical value, e.g. lycopene concentration, and overcome the challenges of sustainable production and food security. The use of arbuscular mycorrhizal fungi (AMF)-based biostimulants represents one of the most promising tools for sustainable management of agricultural soils, being fundamental for organic food production, reducing fertilizers and pesticides use, and decreasing environmental damage. This study aimed at elucidating whether native arbuscular mycorrhizal fungi (AMF) could positively affect tomato yield and lycopene concentration. RESULTS: Native AMF inoculum consisted of two inoculum types: the single species Claroideoglomus claroideum, and a mix of Scutellospora calospora, Acaulospora laevis, Claroideoglomus claroideum, and Claroideoglomus etunicatum. At the end of the study up to 78% of the root system was colonized by single inoculum. Tomato diameters in single and mix mycorrhizal plants showed increases of 80% and 35% respectively. Fresh weights were 84% and 38% higher with single and mix inocula compared with the controls, respectively. The lycopene concentration in tomato fruits of plants with single and mix inoculum was higher than controls. The lycopene concentration was 124.5% and 113.9% greater in single and mix than non-inoculated plants. CONCLUSION: Tomato diameters, fresh weight and lycopene concentration was significantly higher in plants colonized by AMF compared with uninoculated plants. Results suggest that the role of single species Claroideoglomus claroideum could generate better plant performance due to its high production of extraradical mycelium. © 2021 Society of Chemical Industry.

Ancient lineages of arbuscular mycorrhizal fungi provide little plant benefit.

Almost all land plants form symbiotic associations with arbuscular mycorrhizal fungi (AMF). Individual plants usually are colonized by a wide range of phylogenetically diverse AMF species. The impact that different AMF taxa have on plant growth is only partly understood. We screened 44 AMF isolates for their effect on growth promotion and nutrient uptake of leek plants (Allium porrum), including isolates that have not been tested previously. In particular, we aimed to test weather AMF lineages with an ancient evolutionary age differ from relatively recent lineages in their effects on leek plants. The AMF isolates that were tested covered 18 species from all five AMF orders, eight families, and 13 genera. The experiment was conducted in a greenhouse. A soil-sand mixture was used as substrate for the leek plants. Plant growth response to inoculation with AMF varied from - 19 to 232% and depended on isolate, species, and family identity. Species from the ancient families Archaeosporaceae and Paraglomeraceae tended to be less beneficial, in terms of stimulation plant growth and nutrient uptake, than species of Glomeraceae, Entrophosporaceae, and Diversisporaceae, which are considered phylogenetically more recent than those ancient families. Root colonization levels also depended on AMF family. This study indicates that plant benefit in the symbiosis between plants and AMF is linked to fungal identity and phylogeny and it shows that there are large differences in effectiveness of different AMF.

Root metabolic plasticity underlies functional diversity in mycorrhiza-enhanced stress tolerance in tomato.

Arbuscular mycorrhizal (AM) symbioses can improve plant tolerance to multiple stresses. We compared three AM fungi (AMF) from different genera, one of them isolated from a dry and saline environment, in terms of their ability to increase tomato tolerance to moderate or severe drought or salt stress. Plant physiological parameters and metabolic profiles were compared in order to find the molecular mechanisms underlying plant protection against stress. Mycorrhizal growth response was determined, and ultrahigh-performance LC-MS was used to compare the metabolic profile of plants under the different treatments. All AMF increased plant tolerance to stress, and the positive effects of the symbiosis were correlated with the severity of the stress. The AMF isolated from the stressful environment was the most effective in improving plant tolerance to salt stress. Differentially accumulated compounds were identified and the antistress properties of some of them were confirmed. We demonstrate that AM symbioses increase plant metabolic plasticity to cope with stress. Some responses were common to all AMF tested, while others were specifically related to particular isolates. Important metabolism reprograming was evidenced upon salt stress, and we identified metabolic pathways and compounds differentially accumulated in mycorrhizas that may underlie their enhanced tolerance to stress.

Plant traits determine the phylogenetic structure of arbuscular mycorrhizal fungal communities.

Functional diversity in ecosystems has traditionally been studied using aboveground plant traits. Despite the known effect of plant traits on the microbial community composition, their effects on the microbial functional diversity are only starting to be assessed. In this study, the phylogenetic structure of arbuscular mycorrhizal (AM) fungal communities associated with plant species differing in life cycle and growth form, that is, plant life forms, was determined to unravel the effect of plant traits on the functional diversity of this fungal group. The results of the 454 pyrosequencing showed that the AM fungal community composition differed across plant life forms and this effect was dependent on the soil collection date. Plants with ruderal characteristics tended to associate with phylogenetically clustered AM fungal communities. By contrast, plants with resource-conservative traits associated with phylogenetically overdispersed AM fungal communities. Additionally, the soil collected in different seasons yielded AM fungal communities with different phylogenetic dispersion. In summary, we found that the phylogenetic structure, and hence the functional diversity, of AM fungal communities is dependent on plant traits. This finding adds value to the use of plant traits for the evaluation of belowground ecosystem diversity, functions and processes.

Differences in the composition of arbuscular mycorrhizal fungal communities promoted by different propagule forms from a Mediterranean shrubland.

As it is well known, arbuscular mycorrhizal (AM) colonization can be initiated from the following three types of fungal propagules: spores, extraradical mycelium (ERM), and mycorrhizal root fragments harboring intraradical fungal structures. It has been shown that biomass allocation of AM fungi (AMF) among these three propagule types varies between fungal taxa, as also differs the ability of the different AMF propagule fractions to initiate new colonizations. In this study, the composition of the AMF community in the roots of rosemary (Rosmarinus officinalis L., a characteristic Mediterranean shrub), inoculated with the three different propagule types, was analyzed. Accordingly, cuttings from this species were inoculated with either AMF spores, ERM, or colonized roots extracted from a natural soil. The AMF diversity within the rosemary roots was characterized using terminal restriction fragment length polymorphism (T-RFLP) of the small subunit (SSU) rDNA region. The AMF community established in the rosemary plants was significantly different according to the type of propagule used as inoculum. AMF taxa differed in their ability to initiate new colonizations from each propagule type. Results suggest different colonization strategies for the different AMF families involved, Glomeraceae and Claroideoglomeraceae colonizing mainly from colonized roots whereas Pacisporaceae and Diversisporaceae from spores and ERM. This supports that AMF taxa show contrasting life-history strategies in terms of their ability to initiate new colonizations from the different propagule types. Further research to fully understand the colonization and dispersal abilities of AMF is essential for their rational use in ecosystem restoration programs.

The composition of arbuscular mycorrhizal fungal communities differs among the roots, spores and extraradical mycelia associated with five Mediterranean plant species.

Arbuscular mycorrhizal fungi (AMF) are essential constituents of most terrestrial ecosystems. AMF species differ in terms of propagation strategies and the major propagules they form. This study compared the AMF community composition of different propagule fractions - colonized roots, spores and extraradical mycelium (ERM) - associated with five Mediterranean plant species in Sierra de Baza Natural Park (Granada, Spain). AMF were identified using 454 pyrosequencing of the SSU rRNA gene. A total of 96 AMF phylogroups [virtual taxa (VT)] were detected in the study site, including 31 novel VT. After per-sample sequencing depth standardization, 71 VT were recorded from plant roots, and 47 from each of the spore and ERM fractions. AMF communities differed significantly among the propagule fractions, and the root-colonizing fraction differed among host plant species. Indicator VT were detected for the root (13 Glomus VT), spore (Paraglomus VT281, VT336, Pacispora VT284) and ERM (Diversispora VT62) fractions. This study provides detailed evidence from a natural system that AMF taxa are differentially allocated among soil mycelium, soil spores and colonized root propagules. This has important implications for interpreting AMF diversity surveys and designing applications of AMF in vegetation restoration.

Phytohormones as integrators of environmental signals in the regulation of mycorrhizal symbioses.

For survival, plants have to efficiently adjust their phenotype to environmental challenges, finely coordinating their responses to balance growth and defence. Such phenotypic plasticity can be modulated by their associated microbiota. The widespread mycorrhizal symbioses modify plant responses to external stimuli, generally improving the resilience of the symbiotic system to environmental stresses. Phytohormones, central regulators of plant development and immunity, are instrumental in orchestrating plant responses to the fluctuating environment, but also in the regulation of mycorrhizal symbioses. Exciting advances in the molecular regulation of phytohormone signalling are providing mechanistic insights into how plants coordinate their responses to environmental cues and mycorrhizal functioning. Here, we summarize how these mechanisms permit the fine-tuning of the symbiosis according to the ever-changing environment.

The interactions between plant life form and fungal traits of arbuscular mycorrhizal fungi determine the symbiotic community.

Arbuscular mycorrhizal (AM) fungi have traditionally been considered generalist symbionts. However, an increasing number of studies are pointing out the selectivity potential of plant hosts. Plant life form, determined by plant life history traits, seems to drive the AM fungal community composition. The AM fungi also exhibit a wide diversity of functional traits known to be responsible for their distribution in natural ecosystems. However, little is known about the role of plant and fungal traits driving the resultant symbiotic assemblages. With the aim of testing the feedback relationship between plant and fungal traits on the resulting AM fungal community, we inoculated three different plant life forms, i.e. annual herbs, perennial herbs and perennial semi-woody plants, with AM fungal communities sampled in different seasons. We hypothesized that the annual climate variation will induce changes in the mean traits of the AM fungal communities present in the soil throughout the year. Furthermore, the association of plants with different life forms with AM fungi with contrasting life history traits will show certain preferences according to reciprocal traits of the plants and fungi. We found changes in the AM fungal community throughout the year, which were differentially disrupted by disturbance and altered by plant growth form and plant biomass. Both plant and fungal traits clearly contributed to the resultant AM fungal communities. The revealed process can have implications for the functioning of ecosystems since changes in dominant plant life forms or climatic variables could influence the traits of AM fungal communities in soil and hence ecosystem processes.

GintAMT2, a new member of the ammonium transporter family in the arbuscular mycorrhizal fungus Glomus intraradices.

In the symbiotic association of plants and arbuscular mycorrhizal (AM) fungi, the fungus delivers mineral nutrients, such as phosphate and nitrogen, to the plant while receiving carbon. Previously, we identified an NH(4)(+) transporter in the AM fungus Glomus intraradices (GintAMT1) involved in NH(4)(+) uptake from the soil when preset at low concentrations. Here, we report the isolation and characterization of a new G. intraradicesNH(4)(+) transporter gene (GintAMT2). Yeast mutant complementation assays showed that GintAMT2 encodes a functional NH(4)(+) transporter. The use of an anti-GintAMT2 polyclonal antibody revealed a plasma membrane location of GintAMT2. GintAMT1 and GintAMT2 were differentially expressed during the fungal life cycle and in response to N. In contrast to GintAMT1, GintAMT2 transcript levels were higher in the intraradical than in the extraradical fungal structures. However, transcripts of both genes were detected in arbuscule-colonized cortical cells. GintAMT1 expression was induced under low N conditions. Constitutive expression of GintAMT2 in N-limiting conditions and transitory induction after N re-supply suggests a role for GintAMT2 to retrieve NH(4)(+) leaked out during fungal metabolism.

Mycorrhiza-Induced Resistance against Foliar Pathogens Is Uncoupled of Nutritional Effects under Different Light Intensities.

The use of microbial inoculants, particularly arbuscular mycorrhizal fungi, has great potential for sustainable crop management, which aims to reduce the use of chemical fertilizers and pesticides. However, one of the major challenges of their use in agriculture is the variability of the inoculation effects in the field, partly because of the varying environmental conditions. Light intensity and quality affect plant growth and defense, but little is known about their impacts on the benefits of mycorrhizal symbioses. We tested the effects of five different light intensities on plant nutrition and resistance to the necrotrophic foliar pathogen Botrytis cinerea in mycorrhizal and non-mycorrhizal lettuce plants. Our results evidence that mycorrhiza establishment is strongly influenced by light intensity, both regarding the extension of root colonization and the abundance of fungal vesicles within the roots. Light intensity also had significant effects on plant growth, nutrient content, and resistance to the pathogen. The effect of the mycorrhizal symbiosis on plant growth and nutrient content depended on the light intensity, and mycorrhiza efficiently reduced disease incidence and severity under all light intensities. Thus, mycorrhiza-induced resistance can be uncoupled from mycorrhizal effects on plant nutrition. Therefore, mycorrhizal symbioses can be beneficial by providing biotic stress protection even in the absence of nutritional or growth benefits.

Characterization of a CuZn superoxide dismutase gene in the arbuscular mycorrhizal fungus Glomus intraradices.

To gain further insights into the mechanisms of redox homeostasis in arbuscular mycorrhizal fungi, we characterized a Glomus intraradices gene (GintSOD1) showing high similarity to previously described genes encoding CuZn superoxide dismutases (SODs). The GintSOD1 gene consists of an open reading frame of 471 bp, predicted to encode a protein of 157 amino acids with an estimated molecular mass of 16.3 kDa. Functional complementation assays in a CuZnSOD-defective yeast mutant showed that GintSOD1 protects the yeast cells from oxygen toxicity and that it, therefore, encodes a protein that scavenges reactive oxygen species (ROS). GintSOD1 transcripts differentially accumulate during the fungal life cycle, reaching the highest expression levels in the intraradical mycelium. GintSOD1 expression is induced by the well known ROS-inducing agents paraquat and copper, and also by fenpropimorph, a sterol biosynthesis inhibitor (SBI) fungicide. These results suggest that GintSOD1 is involved in the detoxification of ROS generated from metabolic processes and by external agents. In particular, our data indicate that the antifungal effects of fenpropimorph might not be only due to the interference with sterol metabolism but also to the perturbation of other biological processes and that ROS production and scavenging systems are involved in the response to SBI fungicides.

Hormonal and transcriptional profiles highlight common and differential host responses to arbuscular mycorrhizal fungi and the regulation of the oxylipin pathway.

Arbuscular mycorrhizal (AM) symbioses are mutualistic associations between soil fungi and most vascular plants. The symbiosis significantly affects the host physiology in terms of nutrition and stress resistance. Despite the lack of host range specificity of the interaction, functional diversity between AM fungal species exists. The interaction is finely regulated according to plant and fungal characters, and plant hormones are believed to orchestrate the modifications in the host plant. Using tomato as a model, an integrative analysis of the host response to different mycorrhizal fungi was performed combining multiple hormone determination and transcriptional profiling. Analysis of ethylene-, abscisic acid-, salicylic acid-, and jasmonate-related compounds evidenced common and divergent responses of tomato roots to Glomus mosseae and Glomus intraradices, two fungi differing in their colonization abilities and impact on the host. Both hormonal and transcriptional analyses revealed, among others, regulation of the oxylipin pathway during the AM symbiosis and point to a key regulatory role for jasmonates. In addition, the results suggest that specific responses to particular fungi underlie the differential impact of individual AM fungi on plant physiology, and particularly on its ability to cope with biotic stresses.

Entrophospora nevadensis, a new arbuscular mycorrhizal fungus from Sierra Nevada National Park (southeastern Spain).

A new fungal species in the arbuscular mycorrhiza-forming Glomeromycetes, Entrophospora nevadensis, was isolated from soil near the roots of several endemic and endangered plant species (e.g. Plantago nivalis and Alchemilla fontqueri) growing in Sierra Nevada National Park (Granada, Andalucia, Spain). The fungus was propagated in trap cultures on Plantago nivalis and Sorbus hybrida and in pure cultures on Trifolium pratense and Sorghum vulgare. Spores are yellow brown to brown, 90-115 .m diam and form singly in soil, in the neck of adherent sporiferous saccules that form either terminally or intercalary on mycelial hyphae. Spores have two three-layered walls and conspicuous, 6-12 microm long, spiny, thorn-like projections on the outer wall consisting of hyaline to subhyaline, evanescent tips and yellow brown to brown, persistent bases. In aging spores these projections are usually shorter (1-2.8 microm) and dome-shaped or rounded, sometimes with a central pit on top where the evanescent tip has sloughed off. Molecular analysis with partial sequences of the 18S ribosomal gene places the fungus within the Diversisporales. The new fungus was found in soil near plants with different living strategies but growing in high altitude soils with acidic pH, high soil moisture and organic carbon content, and close to streams.

GintABC1 encodes a putative ABC transporter of the MRP subfamily induced by Cu, Cd, and oxidative stress in Glomus intraradices.

A full-length cDNA sequence putatively encoding an ATP-binding cassette (ABC) transporter (GintABC1) was isolated from the extraradical mycelia of the arbuscular mycorrhizal fungus Glomus intraradices. Bioinformatic analysis of the sequence indicated that GintABC1 encodes a 1513 amino acid polypeptide, containing two six-transmembrane clusters (TMD) intercalated with sequences characteristics of the nucleotide binding domains (NBD) and an extra N-terminus extension (TMD0). GintABC1 presents a predicted TMD0-(TMD-NBD)(2) topology, typical of the multidrug resistance-associated protein subfamily of ABC transporters. Gene expression analyses revealed no difference in the expression levels of GintABC1 in the extra- vs the intraradical mycelia. GintABC1 was up-regulated by Cd and Cu, but not by Zn, suggesting that this transporter might be involved in Cu and Cd detoxification. Paraquat, an oxidative agent, also induced the transcription of GintABC1. These data suggest that redox changes may be involved in the transcriptional regulation of GintABC1 by Cd and Cu.

Effect of Arbuscular Mycorrhizal Colonization on Cadmium-Mediated Oxidative Stress in Glycine max (L.) Merr.

Cadmium is a heavy metal (HM) that inhibits plant growth and leads to death, causing great losses in yields, especially in Cd hyperaccumulator crops such as Glycine max (L.) Merr. (soybean), a worldwide economically important legume. Furthermore, Cd incorporation into the food chain is a health hazard. Oxidative stress (OS) is a plant response to abiotic and biotic stresses with an intracellular burst of reactive oxygen species (ROS) that causes damage to lipids, proteins, and DNA. The arbuscular mycorrhizal fungal (AMF) association is a plant strategy to cope with HM and to alleviate OS. Our aim was to evaluate the mitigation effects of mycorrhization with AMF Rhizophagus intraradices on soybean growth, nutrients, Cd accumulation, lipid peroxidation, and the activity of different antioxidant agents under Cd (0.7-1.2 mg kg-1 bioavailable Cd) induced OS. Our results suggest that glutathione may act as a signal molecule in a defense response to Cd-induced OS, and mycorrhization may avoid Cd-induced growth inhibition and reduce Cd accumulation in roots. It is discussed that R. intraradices mycorrhization would act as a signal, promoting the generation of a soybean cross tolerance response to Cd pollution, therefore evidencing the potential of this AMF association for bioremediation and encouragement of crop development, particularly because it is an interaction between a worldwide cultivated Cd hyperaccumulator plant and an AMF-HM-accumulator commonly present in soils.

Unraveling mycorrhiza-induced resistance.

Arbuscular mycorrhizal symbioses have a significant impact on plant interactions with other organisms. Increased resistance to soil-borne pathogens has been widely described in mycorrhizal plants. By contrast, effects on shoot diseases largely rely on the lifestyle and challenge strategy of the attacker. Among the potential mechanisms involved in the resistance of mycorrhizal systems, the induction of plant defenses is the most controversial. During mycorrhiza formation, modulation of plant defense responses occurs, potentially through cross-talk between salicylic acid and jasmonate dependent signaling pathways. This modulation may impact plant responses to potential enemies by priming the tissues for a more efficient activation of defense mechanisms.

GintGRX1, the first characterized glomeromycotan glutaredoxin, is a multifunctional enzyme that responds to oxidative stress.

Glutaredoxins (GRXs) are small proteins with glutathione-dependent disulfide oxidoreductase activity involved in cellular defense against oxidative stress. This work reports the identification and characterization of the first glomeromycotan dithiol glutaredoxin gene from the fungus Glomus intraradices. The corresponding gene, named GintGRX1, shares high sequence similarity with previously described fungal GRXs. GintGRX1 contains the characteristic dithiol active site CPYC. By using a yeast expression system, we found that GintGRX1 encodes a multifunctional protein with oxidoreductase, peroxidase and glutathione S-transferase activity. GintGRX1 partially reverted sensitivity to superoxide radicals of the Deltagrx1Deltagrx2Saccharomyces cerevisiae strain. GintGRX1 was transcriptionally regulated by paraquat but not by hydrogen peroxide. Copper induced an accumulation of reactive oxygen species in the extraradical mycelium of G. intraradices and up-regulation of GintGRX1 transcript levels. These data suggest a role for GintGRX1 in protecting the fungus against the oxidative damage induced directly by the superoxide anion or indirectly by copper.

GintPDX1 encodes a protein involved in vitamin B6 biosynthesis that is up-regulated by oxidative stress in the arbuscular mycorrhizal fungus Glomus intraradices.

Vitamin B6 is an essential metabolite that has recently been implicated in defense against cellular oxidative stress. In fungi, the de novo biosynthetic pathway of vitamin B6 involves two genes, PDX1 and PDX2. Here, we report a component of the PDX1/PDX2 vitamin B6 biosynthetic pathway in an arbuscular mycorrhizal (AM) fungus. Using rapid amplification of cDNA ends, we isolated the full-length cDNA of a PDX-like gene, GintPDX1, from Glomus intraradices. GintPDX1 expression was analysed by real-time reverse transcription-polymerase chain reaction (RT-PCR). GintPDX1 activity and function were investigated by heterologous complementation of the yeast strainDeltasnz1, which is deficient in vitamin B6 biosynthesis. Sequence data revealed that GintPDX1 is highly homologous to other identified PDX1 proteins. GintPDX1 restores prototrophy to the vitamin B6 auxotrophic yeast mutant and reverts its superoxide sensitivity. GintPDX1 is expressed throughout the fungal life cycle, with the highest transcription levels found in the intraradical fungal structures. GintPDX1 expression was induced in response to hydrogen peroxide, paraquat and copper. The results demonstrate that AM fungi possess at least one component of the machinery necessary for vitamin B6 biosynthesis. Transcriptional regulation of GintPDX1 suggests a role for vitamin B6 as an antioxidant and modulator of reactive oxygen species in G. intraradices.

Review: Arbuscular mycorrhizas as key players in sustainable plant phosphorus acquisition: An overview on the mechanisms involved.

Phosphorus (P) is a poorly available macronutrient essential for plant growth and development and consequently for successful crop yield and ecosystem productivity. To cope with P limitations plants have evolved strategies for enhancing P uptake and/or improving P efficiency use. The universal 450-million-yr-old arbuscular mycorrhizal (AM) (fungus-root) symbioses are one of the most successful and widespread strategies to maximize access of plants to available P. AM fungi biotrophically colonize the root cortex of most plant species and develop an extraradical mycelium which overgrows the nutrient depletion zone of the soil surrounding plant roots. This hyphal network is specialized in the acquisition of low mobility nutrients from soil, particularly P. During the last years, molecular biology techniques coupled to novel physiological approaches have provided fascinating contributions to our understanding of the mechanisms of symbiotic P transport. Mycorrhiza-specific plant phosphate transporters, which are required not only for symbiotic P transfer but also for maintenance of the symbiosis, have been identified. The present review provides an overview of the contribution of AM fungi to plant P acquisition and an update of recent findings on the physiological, molecular and regulatory mechanisms of P transport in the AM symbiosis.