Different levels of hyphal self-incompatibility modulate interconnectedness of mycorrhizal networks in three arbuscular mycorrhizal fungi within the Glomeraceae

Arbuscular mycorrhizal fungi (AMF) live in symbiosis with most plant species and produce underground extraradical hyphal networks functional in the uptake and translocation of mineral nutrients from the soil to host plants. This work investigated whether fungal genotype can affect patterns of interconnections and structural traits of extraradical mycelium (ERM), by comparing three Glomeraceae species growing in symbiosis with five plant hosts. An isolate of Funneliformis coronatus consistently showed low ability to form interconnected ERM and self-incompatibility that represented up to 21 % of hyphal contacts. The frequency of post-fusion self-incompatible interactions, never detected before in AMF extraradical networks, was 8.9 %. In F. coronatus ERM, the percentage of hyphal contacts leading to perfect hyphal fusions was 1.2–7.7, while it ranged from 25.8–48 to 35.6–53.6 in Rhizophagus intraradices and Funneliformis mosseae, respectively. Low interconnectedness of F. coronatus ERM resulted also from a very high number of non-interacting contacts (83.2 %). Such findings show that AMF genotypes in Glomeraceae can differ significantly in anastomosis behaviour and that ERM interconnectedness is modulated by the fungal symbiont, as F. coronatus consistently formed poorly interconnected networks when growing in symbiosis with five different host plants and in the asymbiotic stage. Structural traits, such as extent, density and hyphal self-compatibility/incompatibility, may represent key factors for the differential performance of AMF, by affecting fungal absorbing surface and foraging ability and thus nutrient flow from soil to host roots

Lifespan and functionality of mycorrhizal fungal mycelium are uncoupled from host plant lifespan

Arbuscular mycorrhizal fungi (AMF) are obligate symbionts, living in associations with the roots of most land plants. AMF produce wide networks of extraradical mycelium (ERM) of indeterminate length, spreading from host roots into the surrounding soil and establishing belowground interconnections among plants belonging to the same or to different taxa. Whether their lifespan and functionality are limited by host plant viability or can be extended beyond this limit is unknown. To address this issue, we performed time-course studies to investigate viability and functionality of ERM produced in an in vivo whole-plant system by Funneliformis mosseae and Rhizoglomus irregulare, after shoot detachment. Our data revealed that viability and functionality of F. mosseae and R. irregulare extraradical hyphae were uncoupled from host plant lifespan. Indeed, ERM spreading from roots of intact or shootless plants showed comparable levels of viability, similar structural traits and ability to establish mycorrhizal symbioses with new plants, as long as five months after shoot removal. Our findings expand the current knowledge on AMF biology and life cycle, providing data on ERM long-term survival in the soil of two Glomeracean species, functional to the prompt establishment of mycorrhizal symbioses and to the maintenance of soil biological fertility

Health-Promoting Properties of Plant Products: The Role of Mycorrhizal Fungi and Associated Bacteria

The concept of food quality, traditionally based on nutritional and sensory properties, has recently acquired an additional meaning, referring to the health-promoting properties of plant products, that are ascribed to plant secondary metabolites called phytochemicals, primarily represented by polyphenolic compounds and glucosinolates. The diversity and content of phytochemicals in plant products are affected by different variables, such as plant genotype, agronomic factors, and arbuscular mycorrhizal fungi (AMF), which establish mycorrhizal symbioses with most crops, including cereals, legumes, vegetables, fruit trees, sunflower, cotton, and sugarcane. AMF and associated bacteria enhance plant growth and health, and affect the production of polyphenols and carotenoids, and the activity of antioxidant enzymes. The production of health-promoting phytochemicals was shown to be differentially modulated by different AMF isolates and bacterial strains, in several food plants, i.e., tomato, lettuce, strawberry, artichoke, maize, grapevine, sunflower. Here, we provide an overview of recent studies concerning the multiple roles played by AMF and associated bacteria in the modulation of the biosynthesis of plant secondary metabolites with health-promoting activity, and discuss the development of designed multifunctional consortia to be used in sustainable agriculture

At the root of the wood wide web: self recognition an nonself incompatibility in mycorrhizal networks.

Arbuscular mycorrhizal (AM) fungi are mutualistic symbionts living in the roots of 80% of land plant species, and developing extensive, below-ground extraradical hyphae fundamental for the uptake of soil nutrients and their transfer to host plants. Since AM fungi have a wide host range, they are able to colonize and interconnect contiguous plants by means of hyphae extending from one root system to another. Such hyphae may fuse due to the widespread occurrence of anastomoses, whose formation depends on a highly regulated mechanism of self recognition. Here, we examine evidences of self recognition and non-self incompatibility in hyphal networks formed by AM fungi and discuss recent results showing that the root systems of plants belonging to different species, genera and families may be connected by means of anastomosis formation between extraradical mycorrhizal networks, which can create indefinitely large numbers of belowground fungal linkages within plant communities

Divergence of Funneliformis mosseae populations over 20 years of laboratory cultivation, as revealed by vegetative incompatibility and molecular analysis

Arbuscular mycorrhizal fungi (AMF) are widespread, important plant symbionts. They absorb and translocate mineral nutrients from the soil to host plants through an extensive extraradical mycelium, consisting of indefinitely large networks of nonseptate, multinucleated hyphae which may be interconnected by hyphal fusions (anastomoses). This work investigated whether different lineages of the same isolate may lose the ability to establish successful anastomoses, becoming vegetatively incompatible, when grown separately. The occurrence of hyphal incompatibility among five lineages of Funneliformis mosseae, originated from the same ancestor isolate and grown in vivo for more than 20 years in different European locations, was assessed by systematic detection of anastomosis frequency and cytological studies. Anastomosis frequencies ranged from 60 to 80% within the same lineage and from 17 to 44% among different lineages. The consistent detection of protoplasm continuity and nuclei in perfect fusions showed active protoplasm flow both within and between lineages. In pairings between different lineages, post-fusion incompatible reactions occurred in 6–48% of hyphal contacts and pre-fusion incompatibility in 2–17%. Molecular fingerprinting profiles showed genetic divergence among lineages, with overall Jaccard similarity indices ranging from 0.85 to 0.95. Here, phenotypic divergence among the five F. mosseae lineages was demonstrated by the reduction of their ability to form anastomosis and the detection of high levels of vegetative incompatibility. Our data suggest that potential genetic divergence may occur in AMF over only 20 years and represent the basis for detailed studies on the relationship between genes regulating anastomosis formation and hyphal compatibility in AMF

Genetic variability assessment of 127 Triticum turgidum L. accessions for mycorrhizal susceptibility-related traits detection

Positive effects of arbuscular mycorrhizal fungi (AMF)—wheat plant symbiosis have been well discussed by research, while the actual role of the single wheat genotype in establishing this type of association is still poorly investigated. In this work, the genetic diversity of Triticum turgidum wheats was exploited to detect roots susceptibility to AMF and to identify genetic markers in linkage with chromosome regions involved in this symbiosis. A tetraploid wheat collection of 127 accessions was genotyped using 35K single-nucleotide polymorphism (SNP) array and inoculated with the AMF species Funneliformis mosseae (F. mosseae) and Rhizoglomus irregulare (R. irregulare), and a genome‐wide association study (GWAS) was conducted. Six clusters of genetically related accessions were identified, showing a different mycorrhizal colonization among them. GWAS revealed four significant quantitative trait nucleotides (QTNs) involved in mycorrhizal symbiosis, located on chromosomes 1A, 2A, 2B and 6A. The results of this work enrich future breeding activities aimed at developing new grains on the basis of genetic diversity on low or high susceptibility to mycorrhization, and, possibly, maximizing the symbiotic effects

Smart use of microbial-rich vermicomposting to enhance tripartite plant-microbe-soil interactions

Vermicomposting is a natural process that utilizes earthworms and associated microbiome to transform organic wastes into vermicompost by-products that are rich in beneficial microorganisms and nutrients such as carbon, phosphorus, nitrogen, magnesium, and calcium. Liquid vermicompost extract (LVE), a derivative of the vermicomposting process, has recently gained interest among scientists and organic farmers due to their potential ability to enhance tripartite plant-microbe-soil interactions that would lead to improved plant and root growth, soil health and overall crop yield productivity. To investigate the short-term effect of LVE on soil mycorrhizal inoculum potential (MIP) and plant-mycobiome interactions, a field trial was carried out at CiRAA E. Avanzi, San Piero a Grado, Pisa, Italy. The effect of LVE and its associated microbial and chemical components on soil MIP and AMF root colonization was evaluated on five summer crops, i.e. chickpea (Cicer arietinum L.), berseem clover (Trifolium alexandrinum L.), lentil (Lens culinaris L.), soybean (Glycine max L. Merrill), and sunflower (Helianthus annuus L.). The test plants were grown with or without the application of LVE in a split-plot trial with five replicates. Freshly made LVE from vermicomposting of wheat straws mixed with horse manure was screened for microbial properties using the Illumina Miseq sequencing platform. Seed inoculation with LVE was done before planting while field inoculation was done at the stem-elongation stage. Un-inoculated seeds and plots were used as controls. Soil MIP was assessed before planting and after harvesting, while AMF root colonization was evaluated at the mid-flowering stage of each crop. The bacterial 16S and fungal ITS sequence analyses showed a high bacteria and fungal abundance and taxonomic alpha diversity present in the LVE. The most dominant taxa included Mucor, Citrobacter, Pseudomonas, Arcobacter, Azomonas and Clostridium. These microbes are commonly found in agricultural soil and are linked to the hydrolysis of complex organic matter, nutrient recycling, production of growth-promoting factors and siderophores, while others are known to produce peptide antimycotics and antibiotics that protect plants against pathogenic soil microorganisms. The soil MIP significantly (p < 0.0001) differed between the two soil sampling times (before planting and after harvesting). It was evident that both seed and field inoculation with LVE significantly enhanced the soil MIP and this could benefit the next crop under rotation. AMF root colonization varied significantly across the crop species (p < 0.0001) and LVE treatment (p = 0.006). Highly nodulated lentils and berseem clover roots recorded significantly higher AMF root colonization than all the other crops. LVE inoculation had an overall positive effect on AMF root colonization with an average increase of 6.2% compared to the un-inoculated crops. These short-term results indicate that there could be a positive effect of the LVE inoculation on the soil MIP and AMF root colonization of our test crops, which could be attributed to the beneficial additive effects of the LVE that enhanced the tripartite plant-microbe-soil interactions

Arbuscular Mycorrhizal Fungi and Associated Microbiota as Plant Biostimulants: Research Strategies for the Selection of the Best Performing Inocula

Arbuscular mycorrhizal fungi (AMF) are beneficial soil microorganisms establishing mutualistic symbioses with the roots of the most important food crops and playing key roles in the maintenance of long-term soil fertility and health. The great inter- and intra-specific AMF diversity can be fully exploited by selecting AMF inocula on the basis of their colonization ability and efficiency, which are aaffected by fungal and plant genotypes and diverse environmental variables. The multiple services provided by AMF are the result of the synergistic activities of the bacterial communities living in the mycorrhizosphere, encompassing nitrogen fixation, P solubilization, and the production of phytohormones, siderophores, and antibiotics. The tripartite association among host plants, mycorrhizal symbionts, and associated bacteria show beneficial emerging properties which could be efficiently exploited in sustainable agriculture. Further in-depth studies, both in microcosms and in the field, performed on different AMF species and isolates, should evaluate their colonization ability, efficiency, and resilience. Transcriptomic studies can reveal the expression levels of nutrient transporter genes in fungal absorbing hyphae in the presence of selected bacterial strains. Eventually, newly designed multifunctional microbial consortia can be utilized as biofertilizers and biostimulants in sustainable and innovative production systems

Local diversity of native arbuscular mycorrhizal symbionts differentially affects growth and nutrition of three crop plant species

Intact whole native AMF communities occurring across a 100-m-long field were used for the evaluation of plant performance, as determined by the actual fungal species colonizing host roots. The soil from distinct plots within a "hot spot" field was collected to set up 54 experimental units where three different plant species were grown, in order to test whether the whole native AMF communities were able to differentially affect plant growth, to assess the genetic identity of the AMF actually colonizing the tested plants and to analyse their community composition in the different hosts. Molecular analyses revealed that plant growth and nutrition of the crop plants were differentially affected by the diverse native arbuscular mycorrhizal communities colonizing the roots of the three plants, whose performance varied depending on the identity of plant hosts and fungal symbionts, more than on a rich and diversified AMF community. Such results, improving our understanding of AMF distribution at the local scale, represent a starting point allowing the selection, isolation and characterization of the most efficient AMF assemblages to be used as inoculants in sustainable food production systems

Genetic markers associated to arbuscular mycorrhizal colonization in durum wheat

In this work we investigated the variability and the genetic basis of susceptibility to arbuscular mycorrhizal (AM) colonization of wheat roots. The mycorrhizal status of wild, domesticated and cultivated tetraploid wheat accessions, inoculated with the AM species Funneliformis mosseae, was evaluated. In addition, to detect genetic markers in linkage with chromosome regions involved in AM root colonization, a genome wide association analysis was carried out on 108 durum wheat varieties and two AM fungal species (F. mosseae and Rhizoglomus irregulare). Our findings showed that a century of breeding on durum wheat and the introgression of Reduced height (Rht) genes associated with increased grain yields did not select against AM symbiosis in durum wheat. Seven putative Quantitative Trait Loci (QTLs) linked with durum wheat mycorrhizal susceptibility in both experiments, located on chromosomes 1A, 2B, 5A, 6A, 7A and 7B, were detected. The individual QTL effects (r2) ranged from 7 to 16%, suggesting a genetic basis for this trait. Marker functional analysis identified predicted proteins with potential roles in host-parasite interactions, degradation of cellular proteins, homeostasis regulation, plant growth and disease/defence. The results of this work emphasize the potential for further enhancement of root colonization exploiting the genetic variability present in wheat