V. Bianciotto

The impact of tillage practices on arbuscular mycorrhizal fungal diversity in subtropical crops

Arbuscular mycorrhizal fungi (AMF) are a main component of soil microbiota in most agrosystems. As obligately mutualistic symbionts, they colonize the roots of the majority of plants, including crop plants. We used molecular techniques to investigate how different tillage systems (moldboard, shred-bedding, subsoil-bedding, and no tillage) can influence the AM fungal community colonizing maize, bean, and sorghum roots in an experimental site located in northern Tamaulipas, Mexico. Roots from 36 plants were analyzed using AM fungal-specific primers to partially amplify the small subunit (SSU) of the ribosomal DNA genes. More than 880 clones were screened for restriction fragment length polymorphism (RFLP) variation, and 173 of these were sequenced. Ten AM fungal types were identified and clustered into three AM fungal families: Gigasporaceae, Glomaceae, and Paraglomaceae. Glomus was the dominating taxon in all the samples. Four of the 10 identified types were distinct from any previously published sequences and could correspond to either known unsequenced species or unknown species. The fungal diversity was low in the four agriculture management systems, but the multidimensional scaling (MDS) analysis and log-linear-saturated model indicated that the composition of the AMF community was significantly affected by the tillage system. In conclusion, since some fungal types were treatment specific, agricultural practices could directly or indirectly influence AM biodiversity.

Vertical Transmission of Endobacteria in the Arbuscular Mycorrhizal Fungus Gigaspora margarita through Generation of Vegetative Spores

ABSTRACT Arbuscular mycorrhizal (AM) fungi living in symbiotic association with the roots of vascular plants have also been shown to host endocellular rod-shaped bacteria. Based on their ribosomal sequences, these endobacteria have recently been identified as a new taxon, Candidatus Glomeribacter gigasporarum. In order to investigate the cytoplasmic stability of the endobacteria in their fungal host and their transmission during AM fungal reproduction (asexual), a system based on transformed carrot roots and single-spore inocula of Gigaspora margarita was used. Under these in vitro sterile conditions, with no risk of horizontal contamination, the propagation of endobacteria could be monitored, and it was shown, by using primers designed for both 16S and 23S ribosomal DNAs, to occur through several vegetative spore generations (SG0 to SG4). A method of confocal microscopy for quantifying the density of endobacteria in spore cytoplasm was designed and applied; endobacteria were consistently found in all of the spore generations, although their number rapidly decreased from SG0 to SG4. The study demonstrates that a vertical transmission of endobacteria takes place through the fungal vegetative generations (sporulation) of an AM fungus, indicating that active bacterial proliferation occurs in the coenocytic mycelium of the fungus, and suggests that these bacteria are obligate endocellular components of their AM fungal host.

Arbuscular Mycorrhizal Fungi as Natural Biofertilizers: Let's Benefit from Past Successes

Arbuscular Mycorrhizal Fungi (AMF) constitute a group of root obligate biotrophs that exchange mutual benefits with about 80% of plants. They are considered natural biofertilizers, since they provide the host with water, nutrients, and pathogen protection, in exchange for photosynthetic products. Thus, AMF are primary biotic soil components which, when missing or impoverished, can lead to a less efficient ecosystem functioning. The process of re-establishing the natural level of AMF richness can represent a valid alternative to conventional fertilization practices, with a view to sustainable agriculture. The main strategy that can be adopted to achieve this goal is the direct re-introduction of AMF propagules (inoculum) into a target soil. Originally, AMF were described to generally lack host- and niche-specificity, and therefore suggested as agriculturally suitable for a wide range of plants and environmental conditions. Unfortunately, the assumptions that have been made and the results that have been obtained so far are often worlds apart. The problem is that success is unpredictable since different plant species vary their response to the same AMF species mix. Many factors can affect the success of inoculation and AMF persistence in soil, including species compatibility with the target environment, the degree of spatial competition with other soil organisms in the target niche and the timing of inoculation. Thus, it is preferable to take these factors into account when “tuning” an inoculum to a target environment in order to avoid failure of the inoculation process. Genomics and transcriptomics have led to a giant step forward in the research field of AMF, with consequent major advances in the current knowledge on the processes involved in their interaction with the host-plant and other soil organisms. The history of AMF applications in controlled and open-field conditions is now long. A review of biofertilization experiments, based on the use of AMF, has here been proposed, focusing on a few important factors that could increase the odds or jeopardize the success of the inoculation process.

Unravelling Soil Fungal Communities from Different Mediterranean Land-Use Backgrounds

Background Fungi strongly influence ecosystem structure and functioning, playing a key role in many ecological services as decomposers, plant mutualists and pathogens. The Mediterranean area is a biodiversity hotspot that is increasingly threatened by intense land use. Therefore, to achieve a balance between conservation and human development, a better understanding of the impact of land use on the underlying fungal communities is needed. Methodology/Principal Findings We used parallel pyrosequencing of the nuclear ribosomal ITS regions to characterize the fungal communities in five soils subjected to different anthropogenic impact in a typical Mediterranean landscape: a natural cork-oak forest, a pasture, a managed meadow, and two vineyards. Marked differences in the distribution of taxon assemblages among the different sites and communities were found. Data analyses consistently indicated a sharp distinction of the fungal community of the cork oak forest soil from those described in the other soils. Each soil showed features of the fungal assemblages retrieved which can be easily related to the above-ground settings: ectomycorrhizal phylotypes were numerous in natural sites covered by trees, but were nearly completely missing from the anthropogenic and grass-covered sites; similarly, coprophilous fungi were common in grazed sites. Conclusions/Significance Data suggest that investigation on the below-ground fungal community may provide useful elements on the above-ground features such as vegetation coverage and agronomic procedures, allowing to assess the cost of anthropogenic land use to hidden diversity in soil. Datasets provided in this study may contribute to future searches for fungal bio-indicators as biodiversity markers of a specific site or a land-use degree.

Interactions of fungi with other organisms

Living organisms establish complex networks of mutualistic and antagonistic interactions in nature, which impact strongly on their own survival and on the stability of the whole population. Fungi, in particular, can shape natural as well as man-managed ecosystems due to their ubiquitous occurrence and the range of interactions they establish with plants, animals and other microbes. This review describes some examples of mutualistic and antagonistic fungal interactions that are of particular interest for their ecological role, or because they can be exploited by man to improve plant health and/or productivity in sustainable agriculture and forestry.

Arbuscular Mycorrhizal Fungi and their Value for Ecosystem Management

Arbuscular Mycorrhizal Fungi (AMF) are a group of obligate biotrophs, to the extent that they must develop a close symbiotic association with the roots of a living host plant in order to grow and complete their life cycle [1]. The term “mycorrhiza” literally derives from the Greek mykes and rhiza, meaning fungus and root, respectively. AMF can symbiotically interact with almost all the plants that live on the Earth. They are found in the roots of about 80-90% of plant species (mainly grasses, agricultural crops and herbs) and exchange benefits with their partners, as is typical of all mutual symbiotic relationships [2]. They represent an interface between plants and soil, growing their mycelia both inside and outside the plant roots. AMF provide the plant with water, soil mineral nutrients (mainly phosphorus and nitrogen) and pathogen protection. In exchange, photosynthetic compounds are transferred to the fungus [3].

Sequencing and comparison of the mitochondrial COI gene from isolates of Arbuscular Mycorrhizal Fungi belonging to Gigasporaceae and Glomeraceae families.

Arbuscular Mycorrhizal Fungi (AMF) are well known for their ecological importance and their positive influence on plants. The genetics and phylogeny of this group of fungi have long been debated. Nuclear markers are the main tools used for phylogenetic analyses, but they have sometimes proved difficult to use because of their extreme variability. Therefore, the attention of researchers has been moving towards other genomic markers, in particular those from the mitochondrial DNA. In this study, 46 sequences of different AMF isolates belonging to two main clades Gigasporaceae and Glomeraceae have been obtained from the mitochondrial gene coding for the Cytochrome c Oxidase I (COI), representing the largest dataset to date of AMF COI sequences. A very low level of divergence was recorded in the COI sequences from the Gigasporaceae, which could reflect either a slow rate of evolution or a more recent evolutionary divergence of this group. On the other hand, the COI sequence divergence between Gigasporaceae and Glomeraceae was high, with synonymous divergence reaching saturated levels. This work also showed the difficulty in developing valuable mitochondrial markers able to effectively distinguish all Glomeromycota species, especially those belonging to Gigasporaceae, yet it represents a first step towards the development of a full mtDNA-based dataset which can be used for further phylogenetic investigations of this fungal phylum.

AMF components from a microbial inoculum fail to colonize roots and lack soil persistence in an arable maize field

Arbuscular mycorrhizal fungi (AMF) are root obligate biotrophs that provide the host with nutrients and pathogen protection, in exchange of photosynthetic products. A decline in AMF diversity can reduce the overall benefit for host plants. A sustainable strategy to re-establish AMF diversity is to supply the target soil with AMF inoculants. After inoculation, it is essential to verify whether the inoculants successfully colonize the host plant and persist, and if the resident AMF community is affected. The AMF components of a microbial inoculum (including other saprotrophs) that was applied to maize were identified and traced in field by 454-pyrosequencing of the partial rRNA 18S gene. In addition, mycorrhizal colonization and plant biomass were monitored in inoculated and non-inoculated maize. The inoculated AMF taxa failed to colonize roots and lacked soil persistence. Nevertheless, the inoculation process reduced species dominance and increased diversity in the pre-existing AMF community. No differences were seen between mycorrhizal colonization in treated and control maize. We suggest that the slightly significant increase in treated plant biomass was potentially due to (i) marginally colonizing inoculated AMF that remained unseen and other saprotroph inoculants applied and/or (ii) the effect of inoculation on the pre-existing AMF community in treated maize roots.