Application of laser microdissection to identify the mycorrhizal fungi that establish arbuscules inside root cells.

Obligate symbiotic fungi that form arbuscular mycorrhizae (AMF; belonging to the Glomeromycota phylum) are some of the most important soil microorganisms. AMFs facilitate mineral nutrient uptake from the soil, in exchange for plant-assimilated carbon, and promote water-stress tolerance and resistance to certain diseases. AMFs colonize the root by producing inter- and intra-cellular hyphae. When the fungus penetrates the inner cortical cells, it produces a complex ramified structure called arbuscule, which is considered the preferential site for nutrient exchange. Direct DNA extraction from the whole root and sequencing of ribosomal gene regions are commonly carried out to investigate intraradical AMF communities. Nevertheless, this protocol cannot discriminate between the AMFs that actively produce arbuscules and those that do not. To solve this issue, the authors have characterized the AMF community of arbusculated cells (AC) through a laser microdissection (LMD) approach, combined with sequencing-based taxa identification. The results were then compared with the AMF community that was found from whole root DNA extraction. The AMF communities originating from the LMD samples and the whole root samples differed remarkably. Five taxa were involved in the production of arbuscules, while two taxa were retrieved inside the root but not in the AC. Unexpectedly, one taxon was found in the AC, but its detection was not possible when extracting from the whole root. Thus, the LMD technique can be considered a powerful tool to obtain more precise knowledge on the symbiotically active intraradical AMF community

Arbuscular Mycorrhizal Fungi and their Value for Ecosystem Management

Arbuscular Mycorrhizal Fungi (AMF) are root obligate symbionts of nearly all the plants living on Earth. They are considered as living fossils: there are evidences that date back to 460 million years ago their presence on our planet. Taxonomically, they belong to the Glomeromycota phylum. They are found in roots of 80% of plant species and give and get back benefits to their partners, as happens in all mutual symbiotic relationships. They build up a bridge between plant and soil, growing their mycelia both inside and outside 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. Besides physiological benefits to the host-plant, AMF play an important ecological role. They are important in soil structuring thanks to their thick extraradical hyphal network able to aggregate soil particles. They help plants establish in degraded ecosystems (e.g. desert areas and mine spoils) and positively affect phytoremediation. They can influence processes (i.e. soil carbon sequestration) related to climate change. Last but not least, their belowground presence and diversity can positively affect the aboveground plant biodiversity and productivity. These roles played by AMF for the ecosystem functioning lead to consider them as key soil organisms. Therefore, every AMF aspect is extensively studied: from biological features, through biogeography and biodiversity, to phylogeny. [...]JRC.H.5-Land Resources Managemen

Unique arbuscular mycorrhizal fungal communities uncovered in date palm plantations and surrounding desert habitats of Southern Arabia

The main objective of this study was to shed light on the previously unknown arbuscular mycorrhizal fungal (AMF) communities in Southern Arabia. We explored AMF communities in two date palm (Phoenix dactylifera) plantations and the natural vegetation of their surrounding arid habitats. The plantations were managed traditionally in an oasis and according to conventional guidelines at an experimental station. Based on spore morphotyping, the AMF communities under the date palms appeared to be quite diverse at both plantations and more similar to each other than to the communities under the ruderal plant, Polygala erioptera, growing at the experimental station on the dry strip between the palm trees, and to the communities uncovered under the native vegetation (Zygophyllum hamiense, Salvadora persica, Prosopis cineraria, inter-plant area) of adjacent undisturbed arid habitat. AMF spore abundance and species richness were higher under date palms than under the ruderal and native plants. Sampling in a remote sand dune area under Heliotropium kotschyi yielded only two AMF morphospecies and only after trap culturing. Overall, 25 AMF morphospecies were detected encompassing all study habitats. Eighteen belonged to the genus Glomus including four undescribed species. Glomus sinuosum, a species typically found in undisturbed habitats, was the most frequently occurring morphospecies under the date palms. Using molecular tools, it was also found as a phylogenetic taxon associated with date palm roots. These roots were associated with nine phylogenetic taxa, among them eight from Glomus group A, but the majority could not be assigned to known morphospecies or to environmental sequences in public databases. Some phylogenetic taxa seemed to be site specific. Despite the use of group-specific primers and efficient trapping systems with a bait plant consortium, surprisingly, two of the globally most frequently found species, Glomus intraradices and Glomus mosseae, were not detected neither as phylogenetic taxa in the date palm roots nor as spores under the date palms, the intermediate ruderal plant, or the surrounding natural vegetation. The results highlight the uniqueness of AMF communities inhabiting these diverse habitats exposed to the harsh climatic conditions of Southern Arabi

Arbuscular mycorrhizal fungi from argentinean highland puna soils unveiled by propagule multiplication

Low arbuscular-mycorrhizal (AM) sporulation in arid field soils limits our knowledge of indigenous species when diversity studies are based only on spore morphology. Our aim was to use different approaches (i.e., spore morphological approach and PCR–SSCP (single-strand-conformation-polymorphism) analysis after trap plant multiplication strategies to improve the knowledge of the current richness of glomalean AM fungi (Glomerales; Glomeromycota) from the Argentine Puna. Indigenous propagules from two pristine sites at 3870 and 3370 m of elevation were multiplied using different host plants; propagation periods (2–6 months), and subculture cycles (1; 2; or 3) from 5 to 13 months. The propagule multiplication experiment allowed the detection of different glomoid taxa of Funneliformis spp. and Rhizoglomus spp., which were considered cryptic species since they had never been found in Puna soils before. On the other hand; almost all the generalist species previously described were recovered from cultures; except for Glomus ambisporum. Both plant host selection and culture times are critical for Glomerales multiplication. The SSCP analysis complemented the morphological approach and showed a high variability of Glomus at each site; revealing the presence of Funneliformis mosseae. This study demonstrates that AMF trap culture (TC) is a useful strategy for improving the analysis of AM fungal diversity/richness in the Argentinean highlands.EEA BalcarceFil: Covacevich, Fernanada. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Fundación para la Investigación Biológicas Aplicadas. Instituto de Investigaciones en Biodiversidad y Biotecnología; Argentina.Fil: Hernández Guijarro, Keren. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina.Fil: Crespo, Esteban. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia; Argentina.Fil: Lumini, Erica. Institute for Sustainable Plant Protection of National Research Council; Italia.Fil: Rivero Mega, María Soledad. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia; Argentina.Fil: Lugo, Mónica. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia; Argentina.Fil: Lugo, Mónica. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de San Luis; Argentina. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis; Argentina