Genome-wide analysis of copper, iron and zinc transporters in the arbuscular mycorrhizal fungus Rhizophagus irregularis

Arbuscular mycorrhizal fungi (AMF), belonging to the Glomeromycota, are soil microorganisms that establish mutualistic symbioses with the majority of higher plants. The efficient uptake of low mobility mineral nutrients by the fungal symbiont and their further transfer to the plant is a major feature of this symbiosis. Besides improving plant mineral nutrition, AMF can alleviate heavy metal toxicity to their host plants and are able to tolerate high metal concentrations in the soil. Nevertheless, we are far from understanding the key molecular determinants of metal homeostasis in these organisms. To get some insights into these mechanisms, a genome-wide analysis of Cu, Fe and Zn transporters was undertaken, making use of the recently published whole genome of the AMF Rhizophagus irregularis. This in silico analysis allowed identification of 30 open reading frames in the R. irregularis genome, which potentially encode metal transporters. Phylogenetic comparisons with the genomes of a set of reference fungi showed an expansion of some metal transporter families. Analysis of the published transcriptomic profiles of R. irregularis revealed that a set of genes were up-regulated in mycorrhizal roots compared to germinated spores and extraradical mycelium, which suggests that metals are important for plant colonization.This research was supported by the Spanish Ministry of Economy and Competitivity (Project AGL2012-35611). Elisabeth Tamayo was supported by a Ph.D. contract (I3P) from the Spanish National Research Council (CSIC) and Tamara Gómez-Gallego by a Ph.D. contract from the Spanish Ministry of Economy and Competitivity.Peer reviewedPeer Reviewe

The potential of arbuscular mycorrhizal fungi to enhance metallic micronutrient uptake and mitigate food contamination in agriculture: prospects and challenges

Optimizing agroecosystems and crops for micronutrient uptake while reducing issues with inorganic contaminants (metal(loid)s) is a challenging task. One promising approach is to use arbuscular mycorrhizal fungi (AMF) and investigate the physiological, molecular and epigenetic changes that occur in their presence and that lead to changes in plant metal(loid) concentration (biofortification of micronutrients or mitigation of contaminants). Moreover, it is important to understand these mechanisms in the context of the soil microbiome, particularly those interactions of AMF with other soil microbes that can further shape crop nutrition. To address these challenges, a two-pronged approach is recommended: exploring molecular mechanisms and investigating microbiome management and engineering. Combining both approaches can lead to benefits in human health by balancing nutrition and contamination caused by metal(loid)s in the agro-ecosystemThis work was supported by grant PID2021-1255210B-I00 funded by MCIN/AEI/10.13039/ 501100011033 and by ‘ERDF A way of making Europe’, by the ‘European Union’. NC is a University of Ottawa Research Chair in Microbial Genomics, and his research on AMF genetics and genomics is supported by the Discovery Program of the Natural Sciences and Engineering Research Council (RGPIN2020-05643) and a Discovery Accelerator Supplements Program (RGPAS2020-00033

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.Fil: Molina, Alicia Susana. Universidad Nacional de San Luis; ArgentinaFil: Lugo, Mónica Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis. Universidad Nacional de San Luis. Facultad de Ciencias Físico Matemáticas y Naturales. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis; ArgentinaFil: Pérez Chaca, María V.. Universidad Nacional de San Luis; ArgentinaFil: Vargas Gil, Silvina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigaciones Agropecuarias. Instituto de Patología Vegetal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Zirulnik, Fanny. Universidad Nacional de San Luis; ArgentinaFil: Leporati, Jorge Leandro. Universidad Nacional de San Luis; ArgentinaFil: Ferrol, Nuria. Consejo Superior de Investigaciones Científicas. Estación Experimental del Zaidín; EspañaFil: Azcón González de Aguilar, Concepción. Consejo Superior de Investigaciones Científicas. Estación Experimental del Zaidín; Españ

The Rhizophagus irregularis Genome Encodes Two CTR Copper Transporters That Mediate Cu Import Into the Cytosol and a CTR-Like Protein Likely Involved in Copper Tolerance

Arbuscular mycorrhizal fungi increase fitness of their host plants under Cu deficient and toxic conditions. In this study, we have characterized two Cu transporters of the CTR family (RiCTR1 and RiCTR2) and a CTR-like protein (RiCTR3A) of Rhizophagus irregularis. Functional analyses in yeast revealed that RiCTR1 encodes a plasma membrane Cu transporter, RiCTR2 a vacuolar Cu transporter and RiCTR3A a plasma membrane protein involved in Cu tolerance. RiCTR1 was more highly expressed in the extraradical mycelia (ERM) and RiCTR2 in the intraradical mycelia (IRM). In the ERM, RiCTR1 expression was up-regulated by Cu deficiency and down-regulated by Cu toxicity. RiCTR2 expression increased only in the ERM grown under severe Cu-deficient conditions. These data suggest that RiCTR1 is involved in Cu uptake by the ERM and RiCTR2 in mobilization of vacuolar Cu stores. Cu deficiency decreased mycorrhizal colonization and arbuscule frequency, but increased RiCTR1 and RiCTR2 expression in the IRM, which suggest that the IRM has a high Cu demand. The two alternatively spliced products of RiCTR3, RiCTR3A and RiCTR3B, were more highly expressed in the ERM. Up-regulation of RiCTR3A by Cu toxicity and the yeast complementation assays suggest that RiCTR3A might function as a Cu receptor involved in Cu tolerance

Arbuscular Mycorrhizal Fungi. Methods and Protocols.

Ferrol, N.; Lanfranco, L. (Eds.) 2020 Arbuscular Mycorrhizal Fungi. Methods and Protocols. Vol 2146. pp. 257. ISBN 978 1 0716-0602-

Regulación por el boro del transporte de H+ en suspensiones celulares y en membranas de girasol (Helianthus annuus, L.)

Universidad de Granada, Facultad de Ciencias. Leída en Julio de 199

GintAMT3 – a Low-Affinity Ammonium Transporter of the Arbuscular Mycorrhizal Rhizophagus irregularis

Nutrient acquisition and transfer are essential steps in the arbuscular mycorrhizal (AM) symbiosis, which is formed by the majority of land plants. Mineral nutrients are taken up by AM fungi from the soil and transferred to the plant partner. Within the cortical plant root cells the fungal hyphae form tree-like structures (arbuscules) where the nutrients are released to the plant-fungal interface, i.e., to the periarbuscular space, before being taken up by the plant. In exchange, the AM fungi receive carbohydrates from the plant host. Besides the well-studied uptake of phosphorus (P), the uptake and transfer of nitrogen (N) plays a crucial role in this mutualistic interaction. In the AM fungus Rhizophagus irregularis (formerly called Glomus intraradices), two ammonium transporters (AMT) were previously described, namely GintAMT1 and GintAMT2. Here, we report the identification and characterization of a newly identified R. irregularis AMT, GintAMT3. Phylogenetic analyses revealed high sequence similarity to previously identified AM fungal AMTs and a clear separation from other fungal AMTs. Topological analysis indicated GintAMT3 to be a membrane bound pore forming protein, and GFP tagging showed it to be highly expressed in the intraradical mycelium of a fully established AM symbiosis. Expression of GintAMT3 in yeast successfully complemented the yeast AMT triple deletion mutant (MATa ura3 mep1Δ mep2Δ::LEU2 mep3Δ::KanMX2). GintAMT3 is characterized as a low affinity transport system with an apparent Km of 1.8 mM and a Vmax of 240 nmol-1 min-1 108 cells-1, which is regulated by substrate concentration and carbon supply.This project was supported by the Swiss National Science Foundation (grant no. PZ00P3_136651 to P-EC., grant No. 127563 to TB) the Conseil Régional de Bourgogne PARI AGREE grant to DW and the Spanish Ministry of Economy and Competitivity (Projects AGL2012-35611 and AGL2015-67098-R).Peer reviewedPeer Reviewe

Characterization of Three New Glutaredoxin Genes in the Arbuscular Mycorrhizal Fungus Rhizophagus irregularis: Putative Role of RiGRX4 and RiGRX5 in Iron Homeostasis.

Glutaredoxins (GRXs) are small ubiquitous oxidoreductases involved in the regulation of the redox state in living cells. In an attempt to identify the full complement of GRXs in the arbuscular mycorrhizal (AM) fungus Rhizophagus irregularis, three additional GRX homologs, besides the formerly characterized GintGRX1 (renamed here as RiGRX1), were identified. The three new GRXs (RiGRX4, RiGRX5 and RiGRX6) contain the CXXS domain of monothiol GRXs, but whereas RiGRX4 and RiGRX5 belong to class II GRXs, RiGRX6 belongs to class I together with RiGRX1. By using a yeast expression system, we observed that the newly identified homologs partially reverted sensitivity of the GRX deletion yeast strains to external oxidants. Furthermore, our results indicated that RiGRX4 and RiGRX5 play a role in iron homeostasis in yeast. Gene expression analyses revealed that RiGRX1 and RiGRX6 were more highly expressed in the intraradical (IRM) than in the extraradical mycelium (ERM). Exposure of the ERM to hydrogen peroxide induced up-regulation of RiGRX1, RiGRX4 and RiGRX5 gene expression. RiGRX4 expression was also up-regulated in the ERM when the fungus was grown in media supplemented with a high iron concentration. These data indicate the two monothiol class II GRXs, RiGRX4 and RiGRX5, might be involved in oxidative stress protection and in the regulation of fungal iron homeostasis. Increased expression of RiGRX1 and RiGRX6 in the IRM suggests that these GRXs should play a key role in oxidative stress protection of R. irregularis during its in planta phase

Testing the trade‐balance model: resource stoichiometry does not sufficiently explain AM effects

Variations in arbuscular mycorrhizae (AM) effects on plant growth (MGR) are commonly assumed to result from cost : benefit balances, with C as the cost and, most frequently, P as the benefit. The trade-balance model (TBM) adopts these assumptions and hypothesizes that mycorrhizal benefit depends on C : N : P stoichiometry. Although widely accepted, the TBM has not been experimentally tested. We isolated the parameters included in the TBM and tested these assumptions using it as framework. Oryza sativa plants were supplied with different N : P ratios at low light level, establishing different C : P and C : N exchange rates, and C, N or P limitation. MGR and effects on nutrient uptake, %M, ERM, photosynthesis and shoot starch were measured. C distribution to AM fungi played no role in MGR, and N was essential for all AM effects, including on P nutrition. C distribution to AM and MGR varied with the limiting nutrient (N or P), and evidence of extensive interplay between N and P was observed. The TBM was not confirmed. The results agreed with the exchange of surplus resources and source–sink regulation of resource distribution among plants and AMF. Rather than depending on exchange rates, resource exchange may simply obey both symbiont needs, not requiring further regulation.info:eu-repo/semantics/acceptedVersio