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ABSTRACT In vitro root cultivation techniques based on modified root systems are often used in studies on Arbuscular Mycorrhizal Fungi (AMF). It is a simplified but powerful tool to investigate AMF root colonization and development of the extraradical mycelium. The aim of this study was to establish and characterize the in vitro culture of a Cuban strain of Rhizophagus irregularis (INCAM 11) by using transformed chicory roots. For that, superficially disinfected propagules of R. irregularis were co-culture with the hairy transformed chicory roots on Modified Strullu and Romand (MSR) medium during five months. Spore germination was observed 3-5 days after surface disinfection. The first contact between AMF hyphae and roots occurred 1 - 3 days after germination and a significant production of extensive extraradical mycelium was observed. New spore formation started within 21 - 25 days. After 5 months, 2000 spores could be observed per plate which were able to germinate, colonize, establish and reproduce again spores when associated to young transformed roots of chicory. The most frequent associated microorganism to the in vitro culture of INCAM 11 was isolated and identified as Paenibacillus sp.

RESUMEN Las técnicas de cultivo de raíces in vitro basadas en sistemas de raíces modificadas se utilizan a menudo en los estudios sobre hongos micorrízicos arbusculares (HMA). Es una herramienta simplificada pero poderosa para investigar la colonización de las raíces de los HMA y el desarrollo del micelio extrarradical. El objetivo de este estudio fue establecer y caracterizar el cultivo in vitro de una cepa cubana de Rhizophagus irregularis (INCAM 11) utilizando raíces transformadas de achicoria. Para ello, propágulos de R. irregularis desinfectados superficialmente fueron co-cultivados con las raíces transformadas de achicoria en medio Strullu y Romand modificado (SRM) durante cinco meses. La germinación de las esporas se observó 3-5 días después de la desinfección superficial. El primer contacto entre las hifas y las raíces se produjo entre 1 y 3 días después de la germinación y se observó una producción significativa de micelio extrarradical. La formación de nuevas esporas comenzó entre 21 - 25 días. Después de 5 meses, se pudieron observar 2000 esporas por placa que fueron capaces de germinar, colonizar, establecer y reproducir nuevas esporas cuando se asociaron a raíces jóvenes transformadas de achicoria. El microorganismo asociado frecuentemente al cultivo in vitro de INCAM 11 fue aislado e identificado como Paenibacillus sp.

RESUMO

Sucrose non-fermentation-related protein kinase 1 (SnRK1) a Ser/Thr protein kinase, is known to play a crucial role in plants during biotic and abiotic stress responses by activating protein phosphorylation pathways. SnRK1 and some members of the plant-specific SnRK2 and SnRK3 sub-families have been studied in different plant species. However, a comprehensive study of the SnRK gene family in Phaseolus vulgaris is not available. Symbiotic associations of P. vulgaris with Rhizobium and/or mycorrhizae are crucial for the growth and productivity of the crop. In the present study, we identified PvSnRK genes and analysed their expression in response to the presence of the symbiont. A total of 42 PvSnRK genes were identified in P. vulgaris and annotated by comparing their sequence homology to Arabidopsis SnRK genes. Phylogenetic analysis classified the three sub-families into individual clades, and PvSnRK3 was subdivided into two groups. Chromosome localization analysis showed an uneven distribution of PvSnRK genes on 10 of the 11 chromosomes. Gene structural analysis revealed great variation in intron number in the PvSnRK3 sub-family, and motif composition is specific and highly conserved in each sub-family of PvSnRKs. Analysis of cis-acting elements suggested that PvSnRK genes respond to hormones, symbiosis and other abiotic stresses. Furthermore, expression data from databases and transcriptomic analyses revealed differential expression patterns for PvSnRK genes under symbiotic conditions. Finally, an in situ gene interaction network of the PvSnRK gene family with symbiosis-related genes showed direct and indirect interactions. Taken together, the present study contributes fundamental information for a better understanding of the role of the PvSnRK gene family not only in symbiosis but also in other biotic and abiotic interactions in P. vulgaris.

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Drought generates a complex scenario worldwide in which agriculture should urgently be reframed from an integrative point of view. It includes the search for new water resources and the use of tolerant crops and genotypes, improved irrigation systems, and other less explored alternatives that are very important, such as biotechnological tools that may increase the water use efficiency. Currently, a large body of evidence highlights the role of specific strains in the main microbial rhizosphere groups (arbuscular mycorrhizal fungi, yeasts, and bacteria) on increasing the drought tolerance of their host plants through diverse plant growth-promoting (PGP) characteristics. With this background, it is possible to suggest that the joint use of distinct PGP microbes could produce positive interactions or additive beneficial effects on their host plants if their co-inoculation does not generate antagonistic responses. To date, such effects have only been partially analyzed by using single omics tools, such as genomics, metabolomics, or proteomics. However, there is a gap of information in the use of multi-omics approaches to detect interactions between PGP and host plants. This approach must be the next scale-jump in the study of the interaction of soil-plant-microorganism. In this review, we analyzed the constraints posed by drought in the framework of an increasing global demand for plant production, integrating the important role played by the rhizosphere biota as a PGP agent. Using multi-omics approaches to understand in depth the processes that occur in plants in the presence of microorganisms can allow us to modulate their combined use and drive it to increase crop yields, improving production processes to attend the growing global demand for food.

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Arbuscular mycorrhiza (AM) and ectomycorrhiza (EcM) are the most abundant and widespread types of mycorrhizal symbiosis, but there is little and sometimes conflicting information regarding the interaction between AM fungi (AMF) and EcM fungi (EcMF) in soils. Their competition for resources can be particularly relevant in successional ecosystems, which usually present a transition from AM-forming herbaceous vegetation to EcM-forming woody species. The aims of this study were to describe the interaction between mycorrhizal fungal communities associated with AM and EcM hosts naturally coexisting during primary succession on spoil banks and to evaluate how this interaction affects growth and mycorrhizal colonization of seedlings of both species. We conducted a greenhouse microcosm experiment with Betula pendula and Hieracium caespitosum as EcM and AM hosts, respectively. They were cultivated in three-compartment rhizoboxes. Two lateral compartments contained different combinations of both host plants as sources of fungal mycelia colonizing the middle compartment, where fungal biomass, diversity, and community composition as well as the growth of each host plant species' seedlings were analyzed. The study's main finding was an asymmetric outcome of the interaction between the two plant species: while H. caespitosum and associated AMF reduced the abundance of EcMF in soil, modified the composition of EcMF communities, and also tended to decrease growth and mycorrhizal colonization of B. pendula seedlings, the EcM host did not have such effects on AM plants and associated AMF. In the context of primary succession, these findings suggest that ruderal AM hosts could hinder the development of EcM tree seedlings, thus slowing the transition from AM-dominated to EcM-dominated vegetation in early successional stages.

RESUMO

BACKGROUND: Hydric stress affects the production of wheat (Triticum aestivum L.) worldwide, making some tools necessary to cope with the decrease in rainfall. A sustainable alternative is the use of arbuscular mycorrhizal fungi (AMF) as biofertilisers. Here, we analysed the effects of AMF strains adapted or non-adapted to hyper-arid conditions on the phenolic profiles and antioxidant activities of wheat grains from two cultivars with contrasting tolerance to osmotic stress (Ilustre, moderately tolerant; and Maxi, tolerant) grown with and without hydric stress. RESULTS: Eight phenolic compounds were detected, apigenin-C-pentoside-C-hexoside I being the most abundant and showing an increase of 80.5% when inoculated with the fungus Funneliformis mosseae (FM) obtained from Atacama Desert under normal irrigation with respect to non-mycorrhizal (NM) plants. NM treatments were associated with higher grain yields. FM showed a noticeable effect on most phenolic compounds, with an increase up to 30.2% in apigenin-C-pentoside-C-hexoside III concentration under hydric stress with respect to normal irrigation, being also responsible for high antioxidant activities such as ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) and DPPH (2,2-diphenyl-1-picrylhydrazyl) activities. CONCLUSION: Inoculation with FM adapted to hydric stress produced improvements in phenolics composition and antioxidant activities in grains from wheat plants growing under hydric stress conditions, improving their food quality and supporting the development of further studies to determine whether the use of adapted AMF could be a realistic tool to improve grain quality in a scenario of increasing hydric stress conditions. © 2021 Society of Chemical Industry.

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At high soil concentrations, copper (Cu) is toxic to plant development. Symbiosis carried out between microorganisms and plant species are alternatives to minimize plant toxicity in copper contaminated soil. The present study aimed to select species of arbuscular mycorrhizal fungi for the development of wheat plants in copper contaminated soil. The experimental design was completely randomized in a 4 × 6 factorial arrangement, with four sources of inocula: three species of arbuscular mycorrhizae (Acaulospora colombiana, Gigaspora Margarita and Rhizophagus clarus) and one without inoculum (control), and six doses of copper in the soil (0, 80, 160, 240, 320, and 400 mg kg-1), with seven replications. Plant height, root length, root volume, root dry mass, specific surface area, average root diameter, root, shoot and grain Cu content and mycorrhizal colonization rate were evaluated. Root dry mass and mycorrhizal colonization were negatively affected by high Cu doses in the soil. The mycorrhizal isolates Acaulospora Colombiana and Gigaspora Margarita provide greater wheat height, root length and specific surface area and lower Cu content. Mycorrhizal colonization with the fungi Acaulospora Colombiana, Gigaspora Margarita and Rhizophagus clarus results in lower copper content in wheat shoots and grains.(AU)

Em elevadas concentrações no solo, o cobre (Cu) é tóxico para o desenvolvimento dos vegetais. Simbioses realizadas entre microrganismos e espécies vegetais são alternativas para minimizar a toxidez das plantas em solo contaminado com cobre. O trabalho teve como objetivo selecionar espécies de fungos micorrízicos arbusculares para o desenvolvimento de plantas de trigo em solo contaminado com cobre. O delineamento experimental foi inteiramente casualizado em arranjo fatorial 4 × 6, sendo quatro fontes de inóculos: três espécies de micorrizas arbusculares (Acaulospora colombiana, Gigaspora margarita e Rhizophagus clarus) e sem inóculo (testemunha) e seis doses de cobre no solo (0, 80, 160, 240, 320 e 400 mg kg-1), com sete repetições. Avaliou-se a altura de plantas, comprimento de raiz, volume de raízes, massa seca de raiz, área superficial específica, diâmetro médio de raiz, teor de cobre na raiz, na parte aérea e no grão e porcentagem de colonização micorrízica. A massa seca de raiz e a colonização micorrízica foram afetadas negativamente com altas doses de Cu no solo. Os isolados micorrízicos Acaulospora Colombiana e Gigaspora Margarita proporcionam maior altura, comprimento de raiz e área superficial específica e menor teor de Cu no grão na cultura do trigo. A colonização micorrízica com os fungos Acaulospora Colombiana, Gigaspora Margarita e Rhizophagus clarus resulta em menor teor de cobre na parte aérea e nos grãos do trigo.(AU)

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High concentrations of copper in the soil are toxic to the development of plants and microorganisms. The aim of this study was to select arbuscular mycorrhizal fungi efficient for the development and yield of soybeangrown in copper-contaminated soil. The experiment was laid out in a completely randomized design with a 7 × 4 factorial arrangement corresponding to seven rates of copper (0, 80, 160, 240, 320, 400, and 480 mg kg-1 of soil) and four inocula (uninoculated control and three mycorrhizal fungi, namely, Acaulospora colombiana, Dentiscutata heterogama and Rhizophagus clarus), in seven replicates. Shoot height; collar diameter; number of grains per plant; shoot and root-system dry mass; leaf area; specific root surface; copper content and accumulation in the shoots, roots, and grain; chlorophyll parameters; and mycorrhizal colonization percentage were evaluated. Inoculation with the arbuscular mycorrhizal fungi Acaulospora colombiana, Dentiscutata heterogama and Rhizophagus clarus increases the phenological and physiological parameters of soybean and its yield when grown in soil contaminated with up to 480 mg kg-1 of copper applied to the soil. The Rhizophagus clarus isolate provides greater development and yield in soybean grown in soil contaminated with up to 480 mg kg-1 of copper applied to the soil, as compared with the other isolates.(AU)

O cobre em elevada concentração no solo torna-se tóxico ao desenvolvimento das plantas e microrganismo. O trabalho objetivou selecionar fungos micorrízicos arbusculares eficientes para o desenvolvimento e produtividade da soja cultivada em solo contaminado com cobre. O delineamento experimental foi em Inteiramente casualizado em arranjo fatorial 7 × 4, sendo, sete doses de cobre (0, 80, 160, 240, 320, 400 e 480 mg de cobre kg-1 de solo), quatro inóculos (testemunha sem inoculação e três fungos micorrízicos: Acaulospora colombiana, Dentiscutata heterogama e Rhizophagus clarus), com 7 repetições. Avaliou-se a altura da parte aérea, diâmetro do colo, número de grão por planta, massa seca da parte aérea e sistema radicular, área foliar, área superficial específica de raízes, teores e acúmulo de cobre na parte aérea, radicular e no grão, parâmetros da clorofila, e porcentagem de colonização micorrízica. A inoculação com fungos micorrízicos arbusculares Acaulospora colombiana, Dentiscutata heterogama, Rhizophagus clarus aumenta os parâmetros fenológicos e fisiológicos da soja e a sua produtividade soja em solo contaminado com até 480 mg kg-1 de cobre aplicado no solo. O isolado Rhizophagus clarus promove maior desenvolvimento da soja e aumenta sua produtividade em solo contaminado com até 480 mg kg-1 de cobre aplicado no solo em relação aos demais isolados.(AU)

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Arbuscular mycorrhizal fungi (AMF) play an important role in plant growth. However, there are no reports of legally commercialized AMF-based inoculants for agricultural crops in Brazil. The objective of this research was to evaluate the agronomic efficiency of a Rhizophagus intraradices inoculant in combination with phosphate fertilization in grain yield of corn under different edaphoclimatic conditions in Brazil. Experiments were conducted in five Brazilian states (Goiás, Mato Grosso, Minas Gerais, Rio Grande do Sul and Santa Catarina) in a 2 x 3 factorial scheme, with two inoculation treatments (inoculated and non-inoculated seeds) and three doses of phosphate fertilization (0, 50 and 100% of the recommended P). At the end of the crop cycle (stages R4-R5), inoculation provided increases in biomass (average of 48%) regardless of the applied dose of P, higher P absorption, and 54% average increase in grain yield. In conclusion, the mycorrhizal inoculant increases biomass yield, P uptake and corn grain yield under different edaphoclimatic conditions in Brazil, especially in soils that originally had low or medium levels of available P.(AU)

Os fungos micorrízicos arbusculares (FMA) desempenham um papel importante no crescimento das plantas, no entanto, não há relatos de inoculantes legais baseados em AMF para culturas agrícolas no Brasil. O objetivo deste trabalho foi avaliar a eficiência agronômica de um inoculante a base de Rhizophagus intraradices em combinação com a adubação fosfatada no rendimento de grãos de milho sob diferentes condições edafoclimáticas no Brasil. Os experimentos foram conduzidos em cinco estados brasileiros (Goiás, Mato Grosso, Minas Gerais, Rio Grande do Sul e Santa Catarina) em esquema fatorial 2 x 3, com dois tratamentos de inoculação (sementes inoculadas e não inoculadas) e três doses de adubação fosfatada (0, 50 e 100% do P recomendado). No final do ciclo da cultura (estágios R4-R5), a inoculação proporcionou aumentos na biomassa (média de 48%), independentemente da dose aplicada de P, maior absorção de P e aumento médio de 54% no rendimento de grãos. Conclui-se que o inoculante micorrízico arbuscular aumenta a produção de biomassa, a absorção de P e a produtividade de grãos de milho em diferentes condições edafoclimáticas no Brasil, especialmente em solos que originalmente apresentavam níveis baixos ou médios de P.(AU)

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Septoglomus mexicanum is here described as a new species of arbuscular mycorrhizal fungi (AMF; Glomeromycota) based on morphological and phylogenetic analyses. It was isolated from rhizospheric soil of two endemic Mexican legumes: Prosopis laevigata and Mimosa luisana, which grow in semiarid regions of central Mexico. Septoglomus mexicanum is characterized by forming globose spores of (154.5-)202.8(-228.9) µm diam and a spore wall consisting of four layers (SWL1-SWL4): outer wall layer (SWL1) hyaline, evanescent, (1.7-)3.2(-4.3) µm thick; SWL2 laminate and smooth, orange to reddish orange, (3.1-)4.5(-6.1) µm thick; SWL3 laminate, smooth, reddish orange to reddish brown, (4.1-)5.1(-5.7) µm thick; and SWL4 hyaline, semiflexible, (0.93-)1.2(-1.4) µm thick. None of the spore wall layers stain with Melzer's reagent. The subtending hypha has a color from yellowish to golden and presents a septum on spore base. Septoglomus mexicanum can be distinguished from all other Septoglomus species by spore size and color, by spore wall structure (four layers), and by color change of the subtending hypha. Phylogenetic analysis based on the AMF extended DNA barcode covering a 1.5-kb fragment of the small subunit (SSU), internal transcribed spacer region (ITS1-5.8S-ITS2), and the large subunit (LSU) of rRNA genes places S. mexicanum in the genus Septoglomus, separated from other described Septoglomus species, especially S. turnauae, with whom it could be confused morphologically. All available sequences in public databases suggest that this new fungal species has not yet been previously detected. Thus, there are currently 149 Glomeromycota species registered in Mexico, representing 47.4% of the known species worldwide.

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ABSTRACT: Arbuscular mycorrhizal fungi (AMF) play an important role in plant growth. However, there are no reports of legally commercialized AMF-based inoculants for agricultural crops in Brazil. The objective of this research was to evaluate the agronomic efficiency of a Rhizophagus intraradices inoculant in combination with phosphate fertilization in grain yield of corn under different edaphoclimatic conditions in Brazil. Experiments were conducted in five Brazilian states (Goiás, Mato Grosso, Minas Gerais, Rio Grande do Sul and Santa Catarina) in a 2 x 3 factorial scheme, with two inoculation treatments (inoculated and non-inoculated seeds) and three doses of phosphate fertilization (0, 50 and 100% of the recommended P). At the end of the crop cycle (stages R4-R5), inoculation provided increases in biomass (average of 48%) regardless of the applied dose of P, higher P absorption, and 54% average increase in grain yield. In conclusion, the mycorrhizal inoculant increases biomass yield, P uptake and corn grain yield under different edaphoclimatic conditions in Brazil, especially in soils that originally had low or medium levels of available P.

RESUMO: Os fungos micorrízicos arbusculares (FMA) desempenham um papel importante no crescimento das plantas, no entanto, não há relatos de inoculantes legais baseados em AMF para culturas agrícolas no Brasil. O objetivo deste trabalho foi avaliar a eficiência agronômica de um inoculante a base de Rhizophagus intraradices em combinação com a adubação fosfatada no rendimento de grãos de milho sob diferentes condições edafoclimáticas no Brasil. Os experimentos foram conduzidos em cinco estados brasileiros (Goiás, Mato Grosso, Minas Gerais, Rio Grande do Sul e Santa Catarina) em esquema fatorial 2 x 3, com dois tratamentos de inoculação (sementes inoculadas e não inoculadas) e três doses de adubação fosfatada (0, 50 e 100% do P recomendado). No final do ciclo da cultura (estágios R4-R5), a inoculação proporcionou aumentos na biomassa (média de 48%), independentemente da dose aplicada de P, maior absorção de P e aumento médio de 54% no rendimento de grãos. Conclui-se que o inoculante micorrízico arbuscular aumenta a produção de biomassa, a absorção de P e a produtividade de grãos de milho em diferentes condições edafoclimáticas no Brasil, especialmente em solos que originalmente apresentavam níveis baixos ou médios de P.