Fungal diversity and Fusarium oxysporum pathogenicity associated with coffee corky-root disease in Mexico

The disease known as coffee corky-roots associated to the infection by the root-knot nematode Meloidogyne paranaensis is an important issue for coffee crop in several countries. In Mexico, particularly in the Veracruz state, considerable loses are recorded annually in Coffea arabica plantations by corky-root disease. Previous studies have revealed the presence of fungi in coffee corky-root tissues. However, these fungi have not been yet identified. This work aimed to identify at species level the fungi associated to the coffee corky-root symptoms and determine their pathogenicity on coffee plants. Coffee roots with corky-root symptoms were collected in eight sites distributed through the major coffee growing region of Veracruz. Observations of inside cortical root tissues under scanning electron microscope revealed abundant mycelium and conidia incorky-root samples in contrast with absence of any fungi development in healthy roots. Forty-nine fungi strains from internal corky-root tissue were isolated and identified at species level by ITS sequences. Fusarium oxysporum was the most frequent species and the only present in all of the corky-root samples. These strains were selected for the pathogenicity test. All F. oxysporum strains colonized the vascular system of coffee plants although none caused wilting symptoms. Highlights Fusarium oxysporum is a major fungus associated with coffee corky-root disease. None of the oxysporum strains inoculated into the roots of coffee plants caused vascular withering symptoms In the tissues damaged by coffee corky-root disease interact nematodes, fungi and bacteria.The disease known as coffee corky-roots associated to the infection by the root-knot nematode Meloidogyne paranaensis is an important issue for coffee crop in several countries. In Mexico, particularly in the Veracruz state, considerable loses are recorded annually in Coffea arabica plantations by corky-root disease. Previous studies have revealed the presence of fungi in coffee corky-root tissues. However, these fungi have not been yet identified. This work aimed to identify at species level the fungi associated to the coffee corky-root symptoms and determine their pathogenicity on coffee plants. Coffee roots with corky-root symptoms were collected in eight sites distributed through the major coffee growing region of Veracruz. Observations of inside cortical root tissues under scanning electron microscope revealed abundant mycelium and conidia incorky-root samples in contrast with absence of any fungi development in healthy roots. Forty-nine fungi strains from internal corky-root tissue were isolated and identified at species level by ITS sequences. Fusarium oxysporum was the most frequent species and the only present in all of the corky-root samples. These strains were selected for the pathogenicity test. All F. oxysporum strains colonized the vascular system of coffee plants although none caused wilting symptoms. Highlights Fusarium oxysporum is a major fungus associated with coffee corky-root disease. None of the oxysporum strains inoculated into the roots of coffee plants caused vascular withering symptoms In the tissues damaged by coffee corky-root disease interact nematodes, fungi and bacteria

Functional Signatures of the Epiphytic Prokaryotic Microbiome of Agaves and Cacti.

Microbial symbionts account for survival, development, fitness and evolution of eukaryotic hosts. These microorganisms together with their host form a biological unit known as holobiont. Recent studies have revealed that the holobiont of agaves and cacti comprises a diverse and structured microbiome, which might be important for its adaptation to drylands. Here, we investigated the functional signatures of the prokaryotic communities of the soil and the episphere, that includes the rhizosphere and phyllosphere, associated with the cultivated Agave tequilana and the native and sympatric Agave salmiana, Opuntia robusta and Myrtillocactus geometrizans by mining shotgun metagenomic data. Consistent with previous phylogenetic profiling, we found that Proteobacteria, Actinobacteria and Firmicutes were the main represented phyla in the episphere of agaves and cacti, and that clustering of metagenomes correlated with the plant compartment. In native plants, genes related to aerobic anoxygenic phototrophy and photosynthesis were enriched in the phyllosphere and soil, while genes coding for biofilm formation and quorum sensing were enriched in both epiphytic communities. In the episphere of cultivated A. tequilana fewer genes were identified, but they belonged to similar pathways than those found in native plants. A. tequilana showed a depletion in several genes belonging to carbon metabolism, secondary metabolite biosynthesis and xenobiotic degradation suggesting that its lower microbial diversity might be linked to functional losses. However, this species also showed an enrichment in biofilm and quorum sensing in the epiphytic compartments, and evidence for nitrogen fixation in the rhizosphere. Aerobic anoxygenic phototrophic markers were represented by Rhizobiales (Methylobacterium) and Rhodospirillales (Belnapia) in the phyllosphere, while photosystem genes were widespread in Bacillales and Cyanobacteria. Nitrogen fixation and biofilm formation genes were mostly related to Proteobacteria. These analyses support the idea of niche differentiation in the rhizosphere and phyllosphere of agaves and cacti and shed light on the potential mechanisms by which epiphytic microbial communities survive and colonize plants of arid and semiarid ecosystems. This study establishes a guideline for testing the relevance of the identified functional traits on the microbial community and the plant fitness

Influence of phylogenetic, environmental, and behavioral factors on the gut bacterial community structure of dung beetles (Scarabaeidae: Scarabaeinae) in a Neotropical Biosphere Reserve

Gut bacteria help dung beetles metabolize nutrients contained and synthesize those unavailable in their food, depending on the ecological scenario in which they develop. However, less is known about the influence of environmental and behavioral factors on the taxonomic composition of bacterial gut communities in Scarabaeinae beetles. To address this research topic, we analyzed 13 tropical dung beetle species in the Los Tuxtlas Biosphere Reserve, Mexico, to understand how the beetle tribe, habitat, food preference, food relocation, and parental care influence the composition of gut bacterial communities. We found that the beetle tribe is the primary factor impacting the taxonomic composition of gut bacterial communities. Among them, Deltochilini displayed the highest variability in diversity due to the different combinations of habitat and food preferences among its species. On the other hand, the other tribes studied did not exhibit such variable combinations. Habitat emerged as the second most influential factor, with forest-dwelling beetles displaying higher diversity. This can be attributed to the heterogeneous environments within tropical forests, which offer a greater diversity of food resources. In contrast, grassland beetles, living in more homogeneous environments and relying on cow feces as their main food source, exhibited lower diversity. Our findings suggest a correlation between bacterial diversity and food resource availability in complex habitats, such as tropical forests, which offer a wider array of food sources compared to simpler environments like grasslands

Evidence for succession and putative metabolic roles of fungi and bacteria in the farming mutualism of the ambrosia beetle Xyleborus affinis

The bacterial and fungal community involved in ambrosia beetle fungiculture remains poorly studied compared to the famous fungus-farming ants and termites. Here we studied microbial community dynamics of laboratory nests, adults, and brood during the life cycle of the sugarcane shot hole borer, Xyleborus affinis. We identified a total of 40 fungal and 428 bacterial operational taxonomic units (OTUs), from which only five fungi (a Raffaelea fungus and four ascomycete yeasts) and four bacterial genera (Stenotrophomonas, Enterobacter, Burkholderia, and Ochrobactrum) can be considered the core community playing the most relevant symbiotic role. Both the fungal and bacterial populations varied significantly during the beetle’s life cycle. While the ascomycete yeasts were the main colonizers of the gallery early on, the Raffaelea and other filamentous fungi appeared after day 10, at the time when larval hatching happened. Regarding bacteria, Stenotrophomonas and Enterobacter dominated overall but decreased in foundresses and brood with age. Finally, inferred analyses of the putative metabolic capabilities of the bacterial microbiome revealed that they are involved in (i) degradation of fungal and plant polymers, (ii) fixation of atmospheric nitrogen, and (iii) essential amino acid, cofactor, and vitamin provisioning. Overall, our results suggest that yeasts and bacteria are more strongly involved in supporting the beetle-fungus farming symbiosis than previously thought.Research reported in this publication was supported by CONACyT-FORDECYT number 292399, Spanish Ministry of Economy and Competitiveness under award numbers SAF2015-65878-R and PGC2018-099344-B-I00, cofinanced by the European Regional Development Fund (ERDF), and from Generalitat Valenciana (project Prometeo/2018/A133). P.H.W.B. was supported by the German Research Foundation (DFG Emmy Noether grant BI 1956/1-1).Peer reviewe

Nematofauna asociada a la rizosfera de papas (Solanum tuberosum) cultivadas en la zona productora del Cofre de Perote, Veracruz, México

Se determinaron y clasificaron en grupos tróficos las especies de nematodos asociados a la rizosfera de Solanum tuberosum en suelos altamente infectados por el nematodo dorado de la papa (Globodera rostochiensis) en un ciclo de cultivo, en la zona productora del Cofre de Perote, Veracruz, México. Se identificaron 7 géneros (Aphelenchoides, Aphelenchus, Crassolabium, Mesodorylaimus, Plectus y Steinernema) y 8 especies (Aporcelaimellus obtusicaudatus, Ecumenicus monohystera, Acrobeles mariannae, Acrobeles singulus, Acrobeloides nanus, Cruznema tripartitum, Eucephalobus oxyuroides, y Globodera rostochiensis) en asociación con la rizosfera de papas cultivadas. Crassolabium sp. y E. monohystera se registran por primera vez para México

Nematofauna asociada a la rizosfera de papas (Solanum tuberosum) cultivadas en la zona productora del Cofre de Perote, Veracruz, México Nematode fauna associated with the rhizosphere of potato crop (Solanum tuberosum) grown in the region of Cofre de Perote, Veracruz, Mexico

Se determinaron y clasificaron en grupos tróficos las especies de nematodos asociados a la rizosfera de Solanum tuberosum en suelos altamente infectados por el nematodo dorado de la papa (Globodera rostochiensis) en un ciclo de cultivo, en la zona productora del Cofre de Perote, Veracruz, México. Se identificaron 7 géneros (Aphelenchoides, Aphelenchus, Crassolabium, Mesodorylaimus, Plectus y Steinernema) y 8 especies (Aporcelaimellus obtusicaudatus, Ecumenicus monohystera, Acrobeles mariannae, Acrobeles singulus, Acrobeloides nanus, Cruznema tripartitum, Eucephalobus oxyuroides, y Globodera rostochiensis) en asociación con la rizosfera de papas cultivadas. Crassolabium sp. y E. monohystera se registran por primera vez para México.Nematode species associated with the rhizosphere of Solanum tuberosum were identified and classified into trophic groups from soils highly infected by the golden potato cyst nematode (Globodera rostochiensis) in a single crop cycle in the producing zone on the Cofre de Perote, Veracruz, Mexico. Seven genera (Aphelenchoides, Aphelenchus, Crassolabium, Mesodorylaimus, Plectus and Steinernema) and 8 species (Aporcelaimellus obtusicaudatus, Ecumenicus monohystera, Acrobeles mariannae, Acrobeles singulus, Acrobeloides nanus, Cruznema tripartitum, Eucephalobus oxyuroides, and Globodera rostochiensis) were found in association with the rhizosphere of cultivated potatoes. Crassolabium sp. and E. monohystera are recorded for the first time in México

Diversidad de hongos y patogenicidad de Fusarium oxysporum asociados a la corchosis de la raíz del cafeto en México

International audienceThe disease known as coffee corky-roots associated to the infection by the root-knot nematode Meloidogyne paranaensis is an important issue for coffee crop in several countries. In Mexico, particularly in the Veracruz state, considerable loses are recorded annually in Coffea arabica plantations by corky-root disease. Previous studies have revealed the presence of fungi in coffee corky-root tissues. However, these fungi have not been yet identified. This work aimed to identify at species level the fungi associated to the coffee corky-root symptoms and determine their pathogenicity on coffee plants. Coffee roots with corky-root symptoms were collected in eight sites distributed through the major coffee growing region of Veracruz. Observations of inside cortical root tissues under scanning electron microscope revealed abundant mycelium and conidia incorky-root samples in contrast with absence of any fungi development in healthy roots. Forty-nine fungi strains from internal corky-root tissue were isolated and identified at species level by ITS sequences. Fusarium oxysporum was the most frequent species and the only present in all of the corky-root samples. These strains were selected for the pathogenicity test. All F. oxysporum strains colonized the vascular system of coffee plants although none caused wilting symptoms.La corchosis de la raíz del café asociada a la infección del nematodo agallador de la raíz Meloidogyne paranaensis es un importante problema para el cultivo de café en varios países. En México, particularmente en el estado de Veracruz, se registran considerables pérdidas anuales en las plantaciones de Coffea arabica por esta enfermedad. Estudios anteriores han revelado la presencia de hongos en los tejidos afectados con corchosis de la raíz del café. Sin embargo, estos hongos aún no han sido identificados. El objetivo de este trabajo fue identificar a nivel de especie los hongos asociados a la corchosis de la raíz y determinar su patogenicidad en plantas de café. Se recolectaron raíces de cafetos con síntomas de corchosis en ocho sitios distribuidos a través de la principal región cafetalera de Veracruz. Las observaciones de los tejidos internos de las raíces bajo el microscopio electrónico de barrido revelaron abundante micelio y conidios en muestras de raíz con corchosis, en contraste con su ausencia en raíces sanas. Se aislaron 49 hongos de los tejidos internos afectados con corchosis y se identificaron a nivel de especie mediante secuencias de ITS. Fusarium oxysporum fue la especie más frecuente y la única presente en todos los sitios de muestreo, por lo que estas cepas fueron seleccionadas para la prueba de patogenicidad. Todas las cepas de F. oxysporum fueron capaces de colonizar el sistema vascular de las plantas de café, aunque ninguna causó síntomas de marchitez

Molecular and Environmental Triggering Factors of Pathogenicity of Fusarium oxysporum and F. solani Isolates Involved in the Coffee Corky-Root Disease

Coffee corky-root disease causes serious damages to coffee crop and is linked to combined infection of Fusarium spp. and root-knot nematodes Meloidogyne spp. In this study, 70 Fusarium isolates were collected from both roots of healthy coffee plants and with corky-root disease symptoms. A phylogenetic analysis, and the detection of pathogenicity SIX genes and toxigenicity Fum genes was performed for 59 F. oxysporum and 11 F. solani isolates. Based on the molecular characterization, seven F. oxysporum and three F. solani isolates were assessed for their pathogenicity on coffee seedlings under optimal watering and water stress miming root-knot nematode effect on plants. Our results revealed that a drastic increment of plant colonization capacity and pathogenicity on coffee plants of some Fusarium isolates was caused by water stress. The pathogenicity on coffee of F. solani linked to coffee corky-root disease and the presence of SIX genes in this species were demonstrated for the first time. Our study provides evidence for understanding the pathogenic basis of F. oxysporum and F. solani isolates on coffee and revealed the presence of SIX and Fum genes as one of their pathogenicity-related mechanisms. We also highlight the relevance of chlorophyll, a fluorescence as an early and high-throughput phenotyping tool in Fusarium pathogenicity studies on coffee

Functional Signatures of the Epiphytic Prokaryotic Microbiome of Agaves and Cacti.

Microbial symbionts account for survival, development, fitness and evolution of eukaryotic hosts. These microorganisms together with their host form a biological unit known as holobiont. Recent studies have revealed that the holobiont of agaves and cacti comprises a diverse and structured microbiome, which might be important for its adaptation to drylands. Here, we investigated the functional signatures of the prokaryotic communities of the soil and the episphere, that includes the rhizosphere and phyllosphere, associated with the cultivated Agave tequilana and the native and sympatric Agave salmiana, Opuntia robusta and Myrtillocactus geometrizans by mining shotgun metagenomic data. Consistent with previous phylogenetic profiling, we found that Proteobacteria, Actinobacteria and Firmicutes were the main represented phyla in the episphere of agaves and cacti, and that clustering of metagenomes correlated with the plant compartment. In native plants, genes related to aerobic anoxygenic phototrophy and photosynthesis were enriched in the phyllosphere and soil, while genes coding for biofilm formation and quorum sensing were enriched in both epiphytic communities. In the episphere of cultivated A. tequilana fewer genes were identified, but they belonged to similar pathways than those found in native plants. A. tequilana showed a depletion in several genes belonging to carbon metabolism, secondary metabolite biosynthesis and xenobiotic degradation suggesting that its lower microbial diversity might be linked to functional losses. However, this species also showed an enrichment in biofilm and quorum sensing in the epiphytic compartments, and evidence for nitrogen fixation in the rhizosphere. Aerobic anoxygenic phototrophic markers were represented by Rhizobiales (Methylobacterium) and Rhodospirillales (Belnapia) in the phyllosphere, while photosystem genes were widespread in Bacillales and Cyanobacteria. Nitrogen fixation and biofilm formation genes were mostly related to Proteobacteria. These analyses support the idea of niche differentiation in the rhizosphere and phyllosphere of agaves and cacti and shed light on the potential mechanisms by which epiphytic microbial communities survive and colonize plants of arid and semiarid ecosystems. This study establishes a guideline for testing the relevance of the identified functional traits on the microbial community and the plant fitness