In Colombia, tomato production under protected conditions represents an important economic contribution to the agricultural sector. Fusarium wilt diseases, caused by pathogenic formae speciales of the soil-borne fungus Fusarium oxysporum Schltdl., cause significant yield losses in tomatoes throughout the world. Investigation of the F. oxysporum–tomato pathosystem in Colombia is required to develop appropriate alternative disease management. In this study, 120 fungal isolates were obtained from four different departments in the Central Andean Region in Colombia from tomato crops with symptoms of wilt disease. A molecular characterization of the fungal isolates was performed using the SIX1, SIX3, and SIX4 effector genes of Fusarium oxysporum f. sp. lycopersici W.C. Snyder & H.N. Hansen (Fol). Additionally, we developed a new specific marker to distinguish between Fusarium oxysporum f. sp. radicis-lycopersici Jarvis & Shoemaker (Forl) and Fol isolates. Furthermore, a phylogenetic analysis using the Translation Elongation Factor 1-alpha (EF1a) gene was performed with the collected isolates. Two isolates (named Fol59 and Fol-UDC10) were identified as Fol race 2, four isolates were identified as Forl, six isolates were identified as F. solani, and most of the isolates were grouped within the F. oxysporum species complex. The phylogenetic tree of EF1a showed that most of the isolates could potentially correspond to nonpathogenic strains of F. oxysporum. Additional pathogenicity assays carried out with Fol59 and Fol-UDC10 confirmed that both isolates were highly virulent strains. This study represents a contribution to the understanding of the local interaction between tomatoes and F. oxysporum in Colombia.
The vascular wilt disease caused by the fungus Fusarium oxysporum f. sp. physali (Foph) is one of the most limiting factors for the production and export of cape gooseberry (Physalis peruviana) in Colombia. A transcriptomic analysis of a highly virulent strain of F. oxysporum in cape gooseberry plants, revealed the presence of secreted in the xylem (SIX) effector genes, known to be involved in the pathogenicity of other formae speciales (ff. spp.) of F. oxysporum. This pathogenic strain was classified as a new f. sp. named Foph, due to its specificity for cape gooseberry hosts. Here, we sequenced and assembled the genome of five strains of F. oxysporum from a fungal collection associated to the cape gooseberry crop (including Foph), focusing on the validation of the presence of SIX homologous and on the identification of putative effectors unique to Foph. By comparative and phylogenomic analyses based on single-copy orthologous, we found that Foph is closely related to F. oxysporum ff. spp., associated with solanaceous hosts. We confirmed the presence of highly identical homologous genomic regions between Foph and Fol that contain effector genes and identified six new putative effector genes, specific to Foph pathogenic strains. We also conducted a molecular characterization using this set of putative novel effectors in a panel of 36 additional stains of F. oxysporum including two of the four sequenced strains, from the fungal collection mentioned above. These results suggest the polyphyletic origin of Foph and the putative independent acquisition of new candidate effectors in different clades of related strains. The novel effector candidates identified in this genomic analysis, represent new sources involved in the interaction between Foph and cape gooseberry, that could be implemented to develop appropriate management strategies of the wilt disease caused by Foph in the cape gooseberry crop.
The global banana industry is threatened by one of the most devastating diseases: Fusarium wilt (FWB). FWB is caused by the soil-borne fungus Fusarium oxysporum f. sp. cubense (Foc), which almost annihilated the banana production in the late 1950s. A new strain of Foc, known as tropical race 4 (TR4), attacks a wide range of banana varieties including Cavendish clones which are the source of 99% of banana exports. In 2019, Foc TR4 was reported in Colombia, and more recently (2021) in Peru. In this study, we sequenced three fungal isolates identified as Foc TR4 from La Guajira (Colombia) and compared them against 19 whole-genome sequences of Foc TR4 publicly available, including four genome sequences recently released from Peru. To understand the genetic relatedness of the Colombian Foc TR4 isolates and those from Peru, we conducted a phylogenetic analysis based on a genome-wide set of single nucleotide polymorphisms (SNPs). Additionally, we compared the genomes of the 22 available Foc TR4 isolates looking for the presence-absence of gene polymorphisms and genomic regions. Our results reveal that (i) the Colombian and Peruvian isolates are genetically distant, which could be better explained by independent incursions of the pathogen to the continent, and (ii) there is a high correspondence between the genetic relatedness and geographic origin of Foc TR4. The profile of present/absent genes and the distribution of missing genomic regions showed a high correspondence to the clades recovered in the phylogenetic analysis, supporting the results obtained by SNP-based phylogeny.
Introducción. Phytophthora infestans (Mont.) de Bary, es considerado el agente causal de una de las enfermedades más devastadoras y limitantes de cultivos de papa (Solanum tuberosum L.) en el departamento de Nariño (Colombia). Esta enfermedad es controlada con fungicidas sistémicos, los cuales tienen un impacto negativo sobre el ambiente e incrementan los costos de producción, sin embargo, existen alternativas como los denominados fungicidas orgánicos, los cuales se basan en el uso de extractos de plantas con propiedades antifúngicas para el control del patógeno. Objetivo. Evaluar extractos acuosos de O. vulgare y L. origanoides para el control de P. infestans bajo condiciones in vitro. Materiales y métodos. La presente investigación se desarrolló en el Laboratorio de microbiología del grupo de investigación en Genética y Evolución de Organismos Tropicales (GENPAT) de la Universidad de Nariño, durante el año 2017. Se evaluó la sensibilidad in vitro de P. infestans frente a extractos de las especies botánicas Lippia origanoides (orégano silvestre) y Origanum vulgare (orégano común). Resultados. Los datos obtenidos permitieron el establecimiento de la línea base de comportamiento de las poblaciones del patógeno frente a los compuestos evaluados, hubo una reducción gradual tanto en el crecimiento como en la esporulación cuando se empleó el extracto de Lippia origanoides y que este mismo tiene la capacidad de controlar el patógeno en concentraciones de 100 mg ml-1. Con el empleo de Origanum vulgare los aislamientos de P. infestans presentaron diferentes niveles de sensibilidad al compuesto con bajos niveles de inhibición y altos valores de EC-50. Conclusión. Los aislamientos de P. infestans presentaron diferentes niveles de sensibilidad frente a los dos extractos acuosos evaluados, con una reducción del 100 % en la inhibición del crecimiento radial y la esporulación a partir de la concentración de 100 mg ml-1, cuando se empleó el extracto de L. origanoides.
Physiological processes of plants infected by vascular pathogens are mainly affected by vascular bundle obstruction, decreasing the absorption of water and nutrients and gas exchange by stomatal closure, and inducing oxidative cascades and PSII alterations. Chitosan, a derivative of chitin present in the cell wall of some organisms including fungi, induces plant defense responses, activating systemic resistance. In this study, the effect of chitosan on the physiological and molecular responses of tomato plants infected with Fusarium oxysporum f. sp. lycopersici (Fol) was studied, evaluating the maximum potential quantum efficiency of PSII photochemistry (Fv/Fm), photochemical efficiency of PSII (Y(II)), photochemical quenching (qP), stomatal conductance (gs), relative water content (RWC), proline content, photosynthetic pigments, dry mass, and differential gene expression (PAL, LOXA, ERF1, and PR1) of defense markers. A reduction of 70% in the incidence and 91% in the severity of the disease was achieved in plants treated with chitosan, mitigating the damage caused by Fol on Fv/Fm, Y(II), and chlorophyll contents by 23%, 36%, and 47%, respectively. Less impact was observed on qP, gs, RWC, and dry mass (16%, 11%, and 26%, respectively). Chitosan-treated and Fol-infected plants over-expressed PR1a gene suggesting a priming-associated response. These results demonstrate the high potential of chitosan to protect tomato plants against Fol by regulating physiological and molecular responses in tomato plants.
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