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.

ResumenLa antracnosis es una de las enfermedades que más afecta la producción de frutales tropicales en el mundo. El objetivo del presente trabajo fue identificar a nivel de especie aislamientos de Colletotrichum spp. causantes de la antracnosis en frutos de aguacate, banano, mango y tomate de árbol mediante el análisis filogenético multilocus empleando datos de las secuencias de la región espaciadora interna transcrita (ITS), y de los genes β-tubulina-2 y gliceraldehído-3-fosfato deshidrogenasa. Para cada especie se evaluaron características morfológicas de la colonia, los conidios y los apresorios. Se obtuvieron 16 aislamientos a partir de frutos con síntomas de antracnosis, los cuales se confirmaron como causantes de la enfermedad en frutos de sus respectivos hospedantes con el método de inoculación por herida y goteo. Se identificaron los aislamientos como Colletotrichum asianum, Colletotrichum fructicola, Colletotrichum musae dentro del complejo de especies de Colletotrichum gloeosporioides, o como Colletotrichum tamarilloi dentro del complejo de especies de Colletotrichum acutatum. El presente estudio aporta al conocimiento de las especies causantes de la antracnosis en frutas tropicales en Colombia, lo cual facilitará el diseño de técnicas de diagnóstico y la selección de estrategias óptimas de manejo de la enfermedad.Palabras clave: análisis filogenético multilocus, complejo de especies, diversidad genética, diagnóstico. Genetic identification of Colletotrichum isolates causing anthracnose in fruits of avocado, banana, mango and tamarillo AbstractAnthracnose is a major disease affecting the production of tropical fruit trees on a global level. Our objective was to identify at the species level Colletotrichum isolates causing anthracnose in fruits of avocado, banana, mango, and tamarillo by multilocus phylogenetic analysis based on sequencing data from the ITS region and from the βtubulin-2 and glyceraldehyde-3-phosphate dehydrogenase genes. We evaluated the morphology of colonies, conidia, and appressoria for each species. We obtained 16 isolates from fruits exhibiting symptoms of anthracnose, which were confirmed using the wound and drip inoculation method as causing anthracnose in fruits of their respective hosts. The isolates were identified as Colletotrichum asianum, Colletotrichum fructicola, or Colletotrichum musae within the Colletotrichum gloeosporioides species complex, or as Colletotrichum tamarilloi within the Colletotrichum acutatum species complex. This study contributes to the knowledge of species causing anthracnose in tropical fruits in Colombia, which will facilitate the design of diagnosis techniques and the selection of better disease management strategies.

Environmental scanning electron microscopy (ESEM) was performed in seedlings of Passiflora edulis f. flavicarpa inoculated with Fusarium solani f. sp. passiflorae (teleomorph: Haematonectria haematococca) causal agent of passionfruit collar rot. Inoculations were carried out every 24 h until the seventh day and from this day until the fifteenth the interval of inoculation was 72 h. The pathogen isolated in PDA media was placed on the surface of the collar through the modified test-tube screening methodology. Epidermis of collar, stem, and leaves and longitudinal sections of collar and stem were observed. After 24 h of inoculation, conidia and dense septate mycelium were observed over the epidermis of the stem and the collar, respectively. Hypertrophy and cell wall degradation of the vascular tissues were also found during this period. Five days after the inoculation macroconidia were formed from monophialides in the aerial mycelium on the stem. Ten days after inoculation, xylem and pith cells of the collar were colonized by hyphae, inclusions, and mature sporodoquia over the stem surface. Hyphae colonization started six days after inoculation on stomata and thirteen days after inoculation monophialides with in situ microconidia were observed over the surface of the leaves. Based on the evidence and previous studies, internal hyphae colonization of F. solani f. sp. passiflorae concentrates on the collar area and the damages of the cells indicate an extracellular enzymatic activity of the fungus. The incubation and latent periods of F. solani f. sp. passiflorae were 1.4 and 4 days, respectively. © 2017. Acad. Colomb. Cienc. Ex. Fis. Nat.Key Words: Plant diseases; fungal diseases; Passifloraceae; collar rot; scanning microscopy. Microscopía electrónica de barrido ambiental del proceso de infección de ResumenSe realizó microscopía electrónica de barrido ambiental (ESEM) en plántulas de maracuyá (Passiflora edulis f. flavicarpa) inoculadas con Fusarium solani f. sp. passiflorae (teleomorfo: Haematonectria haematococca) agente causal de la secadera. Las inoculaciones se realizaron cada 24 h hasta el séptimo día y hasta el día quince el intervalo fue de 72 h. El aislamiento en medio de papa (PDA) se colocó sobre el cuello de la raíz mediante la metodología de cribado en tubos de ensayo modificada. La superficie del cuello, tallo, hojas y secciones longitudinales del cuello y tallo fueron observadas. Después de 24 h de la inoculación, se observaron conidios y micelio denso septado sobre la epidermis del tallo y el cuello e hipertrofia y degradación de la pared celular de los tejidos vasculares. Al quinto día se formaron macroconidios a partir de monofiálides en el micelio aéreo en el tallo. Diez días después, las células del xilema y médula del cuello fueron colonizadas por hifas, esporodoquios maduros e inclusiones. La colonización de las estomas por las hifas comenzó seis días después de la inoculación y trece días después de la inoculación se observaron monofiálides con microconidios in situ sobre la superfi...

En años recientes una enfermedad caracterizada por la necrosis y el hundimiento de la epidermis de las bayas de la vid (Vitis viinifera L.) se ha venido presentando en la variedad Red Globe, cultivada en predios vitícolas del municipio de La Unión, Valle del Cauca, lo que ha resultado en pérdidas de rendimiento y calidad. Para determinar la etiología de la enfermedad, se recolectaron y procesaron frutos que mostraban los signos típicos de decoloración y posterior necrosis. Con mayor prevalencia se encontró un hongo, que fue sometido a pruebas de patogenicidad y caracterización morfológica mediante microscopía de luz y electrónica (Environmental scanning electron microscopy, ESEM), complementadas con pruebas moleculares. Los postulados de Koch se cumplieron mediante la inoculación de una suspensión conidial de 1x106 conidios por mL-1 de agua en bayas sanas de la misma variedad. Al cabo una semana se empezaron a observar signos similares a los observados en campo. Las búsquedas de similitud con la herramienta BLAST mostraron una identidad del 100 % entre las secuencias del espaciador transcribible interno (internal transcribed spacer, ITS) y Colletotrichum aenigma y C. siamense, pertenecientes al complejo de especies Colletotrichum gloeosporioides, lo que proporciona información útil para entender la enfermedad de las bayas de la vid y poder diseñar estrategias de manejo. © 2019. Acad. Colomb. Cienc. Ex. Fis. Nat.

Anthracnose is considered one of the most important diseases of grape crops. In this study, we evaluated the in vitro effect on potato dextrose agar (PDA) of four fungicides: benomyl (Benlate® 50 WP), carbendazim (Belico ® 500 SC), chlorothalonil (Odeon® 720 SC), and dodine (Syllit ® 400 SC) using three concentrations on a Colletotrichum gloeosporioides isolate. This species complex causes anthracnose on the Red Globe grape variety. The concentrations of each fungicide were: benomyl at 0.5, 0.25, and 1 g.L-1; carbendazim at  0.6, 0.3, and 1.2 mL.L-1; chlorothalonil at 2.4, 1.2, and 4.8 mL.L-1, and dodine at 1.6, 0.8, and 3.2 mL.L-1. Evaluated the inhibition of mycelial growth and the sporulation of the fungus. The fungus mycelium exposed to the commercial dose of the fungicides was observed using environmental scanning electron microscopy (ESEM). We used a completely randomized design with 13 treatments and five repetitions. Our results showed a degree of differential efficacy for the four fungicides and their effects were directly proportional to the dose used, benomyl and chlorothalonil being the most efficient in controlling the mycelial growth of the fungus. At the commercial dose, sporulation was inhibited at a higher rate by benomyl showing no statistically significant differences with the other products except chlorothalonil and dodine when the recommended dose was reduced by half. The damage observed by ESEM on the fungus mycelium ratified the effect obtained in vitro, i.e., deformed hyphae and irregular growth. The results of this study, complemented by electron microscopy, are promising for the appropriate selection of the best fungicides for controlling anthracnose on grape fruits, subject to the timely and correct diagnosis of the disease.