RESUMEN
The increasing tolerance to currently used fungicides and the need for environmentally friendly antimicrobial approaches have stimulated the development of novel strategies to control plant-pathogenic fungi such as antimicrobial phototreatment (APT). We investigated the in vitro APT of the plant-pathogenic fungus Colletotrichum acutatum with furocoumarins and coumarins and solar radiation. The compounds used were: furocoumarins 8-methoxypsoralen (8-MOP) and 5,8-dimethoxypsoralen (isopimpinellin), coumarins 2H-chromen-2-one (coumarin), 7-hydroxycoumarin, 5,7-dimethoxycoumarin (citropten) and a mixture (3:1) of 7-methoxycoumarin and 5,7-dimethoxycoumarin. APT of conidia with crude extracts from 'Tahiti' acid lime, red and white grapefruit were also performed. Pure compounds were tested at 50µM concentration and mixtures and extracts at 12.5mgL(-1). The C. acutatum conidia suspension with or without the compounds was exposed to solar radiation for 1h. In addition, the effects of APT on the leaves of the plant host Citrus sinensis were determined. APT with 8-MOP was the most effective treatment, killing 100% of the conidia followed by the mixture of two coumarins and isopimpinellin that killed 99% and 64% of the conidia, respectively. APT with the extracts killed from 20% to 70% of the conidia, and the extract from 'Tahiti' lime was the most effective. No damage to sweet orange leaves was observed after APT with any of the compounds or extracts.
Asunto(s)
Citrus sinensis/microbiología , Colletotrichum/efectos de los fármacos , Colletotrichum/efectos de la radiación , Furocumarinas/farmacología , Viabilidad Microbiana/efectos de los fármacos , Viabilidad Microbiana/efectos de la radiación , Fármacos Fotosensibilizantes/farmacología , Citrus sinensis/química , Colletotrichum/fisiología , Estabilidad de Medicamentos , Furocumarinas/química , Furocumarinas/aislamiento & purificación , Fármacos Fotosensibilizantes/química , Fármacos Fotosensibilizantes/aislamiento & purificaciónRESUMEN
Photodynamic inactivation (PDI) is an efficient approach for the elimination of a series of microorganisms; however, PDI involving phytopathogenic filamentous fungi is scarce in the literature. In the present study, we have demonstrated the photoinactivating properties of five cationic meso-(1-methyl-4-pyridinio)porphyrins on conidia of the phytopathogen Colletotrichum graminicola. For this purpose, photophysical properties (photostability and (1)O2 singlet production) of the porphyrins under study were first evaluated. PDI assays were then performed with a fluence of 30, 60, 90 and 120 J cm(-2) and varying the porphyrin concentration from 1 to 25 µmol L(-1). Considering the lowest concentration that enabled the best photoinactivation, with the respective lowest effective irradiation time, the meso-(1-methyl-4-pyridinio)porphyrins herein studied could be ranked as follows: triple-charged 4 (1 µmol L(-1) with a fluence of 30 J cm(-2)) > double-charged-trans2 (1 µmol L(-1) with 60 J cm(-2)) > tetra-charged 5 (15 µmol L(-1) with 90 J cm(-2)) > mono-charged 1 (25 µmol L(-1) with 120 J cm(-2)). Double-charged-cis-porphyrin 3 inactivated C. graminicola conidia in the absence of light. Evaluation of the porphyrin binding to the conidia and fluorescence microscopic analysis were also performed, which were in agreement with the PDI results. In conclusion, the cationic porphyrins herein studied were considered efficient photosensitizers to inactivate C. graminicola conidia. The amount and position of positive charges are related to the compounds' amphiphilicity and therefore to their photodynamic activity.
Asunto(s)
Colletotrichum/efectos de los fármacos , Colletotrichum/efectos de la radiación , Fármacos Fotosensibilizantes/farmacología , Porfirinas/farmacología , Cationes/química , Cationes/farmacología , Colletotrichum/metabolismo , Luz , Fármacos Fotosensibilizantes/química , Plantas/microbiología , Porfirinas/química , Oxígeno Singlete/metabolismoRESUMEN
Light conditions can influence fungal development. Some spectral wavebands can induce conidial production, whereas others can kill the conidia, reducing the population size and limiting dispersal. The plant pathogenic fungus Colletotrichum acutatum causes anthracnose in several crops. During the asexual stage on the host plant, Colletototrichum produces acervuli with abundant mucilage-embedded conidia. These conidia are responsible for fungal dispersal and host infection. This study examined the effect of visible light during C. acutatum growth on the production of conidia and mucilage and also on the UV tolerance of these conidia. Conidial tolerance to an environmentally realistic UV irradiance was determined both in conidia surrounded by mucilage on sporulating colonies and in conidial suspension. Exposures to visible light during fungal growth increased production of conidia and mucilage as well as conidial tolerance to UV. Colonies exposed to light produced 1.7 times more conidia than colonies grown in continuous darkness. The UV tolerances of conidia produced under light were at least two times higher than conidia produced in the dark. Conidia embedded in the mucilage on sporulating colonies were more tolerant of UV than conidia in suspension that were washed free of mucilage. Conidial tolerance to UV radiation varied among five selected isolates.
Asunto(s)
Colletotrichum/efectos de la radiación , Polisacáridos Fúngicos/agonistas , Tolerancia a Radiación , Esporas Fúngicas/efectos de la radiación , Adaptación Fisiológica , Citrus/microbiología , Colletotrichum/fisiología , Polisacáridos Fúngicos/biosíntesis , Fotoperiodo , Esporas Fúngicas/fisiología , Rayos UltravioletaRESUMEN
To study microwave heating for potential postharvest treatments against anthracnose disease, Colletotrichum gloeosporioides growth-no-growth response after selected microwave treatments (2,450 MHz) was fitted by using a logistic regression model. Evaluated variables were power level, exposure time, presence or absence of water in the medium during treatment, and incubation-observation time. Depending on the setting, the applied power ranged from 77.2 to 435.6 W. For the experiments on dry medium (mold spores over filter paper), exposure times were 1, 2, 3, or 4 min, whereas spores dispersed in potato dextrose agar, a wet medium, had exposure times of 3, 6, or 9 s. Growth (response = 1) or no growth (response = 0) was observed after two different incubation-observation times (4 or 10 days). As expected, high power levels and long exposure times resulted in complete inhibition of C. gloeosporioides spore germination. In a number of cases (such as low power levels and short treatment times), only a delay in mold growth was observed. Scanning electron micrographs showed signs of mycelia dehydration and structural collapse in the spores of the studied mold. Cell damage was attributed to heating during microwave exposure. Reduced logistic models included variables and interactions that significantly (P < 0.05) affected mold growth, and were able to predict the growth-no-growth response in at least 83% of the experimental conditions. Microwave treatments (4 min at any of the studied power levels in dry medium, and 9 s at power levels of 30% or more for wet medium) proved effective in the inhibition of C. gloeosporioides in model systems. These no-growth conditions will be tested further on fresh fruits in order to develop feasible postharvest microwave treatments.