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Antifungal and anti-aflatoxigenic activity of Heliopsis longipes roots and affinin/spilanthol against Aspergillus parasiticus by downregulating the expression of alfD and aflR genes of the aflatoxins biosynthetic pathway
Authors:Génesis V Buitimea-Cantúa  Nydia E Buitimea-Cantúa  María del Refugio Rocha-Pizaña  Ema Carina Rosas-Burgos  Alejandro Hernández-Morales  Jorge Molina-Torres
Institution:1. Departamento de Biotecnología y Bioquímica, CINVESTAV, Irapuato, Guanajuato, México;2. Tecnologico de Monterrey, Centro de Biotecnología-FEMSA, Escuela de Ingeniería y Ciencias, Monterrey, México;3. genesis.buitimea@cinvestav.mx;5. Tecnologico de Monterrey, Centro de Biotecnología-FEMSA, Escuela de Ingeniería y Ciencias, Monterrey, México;6. Tecnologico de Monterrey, Escuela de Ingeniería y Ciencias, Puebla, México;7. Unidad Académica Multidisciplinaria Zona Huasteca, Universidad Autónoma de San Luis Potosí, Ciudad Valles S.L.P, Ciudad Valles San Luis Potosí, México;8. Departamento de Investigación y Posgrado en Alimentos, Universidad de Sonora, Hermosillo, Sonora, México
Abstract:Abstract

In the present study, ethanolic extract from Heliopsis longipes roots and affinin/spilanthol against Aspergillus parasiticus growth and aflatoxins production were studied in relation to the expression of aflD and aflR, two key genes of aflatoxins biosynthetic pathway. Phytochemical analysis of the ethanolic extract by GC-EIMS identified affinin/spilanthol (7.84?±?0.27?mg g?1) as the most abundant compounds in H. longipes roots. The antifungal and anti-aflatoxigenic assays showed that affinin/spilanthol at 300?µg mL?1 produced the higher inhibition of radial growth (95%), as well as, the higher aflatoxins production inhibition (61%) in comparison to H. longipes roots (87% and 48%, respectively). qRT-PCR revealed that the expression of aflD and aflR genes showed a higher downregulation in affinin/spilanthol at 300?µg mL?1. The expression ratio of alfD was suppressed by affinin/spilanthol in 79% and aflR in 84%, while, a lower expression ratio suppressed by H. longipes was obtained, alfD (55%) and aflR (59%). Affinin/spilanthol possesses higher antifungal and anti-aflatoxigenic activity against A. parasiticus rather than H. longipes roots, and this anti-aflaxotigenic activity occurring via downregulation of the aflD and aflR genes. Thus, H. longipes roots and affinin/spilanthol can be considered potent antifungal agents against aflatoxigenic fungus, especially, affinin/spilanthol.
Keywords:Alkamides  aflatoxins biosynthetic pathway  natural antifungal compounds  qRT-PCR
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