ABSTRACT
Background Isothiocyanates (ITCs) are natural products obtained from plants of the Brassicas family. They represent an environmentally friendly alternative for the control of phytopathogenic fungi. However, as it has been observed with synthetic fungicides, the possibility of inducing ITC-resistant strains is a major concern. It is, therefore, essential to understanding the molecular mechanisms of fungal resistance to ITCs. We analyzed a subtractive library containing 180 clones of an Alternaria alternata strain resistant to 2-propenyl ITC (2-pITC). After their sequencing, 141 expressed sequence tags (ESTs) were identified using the BlastX algorithm. The sequence assembly was carried out using CAP3 software; the functional annotation and metabolic pathways identification were performed using the Blast2GO program. Results The bioinformatics analysis revealed 124 reads with similarities to proteins involved in transcriptional control, defense and stress pathways, cell wall integrity maintenance, detoxification, organization and cytoskeleton destabilization; exocytosis, transport, DNA damage control, ribosome maintenance, and RNA processing. In addition, transcripts corresponding to enzymes as oxidoreductases, transferases, hydrolases, lyases, and ligases, were detected. Degradation pathways for styrene, aminobenzoate, and toluene were induced, as well as the biosynthesis of phenylpropanoid and several types of N-glycan. Conclusions The fungal response showed that natural compounds could induce tolerance/resistance mechanisms in organisms in the same manner as synthetic chemical products. The response of A. alternata to the toxicity of 2-pITC is a sophisticated phenomenon including the induction of signaling cascades targeting a broad set of cellular processes. Whole-transcriptome approaches are needed to elucidate completely the fungal response to 2-pITC.
Subject(s)
Isothiocyanates , Drug Resistance, Fungal , Alternaria/genetics , Alternaria/metabolism , Fungicides, Industrial , Computational Biology , Subtractive Hybridization Techniques , Hybridization, GeneticABSTRACT
En el presente estudio se evaluó el efecto de extractos fenólicos y de carotenoides procedentes de frutos de chiltepín sobre el crecimiento micelial y la germinación de conidios de Alternaria alternata y Fusarium oxysporum, 2 importantes hongos causantes de podredumbres en frutas y hortalizas. Los extractos fenólicos presentaron una inhibición en el crecimiento micelial de A. alternata del 38,46 %, y redujeron significativamente la germinación de conidios al quinto día después del tratamiento al 92 % en relación al control. No se observaron cambios significativos en el crecimiento micelial de F. oxysporum, pero sí se redujo significativamente al 85 % en relación al control, el número de conidios germinados a los 5 días de tratamiento. Los extractos de carotenoides mostraron una inhibición del 38,5 % en el crecimiento micelial y del 85,3 % en la germinación de conidios de A. alternata, 5 días después del tratamiento. Frente a F. oxysporum, dichos extractos presentaron menor inhibición del crecimiento micelial (20,3 %), mientras que hubo una mayor inhibición en la germinación de conidios (96 %). Los extractos fenólicos y de carotenoides de chiltepín pueden ser una alternativa promisoria de importancia agrícola como fungicidas naturales.
The effect of phenolic and carotenoid extracts from chiltepin fruits on mycelial growth and the inhibition of conidial germination of Alternaria alternata and Fusarium oxysporum were investigated in the present work. Phenolic extracts inhibited mycelial growth of A. alternata by 38.46%, and significantly reduced conidial germination on the fifth day after treatment to 92% in relation to control. No significant changes were observed in the inhibition of mycelial growth in Fusarium oxysporum; however, the number of germinated conidia was reduced, showing 85% inhibition five days after treatment in relation to control. Moreover, carotenoid extracts showed 38.5% inhibition of mycelial growth and 85.3% inhibition of conidial germination of A. alternata, five days after treatment. Carotenoid extracts showed less inhibition of mycelial growth (20.3%) in F. oxysporum, with respect to A. alternata; while there was greater inhibition of conidial germination (96%) on the fifth day after treatment. Phenolic and carotenoid extracts from chiltepin may be a promising alternative as a natural fungicide against fungi of agricultural importance.