Abstract
During the infection of grapevine (Vitis vinifera) by the fungus Botrytis cinerea, the concentration of polyamines, which are toxic substances for the phytopathogen, increases in the grape. Nine NRPS genes have been identified in the genome of B. cinerea, yet the function of five of them remains unknown. For this reason, we have studied the expression of the 9 NRPS genes by RT-qPCR in a medium supplemented with sublethal concentrations of three polyamines (1,3-diaminopropane (1,3-DAP), spermidine (SPD), and spermine (SPM)). Our results show that the presence of polyamines in the culture medium triggered the overexpression of the Bcnrps1 gene in the pathogen. Deleting Bcnrps1 did not affect mycelial growth or adaptation to osmotic stress, and we show that its expression is not essential for the cycle of infection of the B. cinerea. However, mutating the Bcnrps1 gene resulted in overexpression of the Bcnrps6 gene, which encodes for the excretion of siderophores of the coprogen family. Moreover, gene deletion has reduced the tolerance of B. cinerea B05.10 to toxic substances such as the polyamine SPD and the fungicide pyrimethanil, and its virulence has increased. Our findings provide new insights into the function of the Bcnrps1 gene and its involvement in the tolerance of B. cinerea against exogenous toxic compounds.
Highlights
Botrytis cinerea is a phytopathogenic fungus belonging to the phylum Ascomycota and is the causal agent of gray mold disease
We studied the expression of non-ribosomal peptide synthetase (NRPS) genes in B. cinerea by RT-qPCR in the presence and absence of three polyamines
Error bars represent standard deviations of means (n = 3). These results demonstrate that the Bcnrps1 gene is not involved in oxidative stress tolerance, which is a function previously described for this family
Summary
Botrytis cinerea (teleomorph Botryotinia fuckeliana) is a phytopathogenic fungus belonging to the phylum Ascomycota and is the causal agent of gray mold disease. It can infect over 200 ornamental and agriculturally important plant species, resulting in considerable economic losses [1,2,3]. Nine NRPS genes have been identified in the latest update of B. cinerea’s genome [2,5]. Three of these annotated genes (Bcnrps, Bcnrps, Bcnrps7)
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