Abstract
Many phytopathogenic fungi produce necrosis and ethylene inducing peptide 1 (Nep1-like proteins or NLP) that trigger leaf necrosis and the activation of defense mechanisms. These proteins have been widely studied in plant pathogens as Moniliophthora perniciosa or Botrytis cinerea between others, but little is known about their biological roles in grapevine trunk pathogens. Advances in the sequencing of genomes of several fungi involved in grapevine trunk diseases have revealed that these proteins are present in several copies in their genomes. The aim of this project was to analyze the presence of genes encoding NLP proteins in the Diplodia seriata genome and to characterize their putative role as virulence factors associated to grapevine trunk diseases. In this study, we characterized four NLPs from Diplodia seriata. All proteins showed highly similar amino acid sequences and contained the characteristic peptide motifs of NLPs. DserNEPs slightly reduced the viability of Vitis vinifera L. cell cultures. The cytolytic activity from DserNEP1 was stronger than that from DserNEP2, even at low concentrations. Purified DserNEPs also produced necrosis in leaves when they were inoculated into micropropagules of V. vinifera L. This is the first record of Nep1-like proteins from a fungus associated with grapevine trunk diseases and also from a member of the Botryosphaeriaceae family.
Highlights
Grapevines are one of the most important economic crops worldwide
Four bacteriophages containing the whole sequence of four different DserNEP genes were selected: phage λDASH-DsF1 contained a DserNEP1 gene (NCBI GenBank database accession number AKQ49205), λDASH-DsF2 phage contained a DserNEP2 gene (AKQ49206), whereas λDASH-DsF3 phage contained a DserNEP3 gene (MK978328), and λDASH-DsF4 phage contained a DserNEP4 gene (MK978329)
DserNEP1 gene consisted of an open reading frame (ORF) of 877 bp
Summary
Grapevine trunk diseases (GTDs) are a major threat for the wine sector, causing serious economic losses to the wine industry (Siebert, 2001; Gubler et al, 2005). This term encompasses different fungal pathologies such as Botryosphaeria dieback, Esca, Eutypa dieback, Petri disease, or Black foot, to cite the most relevant. Botryosphaeria dieback has been reported since the 1970s as one of the main GTD It is caused by several xylem-inhabiting fungi (Bertsch et al, 2013) which are primarily members of the Botryosphaeriaceae family such as Diplodia seriata De Not. It is caused by several xylem-inhabiting fungi (Bertsch et al, 2013) which are primarily members of the Botryosphaeriaceae family such as Diplodia seriata De Not. (anamorph of Botryosphaeria obtusa, Shoemaker, 1964; Phillips et al, 2007), Diplodia mutila
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