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Olive anthracnose outbreaks caused by the Colletotrichum species complex in the Mediterranean region decrease both fruit yield and olive oil production while also drastically degrading olive oil quality. The presence of various Colletotrichum species able to produce disease symptoms in olive fruits significantly deteriorates the efforts for an efficient crop protection strategy. In this report, the major olive productive area of Peloponnese was screened for Colletotrichum species capable of generating anthracnose symptoms. Olive fruits of 12 different olive cultivars were collected from 60 groves distributed analogously in the Peloponnese. Thirty-two fungal strains isolated from asymptomatic olive drupes were identified morphologically as Colletotrichum spp. and were multilocus genetically analyzed. The 32 isolates were grouped into two primary lineages resembling the previously characterized Colletotrichum acutatum and Colletotrichum nymphaeae based on the conducted genetic analysis for five genetic loci. The virulence of 16 Colletotrichum spp. strains were evaluated in a detached fruit assay of 10 Greek olive cultivars. The results clearly suggested that fungal isolates belonging to both C. acutatum and C. nymphaeae exhibited different levels of pathogenicity in a cultivar-dependent manner. Thus, cultivars examined in terms of the % Disease Index (%DI) were divided into highly tolerant, tolerant, and susceptible, and those analyzed regarding the % Disease Severity Index (%DSI) were divided into tolerant and susceptible. Our results suggest that the Greek cultivars of Athinolia and Megaritiki are highly tolerant to the vast majority of Colletotrichum strains isolated from Peloponnesian groves and consist of a significant genetic material for the future design of crop protection programs against anthracnose breakouts. [ABSTRACT FROM AUTHOR]

Verticillium dahliae is a xylem‐invading fungal pathogen that causes vascular wilt in a wide range of angiosperms. The pathogen uses a variety of virulence factors to invade and colonize its hosts. Here, we report that VdNEP, an NLP (Necrosis and ethylene inducing peptide 1‐Like Protein), functions as one such factor in multiple hosts. Eggplant leaves treated with VdNEP developed necrotic symptoms. Overexpression of VdNEP by incorporating extra copies of the VdNEP gene increased virulence to cotton, eggplant and tomato plants, suggesting its role as a virulence factor in diverse plants. Increased expression of VdNEP among the transformants did not correlate with the number of VdNEP inserts, suggesting that its expression was affected by the genomic context of the insertion sites. Interestingly, a transformant derived from a defoliating strain with high VdNEP transcript levels caused disease symptoms in tomato plants, whereas the corresponding wild‐type strain did not cause visible symptoms. The amount of V. dahliae DNA in plants infected with this VdNEP‐overexpressing transformant was 22 times higher than that in plants infected with the wild‐type isolate, further supporting the critical role of VdNEP in infection. A VdNEP‐EGFP fusion was constructed to follow its localization in fungal cells and during infection. [ABSTRACT FROM AUTHOR]

A comprehensive study was conducted on the prevalence of grapevine trunk diseases in Greece, focusing specifically on contamination of grapevine propagation material by Phaeomoniella chlamydospora and black foot disease‐related species. Additionally, detection of esca pathogen Fomitiporia mediterranea, causing white rot in grapevine and other woody hosts, was assessed using a new PCR‐based assay that distinguishes F. mediterranea from F. punctata. Development of a nested PCR assay, combined with a cost‐effective DNA extraction protocol, revealed a high percentage of infection by P. chlamydospora (51%) and Ilyonectria species associated with black foot disease (28%) in different types of grapevine propagation material (dormant cuttings, field‐rooted benchgrafts and green‐growing plants). Interestingly, black foot disease‐related pathogens were more prevalent in nursery plants grown in the field for 6 months (57%) than in other types of propagation material and compared with P. chlamydospora (43%), indicating increased infection of propagation material during growth in the nursery field. The cost‐effective molecular method developed in this study could be used in mass inspections of propagation material for phytosanitary purposes. Finally, using primers specific for F. mediterranea and F. puncata developed in this study, combined with the universal ITS4 primer, a collection of Fomitiporia isolates from mature grapevine and other woody hosts from Greece and Italy (southern Europe) were characterized as F. mediterranea, whereas German and Swedish isolates from forest plants (central Europe) were classified as F. punctata. The developed primers can discriminate between the two Fomitiporia species, which are indistinguishable based on culture and morphological characteristics alone. [ABSTRACT FROM AUTHOR]

Bacterial speck, caused by Pseudomonas syringae pv. tomato ( Pst), is an economically important disease of tomato, resulting in yield loss of marketable fruit. Management of bacterial speck is a challenge in commercial production fields due to the limited efficacy of current disease management strategies, as the pathogen acquires resistance to antibiotics and fixed copper bactericides and host resistance has not proven durable. Therefore, it is essential to develop alternative disease management strategies, like biological control. In this study, the efficacy of the commercially available biocontrol agent Bacillus subtilis QST 713 along with copper hydroxide was tested against Pst under greenhouse conditions. QST 713 reduced significantly disease severity and incidence compared to control and the copper hydroxide treatment; subsequently, the Pst population was lower in the QST 713-treated plants compared to control. In parallel, QST 713 and copper hydroxide increased plant height compared to control and mock plants. Furthermore, the quantitative PCR analysis of PR1a, PR1b and Pin2 expression suggests a positive role for Pin2 in the plant protective activity of QST 713, as Pin2 expression was significantly higher in the QST 713-treated plants challenged with Pst compared to the control Pst-inoculated plants. [ABSTRACT FROM AUTHOR]

The β-1,6-endoglucanase gene (vegB) of Verticillium dahliae was isolated using a genome walking technique. Nucleotide and deduced amino acid sequences of the gene showed high identity with the PAN1 sequence deposited at the Verticillium genome database (Broad Institute), but significant differences in intron numbers and sites of insertion. Detailed in silico analysis, accompanied by sequencing of both genomic and cDNA, as well as RT-PCR experiments, provided the correct size of the gene and the exact number, length and positions of introns. The putative protein of this gene was compared with corresponding β-1,6-endoglucanases from other fungi, and sequences were used to construct a phylogenetic tree. A clear differentiation between enzymes derived from plant pathogenic and mycoparasitic fungi was observed, fully supported by bootstrap data. An internal fragment (1.2kb) of vegB was used to disrupt the wild-type gene of a V. dahliae tomato race 2 strain, and the mutant strain, vegB-, was tested for pathogenicity on tomato plants. Results showed a small but constant reduction in disease symptoms only on eggplants for the vegB- strain in comparison with the wild type. Growth on minimal medium supplemented with different carbon sources showed reduced ability of the mutant to breakdown cellulose, whereas growth on glucose, pectin and sucrose was similar to the wild type. [ABSTRACT FROM AUTHOR]

Vascular wilts caused by Verticillium spp. are very difficult to control and, as a result, are the cause of severe yield losses in a wide range of economically important crops. The responses of Arabidopsis thaliana mutant plants impaired in known pathogen response pathways were used to explore the components in defence against Verticillium dahliae. Analysis of the mutant responses revealed enhanced resistance in etr1-1[ethylene (ET) receptor mutant] plants, but not in salicylic acid-, jasmonic acid- or other ET-deficient mutants, indicating a crucial role of ETR1 in defence against this pathogen. Quantitative polymerase chain reaction analysis revealed that the decrease in symptom severity shown in etr1-1 plants was associated with significant reductions in the growth of the pathogen in the vascular tissues of the plants, suggesting that impaired perception of ET via ETR1 results in increased disease resistance. Furthermore, the activation and increased accumulation of the PR-1, PR-2, PR-5, GSTF12, GSTU16, CHI-1, CHI-2 and Myb75 genes, observed in etr1-1 plants after V. dahliae inoculation, indicate that the outcome of the induced defence response of etr1-1 plants seems to be dependent on a set of defence genes activated on pathogen attack. [ABSTRACT FROM AUTHOR]