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Eutypa dieback, a devastating disease in grapevines, is caused by the fungal pathogen Eutypa lata, a wood-inhabiting fungus. E. lata acts by degrading wood tissues in the colonisation areas, and produces foliar symptoms. These striking symptoms have been attributed to the production of toxic metabolites by the pathogen, the most widely studied being eutypine. The aims of the study were to compare the effects of E. lata on xylem structure at the site of infection and in remote tissues. In healthy Vitis vinifera, the vessel-associated cells (VACs) in the trunk have a protective layer that covers the entire lignified wall and forms a transfer apparatus in pits located at the VAC / vessel interface. This apparatus occurs similarly in VACs in the basal part of canes but is less developed in the apical part. In the presence of E. lata, which is found only in the trunk and the cordons, the VACs initiated a program of secretory activity that led to the enlargement of the transfer apparatus, which is formed by tightly associated fibrils. This secretory activity was followed by VAC death. Furthermore, the hypertrophy of the transfer apparatus spread according to an acropetal gradient in the canes. Treatment with eutypine also induced the development of the transfer apparatus in VACs of basal and apical parts of canes excised from healthy vines. However, this apparatus was formed by loosely packed fibrils in VACs that were not completely damaged. Therefore, metabolites other than eutypine are expected to be involved in the VAC degeneration observed in infected vines.

Eutypa dieback is a devastating disease induced in vineyards by the fungal pathogen Eutypa lata. The fungus colonizes the xylem tissues of trunk and cordons but is never found in the annual canes. Nevertheless, dwarfed shoots and leaf necrosis observed in diseased plants indicate that a necrotic signal can spread at a distance from the infected area. Eutypine, a small cyclic molecule, and related compounds have been postulated as the toxins inducing these symptoms. In this work, we evidence that E. lata secreted other metabolites of polypeptidic nature which induced toxic effects on canes and leaves of vines, and on leaves of other plant materials. The polypeptide fraction (PF) isolated from culture medium of mycelium induced transitory Hþ fluxes and membrane depolarization of plant cells. Complementary assays with plasma membrane vesicles (PMV) showed that Hþ-ATPase is a primary site of action as indicated by inhibition of the enzyme activity and increase of Hþ conductance of plasma membrane. The toxic effect was also obvious on respiration and photosynthesis. All these impairments led to a hindering in cell energetics and, as a consequence, to an inhibition of uptake of assimilates. Treatment with PF also triggered biological events, characteristics of elicitation as suggested by the early responses on cell membrane described above, the activation of NADPH oxidase and the activation of Phenylalanine ammonia lyase (PAL)

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Eutypa dieback is a devastating disease of Vitis vinifera L. caused by the fungal pathogen Eutypa lata. This wood-inhabiting fungus degrades tissues in the trunk and cordons of infected vines and induces symptoms in the foliage. These symptoms have been attributed to the production of toxic metabolites by the pathogen, in particular eutypine. Recently, we have isolated polypeptide compounds secreted by the fungus in artificial culture. The aims of this study were to examine the effects induced in leaves by applying polypeptides and eutypine to detached canes and to compare this to the changes in leaf structure induced by E. lata in the vineyard. In leaves taken from vines infected with E. lata, the changes in mesophyll cells indicate that the fungus has an effect on tissue remote from the infected area. The size of mesophyll cells decreased by more than half, starch content was reduced and tannins were abundant. Plastids, mitochondria and cell walls were highly modified. In leaves taken from healthy canes treated with polypeptides of E. lata, the structure of mesophyll cells was also modified. The cell size did not change, but the tannin content increased and modifications in plastids and mitochondria were similar to those observed in leaves taken from infected vines. The major effect was the complete disorganisation of cell walls. Eutypine had less effect on organelle structure and did not modify the cell wall. In canes treated with polypeptides, vessel-associated cells (VACs) were also damaged. Abundant tannins occurred in the vacuoles of VACs and marked changes were noted in mitochondria, plastids and the protective layer, in particular in the pit at the vessel interface. In these pits, the protective layer, the primary wall and the middle lamella were all highly modified. In contrast, treatment with eutypine induced the development of a large transfer apparatus bordering the unmodified pectocellulose wall. These results illustrate that treatment with polypeptides produced by E. lata may cause changes in mesophyll cells in leaves and VACs in canes, that resemble changes observed in naturally infected vines. Comparatively, the differences with eutypine action were stressed. Both types of toxins may co-operate in vivo to produce the degeneration observed during the disease.

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Eutypa dieback (dying arm disease, eutypiosis) is a very devastating disease in many grape-producing areas around the world. This vascular disease is induced by the ascomycete Eutypa lata Pers. Fr. Tul & C. Tul. invading the trunk by pruning wounds. The environmental factors and the nutritional requirements regulating fungus development are yet poorly known. This work shows that the isolated strain of E. lata was able to grow in a large temperature range (2-30 • C). However, a higher temperature (35 • C) presented inhibitory effects on mycelial growth. E. lata was able to use various osidic molecules (C5, C6, C12, C18, C24, and starch), showing thus a large adaptation to the carbon source supplied. As nitrogen source, it used salts and numerous natural amino acids. A significant result was obtained with cysteine presenting obvious antifungal properties. This effect can further be used with the aim of setting up a curative treatment of the disease.