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Background and Aims Eutypa dieback is an economically important disease of grapevines and a major threat to vineyard longevity throughout the world. Developing effective preventative strategies offers the best means of control. In this study, pruning wound protectants and various spray applications were evaluated for their ability to prevent infection by ascospores of Eutypa lata. Methods and Results Fungicides and natural alternative treatments were applied by hand to grapevine pruning wounds in winter prior to inoculation with E. lata. In a series of greenhouse and field experiments, tebuconazole, pyraclostrobin, fluazinam, garlic extract and lactoferrin significantly reduced infection. A range of tractor-driven sprayers was used to apply tebuconazole to pruning wounds, and those which provided good coverage reduced infection by E. lata to a level similar to that when tebuconazole was applied with a paintbrush. Conclusion Eutypa dieback can be controlled with several fungicides and natural treatments. Tractor-driven sprayers, which by design or modification can efficiently deliver maximum possible coverage at high output rates, can be used for application of pruning wound treatments. Significance of the Study Results of this study contributed to registration of fungicides for use as pruning wound treatments to control E. lata in Australia. Demonstrating the ability of tractor-driven sprayers to apply treatments effectively has led to greater industry adoption of dormant treatments and may decrease the future impact of Eutypa dieback.

Nitrogen and carbohydrate contents in extracts from Cape Riesling canes/shoots and wood were determined. Nitrogen content from extracts of both canes/shoots and wood was the highest during June, July and August. Sugar content from canes/shoots was highest during July and August, while the sugar content in wood was the highest during July, August and September. Growth of Eutypa lata was determined in these extracts in vitro. Eutypa lata had the highest mycelium yield when grown in canes/shoot extracts collected during July and August. Eutypa lata showed the best growth in extracts from wood collected during July. It is suggested that the high nitrogen and sugar content in grapevine wood could play a role in the susceptibility of pruning wounds during the pruning season (i.e. June to August) to Eutypa lata.

There are presented localities of 100 ascomycetc spccics collected in primeval forest of north-eastern part of Poland. Among them some new species for Poland were collected: Eutypa lata var. aceri, Hypoderma sarmentorum, Lophiotrema curreyi, Massaria sorbi, Massarina chamaecyparisii, Mollisia poaeoides, Mycosphaerella lycopodii-annotini, Phaeosphaeria juncina, Ph. phragmilicola, Plagiosphaera immersa.

UMR AGAP Equipe Diversité, adaptation et amélioration de la vigne (DAAV); Eutypa lata is a fungal pathogen causing severe dieback in vineyards worldwide. This fungus colonizes vines through pruning wounds, eventually causing a brown sectorial necrosis in wood as well as stunted vegetative growth. Several years may pass between infection and the expression of external symptoms, hindering the rapid evaluation of both grapevine cultivars susceptibility and E. lata variation in aggressiveness. We aimed to develop a rapid quantitative method for the assessment of wood colonization after inoculation of cuttings in controlled conditions. We used several grape cultivars varying in susceptibility in the vineyard and fungal isolates with different levels of aggressiveness to monitor wood colonization during a maximum period of 2 months. Re-isolation allowed demonstration of the effects of both cultivars and fungal isolates on the rate of wood colonization. We also developed a real-time PCR method that was efficient in measuring fungal biomass, which was found to be correlated with isolate aggressiveness based on foliar symptom severity. The real-time PCR approach appears to be a useful technology to evaluate grapevine susceptibility to E. lata, and could be adapted to other pathogens associated with grapevine trunk diseases.

Dieback of red currant (Ribes rubrum) and gooseberry (Ribes uva-crispa) is an increasing problem in commercial fields in the Netherlands. Field surveys were done in 2006–2007 and samples with dieback symptoms were analysed. In this study the causal agent was diagnosed as Eutypa lata, based on morphological characteristics and rDNA-ITS sequence data. The field surveys revealed the presence of the anamorph and teleomorph states of the fungus produced on dead infected currant wood. Eutypa lata is a vascular pathogen of many woody plants. Related fungi from the same family Diatrypaceae are difficult to distinguish from E. lata based on morphological features. The genetic variability of E. lata was compared by rDNA-ITS sequencing of isolates from different hosts and origins. Within the E. lata isolates little variability in the ITS sequences was observed. Phylogenetic analysis showed no clear subdivisions within the species. Eutypa lata strains isolated from the different hosts were closely related, indicating that there is no direct evidence for host specificity.

Single-node plantlets were evaluated as a model system for experiments on grapevine trunk diseases. Experiments were conducted to determine how to grow the plantlets, how Botryosphaeria sp. and Eutypa lata symptoms were expressed in the plantlets compared with field-grown plants, and how the detached single-node plantlets (from living canes) responded compared with canes attached to vines under controlled conditions. Single-node plantlets could be grown in distilled water for up to 6 months. Under controlled conditions, significantly longer lesions formed on the single-node plantlets than on attached canes, suggesting that the single-node system may be more sensitive to infection than whole vines. The system shows promise for use in initial experiments on trunk diseases, before field experiments are conducted, but further work is required to validate the method.

Die Eutypa-, die Esca- und die Petrikrankheit haben in den vergangenen 15 Jahren in allen Weinbau betreibenden Bundeslandern Deutschlands zu Absterbeerscheinungen in Rebanlagen gefuhrt. Sie werden durch einen oder mehrere Pilzerreger verursacht. Bisher ist eine direkte Bekampfung im Weinbau nicht moglich. Aus der Literatur bekannte Daten zur Biologie, Epidemiologie und den Schadsymptomen an den Rebstocken werden zusammengefasst und indirekte Bekampfungsempfehlungen vorgeschlagen. Es erfolgt eine Bestandsaufnahme eines im Rahmen des Projekts „Praventiver Pflanzenschutz zur biologischen Bekampfung der Erreger dreier bedeutender Pilzkrankheiten im okologischen Weinbau“ im Jahr 2003 angelegten Freilandversuchs zur biologischen Kontrolle von Pilzkrankheiten im Weinbau mit antagonistischen Mikroorganismen der Gattung Trichoderma. Die Gehalte an Haupt- und Spurennahrelementen in Weinblattern gesunder und erkrankter Rebstocke wurden bestimmt. Die analysierten Elementgehalte in den Blattern des infizierten Bestands differieren von den Richtwerten, die einer ausreichenden Versorgung der Rebblatter mit Nahrstoffen zugrunde gelegt werden. Mikroorganismen der Gattung Trichoderma scheinen die durch die Krankheitserreger gehemmte Aufnahme von Mangan aufzuheben.

Background and Aims: Trichoderma species have potential in biocontrol of eutypa dieback; however, little is known about their ability to colonise grapevine wood. The colonisation of vines by Trichoderma harzianum and its effects on colonisation by Eutypa lata were studied. Methods and Results: T. harzianum colonised canes over a total distance (above and below the inoculation point) of 10 cm or more in 12 weeks after inoculation. Application of T. harzianum to canes prior to E. lata, either at the same inoculation point or at a different point, reduced recovery of the pathogen. In the field, T. harzianum grew 3 cm from the inoculation point in 4 months in all seven of the mature vines treated. The fungus persisted for 20 months in four of the seven vines at 6 cm from the inoculation point and reached the crown of one vine. Prior inoculation of mature vines with Trichodowels, containing T. harzianum, reduced recovery of E. lata 14 months after inoculation. Conclusions: T. harzianum colonised wood of Chardonnay, Shiraz and Nyora and reduced colonisation by E. lata. T. harzianum persisted in wood of mature vines of Nyora for up to 20 months. Significance of the Study: The results support the potential of T. harzianum in the biocontrol of eutypa dieback.

Grapevine cultivar (Vitis vinifera) and isolate of Eutypa lata influence wood and foliar symptoms of Eutypa dieback. Foliar symptoms of Eutypa dieback developed within 8 months of inoculating young grapevines (cvs. Grenache, Cabernet Sauvignon, and Merlot) in a shadehouse. Isolates of E. lata from various wine regions in southern Australia varied in their ability to colonize inoculated grapevines and induce wood and foliar symptoms. Grapevine cultivars varied for wood and foliar symptom expression but not for mycelial colonization. However, the severity of foliar symptoms was not related to the rate of spread of the fungus in the grapevine. Furthermore, the staining of wood typically attributed to E. lata did not reflect the presence of the fungus because the fungus was detected up to 80 mm beyond the stain. A field trial with mature grapevines revealed significant differences in the rate of spread of wood staining due to E. lata among eight cultivars, with up to 50 mm/year detected in Cabernet Sauvignon and Shiraz grapevines. In the shadehouse, the maximum growth rate of E. lata was recorded to be 115 mm/year for Grenache rootlings. Information from this study may help to optimize management strategies for maintaining productivity of grapevines with Eutypa dieback, thus reducing the economic impact of the disease.

Eutypa lata is a significant grapevine pathogen with limited means of prevention and control. The biological control agent Fusarium lateritium can prevent E. lata infection if applied directly onto a vine pruning wound. F. lateritium was suspended and stored in an invert emulsion formulation. A commercially available pruning shear was modified to dispense the formulated F. lateritium onto the cutting blade for direct application onto the pruning wound simultaneously with grapevine cutting. The modified pruner was tested for its ability to cover grapevine pruning wounds using the emulsion formulation. Efficacy of formulated F. lateritium on pruned grapevine canes was also studied using the pruner for application. Addition of grooves in the pruning blade significantly improved wound coverage. Biological efficacy testing determined that applying formulated F. lateritium with the modified pruner was as effective as pipetting formulation directly onto the pruning wound.

The responses of vine to the eutipoze attack were analyzed, depending on the age of the plantation. Of the four varieties studied, regardless of the plantation age, the lowest degree of attack was registered at FeteascAƒ regala (7.0%) and Sauvignon blanc (7.3%), the differences between being insignificant. The results confirm the data from the literature according to which the eutipoza is more widespread in plantations for over 10 years

Eutypa canker and dieback of almonds was first recorded in commercial plantations in Greece in mid-1980 and confirmed as due to the ascomycetous fungus Eutypa lata. The disease manifests itself by the formation of cankers at the point of the junction of the branches and the trunk and is associated with pruning wounds made by the growers in order to confront the‘non-infectious bud failure’disorder or to remove unwanted limbs close to the crotch. Pathogenicity tests made on young trees of cv. Texas with different isolates originated from grapevine, almond, apricot and pear showed significant differences in virulence. Pathogenicity tests carried out on 20 different almond cultivars gave a significant indication of the existence of tolerant and susceptible cultivars.

The vascular pathogen Eutypa lata , which causes Eutypa dieback in grapevines, is a major threat to grape production worldwide. Here, we present the first draft genome sequence of E. lata (UCR-EL1). The computational prediction and annotation of the protein-coding genes of UCR-EL1 provide an initial inventory of its potential virulence factors.

Severe decline of almond trees has recently been observed in several orchards on the island of Mallorca (Balearic Islands, western Mediterranean Sea). However, the identity of the causal agents has not yet been investigated. Between August 2008 and June 2010, wood samples from branches of almond trees showing internal necroses and brown to black vascular streaking were collected in the Llevant region on the island of Mallorca. Several fungal species were subsequently isolated from the margin between healthy and symptomatic tissue. Five species of Botryosphaeriaceae (namely Botryosphaeria dothidea, Diplodia olivarum, D. seriata, Neofusicoccum australe and N. parvum), Eutypa lata, Phaeoacremonium iranianum and Phomopsis amygdali were identified based on morphology, culture characteristics and DNA sequence comparisons. Neofusicoccum parvum was the dominant species, followed by E. lata, D. olivarum and N. australe. First reports from almond include D. olivarum and Pm. iranianum. Two species are newly described, namely Collophora hispanica sp. nov. and Phaeoacremonium amygdalinum sp. nov., We thank 'Conselleria d'Agricultura, Mediambient i Territori' of the Balearic Islands for funding this research. We appreciate the helpful assistance in field works from technicians and land owners: Tomeu Melis, Toni Nicolau, Beatriz Blanquer and Alejandro Aristondo. The authors also wish to thank the staff of 'Conselleria d'Agricultura, Mediambient i Territori': Alicia Nieto, Xavier Nadal and Toni Martorell. We would like to thank Walter Gams for kindly assisting with the nomenclature for the new species.

Une methode a ete mise au point en serre pour etudier le deperissement de la vigne du a Eutypa lata. Des cylindres de mycelium et de milieu PDA ont ete places dans des trous perces 4 cm au-dessous des bourgeons de boutures non racinees. Les pousses montrent les symptomes foliaires caracteristiques de la maladie des le stade F. La severite des symptomes depend de l'isolat d'E lata utilise. Tous les isolats se sont reveles capables de coloniser les boutures mais certains n'ont pas induit de symptomes foliaires. Tous les cepages ont montre les symptomes de la maladie et differents niveaux de sensibilite ont ete mis en evidence

UMR 1334 AGAP : Equipe AFEF ‘Architecture et Fonctionnement des Espèces fruitières’ ; Team AFFS ‘Architecture and Functioning of Fruit Species’ Contact: elisabeth.chevreau@angers.inra.fr Publication Inra prise en compte dans l'analyse bibliométrique des publications scientifiques mondiales sur les Fruits, les Légumes et la Pomme de terre. Période 2000-2012. http://prodinra.inra.fr/record/256699; International audience; The development of alternative selection systems without antibiotic resistance genes is a key issue to produce safer and more acceptable transgenic plants. Eutypine is a toxin produced by Eutypa lata, the causal agent of eutypa dieback of grapevine, which is detoxified in mung bean (Vigna radiata) by the gene Vr-ERE. Many phytotoxic compounds containing an aldehyde group can act as substrates for the Vr-ERE enzyme. The aim of the present work was to evaluate the effects of the overexpression of Vr-ERE in transgenic apple plants, as a first step towards the development of an alternative selection system. Viable transgenic apple clones expressing Vr-ERE were produced from the cultivar Greensleeves under kanamycin selection. Although the Vr-ERE transgene was normally expressed at the RNA and protein levels, the increase in aldehyde reductase activity tested on a range of potential substrates was very low in these clones. None of them revealed a significant increase in tolerance to toxic aldehydes compared to their non-transgenic control. This work with transgenic apple plants overexpressing the detoxifying gene Vr-ERE illustrates some of the difficulties in developing an alternative selection pressure

The synthesis of the novel (1′R*,2′S*,3′R*, 4′S*,5′R*)-1-(3′,4′-epxoy-2′, 5′-dihydroxycyclohexyl)-3-methyl-but-2-enone (2), recently isolated from the culture medium of the fungus Eutypa lata, is described.

International audience; Eutypa dieback is a vascular disease that may severely affect vineyards throughout the world. In the present work, microarrays were made in order (i) to improve our knowledge of grapevine (Vitis vinifera cv. Cabernet-Sauvignon) responses to Eutypa lata, the causal agent of Eutypa dieback; and (ii) to identify genes that may prevent symptom development. Qiagen/Operon grapevine microarrays comprising 14,500 probes were used to compare, under three experimental conditions (in vitro, in the greenhouse, and in the vineyard), foliar material of infected symptomatic plants (S(+)R(+)), infected asymptomatic plants (S(-)R(+)), and healthy plants (S(-)R(-)). These plants were characterized by symptom notation after natural (vineyard) or experimental (in vitro and greenhouse) infection, re-isolation of the fungus located in the lignified parts, and the formal identification of E. lata mycelium by PCR. Semi-quantitative real-time PCR experiments were run to confirm the expression of some genes of interest in response to E. lata. Their expression profiles were also studied in response to other grapevine pathogens (Erysiphe necator, Plasmopara viticola, and Botrytis cinerea). (i) Five functional categories of genes, that is those involved in metabolism, defence reactions, interaction with the environment, transport, and transcription, were up-regulated in S(+)R(+) plants compared with S(-)R(-) plants. These genes, which cannot prevent infection and symptom development, are not specific since they were also up-regulated after infection by powdery mildew, downy mildew, and black rot. (ii) Most of the genes that may prevent symptom development are associated with the light phase of photosynthesis. This finding is discussed in the context of previous data on the mode of action of eutypin and the polypeptide fraction secreted by Eutypa.

International audience; The grapevine (Vitis vinifera) is susceptible to many pathogens such as Botrytis cinerea, Plasmopara viticola, Erysiphe necator, and Eutypa lata. Phytochemicals are used extensively in vineyards to reduce pathogen infections, but the appearance of pesticide-resistant pathogen strains and the need for environmental protection require the use of alternative strategies. The phytohormone ethylene is assumed to play a role in the development of disease resistance. In the present study, we have treated grapevine foliar cuttings (Cabernet Sauvignon) with ethylene-releasing ethephon. This resulted in an increase in the number of pathogenesis-related protein (CHIT4c, PIN, PGIP, and GLU) gene copies and in an enhancement of phytoalexin biosynthesis by inducing the PAL and STS genes that correlated with the accumulation of stilbenes (antimicrobial compounds). Moreover, ethephon treatment triggered the protection of grapevine detached leaves and grapevine foliar cuttings against Erysiphe necator, the causal agent of powdery mildew (64% and 70%, respectively). These studies emphasize the major role of ethylene in grapevine defense.

Structural and synthetic studies of the metabolites isolated from Eutypa lata are reviewed. This fungus is the causative agent of Eutypa dieback disease, also known as eutyposis or dying-arm disease, a perennial canker that affects grapevines and many other woody fruit plants. The review, which encompasses all the literature in this field up to the present and in which 76 references are cited, also includes a detailed study of the biological activity of the metabolites, especially the role of toxins in the development of the plant disease. Some aspects of the synthesis and biosynthesis of these metabolites and related compounds are discussed.

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.

Eutypine (4-hydroxy-3-[3-methyl-3-butene-1-ynyl] benzaldehyde) is a toxin produced by Eutypa lata , the causal agent of eutypa dieback of grapevines. Recently, a eutypine detoxifying gene ( Vr-ERE ) encoding an NADPH-dependent aldehyde reductase which converts eutypine into the corresponding alcohol, eutypinol, a non-toxic form of the toxin, has been cloned. We report here the obtention of transgenic plants from grapevine rootstock 110 Richter Vitis berlandieri × V. rupestris that express the Vr-ERE gene. The over-expression of Vr-ERE gene in grapevine tissues increases their detoxification capacity. The growth and development of the transgenic plants cultured in vitro exhibiting a high Vr-ERE activity were not affected by the presence of 300 μM of toxin whereas the growth of untransformed control plants was highly inhibited. These results suggest that the Vr-ERE gene is an efficient candidate to confer resistance to the toxin and opens new opportunities to study the role of the toxin in the development of the disease.

International audience; Grapevine (Vittis vinifera L.) is susceptible to many pathogens, such as Botrytis cinerea, Plasmopara viticola, Uncinula necator, and Eutypa lata. Phytochemicals are used intensively in vineyards to limit pathogen infections, but the appearance of pesticide-resistant pathogen strains and a desire to protect the environment require that alternative strategies be found. In the present study, the, beta-1,3-glucan laminarin derived from the brown algae Laminaria digitata was shown both to be an efficient elicitor of defense responses in grapevine cells and plants and to effectively reduce B. cinerea and P viticola development on infected grapevine plants. Defense reactions elicited by laminarin in grapevine cells include calcium influx, alkalinization of the extracellular medium, an oxidative burst, activation of two mitogen-activated protein kinases, expression of 10 defense-related genes with, different kinetics and intensities, increases in chitinase and beta-1,3-glucanase activities, and the production of two phytoalexins (resveratrol and epsilon-viniferin). Several of these effects were checked and confirmed in whole plants. Laminarin did not induce cell death. When applied to grapevine plants, laminarin reduced infection by B. cinerea and R viticola by approximately 55 and 75%, respectively. Our data describing a large set of defense reactions in grapevine indicate that the activation of defense responses using elicitors could be a valuable strategy to protect plants against pathogens.

A phytotoxic metabolite has been isolated from liquid cultures of Eutypa lata and structurally characterized as 4-hydroxy-3-(3-methylbut-3-en-1-ynyl)benzaldehyde, named eutypine. It was detected by GC-MS and MS-MS analyses in the crude sap and the inflorescences of diseased plants and was not present in healthy material.

On Cabernet sauvignon, eutypiosis sensible variety, we have compared fatty acids content in healthy Jeaves (S: leaves beared by healthy vine stock), in leaves seeming healthy (Ap.S: leaves beared by arm without symptoms, the other arm is diseased) and in diseased leaves (M: leaves beared by diseased arm). The results show that disease does not alter the fatty acids order. Linolenic acid (CI 8:3) content is always the most important, followed by palmitic (CI6:0) and Iinoleic acids, then by stearic (CI8:0) and oleic acids and at last by arachidic acid (C20:0). During the three stages studied (flowers buds: H, flowering: I and closed cluster: L), only the development of CI 8:3 content was modified by disease. It increases in healthy leaves at stage I, while in Ap.S and M leaves there is a decrease from H to L stages. Otherwise, development of fatty acids proportions does not seem to be changed by eutypiosis.Proportions and content comparison in S, Ap.S and M leaves show a decrease of C 18:3 synthesis benefit of C 18:2.This increases is important as well as the symptoms are evident. These results seem to be in relationship with decrease of leaves size caused by eutypiosis. At the end, the disease induces a desaturation decrease of C18:0 and C18:2 as well as an aliphatic chain lengthening of C16:0. These inhibitions are showed by more increase of CI 6:0 and CI 8:2 in diseased leaves than in healthy leaves. No action is observed in CI 8: 1 desaturation.

(1993). A Hyphomycetous Synanamorph of Eutypa Armeniacae in Artificial Culture. Mycologia: Vol. 85, No. 6, pp. 941-944.

Eight vineyards in northern and central California were surveyed during three consecutive years, 1989-1991, and the presence or absence of Eutypa dieback symptoms was recorded for each vine in contiguous blocks of 1,250-3, 150 vines. The vineyards were located in areas with different levels of mean annual rainfall; some vineyards contained inoculum sources (perithecia) of Eutypa lata; others did not. The spatial patterns of infected vines were examined by ordinary runs, two-dimensional distance class, spatial autocorrelation, and geostatistical analyses. Disease incidence ranged from 3.4% in 1989 to 81.5% in 1991. During the study, disease incidence more than doubled in five of the vineyards. Vineyards with perithecia had higher disease incidence [...]

The fungus Eutypa lata was isolated from diseased walnut trees (Juglans regia) exhibiting small cankers. The morphological characteristics of the culture and the pathogenicity were compared with those of known isolates of the fungus from other hosts. Inoculation tests with walnut isolates on grape, walnut, almond and apricot yielded characteristic cankers. Furthermore, pathogenicity tests on walnut with isolates from other different hosts resulted in differences in virulence. Similar differences in virulence were observed between 13 single ascospore isolates of the fungus ex apricot inoculated on walnut.

The fungus Eutypa lata (syn. E. armeniacae), known as the causal agent of the death of many different woody plants, was found on dead branches of pistachio (Pistacia vera L.) in Greece. Isolations from diseased branches yielded consistently typical colonies of the asexual stage of the fungus (Libertella blepharis, syn. Cytosporina sp.), which proved to be undistinguishable from other cultures of the pathogen obtained from 15 different hosts. Furthermore, all isolates from pistachio tested for pathogenicity on apricot were pathogenic and yielded characteristic cankers.

A manually-operated spraying secateur has been tested for its efficacy in delivering benomyl to apricot trees during pruning for protection against the woundinvading pathogen Eutypa lata. The data indicate that devices of this type, when correctly used, will deliver sufficient biocide suspension to the wound surfaces to confer a high level of protection against the pathogen.