Grapevine Trunk Diseases Research Articles

Cultivar Susceptibility to Natural Infections Caused by Fungal Grapevine Trunk Pathogens in La Mancha Designation of Origin (Spain).

Grapevine trunk diseases (GTDs) are one of the main biotic stress factors affecting this crop. The use of tolerant grapevine cultivars would be an interesting and sustainable alternative strategy to control GTDs. To date, most studies about cultivar susceptibility have been conducted under controlled conditions, and little information is available about tolerance to natural infections caused by GTD fungi. The objectives of this study were: (i) to identify tolerant cultivars to GTD fungi within a Spanish germplasm collection, based on external symptoms observed in the vineyard; and (ii) to characterize the pathogenic mycoflora associated with symptomatic vines. For this purpose, a grapevine germplasm collection including 22 white and 25 red cultivars was monitored along three growing seasons, and their susceptibility for esca foliar symptoms was assessed. Fungi were identified by using morphological and molecular methods. Cultivars such as, ‘Monastrell’, ‘Graciano’, ‘Cabernet Franc’, ‘Cabernet Sauvignon’, ‘Syrah’, ‘Moscatel de Alejandría’, ‘Sauvignon Blanc’, and ‘Airén’ displayed high susceptibility to GTDs, whereas others such as ‘Petit Verdot’, ‘Pinot Noir’, ‘Chardonnay’, and ‘Riesling’ were considered as tolerant. The prevalent fungal species isolated from symptomatic vines were Phaeomoniella chlamydospora (27.9% of the fungal isolates), Cryptovalsa ampelina (24.6%), and Dothiorella sarmentorum (21.3%).

Methyl salicylate, a grape and wine chemical marker and sensory contributor in wines elaborated from grapes affected or not by cryptogamic diseases

Methyl salicylate (MeSA) is a plant metabolite that induces plant defence resistance and an odorous volatile compound presenting green nuances. This volatile compound was shown to be present in wine samples, sometimes at concentrations above its olfactory detection threshold. MeSA is localized in grapes, particularly in the skins and stems, and is extracted during red wine vinification. It was detected at the highest concentrations in wines of several grape varieties, made from grapes affected by cryptogamic diseases, namely downy mildew caused by Plasmopara viticola, and black rot caused by Guignardia bidwellii. It has also been detected in wines from vines affected by Esca, a Grapevine Trunk Disease. MeSA can also be considered to be a chemical marker in grapes and wine indicative of the level of development of several vine cryptogamic diseases.

Assessment of Conjugate Complexes of Chitosan and Urtica dioica or Equisetum arvense Extracts for the Control of Grapevine Trunk Pathogens

In the work presented herein, we analyze the efficacy of three basic substances that comply with European Regulation (EC) No 1107/2009, namely chitosan, horsetail (Equisetum arvense L.) and nettle (Urtica dioica L.), for the control of grapevine trunk diseases (GTDs) in organic farming. The E. arvense and U. dioica aqueous extracts, prepared according to SANCO/12386/2013 and SANTE/11809/2016, have been studied by gas chromatography–mass spectrometry (GC-MS), identifying their main active constituents. The three basic substances, either alone or in combination (forming conjugate complexes), have been tested in vitro against eight Botryosphaeriaceae species, and in vivo, in grafted plants artificially inoculated with Neofusicoccum parvum and Diplodia seriata. A clear synergistic behavior between chitosan and the two plant extracts has been observed in the mycelial growth inhibition tests (resulting in EC90 values as low as 208 μg·mL−1 for some of the isolates), and statistically significant differences have been found in terms of vascular necroses lengths between treated and non-treated plants, providing further evidence of aforementioned synergism in the case of D. seriata. The reported data supports the possibility of extending the applications of these three basic substances in Viticulture beyond the treatment of mildew.

Mycobiota Associated with the Vascular Wilt of Poplar

In 2017, a 560-ha area of hybrid poplar plantation in northern Poland showed symptoms of tree decline. The leaves appeared smaller, yellow-brown, and were shed prematurely. Twigs and smaller branches died without distinct cankers. Trunks decayed from the base. The phloem and xylem showed brown necrosis. Ten percent of the trees died 1–2 months after the first appearance of the symptoms. None of these symptoms were typical for known poplar diseases. The trees’ mycobiota were analysed using Illumina sequencing. A total of 69 467 and 70 218 operational taxonomic units (OTUs) were obtained from the soil and wood. Blastocladiomycota and Chytridiomycota occurred only in the soil, with very low frequencies (0.005% and 0.008%). Two taxa of Glomeromycota, with frequencies of 0.001%, occurred in the wood. In the soil and wood, the frequencies of Zygomycota were 3.631% and 0.006%, the frequencies of Ascomycota were 45.299% and 68.697%, and the frequencies of Basidiomycota were 4.119% and 2.076%. At least 400 taxa of fungi were present. The identifiable Zygomycota, Ascomycota, and Basidiomycota were represented by at least 18, 263 and 81 taxa, respectively. Many fungi were common to the soil and wood, but 160 taxa occurred only in soil and 73 occurred only in wood. The root pathogens included species of Oomycota. The vascular and parenchymal pathogens included species of Ascomycota and of Basidiomycota. The initial endophytic character of the fungi is emphasized. Soil, and possibly planting material, may be the sources of the pathogen inoculum, and climate warming is likely to be a predisposing factor. A water deficit may increase the trees’ susceptibility. The epidemiology of poplar vascular wilt reminds grapevine trunk diseases (GTD), including esca, black foot disease and Petri disease.

Production of Phytotoxic Metabolites by Botryosphaeriaceae in Naturally Infected and Artificially Inoculated Grapevines.

Grapevine trunk diseases (GTDs) are considered a serious problem to viticulture worldwide. Several GTD fungal pathogens produce phytotoxic metabolites (PMs) that were hypothesized to migrate to the foliage where they cause distinct symptoms. The role of PMs in the expression of Botryosphaeria dieback (BD) symptoms in naturally infected and artificially inoculated wood using molecular and analytical chemistry techniques was investigated. Wood samples from field vines naturally infected with BD and one-year-old vines inoculated with Diplodia seriata, Spencermartinsia viticola and Dothiorella vidmadera were analysed by cultural isolations, quantitative PCR (qPCR) and targeted LC-MS/MS to detect three PMs: (R)-mellein, protocatechuic acid and spencertoxin. (R)-mellein was detected in symptomatic naturally infected wood and vines artificially inoculated with D. seriata but was absent in all non-symptomatic wood. The amount of (R)-mellein detected was correlated with the amount of pathogen DNA detected by qPCR. Protocatechuic acid and spencertoxin were absent in all inoculated wood samples. (R)-mellein may be produced by the pathogen during infection to break down the wood, however it was not translocated into other parts of the vine. The foliar symptoms previously reported in vineyards may be due to a combination of PMs produced and climatic and physiological factors that require further investigation.

In planta Activity of Novel Copper(II)-Based Formulations to Inhibit the Esca-Associated Fungus Phaeoacremonium minimum in Grapevine Propagation Material.

Grapevine trunk diseases (GTDs) are a serious and growing threat to vineyards worldwide. The need for innovative control tools persists since pesticides used against some GTDs have been banned and only methods to prevent infections or to reduce foliar symptoms have been developed so far. In this context, the application of imaging methods, already applied to study plant–microbe interactions, represents an interesting approach to understand the effect of experimental treatments applied to reduce fungal colonization, on GTD-related pathogens activity. To this aim, trials were carried out to evaluate the efficacy of copper-based treatments, formulated with hydroxyapatite (HA) as co-adjuvant with innovative delivery properties, loaded with two different copper(II) compounds (tribasic sulfate and sulfate pentahydrate), and applied to grapevine propagation material to inhibit fungal wood colonization. The treated rootstock (Vitis berlandieri × Vitis riparia cv. K5BB) and scion cuttings (Vitis vinifera L., cv. Chardonnay) had been inoculated with a strain of Phaeoacremonium minimum (Pmi) compared to uninoculated rootstocks. Experimental treatments were applied during the water-soaking process, comparing the copper(II) compounds pure or formulated with HA, to hydrate the cuttings. After callusing, grafted vines were grown under greenhouse conditions in a nursery and inoculated with Pmi::gfp7 or with Pmi wild-type. Fifteen weeks post-inoculation, woody tissues close to the inoculation site were sampled to evaluate the efficiency of the treatments by studying the plant–microbe interaction by confocal laser scanning microscopy (CLSM). Copper and further elements were also quantified in the same tissues immediately after the treatments and on the CLSM samples. Finally, the grapevine defense responses were studied in the leaves of cuttings treated with the same formulations. The present investigation confirmed the relevant interaction of Pmi and the related transformed strain on the vascular tissues of grafted vines. Furthermore, in vitro assay revealed (i) the fungistatic effect of HA and the reduced effect of Cu fungicide when combined with HA. In planta assays showed (ii) the reduction of Pmi infection in propagation material treated with HA-Cu formulations, (iii) the movement of HA-Cu formulations inside the plant tissues and their persistence over time, and (iv) the plant defense reaction following the treatment with pure HA or Cu, or combined.

Characterization of the Mycovirome of the Phytopathogenic Fungus, Neofusicoccum parvum.

Neofusicoccum parvum is a fungal plant-pathogen belonging to the family Botryosphaeriaceae, and is considered one of the most aggressive causal agents of the grapevine trunk disease (GTD) Botryosphaeria dieback. In this study, the mycovirome of a single strain of N. parvum (COLB) was characterized by high throughput sequencing analysis of total RNA and subsequent bioinformatic analyses. Contig annotations, genome completions, and phylogenetic analyses allowed us to describe six novel mycoviruses belonging to four different viral families. The virome is composed of two victoriviruses in the family Totiviridae, one alphaendornavirus in the family Endornaviridae, two mitoviruses in the family Mitoviridae, and one narnavirus belonging to the family Narnaviridae. The presence of the co-infecting viruses was confirmed by sequencing the RT-PCR products generated from total nucleic acids extracted from COLB. This study shows that the mycovirome of a single N. parvum strain is highly diverse and distinct from that previously described in N. parvum strains isolated from grapevines.

Incidence of grapevine trunk diseases on four cultivars in Sardinia, Southern Italy

Esca proper and Botryosphaeria dieback are among the most widespread Grapevine trunk diseases (GTDs), characterized by similar decline symptoms. In the present work, chronic, apoplexy and death symptoms were analysed separately in four vineyards and four different cultivars, on more than 1,000 vines per cultivar, taking into account ten-year annual surveys. The cumulative incidence of plants with chronic symptoms (CHR) reached high values on 'Sauvignon Blanc' (81.9 %), 'Cabernet Sauvignon' (79.4 %) and 'Cannonau' (66.5 %), but it was low on 'Merlot' (25.1 %). 'Sauvignon Blanc' showed the highest cumulative incidence of apoplectic events (23.1 %) and dead cordons (49.2 %), while 'Cannonau' had the greatest number of dead plants (28.8 %). In each symptom category, incidence among cultivars differed significantly according to χ2 test at P ≤ 0.05. Annual incidence of foliar symptoms fluctuated over ten years (ranging from 0.9-9.5 % in 'Merlot' to 6.3‑59.1 % in 'Cabernet Sauvignon'), mostly with regard to CHR. On average, every year only 33.9 % of plants showing CHR had expressed symptoms in the previous year, while 48.6 % did not show symptoms the following year. Conversely, most of the plants exhibiting apoplexy or death were symptomatic the previous year. According to Tuckey HSD test (P ≤ 0.05) 'Merlot' had the highest incidence of plants showing CHR symptoms for the first time (72.1 %) and of apparently recovered plants (76.3 %), while 'Cabernet Sauvignon' exhibited the highest incidence of plants showing CHR symptoms also the previous year (50.0 %). The 'Cabernet Sauvignon' attitude to show chronic symptoms with a certain continuity was also confirmed by the low incidence of plants with hidden symptoms (lack of symptoms in previously symptomatic vines). On the contrary, the incidence of acute symptoms (apoplectic events and dead plants) was quite low on 'Cabernet Sauvignon'. The present study confirms that GTD incidence is influenced by cultivar. All the cultivars assessed were susceptible, but with differences in intensity, type (chronic or acute) and fluctuation of symptoms. It cannot be excluded, however, that besides the genotype also external factors, as the vigour conferred by the type of soil or the combination with the rootstock, may have influenced the results.

On the Applicability of Chitosan Oligomers-Amino Acid Conjugate Complexes as Eco-Friendly Fungicides against Grapevine Trunk Pathogens

In a context in which the incidence and severity of grapevine fungal diseases is increasing as a result of both climate change and modern management culture practices, reducing the excessive use of phytosanitary products in viticulture represents a major challenge. Specifically, grapevine trunk diseases (GTDs), caused by several complexes of wood decay or xylem-inhabiting fungi, pose a major challenge to vineyard sustainability. In this study, the efficacy of chitosan oligomers (COS)–amino acid conjugate complexes against three fungal species belonging to the Botryosphaeriaceae family (Neofusicoccum parvum, Diplodia seriata, and Botryosphaeria dothidea) was investigated both in vitro and in planta. In vitro tests led to EC50 and EC90 effective concentrations in the 254.6−448.5 and 672.1−1498.5 µg·mL−1 range, respectively, depending on the amino acid involved in the conjugate complex (viz. cysteine, glycine, proline or tyrosine) and on the pathogen assayed. A synergistic effect between COS and the amino acids was observed against D. seriata and B. dothidea (synergy factors of up to 2.5 and 2.8, respectively, according to Wadley’s method). The formulations based on COS and on the conjugate complex that showed the best inhibition rates, COS−tyrosine, were further investigated in a greenhouse trial on grafted vines of two varieties (”Tempranillo” on 775P and “Garnacha” on 110R rootstock), artificially inoculated with the mentioned three Botryosphaeriaceae species. The in planta bioassay revealed that the chosen formulations induced a significant decrease in disease severity against N. parvum and B. dothidea. In summary, the reported conjugate complexes may be promising enough to be worthy of additional examination in larger field trials.

Plant resilience and physiological modifications induced by curettage of Esca-diseased grapevines

The re-emergence of Grapevine Trunk Diseases (GTDs), mainly Esca, has been observed in most of the world’s vineyards during the last two decades. Development of necrosis in grapevine wood, especially white-rot, is typically associated with Esca-diseased plants. One of the different methods being used in attempts to eradicate GTDs is curettage. This old cultural practice, which consists in surgically removing the necrotic wood, specifically white-rot, retaining only the non-necrotic tissue of Esca-diseased grapevine, is used in some European vineyards (Spain, France, Italy, Portugal), and is being increasingly reintroduced since 10 years ago in France. We, therefore, wanted to study the effect of curettage on vigour, fertility and berry quality, and year after year plant recovery. Our study synthetizes a 3-year experiment on Esca-diseased cv. Sauvignon blanc grapevines curetted in a commercial plot in the Bordeaux region. Asymptomatic control grapevines were compared to Esca-diseased grapevines without curettage (with typical foliar symptoms), and with curetted Esca-diseased grapevines (without foliar symptoms). Even if the curetted grapevines recovered lower vigour and fertility than the control plants, their grape berry quality was comparable, unlike for Esca-diseased grapevines. This cultural practice proved particularly effective in helping Esca-symptomatic grapevines to recover asymptomatic after treatment. Over time, curettage induces the resilience of grapevines, allowing them to recuperate their full physiological functioning, thereby compensating for Esca’s detrimental impact on berry quality.

Paenibacillus alvei K165 and Fusarium oxysporum F2: Potential Biocontrol Agents against Phaeomoniella chlamydospora in Grapevines.

In the last two decades grapevine trunk diseases (GTDs) have emerged as the most significant threat for grapevine sustainability worldwide. The tracheomycotic fungus Phaeomoniella chlamydospora (Pch) is the predominant GTD-associated species and cannot be controlled with available chemicals. In the present study, we evaluated the effectiveness of two microbial strains (Paenibacillus alvei K165 and Fusarium oxysporum F2) against Pch in grapevine. In vitro bioassays, performed in a growth culture medium simulating the xylem environment, indicated that F2 decreased Pch growth and sporulation, whereas K165 did not have any effect on Pch growth. In planta experiments revealed that root-drench and stem-puncture application of K165 and F2 reduced the endophytic relative DNA amount of Pch by 90% and 82%, respectively, compared to controls. However, wood discoloration, the typical symptom of Pch infection, was not reduced in the F2 treated grapevines. Nevertheless, the F2 treated grapevines harbored higher lignin levels compared to mocks, as it was also done by K165. Therefore, F2 and K165 have the potential to be used as biocontrol agents against Pch in grapevines.

Biocontrol Potential of Grapevine Endophytic and Rhizospheric Fungi Against Trunk Pathogens.

Grapevine Trunk Diseases (GTDs) are a major challenge to the grape industry worldwide. GTDs are responsible for considerable loss of quality, production, and vineyard longevity. Seventy-five percent of Chilean vineyards are estimated to be affected by GTDs. GTDs are complex diseases caused by several fungi species, including members of the Botryosphaeriaceae family and Phaeomoniella chlamydospora, considered some of the most important causal agents for these diseases in Chile. In this study, we isolated 169 endophytic and 209 rhizospheric fungi from grapevines grown under organic and conventional farming in Chile. Multiple isolates of Chaetomium sp., Cladosporium sp., Clonostachys rosea, Epicoccum nigrum, Purpureocillium lilacinum, and Trichoderma sp. were evaluated for their potential of biocontrol activity against Diplodia seriata, Neofusicoccum parvum, and Pa. chlamydospora. Tests of antagonism were carried out using two dual-culture-plate methods with multiple media types, including agar containing grapevine wood extract to simulate in planta nutrient conditions. Significant pathogen growth inhibition was observed by all isolates tested. Clonostachys rosea showed 98.2% inhibition of all pathogens in the presence of grapevine wood extract. We observed 100% pathogen growth inhibition when autoclaved lignified grapevine shoots were pre-inoculated with either C. rosea strains or Trichoderma sp. Overall, these results show that C. rosea strains isolated from grapevines are promising biocontrol agents against GTDs.

DNA-based detection of grapevine trunk-disease pathogens from environmental spore samples

In California vineyards, spore dispersal of fungi that cause grapevine trunk diseases Botryosphaeria dieback and Eutypa dieback occurs with winter rains. Spores infect through pruning wounds made to the woody structure of the vine in winter. Better timing of preventative practices that minimize infection may benefit from routine spore-trapping, which could pinpoint site-specific time frames of spore dispersal. To speed pathogen detection from environmental spore samples, we identified species-specific PCR primers and protocols. Then we compared the traditional culture-based method versus our new DNA-based method.•PCR primers for Botryosphaeria-dieback pathogen Neofusicoccum parvum and Eutypa-dieback pathogen Eutypa lata were confirmed species-specific, through extensive testing of related species (in families Botryosphaeriaceae and Diatrypaceae, respectively), other trunk-disease pathogens, and saprophytic fungi that sporulate in vineyards.•Consistent detection of N. parvum was achieved from spore suspensions used fresh or stored at -20°C, whereas consistent detection of E. lata was achieved only with a new spore-lysis method, using zirconia/silica beads in a FastPrep homogenizer (MP Biomedicals; Solon, Ohio, USA), and only from spore suspensions used fresh. Freezing E. lata spores at -20°C made detection inconsistent.•From environmental samples, spores of E. lata were detected only via PCR, whereas spores of N. parvum were detected both via PCR and in culture.

A comparison of the metabolomic response of grapevine to infection with ascomycete wood-infecting fungi

Fungi that cause grapevine trunk diseases, including Diplodia seriata, Eutypa lata, Neofusicoccum parvum, and Phaeomoniella chlamydospora, all colonize woody tissues, but lesions develop to different extents. Such variation is thought to correspond to fungal virulence, but it may also reflect host metabolic responses to limit colonization. From the woody stems of potted Vitis vinifera ‘Cabernet-Sauvignon’ in the greenhouse, we measured wood lesions and quantified metabolites at the lesion margins. Amino acids, sugars, phenolics, and terpenoids were quantified by appropriate chromatography techniques. At 3 months post-inoculation (MPI) lesions from P. chlamydospora and N. parvum were larger than those of D. seriata and E. lata. Among all pathogen treatments, compared to controls, lower concentrations of terpenoids at 0.5 MPI were followed at 2 MPI by higher concentrations of some phenolic compounds. Lesion length did not correlate with primary metabolites, but larger lesions did correlate with higher phenolic concentrations and lower terpenoid concentrations.

Pruning-wound protectants for trunk-disease management in California table grapes

In California's Mediterranean climate, management of grapevine trunk diseases (Botryosphaeria dieback, Eutypa dieback, Phomopsis dieback, Esca) focuses on protecting vines during the dormant season, when pruning coincides with rain-induced spore dispersal of the causal fungi (‘trunk pathogens’). Applying protectants (fungicides, toxic barriers, sealants, biocontrol agents) after pruning can minimize infection of pruning wounds. For the dormant seasons of 2017–2019, in a 2-year-old Vitis vinifera ‘Scarlet Royal’ vineyard in the southern San Joaquin Valley, we evaluated tractor-spray applications of the following protectants labeled for table grapes: fluopyram + tebuconazole, pyraclostrobin + boscalid, thiophanate-methyl + myclobutanil, and Bacillus amyloliquefaciens. A comprehensive set of virulent and widespread trunk pathogens, including Diaporthe ampelina, Eutypa lata, Neofusicoccum parvum, and Phaeomoniella chlamydospora, were inoculated to pruning wounds post-application, with detection attempts after a 3 to 4-week incubation period (before budbreak). Consistently high efficacies of pyraclostrobin + boscalid (68–100% and 56–100%, respectively) and thiophanate-methyl + myclobutanil (64–77% and 91–100%, respectively) against Botryosphaeria-dieback pathogen N. parvum and Phomopsis-dieback pathogen D. ampelina suggest spray applications of these protectants could minimize infection by these common trunk pathogens. Low and variable detection rates of slow-growing Eutypa-dieback pathogen E. lata and Esca pathogen P. chlamydospora suggest that the incubation period should be longer or that more replication is needed. Low efficacy of B. amyloliquefaciens, especially with only 4 days in 2017 before pathogen inoculation, suggests this biological-control agent may need more time and/or repeated applications to become established on pruning wounds before being challenged by a pathogen.

Role of cultivar susceptibility and vineyard age in GTD: examples from the Carpathian Basin

Grapevine trunk diseases (GTDs) are among the most severe problems in viticulture worldwide. The exact etiology and the role of endophytic microorganisms is not known yet and there is no adequate protection or curative treatment against the disease. Hungarian wine regions are also affected by the disease, and there is restricted information about the rate of infection nation-wide and about the susceptibility of the Hungarian cultivars. The main objectives of our research are to measure the symptom expression and the damage caused by GTDs, to understand the epidemiology and etiology of the disease to establish a foundation of a proper disease management. Cultivar susceptibility groups were created with the aim to allocate some Hungarian cultivars and the role of vineyard age was also examined in symptom expression.

Long-Term Benefits of Protecting Table Grape Vineyards against Trunk Diseases in the California Desert

Grapevine trunk diseases (GTD) are caused by several fungal species and are major limiting factors to vineyard productivity and profitability in all viticulture areas. This study is aimed at addressing the gap in the knowledge with regards to measuring the long-term benefits of post-pruning fungicide application on trunk diseases incidence and crop yield in grape production systems. It also calculated the net economic benefit of implementing such practice over the vineyard lifespan. We selected a newly planted commercial table grape vineyard in the California desert and divided it in two blocks. In one block, the registered fungicide thiophanate-methyl was mechanically applied on pruning wounds for six consecutive years, while the other half remained untreated. Our results showed a significant lower GTD incidence and vine replants in treated blocks combined with a significant increase of total and marketable fruit. Potential annual economic benefits of applying fungicide on pruning wounds appear to be in the range of $8500–$12,500 per hectare annually in a 50–75% disease control scenario.

Endophytic Fungi as Potential Biological Control Agents against Grapevine Trunk Diseases in Alentejo Region.

Simple SummaryGrapevine trunk diseases are the most widespread fungal diseases, affecting grapevines in all the major growing regions of the world, and their complete eradication is still not possible. Aiming to search alternatives to avoid the spread and high incidence of these diseases, the present work identified in vineyards within the Alentejo region the grapevine fungal community and among it the fungi responsible for those diseases. Grapevine fungal community showed a wide variety of fungi, nine of them previously described as grapevine trunk diseases-associated fungi. Almost all these fungi were detected in symptomatic and asymptomatic plants, which shows the importance of investigating the interactions of fungal communities and confirms the need for early diagnosis of these diseases. The potential of endophytic fungi as bio-control agents was tested against grapevine trunk diseases-associated fungi. These tests were performed among identified endophytes and grapevine trunk diseases phytopathogenic fungi, and all the endophyte fungi showed potential as biocontrol agents. Our findings suggest that endophytes are promising candidates for their use in biological control due to their antagonistic activity against the mycelia growth of grapevine trunk diseases-associated fungi.Grapevine trunk diseases (GTDs) are the most widespread fungal diseases, affecting grapevines in all the major growing regions of the world, and their complete eradication is still not possible. Aiming to search alternatives to avoid the spread and high incidence of these diseases, the present work intended to molecularly identify the grapevine endophytic community, the phytopathogenic fungi associated with GTDs in vineyards within the Alentejo region, and to test potential antagonist microorganisms as biological control candidates against GTDs-associated fungi. Grapevine endophytic community showed a wide variety of fungi in GTDs’ asymptomatic and symptomatic plants, nine of them previously described as GTDs-associated fungi. GTDs prevalent fungi identified in symptomatic plants were Diaporthe sp., Neofusicoccum sp., and H. viticola. Almost all these fungi were also detected in asymptomatic plants, which shows the importance of investigating the interactions of fungal communities and confirms the need for early diagnosis of these diseases. Direct inhibition antagonism tests were performed among identified endophytes and GTDs phytopathogenic fungi, and all the endophyte fungi showed potential as biocontrol agents. Our findings suggest that endophytes are promising candidates for their use in biological control due to their antagonistic activity against the mycelia growth of some GTDs-associated fungi.

Protection of grapevine pruning wounds against Phaeomoniella chlamydospora and Diplodia seriata by commercial biological and chemical methods

The grapevine trunk diseases (GTD) Botryosphaeria dieback and esca threaten the sustainability of the grapevine industry worldwide. This study aimed to evaluate and compare the efficacy of various liquid (pyraclostrobin + boscalid and thiophanate methyl) and paste (paste + tebuconazole) formulation fungicide treatments as well as biological control agents (Trichoderma atroviride SC1 and T. atroviride I-1237), for their potential to prevent infection of grapevine pruning wounds by Diplodia seriata and Phaeomoniella chlamydospora in two field trials over two growing seasons. Treatments were applied to freshly pruned wounds following label dosages’ recommendations. After 24 h, wounds were artificially inoculated with 400 spores of D. seriata or 800 spores of P. chlamydospora. Isolations were made from the treated pruning wounds after 12 months to evaluate the efficacy of the treatments. Fungicide formulations were superior to Trichoderma-based treatments for the control of both pathogens during both growing seasons, with mean percent disease control of 44–95% for D. seriata and 46–67% for P. chlamydospora. Pyraclostrobin + boscalid was the most effective treatment. Trichoderma atroviride-based treatments did not reduce infection by D. seriata or P. chlamydospora compared to the untreated inoculated control in both vineyards and seasons. This study represents the first scientific comparative vineyard assessment of both commercial and biological treatments to protect pruning wounds against GTD fungi in Europe and provides growers with tangible preventative control practices to minimize yield losses due to GTD.

Esca-affected grapevine leaf metabolome is clone- and vintage-dependent.

Esca is a complex grapevine trunk disease caused by wood-rotting ascomycetes and basidiomycetes and leading to several foliar and wood symptoms. Given that the esca expression can be influenced by several environmental, physiological, and genetic factors, foliar symptoms are inconsistent in incidence and prevalence and may appear 1 year but not the following. We have previously reported a clone-dependent expression of the disease in cv Chardonnay. Owing to metabolome analysis, we could discriminate the metabolite fingerprint of green leaves collected on diseased vines of clones 76 and 95. These clone-dependent fingerprints were year-dependent in intensity and nature. The present work was conducted to determine if the clone-dependent disease expression observed is specific to Chardonnay or if it also occurs in another cultivar. A plot located in the Jura vineyard (France) and planted with both 1004 and 1026 clones of Trousseau, a cultivar highly susceptible to esca, was thus selected and studied during 2017 and 2018. A year-dependent variation of the symptoms expression was first observed and a possible relationship with rainfall is hypothesized and discussed. Moreover, a higher percentage of the clone 1026 vines expressed disease, compared to the 1004 ones, suggesting the higher susceptibility of this clone. Finally, metabolomic analyses of the remaining green leaves (i.e, without symptom expression) of partial esca-apoplectic vines allowed us to confirm a clone-dependent metabolic response to the disease. The metabolite fingerprints obtained differed in nature and intensity to those previously reported for Chardonnay and also between years.