Grapevine Trunk Disease Pathogens Research Articles

Nursery origin and propagation stage influence the endophytic fungal communities inhabiting grapevine planting stocks

Infection of grapevine trunk disease (GTD) pathogens can occur in nursery and may result in young vine decline in newly established vineyards. We aimed at profiling the causal agents of GTD in California nurseries and explore how the vine propagation processes affect pathogens spread and fungal community dynamics. Plant and environmental samples were collected throughout the plant propagation. For grafted vines, wood tissue samples were collected above the graft union (scion), below the graft union (rootstock) and at the crown (rootstock) and wood necrosis was scored using a 0-3 rating system. We used both a metabarcoding approach using PCR primers custom-designed for GTDs and culturing technique coupled with Sanger sequencing for sample diagnosis. Results showed that mycobiome diversity and composition in vines was significantly influenced by the nursery of origin and the propagation stage. We identified 21 GTD-taxa across all plant samples associated with Petri disease, black foot, Botryosphaeria dieback, Phomopsis dieback and Eutypa dieback. Only 10% of the mother vines cuttings were GTD-pathogens free but co-infection with multiple pathogens became more frequent as the vines progressed through the propagation pipeline. Especially, the incidence of pathogens associated with Petri disease (Phaeoacremonium, Cadophora, Pleurostoma) and Fusarium increased during that time span likely because of high contamination levels in soil and water environments. Wood necrosis at the graft union and crown also increased post grafting and Phaeoacremonium and Fusarium correlated with more necrosis whereas Trichoderma with healthier wood ratings. Our study suggests that nursery practices strongly influence GTD pathogen profile.

Trichoderma carraovejensis: a new species from vineyard ecosystem with biocontrol abilities against grapevine trunk disease pathogens and ecological adaptation.

Trichoderma strains used in vineyards for the control of grapevine trunk diseases (GTDs) present a promising alternative to chemical products. Therefore, the isolation and characterization of new indigenous Trichoderma strains for these purposes is a valuable strategy to favor the adaptation of these strains to the environment, thus improving their efficacy in the field. In this research, a new Trichoderma species, Trichoderma carraovejensis, isolated from vineyards in Ribera de Duero (Spain) area, has been identified and phylogenetically analyzed using 20 housekeeping genes isolated from the genome of 24 Trichoderma species. A morphological description and comparison of the new species has also been carried out. In order to corroborate the potential of T. carraovejensis as a biological control agent (BCA), confrontation tests against pathogenic fungi, causing various GTDs, have been performed in the laboratory. The compatibility of T. carraovejensis with different pesticides and biostimulants has also been assessed. This new Trichoderma species demonstrates the ability to control pathogens such as Diplodia seriata, as well as high compatibility with powdered sulfur-based pesticides. In conclusion, the autochthonous species T. carraovejensis can be an effective alternative to complement the currently used strategies for the control of wood diseases in its region of origin.

Arthropods as Vectors of Grapevine Trunk Disease Pathogens: Quantification of Phaeomoniella chlamydospora on Arthropods and Mycobiome Analysis of Earwig Exoskeletons.

Viticulture worldwide is challenged by grapevine trunk diseases (GTDs). Involvement of arthropods in the dissemination process of GTD pathogens, notably esca pathogens, is indicated after detection of associated pathogens on arthropod exoskeletons, and demonstration of transmission under artificial conditions. The present study is the first to quantify spore loads via qPCR of the esca-relevant pathogen Phaeomoniella chlamydospora on arthropods collected in German vineyards, i.e., European earwigs (Forficula auricularia), ants (Formicidae), and two species of jumping spiders (Marpissa muscosa and Synageles venator). Quantification of spore loads showed acquisition on exoskeletons, but most arthropods carried only low amounts. The mycobiome on earwig exoskeletons was described for the first time to reveal involvement of earwigs in the dispersal of GTDs in general. Metabarcoding data support the potential risk of earwigs as vectors for predominantly Pa. chlamydospora and possibly Eutypa lata (causative agent of Eutypa dieback), as respective operational taxonomical unit (OTU) assigned genera had relative abundances of 6.6% and 2.8% in total reads, even though with great variation between samples. Seven further GTD-related genera were present at a very low level. As various factors influence the successful transmission of GTD pathogens, we hypothesize that arthropods might irregularly act as direct vectors. Our results highlight the importance of minimizing and protecting pruning wounds in the field.

First Report of Binucleate Rhizoctonia AG-G Causing Grapevine (Vitis Vinifera) Trunk Diseases in California Nurseries.

Grapevine Trunk Diseases (GTD) are caused by a consortium of fungal pathogens that affect the biological functions of the vascular system of mature and young grapevines (Gramaje et al. 2018). We conducted surveys to profile GTD pathogens in California grapevine nurseries and collected 784 cuttings of cvs. Cabernet Sauvignon and Chardonnay grafted on 1103P rootstock. Several vines exhibited wood necrotic lesions and cankers at the graft union and the root ball (Figure 1A). Symptomatic wood tissues were cultured on PDA medium and after two weeks of incubation at room temperature (22°C), several known GTD pathogens were recovered. We also identified Rhizoctonia from 42 of the 784 vines (5.3% incidence) based on the morphological characteristics of a brown pigmented mycelium (Figure 1B), hyphae branched at a right angle with constrictions at the branch point (Figure 1C) and absence of spores (González García et al., 2006). A subsample of four isolates (DCHG2B, DCSG22R, JCSG9B, and JCHG12B) were randomly selected for further DNA-based taxonomic identification and pathogenicity evaluation to grapevine. The ITS and beta tubulin regions were amplified using the ITS1/ITS4 and B36F/B12R primer sets, respectively (González et al. 2006), and sequences were deposited in the NCBI database (Accession numbers: OR052655, OR052656, OR052657, OR052658 and OR059207, OR059208, OR059209, OR059210). Sequences displayed >99% and >96% identity with the respective ITS and beta tubulin sequences of the binucleate Rhizoctonia AG-G specimen C-653 (González et al. 2006). A phylogenetic tree constructed using the Neighbor-Joining method indicated a 100% bootstrap support with the binucleate Rhizoctonia AG-G (Figure 2). Pathogenicity of the binucleate AG-G Rhizoctonia were confirmed on two separate technical replicates using standard methods. For each replicate, one-year-old rootstock 1103P were wounded with sterile drill bits and inoculated with a single 5 mm diameter agar plug collected from Rhizoctonia growing cultures, while control vines were inoculated with sterile agar. The first replicate lasted 28 weeks with (DCHG2B, DCSG22R) inoculated on seven vines. The second bioassay lasted 24 weeks with two additional isolates (JCSG9B, JCHG12B) inoculated on twelve vines. Rhizoctonia-inoculated vines developed wood symptoms similar to those observed on cuttings in nurseries, with necrotic lesions lengths significantly longer than the controls (First replicate: 3.5  0.4 cm vs. 1.3  0.6 cm; Second replicate: 6.8  0.8 cm vs. 1.1  0.2 cm), based on one-way ANOVA statistical test (P value < 0.05). Rhizoctonia isolates recovery from wood necrotic lesions were confirmed by ITS sequencing, thereby fulfilling Koch's postulate. Several binucleate Rhizoctonia anastomosis groups, including AG-G, have been found to cause root rot and stem necrosis in plant nurseries (Aiello et al., 2017; Rinehart et al., 2007). Rhizoctonia has also been reported to be associated with grapevine nurseries in Europe (Pintos et al., 2018), South Africa (Halleen et al., 2003) and Australia (Walker, 1992). However, the multinucleate Rhizoctonia solani was the only species confirmed to cause root rot on grapevine (Walker, 1992). Our data suggests that the binucleate Rhizoctonia from the AG-G anastomosis group also cause wood necrosis in grapevine. Those findings warrant further studies on the complexity of Rhizoctonia anastomosis groups in nursery and their aggressiveness to grapevine.

In vitro characterization of a Bacillus velezensis isolate as an antagonist of grapevine trunk disease pathogens

AbstractOne of the major and yet unsolved threats for viticulture is the group of vascular fungal infections, the so-called grapevine trunk diseases. Besides their latent nature and the enormous number of associated pathogens, their control is also hampered by the lack of effective fungicides, directing growing attention toward the use of biocontrol agents. In the present study the isolation, identification, and characterization of a bacterial strain are presented, showing biocontrol potential against some main causal agents of grapevine trunk diseases. The strain was isolated from the wood of an asymptomatic grapevine and selected for the fungicidal activity against the pathogen Phaeomoniella chlamydospora. According to 16S rDNA, gyrA, and gyrB sequences, the isolate belongs to Bacillus velezensis species. Confrontation tests with the bacterium or with its fermentation broth further revealed growth inhibition and fungicide activity against Botryosphaeria dothidea, Eutypa lata and Diaporthe ampelina pathogens. Fractionation of the bacterial culture filtrate suggests that the antifungal agents secreted by the B. velezenzis isolate are mainly lipoproteins. Phytotoxicity tests were also carried out with the isolate, showing no harmful effects on grapevine foliar disks.

Prevalence and pathogenicity of fungi associated with grapevine trunk diseases in Jordan

Grapevines (Vitis vinifera) are important fruit producers in Jordan, and grapevine trunk diseases (GTDs) are suspected to cause problems in many Jordanian vineyards. This study aimed to estimate GTDs incidence and severity in selected vineyards, and to isolate and identify the causal agents associated with GTDs in this country. Field surveys were carried out and representative samples of diseased vines showing GTDs symptoms were collected to isolate and identify the causal organisms. Molecular analyses of DNA sequences of the Internal Transcribed Spacer (ITS) regions of fungal ribosomal DNA (rDNA) were used to confirm the morphological identifications of the fungal isolates. GTDs were present in all the surveyed vineyards. Mean GTD incidence was 44% across all the fields evaluated, ranging from 9 to 69% in individual vineyards. Disease severity ranged from 52–74% (mean = 62%) across all vineyards and locations. A total of 325 fungal isolates were recovered from infected grapevines. The most prevalent pathogens identified were those associated with Botryosphaeria dieback, including Diplodia seriata, Lasiodiplodia theobromae and Neofusicoccum parvum, followed by the Esca disease pathogens Phaeoacremonium parasiticum, P. aleophilum, P. rubrigenum, and Fomitiporia spp. Ilyonectria liriodendri and I. spp., known to be associated with black foot of grapevines, were also isolated. Plant pathogens causing vascular wilts and root rots, including Fusarium proliferatum, F. oxysporum, Verticillium spp., and Rhizoctonia solani, were also identified from diseased plant samples, and were found in mixed infections with GTDs pathogens. Most of the identified pathogens, except those associated with vascular wilt and root rot, are reported for the first time in Jordan. Results of this study indicate that GTDs are widespread in Jordan, and that there is urgent need to adopt a “national strategy” for GTD management in this country.

Grapevine dieback caused by Botryosphaeriaceae species, Paraconiothyrium brasiliense, Seimatosporium vitis-viniferae and Truncatella angustata in Piedmont: characterization and pathogenicity

Grapevine Trunk Diseases (GTDs) are major threats in Mediterranean countries, causing economic losses due to reduced grape yields and long-term vine productivity, as well as death of grapevines. A survey was conducted in Piedmont (Northern Italy) during 2021-2022 to investigate the species diversity and distribution of GTD pathogens in this important Italian wine region. Morphological and multi-locus phylogenetic analyses (based on ITS, tef1, tub2, act and rpb2) identified species of Botryosphaeriaceae at high frequency, including Botryosphaeria dothidea, Diplodia mutila, Diplodia seriata and Neofusicoccum parvum. Other pathogens commonly associated with GTDs, including Eutypa lata, Fomitiporia mediterranea and Phaeomoniella chlamydospora, were also isolated. Less commonly isolated species included Neocucurbitaria juglandicola, Paraconiothyrium brasiliense, Seimatosporium vitis-viniferae and Truncatella angustata. Pathogenicity tests with two representative isolates of each species were carried out using one-year-old potted grapevine cuttings (‘Barbera’). All isolates (except N. juglandicola) caused brown wood necrotic vascular discolourations on inoculated plants and were successfully re-isolated. Effects of temperature on colony growth were also assessed. For all tested isolates there was no growth at 5°C, only four isolates (Botryosphaeriaceae) grew at 35°C, and optimum growth temperatures were between 20 and 25°C. This is the first record of Paraconiothyrium brasiliense and Neocucurbitaria juglandicola associated with symptomatic grapevines in Italy.

Susceptibility of pruning wounds to grapevine trunk disease pathogens Eutypa lata and Diplodia seriata in three climatic conditions in Australia

The grapevine trunk diseases Eutypa and Botryosphaeria dieback, caused by fungal species that infect pruning wounds, are a threat to vineyard longevity worldwide. This study evaluated the susceptibility of grapevine pruning wounds in three climatic regions of Australia. In field trials, wounds were made early, mid- and late winter, and inoculated with spores of Eutypa lata or Diplodia seriata at various times, from 1 to 112 days after pruning. For both pathogens, wounds were highly susceptible immediately after pruning, followed by a rapid decrease in susceptibility over the next 14 days in McLaren Vale and Adelaide Hills, South Australia, whereas the period of susceptibility was longer in Big Rivers, New South Wales, where high natural disease pressure of D. seriata confounded results. In the Adelaide Hills, delaying pruning to late winter may reduce the risk of infection by E. lata. A detached cane assay confirmed that the duration of susceptibility of six commonly grown cultivars to E. lata infection was similar.

Susceptibility of Grapevine Sucker and Green Shoot Wounds to Trunk Disease Pathogens

Grapevine trunk disease fungi infect vines through openings, primarily pruning wounds. The main objective of this study was to understand the role of sucker wounds and wounds made by the removal of green shoots from the stems of potted grapevines as potential points of infection for grapevine trunk disease pathogens. Six wine and four table grape vineyards of different ages were sampled in different production areas in the Western Cape grape region of South Africa. Isolations were made from 161 sucker wounds, and fungal pathogens were identified using morphology and DNA sequence analysis of the internal transcribed spacers (ITS1 and ITS2) and the 5.8S ribosomal RNA gene, the translation elongation factor 1alpha or the partial β-tubulin gene. The results show that 62% of the sucker wounds were infected by trunk disease pathogens, including Diaporthe ampelina, Diplodia seriata, Phaeomoniella chlamydospora, Phaeoacremonium minimum, Eutypella microtheca, Cryptovalsa ampelina and Neofusicoccum australe. Diaporthe ampelina was the most common, followed by D. seriata and P. chlamydospora, in both the wine and table grape sucker wounds. Under glasshouse conditions, wounds made by the removal of young green shoots on one-year-old potted grapevine plants were inoculated with spore suspensions of D. ampelina, E. lata, N. parvum, P. minimum and P. chlamydospora. After four months, all the inoculated pathogens could be re-isolated. This study shows that grapevine sucker and green shoot wounds are susceptible to different grapevine trunk disease pathogens and may therefore play a role in the epidemiology of trunk diseases.

Biocontrol Potential of an Endophytic Pseudomonas poae Strain against the Grapevine Trunk Disease Pathogen Neofusicoccum luteum and Its Mechanism of Action.

Grapevine trunk diseases (GTDs) impact the sustainability of vineyards worldwide and management options are currently limited. Biological control agents (BCAs) may offer a viable alternative for disease control. With an aim to develop an effective biocontrol strategy against the GTD pathogen Neofusicoccum luteum, this study investigated the following: (1) the efficacy of the strains in suppressing the BD pathogen N. luteum in detached canes and potted vines; (2) the ability of a strain of Pseudomonas poae (BCA17) to colonize and persist within grapevine tissues; and (3) the mode of action of BCA17 to antagonize N. luteum. Co-inoculations of the antagonistic bacterial strains with N. luteum revealed that one strain of P. poae (BCA17) suppressed infection by 100% and 80% in detached canes and potted vines, respectively. Stem inoculations of a laboratory-generated rifampicin-resistant strain of BCA17 in potted vines (cv. Shiraz) indicated the bacterial strain could colonize and persist in the grapevine tissues, potentially providing some protection against GTDs for up to 6 months. The bioactive diffusible compounds secreted by BCA17 significantly reduced the spore germination and fungal biomass of N. luteum and the other representative GTD pathogens. Complementary analysis via MALDI-TOF revealed the presence of an unknown cyclic lipopeptide in the bioactive diffusible compounds, which was absent in a non-antagonistic strain of P. poae (JMN13), suggesting this novel lipopeptide may be responsible for the biocontrol activity of the BCA17. Our study provided evidence that P. poae BCA17 is a potential BCA to combat N. luteum, with a potential novel mode of action.

Current status of grapevine trunk disease pathogens on asymptomatic nursery-produced grapevines in Türkiye

Good health of grapevine plants is important for productivity and sustainability of newly established vineyards, and accurate detection of bacterial and fungal pathogens is a prerequisite for managing the diseases they cause in nurseries. This study screened marketable, bare-rooted grapevine plants, obtained from different geographical regions of Türkiye, for fungal pathogens associated with grapevine trunk diseases (GTDs). In 2021, 43 grapevine nurseries located in eight provinces were surveyed to reveal the status of GTD pathogens on asymptomatic marketable plants. Fungal pathogens isolated from the roots and basal parts of asymptomatic dormant grapevines were identified using with morphological characteristics and molecular markers, and were subjected to pathogenicity tests. Six species; Cytospora viticola, Diaporthe ampelina, Diplodia seriata, Lasiodiplodia brasiliensis, Neofusicoccum parvum, and Truncatella angustata (associated with dieback), and six species; Cadophora ferruginea, Cadophora luteo-olivacea, Cadophora malorum, Phaeoacremonium minimum, Phaeoacremonium tuscanicum and Phaeomoniella chlamydospora (associated with Petri disease) were identified based on DNA sequencing of ITS and TEF1-α genes. GTD pathogens were detected in 12 and 14 of the 43 nurseries, respectively. Pathogenicity tests on 1103P vines revealed that all species were pathogenic (N. parvum and C. luteo-olivacea being the most virulent), and caused significant wood necroses when compared to non-inoculated experimental controls. This is the first report of C. ferruginea, C. malorum, L. brasiliensis, and P. tuscanicum associated with GTDs in Türkiye.

A systematic survey on prevalence of grapevine trunk disease pathogens in Oregon vineyards.

Grapevine trunk diseases (GTDs) are found in vineyards worldwide and can be caused by different fungal pathogens. To characterize types of GTDs in Oregon vineyards, and how the GTD pathogens' prevalence is affected by two geographical regions, a survey was conducted in which grapevine trunk samples were collected from 15 and 14 wine grape (Vitis vinifera) vineyards in southern and northern Oregon respectively. Fungal species were identified through culture and PCR based methods. GTD pathogens that were identified included Botryosphaeriaceae spp. and Phaeoacremonium spp. from 72 and 21% of the surveyed vineyards respectively; Phaeomoniella chlamydospora, Cryptovalsa ampelina, Truncatella angustata, Seimatosporium lichenicola, Hormonema viticola from 7% of the surveyed vineyards; and Dactylonectria macrodidyma, and Pestaloptiopsis sp. from 3% of the surveyed vineyards. Pathogens were identified in both regions and in young and mature vineyards. The presence of GTD from the Botryosphaeria dieback complex was significantly affected by regions (P=0.021) with pathogens being significantly more abundant in Willamette Valley (northern region) compared to Rogue Valley (southern region) vineyards. Some differences among other tested variables such as vineyard age, cultivars, rootstocks, and pruning methods were observed for all disease complexes; however the differences were not statistically significant. Our study summarizes that Botryosphaeria dieback and Esca disease complexes are the most prevalent diseases infecting grapevines in Oregon vineyards and management practices need to be geared towards these economically important diseases. In addition, pathogens from other disease complexes are also present suggesting a need for regular disease monitoring and following practices to limit the spread of these pathogens.

Healthy Rootstock Cuttings are Essential for the International Nursery Industry

Rootstock cuttings infected with latent Grapevine Trunk Diseases (GTD’s) pathogens are a major problem for nurseries. These fungal pathogens include several species of the genera Phaeoacremonium, Phaeomoniella, Cadophora, Dactylonectria, Ilyonectria, Diaporthe and the family Botryosphaeriaceae. Infected rootstock cuttings can cause losses of up to 60 % during propagation and post planting, and can be a reservoir of disease in the vineyard. This article presents a protocol for sanitary production of rootstock cuttings and is a guide to improving health of the grafted vine, and the profitability of nurseries and vineyards.

Sodium arsenite-induced changes in the wood of esca-diseased grapevine at cytological and metabolomic levels.

In the past, most grapevine trunk diseases (GTDs) have been controlled by treatments with sodium arsenite. For obvious reasons, sodium arsenite was banned in vineyards, and consequently, the management of GTDs is difficult due to the lack of methods with similar effectiveness. Sodium arsenite is known to have a fungicide effect and to affect the leaf physiology, but its effect on the woody tissues where the GTD pathogens are present is still poorly understood. This study thus focuses on the effect of sodium arsenite in woody tissues, particularly in the interaction area between asymptomatic wood and necrotic wood resulting from the GTD pathogens' activities. Metabolomics was used to obtain a metabolite fingerprint of sodium arsenite treatment and microscopy to visualize its effects at the histo-cytological level. The main results are that sodium arsenite impacts both metabolome and structural barriers in plant wood. We reported a stimulator effect on plant secondary metabolites in the wood, which add to its fungicide effect. Moreover, the pattern of some phytotoxins is affected, suggesting the possible effect of sodium arsenite in the pathogen metabolism and/or plant detoxification process. This study brings new elements to understanding the mode of action of sodium arsenite, which is useful in developing sustainable and eco-friendly strategies to better manage GTDs.

Evaluation of Bacillus subtilis PTA-271 and Trichoderma atroviride SC1 to control Botryosphaeria dieback and black-foot pathogens in grapevine propagation material.

Grapevine trunk diseases (GTDs) are a complex group of diseases that lead to major economic losses in all wine-producing countries. The investigation of biocontrol agents (BCAs) capable of forestalling or at least minimizing the development of GTDs has, recently, become a priority. Nursery experiments were set up to (i) assess the biocontrol effect of Trichoderma atroviride (Ta) SC1 and Bacillus subtilis (Bs) PTA-271, alone and in simultaneous application, against Botryosphaeria dieback (BOT)- and black-foot (BF)- associated pathogens during the grapevine propagation process and (ii) evaluate the success of the BCA inoculation during the grapevine propagation process, using quantitative reverse-transcription polymerase chain reaction techniques. The results demonstrated a significant reduction in the percentage of potentially infected plants and the percentage of fungal isolation from wood fragments of BOT and BF pathogens in nursery material treated with Ta SC1 and Bs PTA-271, respectively. In one of the experiments, simultaneous treatments with Bs PTA-271 and Ta SC1 caused a reduction in percentages of potentially infected plants and fungal isolation, from wood fragments containing BOT and BF pathogens. These biological treatments may be relevant components of an integrated approach, using complementary management strategies to limit infection by GTD pathogens, but further research is still needed to elucidate the effectiveness of Bs PTA-271 and the benefits of simultaneous application with Ta SC1 for the control of GTD pathogens in nurseries. © 2022 Society of Chemical Industry.

Diversity and Bioactivity of Endophytic Actinobacteria Associated with Grapevines

Grapevine trunk diseases (GTDs) are a significant problem for New Zealand viticulture. Endophytic actinobacteria are of interest as potential biocontrol agents due to their ability to inhibit plant pathogens and improve plant growth. However, no studies have investigated the diversity of actinobacteria associated with grapevines in New Zealand vineyards and their bioactivity. Actinobacteria diversity in different ‘Sauvignon blanc’ vine tissues from three vineyards (conventional and organic management, and different vine ages) was assessed using different methods and media. Forty-six endophytic actinobacteria were isolated, with more isolates recovered from roots (n = 45) than leaves (n = 1) and shoot internodes (n = 0). More isolates were recovered from the organic (n = 21) than conventional (n = 8) vineyard, mature (25-year old; n = 21) than young (2-year old; n = 2) vines and using a tissue maceration technique (n = 40). Actinomycete Isolation Agar, International Streptomyces Project 2, and Starch Casein media were effective for actinobacteria isolation. Most of the isolates recovered belonged to Streptomyces, with one isolate identified as Mycolicibacterium. Forty isolates were assessed for antifungal activity and plant growth-promoting (PGP) characteristics. Of these, 13 isolates had antifungal activity against test GTD pathogens (Dactylonectria macrodidyma, Eutypa lata, Ilyonectria liriodendri, Neofusicoccum parvum, and N. luteum). Eighteen isolates exhibited more than one PGP trait; 25siderophore production (n = 25), phosphate solubilization (n = 6), and indole acetic acid production (n = 16). Two strains, Streptomyces sp. LUVPK-22 and Streptomyces sp. LUVPK-30, exhibited the best antifungal and PGP properties. This study revealed the diversity of culturable endophytic actinobacteria from grapevines in New Zealand vineyards and their biocontrol potential against GTD pathogens.

Mycoparasitism capability and growth inhibition activity of Clonostachys rosea isolates against fungal pathogens of grapevine trunk diseases suggest potential for biocontrol.

The present study aimed to examine the capability of Clonostachys rosea isolates as a biological control agent against grapevine trunk diseases pathogens. Five C. rosea and 174 pathogenic fungal strains were isolated from grafted grapevines and subjected to in vitro confrontation tests. Efficient antagonism was observed against Eutypa lata and Phaeomoniella chlamydospora while mycoparasitism was observed to the pathogens of Botryosphaeria dothidea and Diaporthe spp. pathogens in in vitro dual culture assays. The conidia production of the C. rosea isolates were also measured on PDA plates. One isolate (19B/1) with high antagonistic capabilities and efficient conidia production was selected for in planta confrontation tests by mixing its conidia with the soil of Cabernet sauvignon grapevine cuttings artificially infected with B. dothidea, E. lata and P. chlamydospora. The length and/or the incidence of necrotic lesions caused by E. lata and P. chlamydospora at the inoculation point were significantly decreased after a three months incubation in the greenhouse on cuttings planted in soils inoculated with the conidia of strain 19B/1, while symptom incidence and severity were unaffected in the case of the pathogen B. dothidea. Based on the above results, we consider C. rosea a promising biological control agent against some grapevine trunk diseases.

Abstracts of oral and poster presentations given at the 12th International Workshop on Grapevine Trunk Diseases, Mikulov, Czech Republic, 10–14 July 2022

The 12th International Workshop on Grapevine Trunk Diseases (12th IWGTD) was held in Mikulov, Czech Republic, from 10 to 14 July, 2022. The Workshop was chaired by Dr Aleš Eichmeier and organized by Mendel University in Brno and Svaz Vinařů České Republiky/Union of Winemakers of the Czech Republic. The ICGTD Council meeting took place on 10 July, and that evening the welcome reception was opened by Rostislav Koštial (Mayor of Mikulov, Senator and viticulturist), Dr Martin Chlad (President of Svaz Vinařů České Republiky/Union of Winemakers of the Czech Republic), and Dr Svatopluk Kapounek (Vice-Rector, Mendel University, Brno).The scientific programme was opened by Dr Alena Salašová (Dean, Faculty of Horticulture, Mendel University, Brno). The Workshop was attended by 133 researchers from 25 countries, presenting 61 oral and 49 poster papers. These presentations were in five sessions, including; Pathogen Characterization and Identification, Epidemiology, Plant-pathogen Interactions, Microbial Ecology, and Disease Management in Nurseries and Vineyards. The Disease Management session aimed to provide grape growers with an overview of practical options for trunk disease control. This session included contributions from researchers who have completed field trials on trunk disease management. David Gramaje (Instituto de Ciencias de la Vid y el Vino (ICVV), Logroño, La Rioja, Spain, outlined an evaluation of treatments for protection of grapevine pruning wounds from natural pathogen infections; Francois Halleen (University of Stellenbosch, South Africa) presented detailed results of efficiency of hot water treatments in nurseries; and Josè Úrbez-Torres (Agriculture and Agri-Food Canada SuRDC) outlined research on biological control of Botryosphaeria dieback in grapevines.A field trip (13 July) visited vineyards in the South Moravia. Delegates were presented with an overview of grapevine production in South Moravia, followed by discussions on the main diseases (including GTDs) affecting grapevines in the Czech Republic. Vineyards affected by GTD pathogens were also visited, where the growers outlined their viewpoints. The field trip focused on presenting the current situation of GTDs in South Moravia, and the management strategies being adopted. &#x0D; Student competitions for best oral and poster presentations included papers presented by 29 postgraduate students. For best posters:1st place went to Catarina Leal (University of Reims Champagne-Ardenne, France, and Polytechnic University of Valencia, Spain), “Evaluation of Trichoderma atroviride SC1 and Bacillus subtilis PTA-271 combination against grapevine trunk diseases pathogens in nursery propagation process”;2nd place went to Marcelo Bustamante (University of California, Davis, United States of America), “Investigating the role of Fusarium spp. in young vine decline in California”;&#x0D; 3rd place went to María Julia Carbone (Universidad de la República, Montevideo, Uruguay), “Interactive effects of Dactylonectria macrodidyma inoculation on the rhizosphere and root microbiome of grapevine”.For best oral presentations:1st place went to Catarina Leal, (University of Reims Champagne-Ardenne, France, and Polytechnic University of Valencia, Spain), “Beneficial effects of Bacillus subtilis PTA-271 and Trichoderma atroviride SC1 against the Botryosphaeria-dieback pathogen Neofusicoccum parvum may vary with grapevine cultivar”;2nd place went to Isidora Silva-Valderama (University of British Columbia, Vancouver, Canada), “Predicting pathogens virulence: linking host breadth and pathogenicity of the Botryosphaeriaceae fungal family in wine grapes (Vitis vinifera)”;3rd place went to Colin Todd (University of California, Riverside, United States of America), “Tracking the fungal pathobiome associated with young grapevine decline in California nurseries”.The 13th IWGTD will be held in Ensenada, Mexico, in 2025.

Lignans Extract from Knotwood of Norway Spruce-A Possible New Weapon against GTDs.

Grapevine trunk diseases (GTDs) pose a major threat to the wine industry worldwide. Currently, efficient biological methods or chemical compounds are not available for the treatment of infected grapevines. In the present study, we used an extract from the knotwood of spruce trees as a biological control against GTDs. Our in vitro trial was focused on the antifungal effects of the extract against the most common GTD pathogens—Cadophora luteo-olivacea, Dactylonectria torresensis, Diaporthe ampelina, Diaporthe bohemiae, Diplodia seriata, Eutypa lata, and Phaeoacremonium minimum. Our in vitro trial revealed a high antifungal effect of the extract against all tested fungi. The inhibition rates varied among the different species from 30% to 100% using 1 mg·mL−1 extract. Subsequently, the efficiency of the extract was supported by an in planta experiment. Commercial grafts of Vitis vinifera were treated with the extract and planted. The total genomic DNA of grapevines was extracted 10 days and 180 days after the treatment. The fungal microbial diversities of the treated/untreated plants were compared using high-throughput amplicon sequencing (HTAS). Treated plants showed 76.9% lower relative abundance of the genus Diaporthe and 70% lower relative abundance of the genus Phaeoacremonium 10 days after treatment. A similar scenario was observed for the genus Cadophora 180 days after treatment, where treated plants showed 76% lower relative abundance of this genus compared with untreated grapevines.

Biological and chemical pruning wound protectants reduce infection of grapevine trunk disease pathogens

Grapevine trunk diseases (GTDs) are currently considered some of the most important challenges for viticulture, curtailing vineyard longevity and productivity in nearly every raisin, table and wine grape production region in California and worldwide. Pruning wounds provide the main entry point for fungal pathogens responsible for these diseases; pathogens enter the wounds following precipitation events. The aim of this study was to evaluate the efficacy of selected chemical and experimental biological fungicides for protection of pruning wounds against two of the most common and virulent fungal pathogens causing GTDs:Eutypa lataandNeofusicoccum parvum. This study was conducted on sauvignon blanc at the UC Davis Department of Plant Pathology Field Station. Results showed that several chemical and biological fungicides, notably the chemical fungicide Luna Sensation, the biofungicide Vintec and a combination of the biofungicides Bio-Tam and CrabLife Powder, provided significant protection against at least one of the two canker pathogens used in this study. However, the majority of products tested did not provide simultaneous control of bothE. lataandN. parvumpathogens, highlighting the continuing challenge of controlling GTDs.