scholarly journals Effectiveness of Natural Antifungal Compounds in Controlling Infection by Grapevine Trunk Disease Pathogens through Pruning Wounds

2015 ◽  
Vol 81 (18) ◽  
pp. 6474-6483 ◽  
Author(s):  
Rebeca Cobos ◽  
Rosa María Mateos ◽  
José Manuel Álvarez-Pérez ◽  
Miguel Angel Olego ◽  
Silvia Sevillano ◽  
...  

ABSTRACTGrapevine trunk fungal pathogens, such asDiplodia seriataandPhaeomoniella chlamydospora, can infect plants through pruning wounds. They cause grapevine trunk diseases and are involved in grapevine decline. Accordingly, the protection of pruning wounds is crucial for the management of grapevine trunk diseases. The efficacy of different natural antifungals in inhibiting the growth of several fungi causing grapevine trunk diseases was evaluatedin vitro. The fungi showing greaterin vitroefficacy were tested on autoclaved grape wood assays againstD. seriataandP. chlamydospora. Based on results from these assays, chitosan oligosaccharide, vanillin, and garlic extract were selected for further evaluation on pruning wounds inoculated withD. seriataandP. chlamydosporain field trials. A significant decrease in plant mortality was observed after 2 years of growth in the plants treated with the different natural antifungals compared to the mortality rate observed in infected plants that were not treated with antifungals. Also, the infection rate for the inoculated pathogens was significantly reduced in plants treated with the selected natural antifungals. Therefore, natural antifungals represent a promising alternative for disease control and could provide significant economic benefits for the grape-growing industry.

2017 ◽  
Vol 83 (24) ◽  
Author(s):  
José Manuel Álvarez-Pérez ◽  
Sandra González-García ◽  
Rebeca Cobos ◽  
Miguel Ángel Olego ◽  
Ana Ibañez ◽  
...  

ABSTRACT Endophytic and rhizosphere actinobacteria isolated from the root system of 1-year-old grafted Vitis vinifera plants were evaluated for their activities against fungi that cause grapevine trunk diseases. A total of 58 endophytic and 94 rhizosphere isolates were tested. Based on an in vitro bioassay, 15.5% of the endophytic isolates and 30.8% of the rhizosphere isolates exhibited antifungal activity against the fungal pathogen Diplodia seriata, whereas 13.8% of the endophytic isolates and 16.0% of the rhizosphere isolates showed antifungal activity against Dactylonectria macrodidyma (formerly Ilyonectria macrodidyma). The strains which showed the greatest in vitro efficacy against both pathogens were further analyzed for their ability to inhibit the growth of Phaeomoniella chlamydospora and Phaeoacremonium minimum (formerly Phaeoacremonium aleophilum). Based on their antifungal activity, three rhizosphere isolates and three endophytic isolates were applied on grafts in an open-root field nursery in a 3-year trial. The field trial led to the identification of one endophytic strain, Streptomyces sp. VV/E1, and two rhizosphere isolates, Streptomyces sp. VV/R1 and Streptomyces sp. VV/R4, which significantly reduced the infection rates produced by the fungal pathogens Dactylonectria sp., Ilyonectria sp., P. chlamydospora, and P. minimum, all of which cause young grapevine decline. The VV/R1 and VV/R4 isolates also significantly reduced the mortality level of grafted plants in the nursery. This study shows that certain actinobacteria could represent a promising new tool for controlling fungal trunk pathogens that infect grapevine plants through the root system in nurseries. IMPORTANCE Grapevine trunk diseases are a major threat to the wine and grape industry worldwide. They cause a significant reduction in yields as well as in grape quality, and they can even cause plant death. Trunk diseases are caused by fungal pathogens that enter through pruning wounds and/or the root system. Although different strategies have recently been developed to protect pruning wounds using antifungal compounds (natural or synthetic) or biocontrol agents, no tools are yet available for controlling soil pathogens that infect plants through their root system. This study shows that different actinobacterial isolates, when applied to grafts in a nursery, can significantly reduce the infection rate caused by fungal pathogens that enter through the root system. This is a new, promising, and green alternative for preventing the decline of young grapevines in nurseries and vineyards.


2021 ◽  
Vol 12 ◽  
Author(s):  
Charis K. Ramsing ◽  
David Gramaje ◽  
Sara Mocholí ◽  
Javier Agustí ◽  
Félix Cabello Sáenz de Santa María ◽  
...  

Fungal grapevine trunk diseases (GTDs) are some of the most pressing threats to grape production worldwide. While these diseases are associated with several fungal pathogens, Phaeomoniella chlamydospora and Phaeoacremonium minimum are important contributors to esca and Petri diseases. Recent research has linked grapevine xylem diameter with tolerance to Pa. chlamydospora in commercial rootstocks. In this study, we screen over 25 rootstocks for xylem characteristics and tolerance to both Pa. chlamydospora and Pm. minimum. Tolerance was measured by fungal incidence and DNA concentration (quantified via qPCR), while histological analyses were used to measure xylem characteristics, including xylem vessels diameter, density, and the proportion of the stem surface area covered by xylem vessels. Rootstocks were grouped into different classes based on xylem characteristics to assess the potential association between vasculature traits and pathogen tolerance. Our results revealed significant differences in all the analyzed xylem traits, and also in DNA concentration for both pathogens among the tested rootstocks. They corroborate the link between xylem vessels diameter and tolerance to Pa. chlamydospora. In Pm. minimum, the rootstocks with the widest xylem diameter proved the most susceptible. This relationship between vasculature development and pathogen tolerance has the potential to inform both cultivar choice and future rootstock breeding to reduce the detrimental impact of GTDs worldwide.


Author(s):  
Robert Blundell ◽  
Akif Eskalen

Grapevine trunk diseases, caused by many different fungal pathogens, are one of the most economically important diseases affecting the grapevine industry worldwide. Pruning wounds are the main point of entry for these fungal pathogens and thus, disease control is focused on preventative pruning wound protection by chemical products and/or biological control agents (BCAs). In this study we evaluated a broad variety of already registered or at the experimental stage of chemical and BCAs in greenhouse and in field trials for the protection of table- and wine-grape vines against infection of Eutypa lata and Neofusicoccum parvum, major pathogens responsible for Eutypa and Botryosphaeria dieback, respectively. Our study showed that Trichoderma asperellum and Trichoderma gamsii consistently provided pruning wound protection in greenhouse and field trials, with a mean percent disease control (MPDC) of 88% and 100% for E. lata and N. parvum, respectively, when compared to the water treated-inoculated positive control (P<0.05). The chemical protectants, thiophanate-methyl + myclobutanil and fluopyram and trifloxystrobin were also able to effectively protect wounds with a MPDC of up to 86% when compared to the water treated-inoculated positive control (P<0.05). When biological treatments were evaluated for recovery from treated canes at the end of the growing season, Trichoderma-based treatments had a rate of recovery between 0 and 100%, Aureobasidium-based treatments had a recovery rate between 25 and 100%, and Bacillus-based treatments had a recovery rate between 0 and 25%.


Plants ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 1527
Author(s):  
Natalia Langa-Lomba ◽  
Eva Sánchez-Hernández ◽  
Laura Buzón-Durán ◽  
Vicente González-García ◽  
José Casanova-Gascón ◽  
...  

In this work, the chemical composition of Rubia tinctorum root hydromethanolic extract was analyzed by GC–MS, and over 50 constituents were identified. The main phytochemicals were alizarin-related anthraquinones and flavoring phenol compounds. The antifungal activity of this extract, alone and in combination with chitosan oligomers (COS) or with stevioside, was evaluated against the pathogenic taxa Diplodia seriata, Dothiorella viticola and Neofusicoccum parvum, responsible for the so-called Botryosphaeria dieback of grapevine. In vitro mycelial growth inhibition tests showed remarkable activity for the pure extract, with EC50 and EC90 values as low as 66 and 88 μg·mL−1, respectively. Nonetheless, enhanced activity was attained upon the formation of conjugate complexes with COS or with stevioside, with synergy factors of up to 5.4 and 3.3, respectively, resulting in EC50 and EC90 values as low as 22 and 56 μg·mL−1, respectively. The conjugate with the best performance (COS–R. tinctorum extract) was then assayed ex situ on autoclaved grapevine wood against D. seriata, confirming its antifungal behavior on this plant material. Finally, the same conjugate was evaluated in greenhouse assays on grafted grapevine plants artificially inoculated with the three aforementioned fungal species, resulting in a significant reduction in the infection rate in all cases. This natural antifungal compound represents a promising alternative for developing sustainable control methods against grapevine trunk diseases.


2020 ◽  
Author(s):  
María del Pilar Martínez-Diz ◽  
Emilia Díaz-Losada ◽  
Ángela Díaz-Fernández ◽  
Yolanda Bouzas-Cid ◽  
David Gramaje

ABSTRACTThe grapevine trunk diseases (GTDs) 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, and 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 their label dosages recommendations. After 24 hours, 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 to 95% for D. seriata and 46 to 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 vineyard assessment of several chemical and biological treatments to protect pruning wounds against GTDs fungi in Europe and provides growers with tangible preventative control practices to minimize yield losses due to GTDs.


Plant Disease ◽  
2016 ◽  
Vol 100 (8) ◽  
pp. 1651-1659 ◽  
Author(s):  
Georgina Elena ◽  
Jordi Luque

Diplodia seriata and Phaeomoniella chlamydospora are two fungal pathogens associated with grapevine trunk diseases worldwide. This study aimed to evaluate the period during which grapevine pruning wounds remain susceptible to fungal infection and to describe the colonization of canes artificially inoculated with these pathogens. In the first experiment, pruning wounds made in either fall or winter were separately inoculated with each pathogen at different times after pruning. Wound susceptibility to both pathogens decreased as the period between pruning and inoculation increased, from high percentages recorded in the first inoculation round (D. seriata, 97.5% and P. chlamydospora, 75%) down to approximately 10% 12 weeks after pruning. Pruning wounds remained more susceptible to D. seriata after a late pruning in winter whereas no overall seasonal changes in wound susceptibility were detected for P. chlamydospora. In the second experiment, canes were pruned by leaving two different lengths between the top node and the pruning wound before inoculations. Pathogens were recovered at different incubation periods and from different sites along the canes to estimate fungal cane colonization. A longer pruned internode made cane colonization by P. chlamydospora difficult, as indicated by fungal recoveries lower than 10% at the lowest recovery site, whereas D. seriata was less inhibited.


2021 ◽  
Author(s):  
Gabriel Perez-Gonzalez ◽  
Dana Sebestyen ◽  
Elsa Petit ◽  
Jody Jellison ◽  
Laura Mugnai ◽  
...  

Eutypa dieback and Esca are serious grapevine trunk diseases (GTDs) caused by fungal consortia causing large economic losses in vineyards. Depending on the disease the species involved include Eutypa lata, Phaeoacremonium minimum, and Phaeomoniella chlamydospora. There is a need to understand the complex pathogenesis mechanisms used by these fungi to develop treatments for the diseases they cause. Low molecular weight metabolites (LMW) are known to be involved in non-enzymatic oxygen radical generation in fungal degradation of wood by some Basidiomycota species, and as part of our work to explore the basis for fungal consortia pathogenesis, LMW metabolite involvement by the causal GTD fungi was explored. The GTD fungal pathogens examined, E. lata, P. minimum and P. chlamydospora, were found to produce low molecular weight iron binding metabolites that preferentially reduced iron or redox cycled to produce hydrogen peroxide. Uniquely, different LMW metabolites isolated from the GTD fungi promoted distinct chemistries that are important in a type of non-enzymatic catalysis known as chelator-mediated Fenton (CMF) reactions. CMF chemistry promoted by LMW metabolites from these fungi allowed for the generation of highly reactive hydroxyl radicals under conditions promoted by the fungi. We hypothesize that this new reported mechanism may help to explain the necrosis of woody grapevine tissue as a causal mechanism important in pathogenesis in these two grapevine trunk diseases.


Plants ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 207
Author(s):  
Fedon-Ioannis Gkikas ◽  
Alexandros Tako ◽  
Danai Gkizi ◽  
Christina Lagogianni ◽  
Emmanouil A. Markakis ◽  
...  

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.


Plants ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 802
Author(s):  
Pierluigi Reveglia ◽  
Regina Billones-Baaijens ◽  
Jennifer Millera Millera Niem ◽  
Marco Masi ◽  
Alessio Cimmino ◽  
...  

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.


Agronomy ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 976
Author(s):  
Natalia Langa-Lomba ◽  
Laura Buzón-Durán ◽  
Pablo Martín-Ramos ◽  
José Casanova-Gascón ◽  
Jesús Martín-Gil ◽  
...  

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.


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