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BACKGROUND: Grapevine downy mildew, caused by Plasmopara viticola, is an economically important disease in Australia and worldwide. The application of fungicides is the main tool to control this disease. Frequent fungicide applications can lead to the selection of resistant P. viticola populations, which has negative impacts on the management of the disease. Identification of resistance and its prevalence is necessary to inform resistance management strategies. RESULTS: A total of 86 P. viticola isolates were collected between 2017 and 2022 from vineyards in 15 growing regions across Australia for four fungicide groups; phenylamide (PA, group 4), carboxylic acid amide (CAA, group 40), quinone outside inhibitor (QoI, group 11) and quinone outside inhibitor stigmatellin binding type (QoSI, group 45). Decreased phenotypic sensitivity was detected for all four groups, and resistance to metalaxyl‐M (PA) and pyraclostrobin (QoI), was detected. Genetic analysis to detect the G143A (QoI) and G1105S (CAA) mutations using amplicon‐based sequencing was performed for 239 and 65 isolates collected in 2014–2017 and 2017–2022, respectively. G143A was detected in 8% and 52% of isolates, respectively, with strong association to phenotypic resistance. However, G1105S was not detected in any isolates. CONCLUSION: Plasmopara viticola isolates in Australia with resistance to at least two fungicide groups have been detected, therefore it is necessary to adopt resistance management strategies where resistance has been detected. Vineyards should continue to be monitored to improve management strategies for downy mildew. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Grapevine trunk diseases cause yield losses and vine mortality in vineyards worldwide. However, there have been few quantitative studies evaluating grapevine dieback on a large spatial and temporal scale. Here, we consolidated and standardized databases from the 13 main wine regions of France, compiling records of leaf symptoms associated with esca and Eutypa dieback from 2082 plots and 36 cultivars over a 20‐year period. This large dataset was used (a) for quantitative analysis of the prevalence (number of plots with at least one symptomatic plant) and incidence (percentage of symptomatic plants) of esca and Eutypa dieback; and (b) to decipher the effects of cultivar, year and plot age on both the prevalence and incidence of esca leaf symptoms by temporal Bayesian modelling. Esca was present on a mean of 74 ± 2% plots annually, with an incidence of 3.1 ± 0.1%. Eutypa dieback occurred in 41 ± 3% of the plots, with an incidence of 1.4 ± 0.1%. Our modelling approach revealed that the cultivar had a significant impact on the prevalence of esca, but not on its incidence when prevalence is greater than zero. Esca prevalence remained stable, whereas esca incidence was higher than the mean value in six of the years after 2012. We also found a significant non‐linear effect of plot age, with 10‐ to 30‐year‐old plots significantly more susceptible, depending on the cultivar. This study clearly illustrates the importance of considering extensive and continuous monitoring to improve our understanding of the impact and evolution of crop diseases. [ABSTRACT FROM AUTHOR]

White rot, caused by the fungus Coniella diplodiella, is an important but poorly studied disease that mainly affects grapevine clusters. White rot control typically involves the repeated application of fungicides, which may be unjustified in some cases given that the key period of berry susceptibility to infection remains unclear and controversial. In this study, germination of C. diplodiella conidia and mycelium growth were investigated on water agar (conidial germination only) and artificial media similar to berry juice at five growth stages from pea‐sized to berries ripe for harvest. On water agar, conidia germinated from 10°C to 35°C (with an optimum of 20–30°C) with >2 h of moisture incubation. Both conidial germination and mycelial growth were higher on agar similar to berry juice at véraison to berry softening than at other stages, with the lowest values detected for the ripe berries. The berries were also artificially inoculated with conidia at the different growth stages, showing that grape clusters were susceptible to infection for a long period, albeit with varying degrees of susceptibility at different stages. This implies that effective disease control requires interventions from early berry development, whenever weather conditions are conducive to infection. Further development of a predictive model accounting for weather conditions and berry susceptibility dynamics would facilitate the dynamic estimation of the disease risk during the grapevine‐growing season and contribute to a risk‐based application of fungicides for white rot control. [ABSTRACT FROM AUTHOR]

Esca is a grapevine trunk disease (GTD) that is caused by filamentous fungi. It is responsible for considerable economic losses in viniculture on a global scale. Despite many unknown factors contributing to the development of symptoms in affected plants, Phaeoacremonium minimum (PMI), Phaeomoniella chlamydospora (PCH) and Fomitiporia mediterranea (FMED) are generally considered as the main causative fungal species. Early detection and specific identification of these pathogens therefore play an important role in disease control and evaluation of suitable countermeasures. In this study, loop‐mediated isothermal amplification (LAMP) assays were developed for each of the three pathogens. A genome‐based approach was applied for detection and selection of unique target DNA sequences. The designed primer sets showed overall good specificities, with some observed cross‐reactions towards closely related Phaeoacremonium species for the PMI primer set. The developed assays had detection limits of 100 pg (FMED, PMI) and 1 pg (PCH) per reaction (corresponding to 1460 [FMED]; 1950 [PMI]; 342 [PCH] genome copies per reaction). The application of the assays to field samples was demonstrated by testing individual infected grapevine trunks from two European viticultural regions using crude DNA obtained in a rapid sample preparation step. LAMP assay results matched those of PCR following a conventional DNA extraction protocol. The study showed that LAMP‐based rapid molecular detection of major Esca agents can serve as a useful tool for further research and surveillance of a highly devastating grapevine disease. The application of computer‐based whole genome comparison between target and non‐target species for the identification of unique target sequences as the basis for LAMP (or PCR) primer design was demonstrated to be a useful approach in species for which scarce sequence information is available. Moreover, the developed method for rapid DNA preparation from grapevine trunks may potentially be adapted to the DNA‐based detection also of other fungal species that cause grapevine trunk diseases. [ABSTRACT FROM AUTHOR]

A comprehensive study was conducted on the prevalence of grapevine trunk diseases in Greece, focusing specifically on contamination of grapevine propagation material by Phaeomoniella chlamydospora and black foot disease‐related species. Additionally, detection of esca pathogen Fomitiporia mediterranea, causing white rot in grapevine and other woody hosts, was assessed using a new PCR‐based assay that distinguishes F. mediterranea from F. punctata. Development of a nested PCR assay, combined with a cost‐effective DNA extraction protocol, revealed a high percentage of infection by P. chlamydospora (51%) and Ilyonectria species associated with black foot disease (28%) in different types of grapevine propagation material (dormant cuttings, field‐rooted benchgrafts and green‐growing plants). Interestingly, black foot disease‐related pathogens were more prevalent in nursery plants grown in the field for 6 months (57%) than in other types of propagation material and compared with P. chlamydospora (43%), indicating increased infection of propagation material during growth in the nursery field. The cost‐effective molecular method developed in this study could be used in mass inspections of propagation material for phytosanitary purposes. Finally, using primers specific for F. mediterranea and F. puncata developed in this study, combined with the universal ITS4 primer, a collection of Fomitiporia isolates from mature grapevine and other woody hosts from Greece and Italy (southern Europe) were characterized as F. mediterranea, whereas German and Swedish isolates from forest plants (central Europe) were classified as F. punctata. The developed primers can discriminate between the two Fomitiporia species, which are indistinguishable based on culture and morphological characteristics alone. [ABSTRACT FROM AUTHOR]

BACKGROUND: Grapevine red blotch virus (GRBV) is a recently discovered virus and a major concern for the wine industry. Prior research indicated that GRBV delays grape ripening by reducing °Brix and anthocyanin concentrations in grapes from infected vines, resulting in higher ethanol concentrations in wines made from healthy fruit compared to diseased vines, which have an impact on sensory properties. In this study, infected fruit (Vitis vinifera L. Merlot) was sequentially harvested (in 2016 and 2017) and chaptalized (in 2017) to ameliorate the impact of GRBV on grape and final wine composition. RESULTS: Chemical parameters including phenolic and volatile profiles of grapes and their subsequent wines were measured. Sensory properties were determined by descriptive analyses. Results demonstrated that GRBV decreased sugar accumulation and anthocyanin synthesis in grapes. Wines from GRBV grapes harvested at later ripening stage produced wines that were more similar chemically and sensorially to wines made from healthy fruit than to wines made from GRBV fruit harvested earlier. CONCLUSION: A longer hang time of GRBV grapes is a potential strategy to mitigate the impacts of GRBV. However, chaptalization of diseased fruit must was inefficient at increasing similarities to wines made from healthy fruit. © 2023 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Botrytis cinerea is the causative agent of grey mould disease in grapes, which was linked to significant postharvest losses. This study examined three grape‐isolated yeasts (Metschnikowia aff. fructicola, Metschnikowia pulcherrima, and Hansenispora uvarum) through in vitro and in vivo tests on detached grape berries against grey mould, as well as the elucidation of their possible mechanisms of action. The antifungal mechanism of action of yeasts was determined by the lytic enzyme activity, inhibition of spore germination, biofilm activity, iron depletion, diffusible metabolites, wound‐site colonisation, mycocin, and volatile organic compounds (VOCs) production. The highest in vitro efficacy (83.13%) was observed on M. aff. fructicola, followed by M. pulcherrima (82.10%) and H. uvarum (71.66%). Metschnikowia yeasts exhibited comparable enzyme activities, including protease, β‐1,3 glucanase, gelatinase chitinase, and cellulase, while H. uvarum had a poor enzymatic activity with chitinase and gelatinase. M. aff. fructicola showed relatively higher iron depletion activity than M. pulcherrima, while M. pulcherrima outperformed via diffusible metabolites. All yeast cultures significantly reduced spore germination by at least 86%. Overall, M. aff. fructicola exhibited the highest biocontrol activity with its iron depletion, inhibition of conidial germination, biofilm formation, VOCs, and well colonisation on grape berries. M. aff. fructicola 1‐UDM outperformed all other yeasts by significantly reducing disease incidence and lesion diameter values (93.4% and 94.3%, respectively). Remarkably, H. uvarum VOCs demonstrated potential as a biofumigant for suppressing grey mould. All yeasts are well adapted to their ecological niche to bio‐protect grapes from grey mould disease. [ABSTRACT FROM AUTHOR]

In a consequence of global warming, grapevine trunk diseases (GTDs) have become a pertinent problem to viticulture, because endophytic fungi can turn necrotrophic upon host stress killing the plant. In Neofusicoccum parvum Bt‐67, plant‐derived ferulic acid makes the fungus release Fusicoccin aglycone triggering plant cell death. Now, we show that the absence of ferulic acid lets the fungus secrete 4‐hydroxyphenylacetic acid (4‐HPA), mimicking the effect of auxins on grapevine defence and facilitating fungal spread. Using Vitis suspension cells, we dissected the mode of action of 4‐HPA during defence triggered by the bacterial cell‐death elicitor, harpin. Early responses (cytoskeletal remodelling and calcium influx) are inhibited, as well as the expression of Stilbene Synthase 27 and phytoalexin accumulation. In contrast to other auxins, 4‐HPA quells transcripts for the auxin conjugating GRETCHEN HAGEN 3. We suggest that 4‐HPA is a key component of the endophytic phase of N. parvum Bt‐67 preventing host cell death. Therefore, our study paves the way to understand how GTDs regulate their latent phase for successful colonisation, before turning necrotrophic and killing the vines. Summary statement: During the asymptomatic phase of grapevine trunk diseases (GTDs), the aggressive Neofusicoccum parvum Bt‐67 secretes 4‐ hydroxyphenylacetic acid which has a similar structure to auxins. This metabolite suppressed plant defence mechanisms and downregulated transcripts of GH3, suggesting a key role in quelling the host response. [ABSTRACT FROM AUTHOR]

For a reliable test result to be obtained the following (external) controls should be included for each series of nucleic acid extraction and amplification of the target organism and target nucleic acid, respectively: Negative isolation control (NIC) to monitor contamination during nucleic acid extraction: nucleic acid extraction and subsequent amplification preferably of a sample of uninfected matrix, or if not available clean extraction buffer. Essential Procedural Information 3.1 Controls For a reliable test result to be obtained, the following (external) controls should be included for each series of nucleic acid extraction and amplification of the target organism and target nucleic acid, respectively Negative isolation control (NIC) to monitor contamination during nucleic acid extraction: nucleic acid extraction and subsequent amplification preferably of a sample of uninfected matrix or if not available clean extraction buffer. Inclusivity 100% evaluated on 55 target strains belonging to I X. fastidiosa i subsp. I fastidiosa i , I X. fastidiosa i subsp. I pauca i , I X. fastidiosa i subsp. I sandyi i , I X. fastidiosa i subsp. I multiplex i . A validation study with non-European I X. fastidiosa i strains (Harper et al., [56]) showed that the RST31/33 primer failed to detect the following American strains from grapes and oaks: I X. fastidiosa i , I Vitis vitifolia i , US (PD0001); I X. fastidiosa i , I V. vitifolia i , US (R. Almeida); I X. fastidiosa i , I Vitis rotundifolia i , US (C. Chang); I X. fastidiosa i , I Quercus laevis i , US (OAK0023); I X. fastidiosa i , I Quercus rubra i , US (OAK0024); I X. fastidiosa i , I Quercus rubra i , US (C. Chang). Inclusivity: 100% tested on 10 target strains ( I X. fastidiosa i subsp. I fastidiosa i , I X. fastidiosa i subsp. I pauca i , I X. fastidiosa i subsp. I sandyi i , I X. fastidiosa i subsp. I multiplex i ). [Extracted from the article]

Summary: Priming is an adaptive mechanism that fortifies plant defense by enhancing activation of induced defense responses following pathogen challenge. Microorganisms have signature microbe‐associated molecular patterns (MAMPs) that induce the primed state. The lipopolysaccharide (LPS) MAMP isolated from the xylem‐limited pathogenic bacterium, Xylella fastidiosa, acts as a priming stimulus in Vitis vinifera grapevines. Grapevines primed with LPS developed significantly less internal tyloses and external disease symptoms than naive vines.Differential gene expression analysis indicated major transcriptomic reprogramming during the priming and postpathogen challenge phases. Furthermore, the number of differentially expressed genes increased temporally and spatially in primed vines, but not in naive vines during the postpathogen challenge phase.Using a weighted gene co‐expression analysis, we determined that primed vines have more genes that are co‐expressed in both local and systemic petioles than naive vines indicating an inherent synchronicity that underlies the systemic response to this vascular pathogen specific to primed plants.We identified a cationic peroxidase, VviCP1, that was upregulated during the priming and postpathogen challenge phases in an LPS‐dependent manner. Transgenic expression of VviCP1 conferred significant disease resistance, thus, demonstrating that grapevine is a robust model for mining and expressing genes linked to defense priming and disease resistance. [ABSTRACT FROM AUTHOR]

Trunk disease is a major problem on grapevine in Zanjan province, causing serious decline, despite which its aetiology remains unknown. The purpose of this study was to investigate factors involved in grapevine decline in vineyards of Zanjan province. Samples were collected from twigs and branches of grapevines in the region between October and November 2018. Fungal isolates were identified based on morphological characteristics and ITS‐rDNA sequence data for selected isolates. The frequency and diversity of the fungal community recovered from grapevines in Darreh Sejin in Zanjan province were higher than in other regions. A total of 112 fungal isolates comprising 22 species were recovered. Phaeoacremonium minimum, Microsphaeropsis olivacea and Kalmusia variispora were identified as the dominant species in the region examined and could be considered the main trunk pathogens of grapevines in Zanjan region. In inoculation tests, M. olivacea was proved to be pathogenic on grapevine for the first time in this study. [ABSTRACT FROM AUTHOR]

Since its discovery in 2012, the role of the trichovirus named grapevine Pinot gris virus (GPGV) as the causative agent of grapevine leaf mottling and deformation (GMLD) syndrome, a grapevine disease for which symptoms had been observed in north Italy and western Slovenia a few years earlier, has been a matter of discussion. In the following 10 years of investigations, evidence has emerged of a role for the genetic diversity of the virus and for the varietal diversity of the host, as well as their interaction during antiviral defence (i.e. RNA silencing) and virus‐mediated defence suppression. In a context of wider research efforts, several relevant aspects of GPGV evolution and GMLD epidemiology were also clarified. Nevertheless, a complete understanding of the molecular events and the environmental conditions that could affect viral infection severity and symptom display remains a challenging puzzle. In the meantime, we are witnessing a large spread of the virus, which has almost completely permeated the regions where it first appeared, while spreading through most areas of grapevine cultivation. A deeper knowledge of the molecular context of the disease is key to establishing whether the impressive potential of GPGV to spread makes it a candidate as the next emerging threat for the grapevine industry. [ABSTRACT FROM AUTHOR]

1.2 Almond leaf scorch The most characteristic symptom associated with I X. fastidiosa i on almond is leaf scorching followed by decreased productivity and general tree decline. EPPO Global Database: Courtesy of M.M. Lopez Instituto Valenciano de Investigaciones Agrarias, Valencia (ES). gl 1.6 Coffee leaf scorch Symptoms of coffee leaf scorch appear on new growth of field plants as large marginal and apical scorched areas on recently developed leaves (Figure 7). The bacterium is the causal agent for Pierce's disease of grapevine, almond leaf scorch, alfalfa dwarf, oak leaf scorch, maple leaf scald, sycamore leaf scorch, mulberry leaf scorch, periwinkle wilt, pecan leaf scorch, elm leaf scorch, oleander leaf scorch, phony peach, plum leaf scald, citrus variegated chlorosis and coffee leaf scorch (Hopkins & Purcell, [28]). Disease symptoms Symptoms depend on the combination of host and I X. fastidiosa i strain and the susceptibility of the host variety. [Extracted from the article]

Coffee leaf scorch Symptoms of coffee leaf scorch appear on new growth of field plants as large marginal and apical scorched areas on recently developed leaves (Figure A7). Almond leaf scorch The most characteristic symptom associated with I X. fastidiosa i on almond is leaf scorching followed by decreased productivity and general decline of the tree. A European Commission database detailing host plants found to be susceptible to I X. fastidiosa i in the European Union is available online.https://ec.europa.eu/food/plants/plant-health-and-biosecurity/legislation/control-measures/xylella-fastidiosa/database-susceptible-host-plants en Plants showing symptoms should be sampled for laboratory testing. Bacterial leaf scorch of shade trees Symptoms of bacterial leaf scorch are similar on different tree hosts such as I Acer i spp., I Cornus florida i , I Celtis occidentalis i , I Liquidambar styraciflua i , I Morus alba i , I Platanus i spp., I Quercus i spp. and I Ulmus americana i (Gould & Lashomb, [28]). [Extracted from the article]

Here, published crop disease risk simulations are summarized, which were driven by climate change scenarios on a timescale terminating between 2001 and 2100. Thirty different agricultural and horticultural crops were specifically considered. Wheat diseases (mainly leaf rust) were most often simulated, followed by rice diseases (mainly leaf blast), grapevine diseases (mainly downy mildew) and potato diseases (mainly late blight). Most simulations suggest that within the projection period simulated, crop disease risk will more often increase (86 simulations) than decrease (45 simulations) or remain similar (12 simulations). The majority of crop disease risk simulations focus on Europe and Brazil. For example, there is agreement across crop disease risk simulations that in Europe the risk of leaf rust of wheat will increase. In other cases, there is disagreement across crop disease risk simulations, for example, due to contemporary climatic differences of locations. Therefore, it is risky to extrapolate results across locations. Mitigation and adaptation methods should be more often integrated in the crop disease risk simulations in order to inform about potential methods to reduce disease risk in future. [ABSTRACT FROM AUTHOR]

Grapevine anthracnose, caused by Elsinöe ampelina, is a fungal disease with no mapped resistance loci. Thus, this study aimed to evaluate different traits used to phenotype grapevine anthracnose looking for correlated traits and reproducibility during evaluations. Phenotyping was conducted with F1 siblings segregating for resistance to grapevine anthracnose. The progenies were inoculated with Elsinoë ampelina. Leaves, canes and shoots were phenotyped three times. Real severity and number of anthracnose spots were determined from digital photographs. Complementary, the number of spots was estimated using a scale from 0 to 9 and the severity using standard area diagram (SAD) developed for leaves and shoots. The results revealed positive correlation between real and estimated number of anthracnose spots, and the real and estimated severity. However, the number of anthracnose spots did not correlate with severity. The real data from severity and number of spots presented higher variation than the estimated data based on the SAD and scale, respectively. The evaluation strategy based on number of anthracnose spots showed reproducibility in the three independent bioassays, being the most accurate phenotyping strategy for grapevine anthracnose symptoms. [ABSTRACT FROM AUTHOR]

Several countries are most reliant on agriculture either in terms of employment opportunities, national income, availability of a raw material, food production, to name but a few. However, it faces a big challenge such as climate changes, diseases, pets, weeds etc. Therefore, last decade has provided a machine learning‐based solution to the agricultural community, which helped farmers to identify the diseases at the early stages. In this article, our focus is on grape diseases, and proposes a novel framework to identify and classify the selected diseases at the early stages. A deep learning‐based solution is embedded into a conventional architecture for optimal performance. Three primary steps are involved; (a) feature extraction after applying transfer learning on pre‐trained deep models, AlexNet and ResNet101, (b) selection of best features using proposed Yager Entropy along with Kurtosis (YEaK) technique, (c) fusion of strong features using proposed parallel approach and later subject to classification step using least squared support vector machine (LS‐SVM). The simulations are performed on infected grape leaves obtained from the plant village dataset to achieving an accuracy of 99%. From the simulation results, we sincerely believe that our proposed approach performed exceptionally compared to several existing methods. [ABSTRACT FROM AUTHOR]

Xylella fastidiosa (Xf) is the bacterial pathogen responsible for Pierce's Disease (PD) in grapevine (Vitis vinifera L.) and numerous diseases in agriculturally and ecologically important species. Current theory suggests that localized inoculations via insect feeding lead to bacterial spread through the xylem, reducing water transport capacity, leading to declines in productivity, and ultimately death. Yet, the underlying mechanisms of Xf‐induced mortality are not fully understood. In this study, we documented the development of PD symptoms over 12–13 weeks postinoculation. Subsequently assessed photosynthetic capacity, starch storage, and stem hydraulics in four grapevine genotypes (two PD‐resistant and two PD‐susceptible), comparing those physiological changes to control plants. PD‐susceptible genotypes showed a coordinated decline in photosynthesis, starch storage, and stem hydraulics, whereas Xf‐inoculation led to no change in starch and stem hydraulics in the PD‐resistant genotypes. Together these data support the idea of a link between loss of hydraulic conductivity due to tylosis production with a downstream photosynthetic decline and starch depletion in the PD‐susceptible genotypes. Our data support the theory that hydraulic failure and carbon starvation underlie plant mortality resulting from PD. [ABSTRACT FROM AUTHOR]

A search for phytoplasma‐associated diseases was conducted for the first time in the main grapevine‐growing localities of the Dukagjini plain in Kosovo. A total of 144 samples were collected from grapevine cultivars displaying leaf yellowing, reddening, discolouration and irregular wood ripening, and analysed using nested and quantitative PCR assays. These assays showed that 35.4% of samples belonging to eight cultivars were positive to the presence of phytoplasmas in the 16SrXII group. The 16S rDNA phytoplasma sequences obtained from 15 samples shared identity greater than 99.5% with 'Candidatus Phytoplasma solani'. Sequence analysis of the tuf gene showed that the strains found in Kosovar grapevines are in the tuf‐type b1 group, sharing 99.6% to 99.8% identity with 'Ca. P. solani'‐related strains associated with the "bois noir" grapevine disease in many European countries; the secY gene sequences, on the other hand, shared 100% identity with 'Ca. P. solani' strains from Bosnia and Herzegovina, Serbia, Croatia and Turkey. This study constitutes the first report on the presence and molecular characterization of phytoplasmas in Kosovar vineyards. Based on these results, it is recommended that testing for phytoplasma be included in the certification program for grapevine in Kosovo. [ABSTRACT FROM AUTHOR]

Summary: Fungi are considered to cause grapevine trunk diseases such as esca that result in wood degradation. For instance, the basidiomycete Fomitiporia mediterranea (Fmed) is overabundant in white rot, a key type of wood‐necrosis associated with esca. However, many bacteria colonize the grapevine wood too, including the white rot. In this study, we hypothesized that bacteria colonizing grapevine wood interact, possibly synergistically, with Fmed and enhance the fungal ability to degrade wood. We isolated 237 bacterial strains from esca‐affected grapevine wood. Most of them belonged to the families Xanthomonadaceae and Pseudomonadaceae. Some bacterial strains that degrade grapevine‐wood components such as cellulose and hemicellulose did not inhibit Fmed growth in vitro. We proved that the fungal ability to degrade wood can be strongly influenced by bacteria inhabiting the wood. This was shown with a cellulolytic and xylanolytic strain of the Paenibacillus genus, which displays synergistic interaction with Fmed by enhancing the degradation of wood structures. Genome analysis of this Paenibacillus strain revealed several gene clusters such as those involved in the expression of carbohydrate‐active enzymes, xylose utilization and vitamin metabolism. In addition, certain other genetic characteristics of the strain allow it to thrive as an endophyte in grapevine and influence the wood degradation by Fmed. This suggests that there might exist a synergistic interaction between the fungus Fmed and the bacterial strain mentioned above, enhancing grapevine wood degradation. Further step would be to point out its occurrence in mature grapevines to promote esca disease development. [ABSTRACT FROM AUTHOR]

Grapevine leafroll‐associated virus (GLRaV) infections are accompanied by symptoms influenced by host genotype, rootstock, environment, and which individual or combination of GLRaVs is present. Using a dedicated experimental vineyard, we studied the responses to GLRaVs in ripening berries from Cabernet Franc grapevines grafted to different rootstocks and with zero, one, or pairs of leafroll infection(s). RNA sequencing data were mapped to a high‐quality Cabernet Franc genome reference assembled to carry out this study and integrated with hormone and metabolite abundance data. This study characterized conserved and condition‐dependent responses to GLRaV infection(s). Common responses to GLRaVs were reproduced in two consecutive years and occurred in plants grafted to different rootstocks in more than one infection condition. Though different infections were inconsistently distinguishable from one another, the effects of infections in plants grafted to different rootstocks were distinct at each developmental stage. Conserved responses included the modulation of genes related to pathogen detection, abscisic acid (ABA) signalling, phenylpropanoid biosynthesis, and cytoskeleton remodelling. ABA, ABA glucose ester, ABA and hormone signalling‐related gene expression, and the expression of genes in several transcription factor families differentiated the effects of GLRaVs in berries from Cabernet Franc grapevines grafted to different rootstocks. These results support that ABA participates in the shared responses to GLRaV infection and differentiates the responses observed in grapevines grafted to different rootstocks. [ABSTRACT FROM AUTHOR]

Parasitoids exhibit distinct behaviours while foraging for their herbivorous hosts. Some are specialised with respect to the host stage they can utilise and even the age of individuals within a single stage. Observing the behavioural response of parasitoids to hosts of varying age may provide more practical understanding of potential biocontrol agents. A wind tunnel experiment was conducted to test for host‐stage dependent foraging behaviour in Goniozus jacintae Farrugia (Hymenoptera: Bethylidae), a common but understudied parasitoid of the light brown apple moth (LBAM), Epiphyas postvittana (Walker) (Lepidoptera: Tortricidae), which is a pest of grapevine, Vitis vinifera L. (Vitaceae), and other crops in Australia. Goniozus jacintae was already known to parasitise instars 3‐6 of LBAM, but most readily parasitises the later instars. Later instars are larger and have a higher rate of feeding, producing more faeces and silk deposits than earlier instars, and are likely to also produce more volatile cues that may be detected by parasitoids. We found that the timing, frequency, and duration of foraging behaviours of G. jacintae varied significantly among host instars. Flight duration was shortest when females were exposed to leaves infested by fifth instars. The profile of behaviours exhibited was different during pre‐ and post‐flight foraging, with the newly described slow walking behaviour only exhibited in close proximity to a potential host. These results contribute to the understanding of bethylid foraging, a relatively unexplored aspect of bethylid reproductive behaviour, and towards enhancing the efficacy of utilising G. jacintae in the control of leafroller pests. [ABSTRACT FROM AUTHOR]

Specific scope: This Standard describes the conduct of trials employing the CIRCE methodology (Cage method for Investigation of pheromone effects by Release and Capture, described by Doye and Koch, 2005) for the evaluation of the potential of products intended for mating disruption of relevant Lepidopteran pests in grapes and pome and stone fruit. Trials following this methodology do not replace the need for large‐scale field trials, but data from such semi‐field studies can be used as part of a registration data package supporting the potential of a given pheromone product to prevent mating in the proposed uses. The Standard should be read in conjunction with the general principles described in PP 1/264 Mating disruption techniques and in PP 1/296 Principles of efficacy evaluation for low‐risk plant protection products. Standard PP 1/314 Evaluation of mating disruption techniques against Lepidopteran pests in grapevine, pome and stone fruits under field conditions describes trials under field conditions. Specific approval and amendment: First approved in 2020–09. [ABSTRACT FROM AUTHOR]

Pierce's disease (PD) in grapevine (Vitis vinifera) is caused by the bacterial pathogen Xylella fastidiosa. X. fastidiosa is limited to the xylem tissue and following infection induces extensive plant‐derived xylem blockages, primarily in the form of tyloses. Tylose‐mediated vessel occlusions are a hallmark of PD, particularly in susceptible V. vinifera. We temporally monitored tylose development over the course of the disease to link symptom severity to the level of tylose occlusion and the presence/absence of the bacterial pathogen at fine‐scale resolution. The majority of vessels containing tyloses were devoid of bacterial cells, indicating that direct, localized perception of X. fastidiosa was not a primary cause of tylose formation. In addition, we used X‐ray computed microtomography and machine‐learning to determine that X. fastidiosa induces significant starch depletion in xylem ray parenchyma cells. This suggests that a signalling mechanism emanating from the vessels colonized by bacteria enables a systemic response to X. fastidiosa infection. To understand the transcriptional changes underlying these phenotypes, we integrated global transcriptomics into the phenotypes we tracked over the disease spectrum. Differential gene expression analysis revealed that considerable transcriptomic reprogramming occurred during early PD before symptom appearance. Specifically, we determined that many genes associated with tylose formation (ethylene signalling and cell wall biogenesis) and drought stress were up‐regulated during both Phase I and Phase II of PD. On the contrary, several genes related to photosynthesis and carbon fixation were down‐regulated during both phases. These responses correlate with significant starch depletion observed in ray cells and tylose synthesis in vessels. [ABSTRACT FROM AUTHOR]

BACKGROUND Black‐foot and Petri diseases are the main fungal diseases associated with young grapevine decline. Two field experiments were established to evaluate the preventive effect of two potential biocontrol agents (BCAs), that is Streptomyces sp. E1 + R4 and Pythium oligandrum Po37, and three BCA‐commercial products containing Trichoderma atroviride SC1, Trichoderma koningii TK7 and Pseudomonas fluorescens + Bacillus atrophaeus on fungal infection in grafted plants and plant growth parameters. RESULTS: The effectiveness of some BCA in reducing the incidence and severity of both diseases was dependent on the plant part analyzed and the plant age. No single BCA application was able to control both diseases. Streptomyces sp. E1 + R4 were able to reduce significantly the infection of the most prevalent black‐foot disease fungi while P. oligandrum Po37 and Trichoderma spp. were able to reduce significantly Phaeomoniella chlamydospora and Phaeoacremonium minimum (Petri disease) infection. BCA treatments had no effect on the shoot weight, and root weight was significantly lower in all BCA treatments with respect to the control. CONCLUSIONS: The combination of the disease‐suppressive activity of two or more beneficial microbes in a biocontrol preparation is required to prevent infection by black‐foot and Petri disease fungi in vineyards. [ABSTRACT FROM AUTHOR]

Summary: Grapevine trunk diseases have devastating consequences on vineyards worldwide. European wild grapevines (Vitis vinifera subs. sylvestris) from the last viable population in Germany along the Rhine river showed variable degrees of resistance against Neofusicoccum parvum (strain Bt‐67), a fungus associated with Botryosphaeriaceae‐related dieback.Representative genotypes from different subclades of this population were mapped with respect to their ability to induce wood necrosis, as well as their defence responses in a controlled inoculation system.The difference in colonization patterns could be confirmed by cryo‐scanning electron microscopy, while there was no relationship between vessel diameter and infection success. Resistant lines accumulated more stilbenes, that were in addition significantly partitioned to nonglycosylated viniferin trimers. By contrast, the susceptible genotypes accumulated less stilbenes with a significantly higher proportion of glycosylated piceid.We suggest a model in which in the resistant genotypes phenylpropanoid metabolism is channelled rapidly and specifically to the bioactive stilbenes. Our study specifies a resistant chemotype against grapevines trunk diseases and paves a way to breed for resistance against grapevine Botryosphaeriaceae‐related dieback. [ABSTRACT FROM AUTHOR]

Understanding how community composition of service‐providing organisms affects ecosystem functioning is a key challenge in ecology. Although it has been proposed that taxonomic diversity and functional traits mediate this relationship, how several facets of community structure affect the delivery of key ecosystem services remains to be explored.In this study, we investigated how abundance, taxonomic richness as well as the shape of biomass distribution in predator communities affect biological pest control services in vineyard landscapes. Our analyses were based on a dataset combining samples of arthropod predators, measures of predation rates of grape pests and characterization of environmental covariables for 42 fields located in South‐Western France.We found that beside the abundance or the taxonomic richness of predators, the shape of biomass distribution (mean, variance, skewness and kurtosis of the distribution) influences the level of biological control. Predator communities largely dominated by low biomass species provided the bulk of biological control services. Lower levels of predation resulted from increased proportions of large biomass species and more evenly distributed biomass values in the communities.Our results indicate that the top‐down control provided by low biomass species decreases as the relative proportion of large biomass species increases in the predator community. This suggests that biological control may be affected by negative interactions (e.g. intraguild predation, behavioural interactions) between predators arising from the recruitment of large individuals in the community.Our study revealed that the shape of biomass distribution is a major aspect of functional diversity in predator communities providing insights into the mechanisms that link biodiversity and ecosystem services. While our study focuses on biomass, considering other traits involved in trophic interactions may increase our ability to predict the level of biological control in ecosystems. A free Plain Language Summary can be found within the Supporting Information of this article. [ABSTRACT FROM AUTHOR]

BACKGROUND Black foot disease (BFD) is one of the main fungal diseases associated with young grapevine decline. Trichoderma holds the potential to be used as biocontrol agent against this disease, though variable success of colonization were found when applied to nursery vines in previous studies. Therefore, field experiments were established to evaluate different methods of application of Trichoderma atroviride, and to evaluate the efficacy of different commercial Trichoderma products on BFD in nursery vines post callusing over two seasons. RESULTS: Only in one season of the trial evaluating different products did all of the Trichoderma treatments significantly lower the black foot infections in the rootstock bases of the vines (mean black foot pathogen incidence of 1.00 to 2.50% in Trichoderma treated vines versus 6.50% in the untreated control). When comparing tissue parts, the base of the vine and collar roots had significantly higher Trichoderma colonization than the middle and root tip parts. Significantly less BFD pathogens were isolated from the base in comparison to the roots. These colonization trends were found for both field trials over both seasons. The different application methods showed that dipping of basal ends in the dry formulation followed by monthly soil drenches consistently gave higher colonization [mean Trichoderma incidence in the bases were 39.20% (2016/2017) and 28.00% (2017/2018)], while the 1 h soak of the bases of vines was not effective [mean Trichoderma incidence in the bases were 8.80% (2016/2017) and 4.00% (2017/2018)] and did not differ from the untreated control. CONCLUSION: Even though Trichoderma spp. were not sufficient to prevent infections by BFD pathogens, a certain degree of protection was obtained in the basal ends, which may contribute to longevity of the vines once planted in the vineyard. [ABSTRACT FROM AUTHOR]

Summary: Esca, a major grapevine trunk disease in old grapevines, is associated with the colonization of woody tissues by a broad range of plant pathogenic fungi. To identify which fungal and bacterial species are involved in the onset of this disease, we analysed the microbiota from woody tissues of young (10‐year‐old) grapevines at an early stage of esca. Using meta‐barcoding, 515 fungal and 403 bacterial operational taxonomic units (OTUs) were identified in woody tissues. In situ hybridization showed that these fungi and bacteria co‐inhabited in grapevine woody tissues. In non‐necrotic woody tissues, fungal and bacterial microbiota varied according to organs and seasons but not diseased plant status. Phaeomoniella chlamydospora, involved in the Grapevine trunk disease, was the most abundant species in non‐necrotic tissues from healthy plants, suggesting a possible non‐pathogenic endophytic behaviour. Most diseased plants (70%) displayed cordons, with their central white‐rot necrosis colonized essentially by two plant pathogenic fungi (Fomitiporia mediterranea: 60%–90% and P. chlamydospora: 5%–15%) and by a few bacterial taxa (Sphingomonas spp. and Mycobacterium spp.). The occurrence of a specific association of fungal and bacterial species in cordons from young grapevines expressing esca‐foliar symptoms strongly suggests that that microbiota is involved in the onset of this complex disease. [ABSTRACT FROM AUTHOR]

Botrytis cinerea, the grey mould agent, is one of the most important pathogens of grapevine, due to the great yield losses caused and the economic costs related to disease control. Ground cover plants are assumed to have a role in the complex epidemiology of the pathogen, even if no information on the genetic variability of the strains is available. In this study, a molecular epidemiology approach, based on the comparative analysis of the nucleotide sequences of multiple genes (ITS1‐ITS2, G3PDH, NEP1, NEP2, BC‐hch, and sdhB), was used to evaluate whether B. cinerea isolated from herbaceous species contributes to grey mould diffusion on grapevines. From 330 samples collected in two vineyards in Lombardy, Italy, 63 B. cinerea strains were isolated from tissues of grapevine with symptoms (50 strains) and spontaneous ground cover plants (13 strains). Capsella bursa‐pastoris, Cardamine impatiens, Lamium purpureum, and Crepis tectorum were identified as novel B. cinerea hosts. Sequence analysis and phylogeny showed that the same genotypes were present on both grapevines and herbaceous plants, with no fitness (estimated from growth and sporulation on potato dextrose agar) or pathogenicity (on grapevine leaves and berries, and tomato leaves) penalties. This confirms that ground cover plants can be a source of inoculum for B. cinerea on grapevine. Moreover, phylogenetic analysis of the BC‐hch gene allowed the identification of two genetically distinct clusters, characterized by vegetative incompatibility and different distributions of the mating types, fitness, and pathogenicity. Therefore, B. cinerea seems to be composed of two diverging subpopulations that do not differ for host specialization. [ABSTRACT FROM AUTHOR]

Ripe rot is one of the most important bunch diseases of grapes in the highlands of southern Brazil and a matter of concern for winegrowers. Sixty‐one strains isolated from berries of Vitis labrusca and V. vinifera varieties with symptoms were classified by phylogenetic analysis of the sequences of β‐tubulin, glyceraldehyde‐3‐phosphate dehydrogenase, D1/D2 domain of 28S rDNA, and rDNA internal transcribed spacer. They were also characterized by morphology, and their pathogenicity was evaluated. The combined molecular data allowed identification of six Colletotrichum species: C. fructicola, C. kahawae and C. viniferum (gloeosporioides complex), C. limitticola, C. nymphaeae (acutatum complex), and C. karstii (boninense complex). This is the first report of C. karstii and C. limitticola associated with the ripe rot of grapes. Morphological characteristics varied within and among species, confirming their separation at the complex level. Pathogenicity tests on V. vinifera berries showed that the most prevalent species, C. viniferum (37.8%) and C. fructicola (36.1%), were more virulent than the less prevalent species C. limitticola and C. karstii. Our findings indicate that there is a high diversity of Colletotrichum species associated with ripe rot disease of grapes in Brazil. There were no clear differences in the distribution of Colletotrichum species between V. labrusca and V. vinifera varieties. The determination of fungal species responsible for grape ripe rot in Brazilian vineyards may contribute to further epidemiological studies and the development of more efficient prophylactic methods for ripe rot management. [ABSTRACT FROM AUTHOR]

Grapevine rupestris stem pitting‐associated virus (GRSPaV) is one of the most widespread grapevine viruses and is transmitted mainly by grafting. GRSPaV presence was tested in 487 samples representative of the Tunisian grapevine germplasm (including autochthonous, table, wine, wild grape, and rootstock varieties) from different Tunisian regions. GRSPaV infection was detected in 51.3% of samples from different Tunisian regions, among which the table grapevine cultivars were the most commonly infected (68.7%). Genetic variability of GRSPaV isolates from wild and cultivated grapevines was assessed by sequencing the partial capsid protein (CP) gene of 19 Tunisian isolates and 1 Italian GRSPaV isolate from Sicily, and the partial RNA‐dependent RNA polymerase (RdRp) gene of 13 Tunisian GRSPaV isolates. According to phylogenetic analysis of CP nucleotide sequences obtained in this study and sequences retrieved from GenBank, Tunisian isolates fell into four phylogenetic groups already described (I, II, III, and IV) and two new phylogenetic groups (VI and VIII). Phylogenetic analysis of the partial RdRp gene revealed that Tunisian isolates of GRSPaV are distributed into four phylogroups. This study highlights the importance of regular monitoring of GRSPaV infections in Tunisia, with special regard to those grapevine accessions employed in conservation and selection programmes. In particular, the presence of new GRSPaV genetic variants and infection of wild grapevines must be taken into account in order to choose a correct control strategy. [ABSTRACT FROM AUTHOR]

Summary: Xylella fastidiosa subsp. fastidiosa causes Pierce's disease of grapevine (PD) and has been present in California for over a century. A singly introduced genotype spread across the state causing large outbreaks and damaging the grapevine industry. This study presents 122 X. fastidiosa subsp. fastidiosa genomes from symptomatic grapevines, and explores pathogen genetic diversity associated with PD in California. A total of 5218 single‐nucleotide polymorphisms (SNPs) were found in the dataset. Strong population genetic structure was found; isolates split into five genetic clusters divided into two lineages. The core/soft‐core genome constituted 41.2% of the total genome, emphasizing the high genetic variability of X. fastidiosa genomes. An ecological niche model was performed to estimate the environmental niche of the pathogen within California and to identify key climatic factors involved in dispersal. A landscape genomic approach was undertaken aiming to link local adaptation to climatic factors. A total of 18 non‐synonymous polymorphisms found to be under selective pressures were correlated with at least one environmental variable highlighting the role of temperature, precipitation and elevation on X. fastidiosa adaptation to grapevines in California. Finally, the contribution to virulence of three of the genes under positive selective pressure and of one recombinant gene was studied by reverse genetics. [ABSTRACT FROM AUTHOR]

Crown gall disease of grapevine, caused by Agrobacterium tumefaciens, often results in severe economic loss to grape production worldwide. This study demonstrated the ability of the endophytic bacteria Pseudomonas sp. Sn48 and Pantoea sp. Sa14 isolated from domesticated and wild grapevines to induce resistance in both above‐ and belowground tissues of grapevines infected with A. tumefaciens. Our results provide evidence that both strains can colonize roots and/or shoots. We showed that the strains Pseudomonas sp. Sn48 and Pantoea sp. Sa14 are capable of inducing stilbenic phytoalexin production in grapevine tissues and to further prime plantlets for enhanced phytoalexin production after A. tumefaciens inoculation. We also showed that in the majority of treatments, polyamine accumulation remained unchanged or slightly increased in plantlets treated with Pseudomonas sp. Sn48 and Pantoea sp. Sa14 compared with the control. Our findings indicated that the levels of polyamines remain unchanged or significantly decrease in plantlets treated with endophytic bacteria after A. tumefaciens challenge compared to the control and plantlets treated with individual endophytic bacterial strains. PR1, PR2, and PR4 gene expression levels of plantlets treated with Pseudomonas sp. Sn48 and Pantoea sp. Sa14 significantly increased after A. tumefaciens inoculation. The findings revealed the efficacy of the selected endophytic bacteria in triggering grapevine resistance against A. tumefaciens and the possible use of these strains as an alternative to chemical control methods in grapevine crown gall disease management. [ABSTRACT FROM AUTHOR]

Aster yellows (AY) phytoplasmas (Candidatus Phytoplasma asteris) are associated with a number of plant diseases throughout the world. Several insect vectors are responsible for spreading AY diseases resulting in wide distribution and low host specificity. Because the role of sucking insects as vectors of phytoplasmas is widely documented, and the citrus flatid planthopper Metcalfa pruinosa is a phloem feeder, it has been incriminated as a possible vector of phytoplasmas. However, its ability to transfer phytoplasma has not been confirmed. The present work shows that M. pruinosa (Hemiptera: Flatidae), a polyphagous planthopper, is able to vector Ca. P. asteris to French marigold (Tagetes patula). Transmission experiments were conducted in 2017 and 2018 in central Hungary by two approaches: (a) AY‐infected M. pruinosa were collected from an area with severe incidence of the disease on T. patula and caged on test plants for an inoculation‐access period of 2 weeks, and (b) presumably phytoplasma‐free insects were collected from apparently healthy grapevines (Vitis vinifera L.) and fed on AY‐infected T. patula plants for 2 weeks prior to being caged on test plants. AY disease symptoms developed on 4 out of 10 and 10 out of 15 test plants, respectively. All phytoplasma‐positive marigold and M. pruinosa samples showed identical RFLP patterns and shared 100% 16S rDNA sequence identity with each other and with the aster yellows phytoplasma strain AJ33 (GenBank accession number MK992774). These results indicated that the phytoplasma belonged to the phytoplasma subgroup 16SrI‐B Ca. P. asteris. Therefore, the work presented here provides experimental evidence that M. pruinosa is a vector of a 16SrI‐B subgroup phytoplasma to T. patula. [ABSTRACT FROM AUTHOR]

Downy mildew, caused by the Oomycete Plasmopara viticola, is one of the most important diseases affecting the Eurasian grapevine, Vitis vinifera. The pathogen originated in Northern America and its presence was signalled for the first time into Europe in 1878. In this study, the genetic variability and structure of Italian P. viticola population was investigated by genotyping 106 P. viticola strains belonging to 12 different regions, at 31 microsatellite markers. As a result of the high percentage of missing data, 96 strains and 19 loci were retained for the data analysis. The overall Italian population presents absence of clones, evidence of sexual and asexual reproduction and a low genetic diversity, as expected for an introduced pathogen, but a slightly higher genetic diversity than that found in other European populations, based on allelic diversity at the investigated microsatellite loci. Out of 19 loci, half shows deviation from Hardy–Weinberg equilibrium and, indeed, structure analysis indicates the presence of two separate genetic clusters, with little but significantly different distribution according to geography (west–east gradient) and climatic conditions. Overall, the analysis of the P. viticola population, 140 years after its appearance in Italy, provides indication on the pathogen adaptability. This should be taken into consideration in the future for preserving the durability of disease resistant varieties in open field. In this view, all the disease control methods available should be integrated in order to reduce the selection of pathogen strains able to overcome plant resistance. [ABSTRACT FROM AUTHOR]

BACKGROUND Fungal contamination in food products leads to mustiness, biochemical changes, and undesirable odors, which result in lower food quality and lower market value. To develop a rapid method for detecting fungi, hyperspectral imaging (HSI) was applied to identify five fungi inoculated on plates (Aspergillus niger, Aspergillus flavus, Penicillium chrysogenum, Aspergillus fumigatus, and Aspergillus ochraceus). Near‐infrared (NIR) spectroscopy, mid‐infrared (MIR) spectroscopy, and an electronic nose (E‐nose) were applied to detect and identify freeze‐dried Agaricus bisporus infected with the five fungi. RESULTS: Partial least squares regression (PLSR) models were used to distinguish the HSI spectra of the five fungi on the plates. The A. ochraceus group had the highest calibration performance: coefficient of calibration (Rc2) = 0.786, root mean‐square error of calibration (RMSEC) = 0.125 log CFU g−1. The A. flavus group had the highest prediction performance: coefficient of prediction (Rp2) = 0.821, root mean‐square error of prediction (RMSEP) = 0.083 log CFU g−1. The ratio of performance deviation (RPD) values of all of the models was higher than 2.0 for the NIR, MIR, and E‐nose results for freeze‐dried A. bisporus infected with different fungi. The fungal species and degree of infection can be distinguished by principal component analysis (PCA) and partial least squares discriminant analysis (PLS‐DA) using NIR, MIR, and E‐nose, as the discrimination accuracy was more than 90%. The NIR methods had a higher recognition rate than the MIR and E‐nose methods. CONCLUSION: Principal component analysis (PCA) and PLSR models based on full spectra of HSI can achieve good discrimination results for these five fungi on plates. Moreover, NIR, MIR, and the E‐nose were proven to be effective in monitoring fungal contamination on freeze‐dried A. bisporus. However, NIR could be a more accurate method. © 2020 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

BACKGROUND: Fungal grapevine trunk diseases (GTDs) represent a threat to viticulture, being responsible for important economic losses worldwide. Nursery and vineyard experiments were set up to evaluate the ability of Trichoderma atroviride SC1 to reduce infections of GTD pathogens in grapevine planting material during the propagation process and to assess the long‐term protection provided by this biocontrol agent on grapevine plants in young vineyards during two growing seasons. RESULTS: Reductions of some GTD pathogen incidence and severity were found on grapevine propagation material after nursery application of T. atroviride SC1 during the grafting process, and also after additional T. atroviride SC1 treatments performed during two growing seasons in young vineyards, when compared with untreated plants. CONCLUSION: Trichoderma atroviride SC1 showed promise to reduce infections caused by some GTD pathogens in nurseries, and also when establishing new vineyards. This biological control agent could possibly be a valuable component in an integrated management approach where various strategies are combined to reduce GTD infections. © 2019 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Phaeomoniella chlamydospora and wood‐rotting basidiomycete fungi, namely Fomitiporia spp., are known esca pathogens. However, the effect of their mixed infections and the sequence of infection on disease development is unclear. To determine the effects of single and co‐inoculations on symptoms, potted Vitis vinifera 'Crimson Seedless' was inoculated with P. chlamydospora either alone or in combination with one of four basidiomycetes: Coprinellus radians, Fomitiporia langloisii, F. polymorpha and the novel species Tropicoporus texanus. Basidiomycetes were isolated from vines with foliar symptoms of esca in California and Texas. In sequential co‐inoculations, the effects of different sequences of infection (P. chlamydospora first, basidiomycete 6 months later; and vice versa) were tested, compared to simultaneous co‐inoculations. Plants inoculated with P. chlamydospora either alone or in combination with a basidiomycete (in any sequence) did not differ significantly in the length of black‐streaking lesions. In plants inoculated only with a basidiomycete, the appearance of large brown lesions, coupled with the absence of this wood symptom from control plants, suggests that C. radians, F. langloisii and T. texanus are pathogenic. Foliar symptoms resembling those of esca in the field (marginal and/or interveinal scorching, combined with red and/or yellow discoloration) were statistically more frequent among plants inoculated with F. polymorpha or T. texanus, either simultaneously or following P. chlamydospora, compared to single inoculations. Sequential co‐inoculations of a basidiomycete before or after P. chlamydospora were associated with similar lesion lengths, suggesting that basidiomycetes may not require infection by P. chlamydospora in order to extensively colonize the wood. [ABSTRACT FROM AUTHOR]

Fungal entomopathogens like Beauveria bassiana (Ascomycota: Hypocreales) are known as antagonists of insects with multiple functional and ecological roles, and have attracted increased attention as biocontrol agents in integrated pest management programmes. For some crop plants, it has been proven that endophytic B. bassiana, besides its entomopathogenic habit, can provide protection against plant pathogens or limit their damaging effects. However, for grapevine, limited knowledge is available on the influence of endophytic B. bassiana on fungal pathogens and about the mechanisms underlying putative protection effects. This study assessed the protective potential of endophytic B. bassiana against grapevine downy mildew Plasmopara viticola in greenhouse experiments. Three and seven days after a B. bassiana treatment, potted grapevine plants were inoculated with P. viticola and the evolving disease severity was assessed. Disease severity was significantly reduced in B. bassiana‐treated plants compared to control plants, depending on the age of leaves. Furthermore, a microarray and an RT‐qPCR analysis were performed to work out fundamental aspects of genes involved in the interaction between grapevine and B. bassiana. The results indicate an up‐regulation of diverse defence‐related genes in grapevine as a response to endophytic establishment of B. bassiana. Thus, endophytic establishment of an entomopathogenic fungus such as B. bassiana in grapevine plants would represent an alternative and sustainable plant protection strategy, with the potential for reducing pesticide applications in viticulture. [ABSTRACT FROM AUTHOR]

Phakopsora meliosmae‐myrianthae, the causal agent of Asian grapevine leaf rust, significantly reduces the photosynthetic efficiency of grapevine leaves in green symptomless tissues surrounding lesions. This study took a close look at grapevine leaf colonization kinetics by P. meliosmae‐myrianthae and compared it to P. pachyrhizi–soybean and Uromyces appendiculatus–bean colonization. It is already known from the literature that soybean rust, similar to grapevine rust, has a negative effect on leaf photosynthesis greater than would be expected based on visual lesions. However, in contrast to soybean and grapevine rusts, the effect of bean rust on leaf photosynthesis is proportional to the diseased leaf area. Colonization progress was monitored by fungal biomass assessed via histological staining and quantitative polymerase chain reaction (qPCR). Individual lesions of P. meliosmae‐myrianthae on grapevine, P. pachyrhizi on soybean and U. appendiculatus on common bean leaves were evaluated every 3–4 days, and the number of uredinia was counted. Staining showed that mycelial colonization did not extend beyond the lesion border. The number of P. pachyrhizi and P. meliosmae‐myrianthae uredinia within the lesions increased over time (on average 14‐fold), whereas the number of U. appendiculatus uredinia remained the same. These findings were corroborated by qPCR, which revealed a greater increase in fungal biomass for Phakopsora spp. than for U. appendiculatus until 12 days post‐inoculation. The high number of satellite uredinia within lesions might be directly related to the impact of this pathogen in photosynthetic efficiency on symptomless areas of diseased grapevine leaves. This study identified accelerated formation of satellite uredinia as an important feature of grapevine colonization by P. meliosmae‐myrianthae. [ABSTRACT FROM AUTHOR]

Bois noir (BN), the most prevalent disease of the grapevine yellows complex, causes considerable yield loss in vineyards. BN is associated with phytoplasma strains of the species 'Candidatus Phytoplasma solani' (taxonomic subgroup 16SrXII‐A). In Europe, the BN phytoplasma is transmitted to grapevine mainly by Hyalesthes obsoletus, a polyphagous cixiid completing its life cycle on stinging nettle and field bindweed. As a result of the complexity of BN epidemiology, no effective control strategies have been developed. In previous studies conducted in the eastern Mediterranean coast of Israel, chaste tree (Vitex agnus‐castus) was found to be the preferred host plant of H. obsoletus but did not harbour BN phytoplasma. Thus, a 'push and pull' strategy was suggested based on the fact that chaste tree plants located at vineyard borders was an effective trap plant for H. obsoletus adults. However, in other studies carried out in the eastern Adriatic coast of Montenegro, chaste tree was found to be a key source plant for BN phytoplasma transmission to grapevine. This study aimed to investigate (i) the interaction between chaste tree and H. obsoletus through survival, attractiveness and oviposition experiments conducted comparing the behaviour of H. obsoletus in chaste tree versus stinging nettle and grapevine and (ii) the capability of chaste tree to harbor 'Ca. P. solani' in northern Italy through transmission trials. H. obsoletus adults were found to survive on chaste tree and grapevine over a 1 week period and prefer chaste tree to grapevine. Moreover, H. obsoletus produced eggs and overwintered as nymphs on chaste tree, even if at a lesser extent than on stinging nettle. H. obsoletus originating from nettle was found able to transmit 'Ca. P. solani' to chaste tree (2 plants of 16 were found infected by the BN phytoplasma strain St5 identified in H. obsoletus specimens). These results increased our knowledge about the role of Vitex agnus‐castus as host plant of H. obsoletus and BN phytoplasma in northern Italy and do not recommend considering chaste tree as trap plant at vineyard borders. [ABSTRACT FROM AUTHOR]

Summary: Europe is the world largest grape producer, but in recent years, the report of diseases due to infection by grapevine trunk pathogens (GTPs) is becoming one of the main constraints for viticulture. Among grapevine trunk diseases (GTDs), esca syndrome is one of the most complex, characterized by simultaneous infection of several fungi, which leads to important reduction in yield and quality. Previous characterization of fungal isolates associated with esca syndrome leads to the recognition of at least three important players: Phaeocremonium minimum, Phaemoniella chlamydospora and Fomitiporia mediterranea. Here we isolated and characterized molecularly fungal endophytes directly from field wood tissues of plants showing or not esca symptoms. In addition, to better characterize such collection, a deep RNA sequencing (100 M reads in paired‐end) to screen for mycovirus presence was performed. Thirty‐nine viral genomes were detected, 38 of which were putative new viral species; some of these viruses infected GTPs, including P. minimum and F. mediterranea. In this work, we reported for the first time a curated collection of grapevine fungal endophytes identifying the associated mycoviruses some of which could be employed in future biotechnological exploitation as biological control agents for sustainable plant protection. [ABSTRACT FROM AUTHOR]

In a collection of 735 Botrytis isolates sampled from Australian wine grape‐growing regions, a single isolate from clade I and group I (based on Bc‐hch RFLP analysis) was found. As many Botrytis species are known to live sympatrically, it was hypothesized that this isolate might be a new Botrytis species. After phenotypic and molecular assays supported this hypothesis, the species was designated B. medusae. Phylogenetic analyses using the nuclear genes G3PDH,HSP60,RPB2,NEP1 and NEP2 consistently placed B. medusae in an early‐diverging clade I Botrytis spp. lineage. Botrytis medusae produced white aerial mycelium, grew faster at 30 °C and produced long‐branched conidiophore extensions, compared with B. cinerea and B. pseudocinerea. Botrytis medusae was only able to infect wounded grape leaves and was significantly less virulent on wounded leaves and berries than B. cinerea. Botrytis medusae also lacked villiform appendages on the conidial surface and long conidiophores, which are defining features of B. sinoviticola and B. californica, respectively. Identification and characterization of new cryptic Botrytis species living in sympatry on grapevines could potentially provide information to assist disease management strategies for B. cinerea. [ABSTRACT FROM AUTHOR]

Downy mildew (Plasmopara viticola) is one of the most important diseases in grape‐growing areas worldwide, including Brazil. To examine pathogen population biology and structure, P. viticola was sampled during the 2015/16 growing season from 516 lesions on nine grape cultivars in 11 locations in subtropical areas of São Paulo State, Brazil. For identification of cryptic species, a subsample of 130 isolates was subjected to cleaved amplified polymorphic sequence (CAPS) analysis, and for 91 of these isolates the ITS1 region was sequenced. These analyses suggest that the population of P. viticola in São Paulo State consists of a single cryptic species, P. viticola clade aestivalis. Seven microsatellite markers were used to determine the genetic structure of all 516 P. viticola isolates, identifying 23 alleles and 55 multilocus genotypes (MLGs). Among these MLGs, 34.5% were clonal and represented 93% of the isolates sampled. Four dominant genotypes were present in at least five different locations, corresponding to 65.7% of the isolates sampled. Genotypic diversity (Ĝ = 0.21–0.89) and clonal fraction (0.58–0.96) varied among locations (populations). Most populations showed significant deviation from Hardy–Weinberg expectations; in addition, excess of heterozygosity was verified for many loci. However, principal coordinate analysis revealed no clusters among locations and no significant isolation by distance was found, suggesting high levels of migration. The results indicate that downy mildew epidemics result from multiple clonal infections caused by a few genotypes of P. viticola, and reproduction of P. viticola in São Paulo State is predominantly asexual. [ABSTRACT FROM AUTHOR]

We describe a droplet‐vitrification cryotherapy method for the eradication of grapevine leafroll‐associated virus‐3 (GLRaV‐3) from diseased in vitro shoots of Vitis plants. The procedure involved pre‐culture of 1.0‐mm shoot tips containing five to six leaf primordia (LPs) for 3 days with a pre‐culture medium containing 0.3 M sucrose, 0.16 mM glutathione and 0.14 mM ascorbic acid, treatment of the pre‐cultured shoot tips for 20 min at room temperature with a loading solution composed of 2 M glycerol and 0.4 M sucrose and exposure to plant vitrification solution 2 (PVS2), prior to freezing in liquid nitrogen of dehydrated shoot tips contained in 2.5‐μL PVS2 droplets on aluminium foil strips. Virus localisation showed GLRaV‐3 was not present in apical dome (AD) and LPs 1–4, but it was detected in the basal shoot tip region, approximately 0.5 mm from the AD, as well as in LP 5 and more mature tissues. Histological observations identify that only freezing in liquid nitrogen results in the death of all cells in areas of shoot tips harbouring virus, whereas PVS2 treatment does not. Thus, freezing in liquid nitrogen is a necessary step that eradicates GLRaV‐3. This cryotherapy procedure produced shoot regrowth levels that ranged from 43% to 59%, and all plants recovered after cryotherapy were free of GLRaV‐3 in two wine, one table and one rootstock cultivars. Thus, this procedure can be considered to be efficient and wildly applicable for eradication of GLRaV‐3 from Vitis spp. A shoot tip cryotherapy was described for efficient eradication of grapevine leafroll‐associated virus‐3. Mechanism for eradication of GLRaV‐3 by shoot tip cryotherapy was provided. [ABSTRACT FROM AUTHOR]

Abstract: Bunch‐zone leaf removal reduced infestation by Lobesia botrana, although the mechanism responsible for this effect is unknown. Based on the mortality of eggs and newly‐hatched larvae exposed to high temperatures (≥37 °C) in the laboratory, the present study aimed to assess the influence of (i) bunch‐zone leaf removal and grapevine‐row orientation on berry surface temperature and (ii) bunch exposure to sunlight on egg and larval mortality. Berry temperatures were measured using a noncontact infrared thermometer in two vineyards and showed that, in direct sunlight, the temperatures of berry surfaces were 9 °C or more above air temperature, and the highest mean temperatures occurred on southwest‐side bunches followed by west‐ and south‐side bunches. The results of four two‐choice field assays, carried out confining fertile females in cages with two bunches, one exposed and one non‐exposed to sunlight, showed that: (i) the females did not avoid laying eggs on sun‐exposed bunches and (ii) the lowest percentages of both egg‐hatching and larval settlement occurred on sun‐exposed bunches. The hypothesis that the high temperatures reached by sun‐exposed berries cause egg and especially larval mortality is confirmed. Bunch‐zone leaf removal combined with optimized grapevine‐row orientations can improve L. botrana control. [ABSTRACT FROM AUTHOR]

Summary: Xylella fastidiosa is a Gram‐negative bacterial plant pathogen with an extremely wide host range. This species has recently been resolved into subspecies that correlate with host specificity. This review focuses on the status of X. fastidiosa pathogenic associations in plant hosts in which the bacterium is either endemic or has been recently introduced. Plant diseases associated with X. fastidiosa have been documented for over a century, and much about what is known in the context of host–pathogen interactions is based on these hosts, such as grape and citrus, in which this pathogen has been well described. Recent attention has focused on newly emerging X. fastidiosa diseases, such as in olives. Taxonomy: Bacteria; Gammaproteobacteria; family Xanthomonadaceae; genus Xylella; species fastidiosa. Microbiological properties: Gram‐negative rod (0.25–0.35 × 0.9–3.5 μm), non‐flagellate, motile via Type IV pili‐mediated twitching, fastidious. Host range: Xylella fastidiosa has a broad host range that includes ornamental, ecological and agricultural plants belonging to over 300 different species in 63 different families. To date, X. fastidiosa has been found to be pathogenic in over 100 plant species. In addition, it can establish non‐symptomatic associations with many plants as a commensal endophyte. Here, we list the four distinct subspecies of X. fastidiosa and some of the agriculturally relevant diseases caused by them: X. fastidiosa ssp. fastidiosa causes Pierce's disease (PD) of grapevine (Vitis vinifera); X. fastidiosa ssp. multiplex causes almond leaf scorch (ALS) and diseases on other nut and shade tree crops; X. fastidiosa ssp. pauca causes citrus variegated chlorosis (CVC) (Citrus spp.), coffee leaf scorch and olive quick decline syndrome (OQDS) (Olea europaea); X. fastidiosa ssp. sandyi causes oleander leaf scorch (OLS) (Nerium oleander). Significant host specificity seemingly exists for some of the subspecies, although this could be a result of technical biases based on the limited number of plants tested, whereas some subspecies are not as stringent in their host range and can infect several plant hosts. Disease symptoms: Most X. fastidiosa‐related diseases appear as marginal leaf necrosis and scorching of the leaves. In the case of PD, X. fastidiosa can also cause desiccation of berries (termed ‘raisining’), irregular periderm development and abnormal abscission of petioles. In olive trees affected with OQDS, leaves exhibit marginal necrosis and defoliation, and overall tree decline occurs. Plants with ALS and OLS also exhibit the characteristic leaf scorch symptoms. Not all X. fastidiosa‐related diseases exhibit the typical leaf scorch symptoms. These include CVC and Phony Peach disease, amongst others. In the case of CVC, symptoms include foliar wilt and interveinal chlorosis on the upper surfaces of the leaves (similar to zinc deficiency), which correspond to necrotic, gum‐like regions on the undersides of the leaves. Additional symptoms of CVC include defoliation, dieback and hardening of fruits. Plants infected with Phony Peach disease exhibit a denser, more compact canopy (as a result of shortened internodes, darker green leaves and delayed leaf senescence), premature bloom and reduced fruit size. Some occlusions occur in the xylem vessels, but there are no foliar wilting, chlorosis or necrosis symptoms . Useful websites: http://www.piercesdisease.org/; https://pubmlst.org/xfastidiosa/; http://www.xylella.lncc.br/; https://nature.berkeley.edu/xylella/; https://ec.europa.eu/food/plant/plant_health_biosecurity/legislation/emergency_measures/xylella-fastidiosa_en [ABSTRACT FROM AUTHOR]

Abstract: Anthracnose caused by Elsinoë ampelina is one of the most important table grape diseases in humid regions in Brazil and Australia. The objective of this study was to characterize E. ampelina isolates from Brazil and Australia by means of phylogenetic analyses, morphological features and pathogenicity tests. Phylogenetic relationships among 35 isolates were determined based on a data set of internal transcribed spacer (ITS), histone H3 (HIS3) and elongation factor 1‐α (TEF) sequences. In phylogenetic tree analyses, using a combined ITS and TEF sequence alignment, all E. ampelina isolates were clustered together in a single well‐supported clade. In contrast to the absence of genetic variability within ITS and TEF sequences, HIS3 sequences showed 54 polymorphic sites. The haplotype network generated from HIS3 data set showed four distinct haplotypes. EA1 was the predominant haplotype including 29 isolates from both countries. High genetic variability was observed in two Brazilian isolates, haplotype EA4, which may have lost the intron region during species evolution. Colony colours differed between Brazilian and Australian isolates, but showed similar wrinkled colony texture, absence of spores, sparse‐to‐absent white aerial mycelium and slow growth (0.049–0.060 mm/day). Brazilian isolates produced conidia of 5.65 × 2.65 μm, larger than conidia from Australian isolates, which measured 5.14 × 2.30 μm. In pathogenicity tests, all nine Australian isolates inoculated were pathogenic on detached canes and potted vines of table grape. [ABSTRACT FROM AUTHOR]

Summary: Grapevines, like other perennial crops, are affected by so‐called ‘trunk diseases’, which damage the trunk and other woody tissues. Mature grapevines typically contract more than one trunk disease and often multiple grapevine trunk pathogens (GTPs) are recovered from infected tissues. The co‐existence of different GTP species in complex and dynamic microbial communities complicates the study of the molecular mechanisms underlying disease development, especially under vineyard conditions. The objective of this study was to develop and optimize a community‐level transcriptomics (i.e. metatranscriptomics) approach that could monitor simultaneously the virulence activities of multiple GTPs in planta. The availability of annotated genomes for the most relevant co‐infecting GTPs in diseased grapevine wood provided the unprecedented opportunity to generate a multi‐species reference for the mapping and quantification of DNA and RNA sequencing reads. We first evaluated popular sequence read mappers using permutations of multiple simulated datasets. Alignment parameters of the selected mapper were optimized to increase the specificity and sensitivity for its application to metagenomics and metatranscriptomics analyses. Initial testing on grapevine wood experimentally inoculated with individual GTPs confirmed the validity of the method. Using naturally infected field samples expressing a variety of trunk disease symptoms, we show that our approach provides quantitative assessments of species composition, as well as genome‐wide transcriptional profiling of potential virulence factors, namely cell wall degradation, secondary metabolism and nutrient uptake for all co‐infecting GTPs. [ABSTRACT FROM AUTHOR]