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Botryosphaeriaceae cause the degenerative disease Botryosphaeria dieback in many woody hosts, including grapevine. These pathogens penetrate host plants through pruning wounds, and colonize vascular tissues causing necrotic lesions, cankers, and eventually plant death. Colonization processes by Botryosphaeriaceae and their interactions with their hosts has been understudied. The colonization mechanisms were examined for Lasiodiplodia brasiliensis, a common pathogen causing Botryosphaeria dieback in Mexican vineyards. Lasiodiplodia brasiliensis MXBCL28 was inoculated onto grapevine ‘Cabernet Sauvignon’ plants, and after 2 months, infected tissues were observed with microscopy using histological techniques. Lasiodiplodia brasiliensis was also cultured on different carbon sources representing cell walls and non-structural plant components, to complement histology data. The host responded to wounding by developing xylem vessel occlusions with tyloses and deposition of suberin in cambium and ray parenchyma. Infection response also included deposition of suberin in pith tissues, reinforcement of cell walls with phenolic compounds, and lignin deposition in xylem vessels and ray parenchyma. The pathogen could overcome host compartmentalization mechanisms and colonize wood tissue causing extensive necrosis. The fungus was visualized in host cambium, vascular bundles, xylem vessels, and pith, and infected tissues were depleted in starch in the ray parenchyma. Cellulose, hemicellulose, and lignin in cell walls were also degraded, supporting in vitro data. [ABSTRACT FROM AUTHOR]

A key determinant of plant resistance to vascular infections lies in the ability of the host to successfully compartmentalize invaders at the xylem level. Growing evidence supports that the structural properties of the vascular system impact host vulnerability towards vascular pathogens. The aim of this study was to provide further insight into the impact of xylem vessel diameter on compartmentalization efficiency and thus vascular pathogen movement, using the interaction between Vitis and Phaeomoniella chlamydospora as a model system. We showed experimentally that an increased number of xylem vessels above 100 μm of diameter resulted in a higher mean infection level of host tissue. This benchmark was validated within and across Vitis genotypes. Although the ability of genotypes to restore vascular cambium integrity upon infection was highly variable, this trait did not correlate with their ability to impede pathogen movement at the xylem level. The distribution of infection severity of cuttings across the range of genotype's susceptibility suggests that a risk‐based mechanism is involved. We used this experimental data to calibrate a mechanistic stochastic model of the pathogen spread and we provide evidence that the efficiency of the compartmentalization process within a given xylem vessel is a function of its diameter. We demonstrate that xylem vessel diameter impacts the compartmentalization efficiency of the vascular pathogen Phaeomoniella chlamydospora within a single, and across many, Vitis genotypes. Our results highlight that structural traits of the plant vascular system shape the capability of plant hosts to respond to pathogen infection. [ABSTRACT FROM AUTHOR]

Trunk diseases are factors that limit sustainability of vineyards worldwide. Botryosphaeria and Eutypa diebacks are caused by several fungi belonging to the Botryosphaeriaceae and Diatrypaceae, respectively, with Diplodia seriata and Eutypa lata being two of the most common species. Previous information indicated that the traditional isolation method used to detect these pathogens from plant samples could underestimate their incidence levels. In the present study, we designed two sets of primers that target the β-tubulin gene and that are amenable for quantitative real-time PCR (qPCR) Sybr-Green assays for the detection and quantification of D. seriata-complex (DseCQF/R) and E. lata (ElQF/R) DNA. The design of a species-specific assay was achieved for E. lata. For D. seriata, a species-specific assay could not be designed. The low interspecific diversity across β-tubulin genes resulted in an assay that could not discriminate D. seriata from some closely related species either not yet reported or presenting a low prevalence on grapevine, such as D. intermedia. We validated our technique on grapevine spur samples naturally and artificially infected with D. seriata and E. lata during the dormant season. Experimental grapevines were located in two counties of northern California where the incidence of both pathogens was previously reported. The qPCR assays revealed that a high frequency of pruning wound infections (65%) was achieved naturally by E. lata, while low infection frequency (less than 5%) was observed using the reisolation method. For D. seriata-complex, low (5%) to no natural infection frequencies were observed by the qPCR and the reisolation method, respectively. These results also provided evidence that our qPCR detection methods were more sensitive to assess the incidence of E. lata and D. seriata-complex in plant samples, than traditional isolation techniques. Benefits of molecular methods for the detection of canker pathogens in the field under natural conditions are discussed. [ABSTRACT FROM AUTHOR]

The fastidious phytopathogenic bacterium, Xylella fastidiosa , poses a substantial threat to many economically important crops, causing devastating diseases including Pierce’s Disease of grapevine. Grapevines ( Vitis vinifera L.) planted in an area under Pierce’s Disease pressure often display differences in disease severity and symptom expression, with apparently healthy vines growing alongside the dying ones, despite the fact that all the vines are genetic clones of one another. Under the hypothesis that endophytic microbes might be responsible for this non-genetic resistance to X. fastidiosa , endophytic fungi were isolated from vineyard cvs. ‘Chardonnay’ and ‘Cabernet Sauvignon’ grown under high Pierce’s Disease pressure. A Cochliobolus sp. isolated from a Cabernet Sauvignon grapevine inhibited the growth of X. fastidiosa in vitro . Bioassay-guided isolation of an organic extract of Cochliobolus sp. yielded the natural product radicinin as the major active compound. Radicinin also inhibited proteases isolated from the culture supernatant of X. fastidiosa . In order to assess structure–activity relationships, three semi-synthetic derivatives of radicinin were prepared and tested for activity against X. fastidiosa in vitro . Assay results of these derivatives are consistent with enzyme inactivation by conjugate addition to carbon-10 of radicinin, as proposed previously. [ABSTRACT FROM AUTHOR]

Eutypa dieback of grapevine is caused by Eutypa lata in production areas with Mediterranean climates in California, Australasia, Europe, and South Africa. Eutypa dieback has also been described in the colder, eastern North American vineyards where cultivars adapted from native Vitis spp. (e.g., Vitis x labruscana 'Concord') are primarily grown. However, the causal agents associated with the diseases in this region have not been conclusively identified. Examination of 48 vine-yards showing symptoms of dieback in the northeastern United States (Connecticut, Massachusetts, Michigan, New York, Ohio, and Rhode Island) and Ontario, Canada revealed that vineyards were mainly in-fected by Eutypa spp. other than E. lata. Multigene phylogenies (inter- nal transcribed spacer ribosomal DNA, ß-tubulin, and RNA polymer-ase II) of isolates recovered from these vineyards indicated that Eutypa dieback is caused primarily by an undescribed Eutypa sp. and E. lae-vata. Eutypa sp. was recovered from 56% of the vineyards examined, whereas E. laevata and E. lata were less far common (17 and 6%, respectively). Fruiting body morphology and spore dimensions supported phylogenetic separation of the three taxa. Pathogenicity tests conducted on Vitis vinifera 'Chardonnay' in the greenhouse and in the field verified that all three species were able to cause wood canker and to infect pruning wounds, respectively. [ABSTRACT FROM AUTHOR]

Cultivars of European grapevine, Vitis vinifera, show varying levels of susceptibility to Eutypa dieback and Esca, in terms of foliar symptoms. However, little is known regarding cultivar susceptibility of their woody tissues to canker formation. Accordingly, we evaluated the relative susceptibility of V. vinifera cultivars ('Cabernet Franc', 'Caber-net Sauvignon', 'Chardonnay', 'Merlot', 'Riesling', 'Petite Syrah', and 'Thompson Seedless') and species or interspecific hybrids of North American Vitis (Vitis hybrid 'Concord', V. arizonica 'b42-26', V. rupestris × V. cinerea '111547-1', and Fennell 6 [V. aestivalis] × Malaga [V. vinifera] 'DVIT0166') to canker formation by seven trunk patho-gens (Neofusicoccum pan'um, Lasiodiplodia theobromae, Phaeomoni-ella chlamydospora, Togninia minima, Phomopsis viticola, Eutypa lata, and an undescribed Eutypa sp.). Susceptibility was based on the length of wood discoloration (LWD) in the woody stems of rooted plants in duplicate greenhouse experiments. Cultivars of V. vinifera and Concord did not vary significantly in susceptibility to N. pan'um or L. theobromae (LWD of 21 to 88 mm at 14 weeks post inoculation; P > 0.16), suggesting that they are similarly susceptible to Botryosphaeria dieback. The table-grape Thompson Seedless was most susceptible to P. viticola (mean LWD of 61 mm at 11 months post inoculation; P < 0.0001). V. vinifera cultivars and Concord showed similar susceptibility to the Esca pathogens, Phaeomoniella chlamydospora and T. minima. Susceptibility to E. lata was greatest in V. arizonica b42-26 (mean LWD of 96 mm at 11 months post inoculation; P < 0.03). In fact, all four American Vitis spp. were more susceptible to Eutypa dieback than the V. vinifera cultivars. Our findings suggest that no one cultivar is likely to provide resistance to the range of trunk pathogens but that certain cultivars may be promising candidates for commercially rele-vant host resistance in grape-production systems where the dominant cultivars are very susceptible. [ABSTRACT FROM AUTHOR]

We sequenced and comprehensively analysed the genomic architecture of 98 fluorescent pseudomonads isolated from different symptomatic and asymptomatic tissues of almond and a few other Prunus spp. Phylogenomic analyses, genome mining, field pathogenicity tests, and in vitro ice nucleation and antibiotic sensitivity tests were integrated to improve knowledge of the biology and management of bacterial blast and bacterial canker of almond. We identified Pseudomonas syringae pv. syringae, P. cerasi, and P. viridiflava as almond canker pathogens. P. syringae pv. syringae caused both canker and foliar (blast) symptoms. In contrast, P. cerasi and P. viridiflava only caused cankers, and P. viridiflava appeared to be a weak pathogen of almond. Isolates belonging to P. syringae pv. syringae were the most frequently isolated among the pathogenic species/pathovars, composing 75% of all pathogenic isolates. P. cerasi and P. viridiflava isolates composed 8.3 and 16.7% of the pathogenic isolates, respectively. Laboratory leaf infiltration bioassays produced results distinct from experiments in the field with both P. cerasi and P. syringae pv. syringae, causing significant necrosis and browning of detached leaves, whereas P. viridiflava conferred moderate effects. Genome mining revealed the absence of key epiphytic fitness-related genes in P. cerasi and P. viridiflava genomic sequences, which could explain the contrasting field and laboratory bioassay results. P. syringae pv. syringae and P. cerasi isolates harboured the ice nucleation protein, which correlated with the ice nucleation phenotype. Results of sensitivity tests to copper and kasugamycin showed a strong linkage to putative resistance genes. Isolates harbouring the ctpV gene showed resistance to copper up to 600 μg/ml. In contrast, isolates without the ctpV gene could not grow on nutrient agar amended with 200 μg/ml copper, suggesting ctpV can be used to phenotype copper resistance. All isolates were sensitive to kasugamycin at the label-recommended rate of 100μg/ml. [ABSTRACT FROM AUTHOR]

Vine training and pruning are cultural strategies that can be deployed to manage grapevine trunk diseases (GTDs). Forty-year-old commercial vineyards in the Cognac region, France, trained to either Guyot-Arcure (severe pruning) or Guyot-Poussard (minimal pruning), were studied to determine how the two systems affected trunk disease symptomatology. Effects of pruning practices on the pathobiome and mycobiome of asymptomatic grapevines were also assessed, using culture-and amplicon-based Illumina sequencing approaches. The hypothesis examined was that severe pruning of Guyot-Arcure increases trunk diseases incidence and severity, and causes higher pathogen load and microbial diversity, compared to Guyot-Poussard. Numbers of symptomatic and asymptomatic vines for the two training systems were recorded over 3 years, including numbers of vines with esca foliar symptoms, and partially unproductive and dead vines. Six asymptomatic vines from each pruning method were selected, and culturing and sequencing data were obtained from 27 samples per vine. Fungi in the Phaeomoniellaceae, Togniniaceae, and Botryosphaeriaceae were the most frequently identified. The data indicated that severe pruning increased risk of pathogen infections, with Phaeomoniella chlamydospora, Phaeoacremonium minimum and Diplodia sp. being the most commonly identified fungi. Greater numbers of dead or dying vines were recorded in the severely pruned vineyard, indicating that this strategy shortens vine longevity. Results also showed that severe pruning increased endophytic microbial diversity, and that the pruning methods influenced mycobiome community composition. This knowledge will improve recommendations to growers for practical and cost-effective ways to manage GTDs. [ABSTRACT FROM AUTHOR]

Botryosphaeria dieback is one of the most prevalent grapevine trunk diseases (GTDs), and is caused by fungi in the Botryosphaeriaceae. Fungi invade grapevine vascular systems mainly through pruning wounds, and cause cankers and necrotic lesions, which lead to grapevine decline and death. Lasiodiplodia theobromae has been reported as a highly virulent pathogen of grapevine, and was previously reported in Mexican vineyards. The taxonomy of Lasiodiplodia was recently revised, adding new species, and some were reduced to synonymy. This study aimed to characterize Lasiodiplodia producing grapevine dieback symptoms in Sonora and Baja California, Mexico. Using the phylogenetic markers tef1-a and ITS regions, Lasiodiplodia brasiliensis, L. crassispora, L. exigua, and L. gilanensis were identified. Lasidiplodia exigua was the most prevalent species. Lasiodiplodia brasiliensis and L. gilanensis were very virulent to 'Cabernet Sauvignon' plants, while L. exigua and L. gilanensis were less virulent, and L. crassispora did not produce lesions at 2 months post-inoculation. The optimum temperature of the Lasiodiplodia spp. was 28°C, but all four species grew up to 37°C, and the isolates of L. exigua grew slowly at 40°C. This is the first report of the four of Lasiodiplodia species in vineyards of Mexico. [ABSTRACT FROM AUTHOR]

Optimizing 'Hass' avocado (Persea americana Mill.) tree nutrient status is essential for maximizing productivity. Leaf nutrient analysis is used to guide avocado fertilization to maintain tree nutrition. The goal of this research was to identify a 'Hass' avocado tissue with nutrient concentrations predictive of yields greater than 40 kg of fruit per tree. This threshold was specified to assist the California avocado industry to increase yields to '11,200 kg·haL1. Nutrient concentrations of cauliflower stage inflorescences (CSI) collected in March proved better predictors of yield than inflorescences collected at full bloom (FBI) in April, fruit pedicels (FP) collected at five different stages of avocado tree phenology from the end of fruit set in June through April the following spring when mature fruit enter a second period of exponential growth, or 6-month-old spring flush leaves (LF) from nonbearing vegetative shoots collected in September (California avocado industry standard). For CSI tissue, concentrations of seven nutrients, nitrogen (N), phosphorus (P), potassium (K), magnesium (Mg), sulfur (S), zinc (Zn), and copper (Cu) were predictive of trees producing greater than 40 kg of fruit annually. Conditional quantile sampling and frequency analysis were used to identify optimum nutrient concentration ranges (ONCR) for each nutrient. Optimum ratios between nutrient concentrations and yields greater than 40 kg per tree were also derived. The high nutrient concentrations characterizing CSI tissue suggest current fertilization practices (timing or amounts) might be causing nutrient imbalances at this stage of avocado tree phenology that are limiting productivity, a possibility that warrants further investigation. Because CSI samples can be collected 4-6 weeks before full bloom, nutritional problems can be addressed before they affect flower retention and fruit set to increase current crop yield, fruit size, and quality. Thus, CSI nutrient analysis warrants further research as a potential supplemental or alternative tool for diagnosing 'Hass' avocado tree nutrient status and increasing yield. [ABSTRACT FROM AUTHOR]

English walnut (Juglans regia) has become an important crop in Chile, representing 11.5% of the total area of fruit trees, surpassed only by grapevine. As the Chilean walnut industry rapidly expands, young orchards are at risk from the emergence of new fungal diseases. Botryosphaeriaceae and Diaporthaceae fungi have been recognized as main causes of wood diseases in walnut, with symptoms of dieback, canker, and blight. In winter 2017, samples were collected from different orchards in Valparaíso and Maule regions. Fungal isolates recovered were cultured, characterized morphologically, and identified using DNA sequence analyses. Three species (Neofusicoccum parvum, Diplodia mutila, Diplodia seriata) were characterized in Botryosphaeriaceae and two (Diaporthe cynaroidis, Diaporthe australafricana) in Diaporthaceae. Pathogenicity tests showed that N. parvum was the most aggressive species to walnut. This study confirmed the presence of pathogenic Botryosphaeriaceae and Diaporthaceae in J. regia that should be considered an increasing risk for the growing Chilean walnut industry. [ABSTRACT FROM AUTHOR]

The number of reports associated with wood dieback caused by fungi in the Botryosphaeriaceae in numerous perennial crops worldwide has significantly increased in the past years. In this study, we investigated the interactions between the canker pathogen Neofusicoccum parvum and the almond tree host (Prunus dulcis), with an emphasis on varietal resistance and host response at the cell wall biochemical and histological levels. Plant bioassays in a shaded house showed that among the four commonly planted commercial almond cultivars ('Butte', 'Carmel', 'Monterey', and 'Nonpareil'), there was no significant varietal difference with respect to resistance to the pathogen. Gummosis was triggered only by fungal infection, not by wounding. A two-dimensional nuclear magnetic resonance and liquid chromatography determination of cell wall polymers showed that infected almond trees differed significantly in their glycosyl and lignin composition compared with healthy, noninfected trees. Response to fungal infection involved a significant increase in lignin, a decrease in glucans, and an overall enrichment in other carbohydrates with a profile similar to those observed in gums. Histological observations revealed the presence of guaiacyl-rich cell wall reinforcements. Confocal microscopy suggested that N. parvum colonized mainly the lumina of xylem vessels and parenchyma cells, and to a lesser extent the gum ducts. We discuss the relevance of these findings in the context of the compartmentalization of decay in trees model in almond and its potential involvement in the vulnerability of the host toward fungal wood canker diseases. [ABSTRACT FROM AUTHOR]

Huanglongbing (HLB) is a destructive citrus disease that is lethal to all commercial citrus making it the most serious citrus disease and one of the most serious plant diseases. Because of the severity of HLB and the paucity of effective control measures, we structured this study to encompass the entirety of the citrus microbiome and the chemistries associated with that microbial community. We describe the spatial niche diversity of bacteria and fungi associated with citrus roots, stems and leaves using traditional microbial culturing integrated with culture- independent m ethods. Using the culturable sector of the citrus microbiome, we created a microbial repository using a high-throughput bulk culturing and microbial identification pipeline. We integrated an in vitro agar-diffusion inhibition bioassay into our culturing pipeline that queried the repository for antimicrobial activity against Liberibacter crescens, a culturable surrogate for the non-culturable Candidatus Liberibacter asiaticus bacterium associated with HLB. We identified microbes with robust inhibitory activity against L. crescens that include the fungi Cladosporium cladosporioides and Epicoccum nigrum and bacteria Pantoea, Bacillus and Curtobacterium. Purified bioactive natural products with anti-CLas activity, were identified from the fungus, C. cladosporiodes. Bioassay-guided fractionation of an organic extract of C. cladosporioides yielded the natural products cladosporols A, C and D as the active agents against L. crescens. This work serves as a foundation for unraveling the complex chemistries associated with the citrus microbiome to begin to understand functional roles of members of the microbiome with the long-term goal of developing anti-CLas bioinoculants that thrive in the citrus holosystem. [ABSTRACT FROM AUTHOR]

Morphological traits of the plant vascular system such as xylem vessel diameter have been implicated in many physiological processes including resistance to drought-induced xylem cavitation and vessel occlusion during infection with vascular wilt diseases. In both events, xylem vessels lose function because they become filled with air or tyloses and gels. Xylem cavitation has been well studied, whereas vessel occlusion remains purely descriptive even though it is a critical response to wounding injuries and compartmentalization of vascular pathogens. The timing of vessel occlusion is a key determinant to a successful compartmentalization of pathogens within the plant vascular system and we hypothesized that xylem vessel diameter is the driving variable. Using a dye injection method coupled with automated image analysis, we parameterized a model to investigate how xylem vessel diameter affects the speed of vessel occlusion in Vitis vinifera L. cv. Cabernet Sauvignon in response to wounding. Our dataset contains observations from 6,646 vessels at five kinetic points following stem pruning, over a time course of 1 week. Using this approach we provide evidence that the diameter of vessels is a key determinant of the timing of their occlusion. We discuss how these findings impact resistance to vascular wilt diseases in perennial woody hosts. [ABSTRACT FROM AUTHOR]

Background: DNA metabarcoding, commonly used in exploratory microbial ecology studies, is a promising method for the simultaneous in planta-detection of multiple pathogens associated with disease complexes, such as the grapevine trunk diseases. Profiling of pathogen communities associated with grapevine trunk diseases is particularly challenging, due to the presence within an individual wood lesion of multiple co-infecting trunk pathogens and other wood-colonizing fungi, which span a broad range of taxa in the fungal kingdom. As such, we designed metabarcoding primers, using as template the ribosomal internal transcribed spacer of grapevine trunk-associated ascomycete fungi (GTAA) and compared them to two universal primer widely used in microbial ecology. Results: We first performed in silico simulations and then tested the primers by high-throughput amplicon sequencing of (i) multiple combinations of mock communities, (ii) time-course experiments with controlled inoculations, and (iii) diseased field samples from vineyards under natural levels of infection. All analyses showed that GTAA had greater affinity and sensitivity, compared to those of the universal primers. Importantly, with GTAA, profiling of mock communities and comparisons with shotgun-sequencing metagenomics of field samples gave an accurate representation of genera of important trunk pathogens, namely Phaeomoniella, Phaeoacremonium, and Eutypa, the abundances of which were over- or under-estimated with universal primers. Conclusions: Overall, our findings not only demonstrate that DNA metabarcoding gives qualitatively and quantitatively accurate results when applied to grapevine trunk diseases, but also that primer customization and testing are crucial to ensure the validity of DNA metabarcoding results. [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]

Background: Trunk diseases threaten the longevity and productivity of grapevines in all viticulture production systems. They are caused by distantly-related fungi that form chronic wood infections. Variation in wood-decay abilities and production of phytotoxic compounds are thought to contribute to their unique disease symptoms. We recently released the draft sequences of Eutypa lata, Neofusicoccum parvum and Togninia minima, causal agents of Eutypa dieback, Botryosphaeria dieback and Esca, respectively. In this work, we first expanded genomic resources to three important trunk pathogens, Diaporthe ampelina, Diplodia seriata, and Phaeomoniella chlamydospora, causal agents of Phomopsis dieback, Botryosphaeria dieback, and Esca, respectively. Then we integrated all currently-available information into a genome-wide comparative study to identify gene families potentially associated with host colonization and disease development. Results: The integration of RNA-seq, comparative and ab initio approaches improved the protein-coding gene prediction in T. minima, whereas shotgun sequencing yielded nearly complete genome drafts of Dia. ampelina, Dip. seriata, and P. chlamydospora. The predicted proteomes of all sequenced trunk pathogens were annotated with a focus on functions likely associated with pathogenesis and virulence, namely (i) wood degradation, (ii) nutrient uptake, and (iii) toxin production. Specific patterns of gene family expansion were described using Computational Analysis of gene Family Evolution, which revealed lineage-specific evolution of distinct mechanisms of virulence, such as specific cell wall oxidative functions and secondary metabolic pathways in N. parvum, Dia. ampelina, and E. lata. Phylogenetically-informed principal component analysis revealed more similar repertoires of expanded functions among species that cause similar symptoms, which in some cases did not reflect phylogenetic relationships, thereby suggesting patterns of convergent evolution. Conclusions: This study describes the repertoires of putative virulence functions in the genomes of ubiquitous grapevine trunk pathogens. Gene families with significantly faster rates of gene gain can now provide a basis for further studies of in planta gene expression, diversity by genome re-sequencing, and targeted reverse genetic approaches. The functional validation of potential virulence factors will lead to a more comprehensive understanding of the mechanisms of pathogenesis and virulence, which ultimately will enable the development of accurate diagnostic tools and effective disease management. [ABSTRACT FROM AUTHOR]

Cadophora species are reported from grapevine ( Vitis vinifera L.) in California, South Africa, Spain, Uruguay, and Canada. Frequent isolation from vines co-infected with the Esca pathogens ( Togninia minima and Phaeomoniella chlamydospora ), and confirmation of its ability to cause wood lesions/discoloration in pathogenicity tests, suggest that C. luteo-olivacea is part of the trunk pathogen complex. In North America, little is known regarding the diversity, geographic distribution, and roles of Cadophora species as trunk pathogens. Accordingly, we characterized 37 Cadophora isolates from ten US states and two Canadian provinces, based on molecular and morphological comparisons, and pathogenicity. Phylogenetic analysis of three loci (ITS, translation elongation factor 1-alpha (TEF1-α) and beta-tubulin (BT)) distinguished two known species ( C. luteo-olivacea and Cadophora melinii ) and three newly-described species ( Cadophora orientoamericana , Cadophora novi-eboraci , and Cadophora spadicis ). C. orientoamericana , C. novi-eboraci , and C. spadicis were restricted to the northeastern US, whereas C. luteo-olivacea was only recovered from California. C. melinii was present in California and Ontario, Canada. Morphological characterization was less informative, due to significant overlap in dimensions of conidia, hyphae, conidiophores, and conidiogenous cells. Pathogenicity tests confirmed the presence of wood lesions after 24 m, suggesting that Cadophora species may have a role as grapevine trunk pathogens. [ABSTRACT FROM AUTHOR]

In eastern North America, Phomopsis cane and leaf spot, caused by Phomopsis viticola, is a foliar disease of grape but, in the Mediterra-nean climate of western North America, P. viticola is primarily associ-ated with wood cankers, along with other Diaporthe spp. To determine the identity of wood-infecting Diaporthe spp. in eastern North Amer-ica, 65 isolates were cultured from 190 wood-canker samples from 23 vineyards with a history of Phomopsis cane and leaf spot. Identifica-tion of 29 representative isolates was based initially on morphology, followed by phylogenetic analyses of DNA sequences of the ribosomal DNA internal transcribed spacer region, elongation factor subunit 1-a, and actin in comparison with those of type specimens. Three species were identified: P. viticola, P. fukushii, and Diaporthe eres. Inoc-ulations onto woody stems of potted Vitis labruscana 'Concord' and V. vinifera 'Chardonnay' showed that D. eres and P. fukushii were pathogenic (mean lesion lengths of 7.4 and 7.1 mm, respectively, compared with 3.5 mm for noninoculated controls) but significantly less so than wood-canker and leaf-spot isolates of P. viticola (13.5 mm). All three species infected pruning wounds of Concord and Char-donnay in the field. Our finding of pathogenic, wood-infecting Di-aporthe spp. in all 23 vineyards suggests a frequent co-occurrence of the foliar symptoms of Phomopsis cane and leaf spot and wood can-kers, although the latter are not always due to P. viticola. [ABSTRACT FROM AUTHOR]

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