The compositional turnover of grapevine-associated plant pathogenic fungal communities are greater among intraindividual microhabitats and terroirs than among healthy and Esca-diseased plants

Grapevine trunk diseases (GTD) are a major threat to the wine industry, causing yield loss and dieback of grapevines. While the increasing damage caused by GTDs in recent decades have spurred several studies on grapevine-associated pathogenic fungi, key questions about the emergence and severity of GTDs remain unanswered, including possible differences in plant pathogenic fungal communities in asymptomatic and symptomatic grapevines. We generated fungal DNA metabarcoding data from soil, bark, and perennial wood samples from asymptomatic and symptomatic grapevines sampled in three terroirs. We observed larger compositional differences in plant pathogenic fungi among different plants parts within grapevine plants than among individual grapevines. This is driven by the dominance of GTD-associated fungi in perennial wood and non-GTD pathogens in soil, as well as by the lack of significant differences among asymptomatic and Esca symptomatic grapevines. These results suggest that fungi generally associated with Esca disease belong to the core grapevine microbiome and likely are commensal endophytes and/or latent saprotrophs, some of which can act as opportunistic pathogens on stressed plants. In addition, we found significant compositional differences among sampling sites, particularly in soil, which suggest a certain influence of local edaphic and mesclimatic factors on plant pathogenic fungal communities. Furthermore, the observed differences among terroirs in plant pathogenic fungal communities in grapevine woody parts indicate that environmental factors likely are important for the development of Esca disease and further studies are needed to investigate the abiotic conditions on fungal compositional dynamics in Esca-affected plants.

Fomitiporia mediterranea M. Fisch., the historical Esca agent: a comprehensive review on the main grapevine wood rot agent in Europe

Fomitiporia mediterranea M. Fisch. (Fmed) is a basidiomycete first described in 2002, and was considered up to then as part of Fomitiporia punctata (P. Karst) Murrill. This fungus can degrade lignocellulosic biomass, causing white rot and leaving bleached fibrous host residues. In Europe Fmed is considered the main grapevine wood rot (Esca) agent within the Esca disease complex, which includes some of the most economically important Grapevine Trunk Diseases (GTDs). This review summarises and evaluates published research on Fmed, on white rot elimination by curettage or management by treatments with specific products applied to diseased grapevines, and on the relationship between wood symptoms and Grapevine Leaf Stripe Disease (GLSD) in the Esca disease complex. Information is also reviewed on the fungus biology, mechanisms of pathogenicity, and their possible relationships with external foliar symptoms of the Esca disease complex. Information on Fmed control strategies is also reviewed.

Phaeoacremonium fusiformostromum sp. nov. and a new record of P. croatiense from China

Phaeoacremonium species are important pathogens with a worldwide distribution and diverse host and are commonly associated with brown wood streaking, dieback and Esca disease of plants and phaeohyphomycosis of humans. In this study, three sexual taxa, representing two Phaeoacremonium species, were collected from dead wood from Guangdong and Guizhou Provinces in China. Based on the morphological characters and phylogenetic analyses of a combined ACT and TUB2 sequence data, they are identified as Phaeoacremonium fusiformostromum sp. nov., and P. croatiense, which is a new record from China with its first sexual morph. Detailed descriptions and illustrations of these two species are provided, and an updated phylogenetic tree for all Phaeoacremonium species is also given.

Fomitiporia punicata and Phaeoacremonium minimum associated with Esca complex of grapevine in China

The Esca disease complex includes some of the most important trunk diseases of grapevines (Vitis spp.) and causes serious yield losses in grape production worldwide. However, there has been no detailed study on its presence and associated pathogens in China. During 2017–2019, a preliminary field survey was conducted in eight vineyards in Hebei and Ningxia provinces, China when unusual foliar symptoms were observed. Symptoms were distinct tiger striped leaves, which are typical of grapevine leaf stripe disease (GLSD), one of the most common diseases in the Esca complex. Tiger-stripe leaf symptoms were found in four vineyards, and incidence was cultivar dependent varying with vineyard and year. A total of 266 fungal isolates were obtained from wood tissues of grapevines with typical foliar symptoms of GLSD. Based on morphological characters and multigene-combined phylogenetic analyses, the Ascomycete Phaeoacremonium minimum, one of the pathogens associated with Esca complex was identified. The basidiomycete Fomitiporia punicata, which has never been reported infecting grapevine, was also identified and found to be associated with wood rot in grapevine. The remaining isolates included some known wood pathogens, such as Neofusicoccum species and Diaporthe species. Koch’s postulates were performed in the greenhouse, confirming that both F. punicata and P. minimum caused leaf interveinal chlorosis and necrosis that resembled the GLSD symptoms of the Esca complex observed in the field. The present study provides the first detailed report of the Esca complex in China. In addition, this is the first record of F. punicata associated with Esca complex of grapevine worldwide. The results of this study provide new insights into the knowledge of the Esca complex.

Changes in rhizosphere bacterial communities associated with tree decline: grapevine esca syndrome case study

An investigation was carried out on rhizosphere bacteria to determine if they may be associated with perennial crops affected by nonspecific decline, a phenomenon that is difficult to diagnose and prevent. Esca disease of grapevine was chosen for this case study because of its easy foliar symptom identification. Ribosomal DNA fingerprint analysis by polymerase chain reaction – denaturing gradient gel electrophoresis (PCR–DGGE), quantitative PCR (qPCR), and rDNA amplicon sequencing by next-generation sequencing (NGS) were adopted to investigate the bacterial communities associated with grapevines, which were selected for the presence and absence of external foliar symptoms in 11 vineyards. According to PCR–DGGE and qPCR, bacterial communities differed in site of origin (vineyards), but not between symptomatic and asymptomatic plants, whereas qPCR gave a significantly higher presence of total bacteria and Pseudomonas spp. in asymptomatic plants. NGS confirmed no difference between symptomatic and asymptomatic plants, apart from a few minor genera (<0.5%) such as Salinibacterium, Flavobacterium, Nocardia, and Janthinobacterium, which were, in all cases, higher in asymptomatic plants and whose functional role should be the object of further investigation. The fact that total bacteria and Pseudomonas were more abundant in the rhizosphere of asymptomatic grapevines and that some bacterial genera were associated with the latter, represents a new element when investigating the multiple-origin phenomenon such as esca disease of grapevine.

Comparison of SIFT Encoded and Deep Learning Features for the Classification and Detection of Esca Disease in Bordeaux Vineyards

Grapevine wood fungal diseases such as esca are among the biggest threats in vineyards nowadays. The lack of very efficient preventive (best results using commercial products report 20% efficiency) and curative means induces huge economic losses. The study presented in this paper is centered around the in-field detection of foliar esca symptoms during summer, exhibiting a typical “striped” pattern. Indeed, in-field disease detection has shown great potential for commercial applications and has been successfully used for other agricultural needs such as yield estimation. Differentiation with foliar symptoms caused by other diseases or abiotic stresses was also considered. Two vineyards from the Bordeaux region (France, Aquitaine) were chosen as the basis for the experiment. Pictures of diseased and healthy vine plants were acquired during summer 2017 and labeled at the leaf scale, resulting in a patch database of around 6000 images (224 × 224 pixels) divided into red cultivar and white cultivar samples. Then, we tackled the classification part of the problem comparing state-of-the-art SIFT encoding and pre-trained deep learning feature extractors for the classification of database patches. In the best case, 91% overall accuracy was obtained using deep features extracted from MobileNet network trained on ImageNet database, demonstrating the efficiency of simple transfer learning approaches without the need to design an ad-hoc specific feature extractor. The third part aimed at disease detection (using bounding boxes) within full plant images. For this purpose, we integrated the deep learning base network within a “one-step” detection network (RetinaNet), allowing us to perform detection queries in real time (approximately six frames per second on GPU). Recall/Precision (RP) and Average Precision (AP) metrics then allowed us to evaluate the performance of the network on a 91-image (plants) validation database. Overall, 90% precision for a 40% recall was obtained while best esca AP was about 70%. Good correlation between annotated and detected symptomatic surface per plant was also obtained, meaning slightly symptomatic plants can be efficiently separated from severely attacked plants.

Alterations in Grapevine Leaf Metabolism Occur Prior to Esca Apoplexy Appearance

Esca disease is one of the major grapevine trunk diseases in Europe and the etiology is complex, since several inhabiting fungi are identified to be associated with this disease. Among the foliar symptom expressions, the apoplectic form may be distinguished and characterized by sudden dieback of shoots, leaf drop, and shriveling of grape clusters in a few days that can ultimately induce the plant death. To further understand this drastic event, we conducted transcriptomic and metabolomic analyses to characterize responses of leaves during the period preceding symptom appearance (20 and 7 days before foliar symptom expression) and at the day of apoplexy expression. Transcriptomic and metabolomic analyses provide signatures for the apoplectic leaves and most changes concerning the metabolism of carbohydrates, amino acids, and phenylpropanoids. In deciphering glutathione-S-transferase (GST), its preferential location in phloem, correlated with the upregulation of GST genes and a decrease of the glutathione level, offers further support to the putative role of glutathione during apoplexy expression.