Grapevine Resistance Research Articles

Breeding for black rot resistance in grapevine: advanced approaches for germplasm screening

Crop improvement by means of traditional or molecular breeding is a key strategy to accomplish the European Green Deal target of reducing pesticides by 50% by 2030. Regarding viticulture, this is exacerbated by the massive use of chemicals to control pathogen infections. Black rot is an emergent disease caused by the ascomycete Phyllosticta ampelicida, and its destructiveness is alarming vine growers. Implementing and improving effective phenotyping strategies are fundamental preliminary steps to breed disease resistant varieties and this work suggests good practices adopted for this purpose. Primarily, the pedigree of black rot resistance donors was reconstructed based on the collection of phenotypic historical data, highlighting unexplored sources of black rot resistance. Strains used for artificial infections were isolated, genetically characterized and mixed to avoid race-specific resistance selection. A new inoculation protocol based on the use of leaf mature lesions was developed. Ex vivo inoculation on detached leaves was effective for the evaluation of conidia germination and hyphal growth, but not for disease progression. Finally, the pedigree was used for the identification of 23 genotypes to be tested. Two breeding selections (NY39 and NY24) resulted symptomless in all assessments and a third one (F25P52) also showed very high resistance, although with a greater variability. Other two genotypes (F12P19 and ‘Charvir’) fell within the medium resistance category, making them good candidates in a regime of well-timed preventive treatments. In conclusion, this work was effective to a comprehensive parental line characterization and preparatory towards grapevine breeding programs for black rot resistance.

Volatile linalool activates grapevine resistance against downy mildew with changes in the leaf metabolome

Volatile organic compounds (VOCs) are produced by plants in response to biotic and abiotic stimuli. In grapevine, volatile terpenoids are triggered by downy mildew infection (caused by Plasmopara viticola), suggesting their involvement in plant defense responses. In particular, linalool was detected in leaves of downy mildew-resistant genotypes, but no information is available on its involvement in the defense mechanisms against P. viticola. The aim of this study was to investigate the defense mechanisms activated by linalool in grapevine leaves against P. viticola and to identify metabolic changes associated with linalool-induced resistance. Linalool treatment reduced downy mildew severity on leaf disks of susceptible grapevines (cultivar Pinot noir) and stimulated callose deposition at the sites of P. viticola infection. Moreover, the upregulation of defense-related genes was found in linalool-treated leaf disks, indicating the activation of grapevine defense mechanisms of salicylic acid and jasmonic acid pathways. Linalool treatment caused changes in the leaf metabolome of mock-inoculated and P. viticola-inoculated samples at one and six days post inoculation, as revealed by ultra-high pressure liquid chromatography-electrospray ionization-high-resolution quadrupole time of flight-mass spectrometry. Pathway analysis of annotated features with significant increases and decreases in abundance revealed the reprogramming of amino acid, phenylpropanoid, and terpenoid metabolisms in response to linalool treatment and P. viticola inoculation. In particular, features with significant increases in abundance in linalool-treated samples mainly belonged to putative phenylpropanoids, putative terpenoids, putative lipids, and lipid-like compounds, including molecules possibly associated with plant defense against pathogens, such as 2-phenylethanol, 2,4-heptadienal, α-terpineol, citral, and geraniol. These results demonstrated that linalool induces grapevine resistance against downy mildew, acting as a signaling molecule for plant resistance induction.

Transcriptomic analysis provides insights into the abscisic acid mediates brassinosteroid-induced cold resistance of grapevine (Vitis vinifera L.)

Transcriptomic analysis provides insights into the abscisic acid mediates brassinosteroid-induced cold resistance of grapevine (Vitis vinifera L.)

VqMAPK3/VqMAPK6, VqWRKY33, and VqNSTS3 constitute a regulatory node in enhancing resistance to powdery mildew in grapevine.

Grapevine powdery mildew is caused by Erysiphe necator, which seriously harms grape production in the world. Stilbene synthase makes phytoalexins that contribute to the resistance of grapevine against powdery mildew. A novel VqNSTS3 was identified and cloned from Chinese wild Vitis quinquangularis accession Danfeng-2. The novel VqNSTS3 was transferred into susceptible 'Thompson Seedless' by Agrobacterium-mediated transformation. The transgenic plants showed resistance to the disease and activated other resistance-related genes. VqNSTS3 expression in grapevine is regulated by VqWRKY33, and which binds to TTGACC in the VqNSTS3 promoter. Furthermore, VqWRKY33 was phosphorylated by VqMAPK3/VqMAPK6 and thus led to enhanced signal transduction and increased VqNSTS3 expression. ProVqNSTS3::VqNSTS3-GFP of transgenic VqNSTS3 in Arabidopsis thaliana was observed to move to and wrap the pathogen's haustoria and block invasion by Golovinomyces cichoracearum. These results demonstrate that stilbene accumulation of novel VqNSTS3 of the Chinese wild Vitis quinquangularis accession Danfeng-2 prevented pathogen invasion and enhanced resistance to powdery mildew. Therefore, VqNSTS3 can be used in generating powdery mildew-resistant grapevines.

Candidate gene discovery of Botrytis cinerea resistance in grapevine based on QTL mapping and RNA-seq.

Grape gray mold disease (Botrytis cinerea) is widespread during grape production especially in Vitis vinifera and causes enormous losses to the grape industry. In nature, the grapevine cultivar 'Beta ' (Vitis riparia × Vitis labrusca) showed high resistance to grape gray mold. Until now, the candidate genes and their mechanism of gray mold resistance were poorly understood. In this study, we firstly conducted quantitative trait locus (QTL) mapping for grape gray mold resistance based on two hybrid offspring populations that showed wide separation in gray mold resistance. Notably, two stable QTL related to gray mold resistance were detected and located on linkage groups LG2 and LG7. The phenotypic variance ranged from 6.86% to 13.70% on LG2 and 4.40% to 11.40% on LG7. Combined with RNA sequencing (RNA-seq), one structural gene VlEDR2 (Vitvi02g00982) and three transcription factors VlERF039 (Vitvi00g00859), VlNAC047 (Vitvi08g01843), and VlWRKY51 (Vitvi07g01847) that may be involved in VlEDR2 expression and grape gray mold resistance were selected. This discovery of candidate gray mold resistance genes will provide an important theoretical reference for grape gray mold resistance mechanisms, research, and gray mold-resistant grape cultivar breeding in the future.

Secondary and primary metabolites reveal putative resistance-associated biomarkers against Erysiphe necator in resistant grapevine genotypes.

Numerous fungicide applications are required to control Erysiphe necator, the causative agent of powdery mildew. This increased demand for cultivars with strong and long-lasting field resistance to diseases and pests. In comparison to the susceptible cultivar 'Teroldego', the current study provides information on some promising disease-resistant varieties (mono-locus) carrying one E. necator-resistant locus: BC4 and 'Kishmish vatkana', as well as resistant genotypes carrying several E. necator resistant loci (pyramided): 'Bianca', F26P92, F13P71, and NY42. A clear picture of the metabolites' alterations in response to the pathogen is shown by profiling the main and secondary metabolism: primary compounds and lipids; volatile organic compounds and phenolic compounds at 0, 12, and 48 hours after pathogen inoculation. We identified several compounds whose metabolic modulation indicated that resistant plants initiate defense upon pathogen inoculation, which, while similar to the susceptible genotype in some cases, did not imply that the plants were not resistant, but rather that their resistance was modulated at different percentages of metabolite accumulation and with different effect sizes. As a result, we discovered ten up-accumulated metabolites that distinguished resistant from susceptible varieties in response to powdery mildew inoculation, three of which have already been proposed as resistance biomarkers due to their role in activating the plant defense response.

Grapevine-Associated Lipid Signalling Is Specifically Activated in an Rpv3 Background in Response to an Aggressive P. viticola Pathovar.

Vitis vinifera L. is highly susceptible to the biotrophic pathogen Plasmopara viticola. To control the downy mildew disease, several phytochemicals are applied every season. Recent European Union requirements to reduce the use of chemicals in viticulture have made it crucial to use alternative and more sustainable approaches to control this disease. Our previous studies pinpoint the role of fatty acids and lipid signalling in the establishment of an incompatible interaction between grapevine and P. viticola. To further understand the mechanisms behind lipid involvement in an effective defence response we have analysed the expression of several genes related to lipid metabolism in three grapevine genotypes: Chardonnay (susceptible); Regent (tolerant), harbouring an Rpv3-1 resistance loci; and Sauvignac (resistant) that harbours a pyramid of Rpv12 and Rpv3-1 resistance loci. A highly aggressive P. viticola isolate was used (NW-10/16). Moreover, we have characterised the grapevine phospholipases C and D gene families and monitored fatty acid modulation during infection. Our results indicate that both susceptible and resistant grapevine hosts did not present wide fatty acid or gene expression modulation. The modulation of genes associated with lipid signalling and fatty acids seems to be specific to Regent, which raises the hypothesis of being specifically linked to the Rpv3 loci. In Sauvignac, the Rpv12 may be dominant concerning the defence response, and, thus, this genotype may present the activation of other pathways rather than lipid signalling.

Black Rot of Grapes (Guignardia bidwellii)—A Comprehensive Overview

The aim of this review is to provide readers with an integrated knowledge on black rot of grapes, based on a critical survey of previous and recent studies of scientific importance. The current state of the art and perspectives of science are presented, not only on the genetic determinants of grapevine resistance to black rot, predictive models of black rot epidemics, but also on the potential of metabolomics to explore black rot-grape interactions and shorten plant breeding processes. Numerous complications of disease management and ambiguities in phenotype-classification are highlighted, and by exploring the limitations and inconsistencies of previous studies, insights into key dilemmas and controversial findings are also provided, suggesting future research directions. Much research has been conducted, but biochemical and molecular studies of the true interactions between grapevine and Guignardia bidwellii are still rarity.

VqWRKY56 interacts with VqbZIPC22 in grapevine to promote proanthocyanidin biosynthesis and increase resistance to powdery mildew.

Powdery mildew (PM) is a severe fungal disease of cultivated grapevine world-wide. Proanthocyanidins (PAs) play an important role in resistance to fungal pathogens; however, little is known about PA-mediated PM resistance in grapevine. We identified a WRKY transcription factor, VqWRKY56, from Vitis quinquangularis, the expression of which was significantly induced by PM. Overexpression (OE) of VqWRKY56 in Vitis vinifera increased PA content and reduced susceptibility to PM. Furthermore, the transgenic plants showed more cell death and increased accumulation of salicylic acid and reactive oxygen species. Transient silencing of VqWRKY56 in V. quinquangularis and V. vinifera reduced PA accumulation and increased the susceptibility to PM. VqWRKY56 interacted with VqbZIPC22 invitro and in planta. The protein VqWRKY56 can bind to VvCHS3, VvLAR1, and VvANR promoters, and VqbZIPC22 can bind to VvANR promoter. Co-expression of VqWRKY56 and VqbZIPC22 significantly increased the transcript level of VvCHS3, VvLAR1, and VvANR genes. Finally, transient OE of VqbZIPC22 in V. vinifera promoted PA accumulation and improved resistance to PM, while transient silencing in V. quinquangularis had the opposite effect. Our study provides new insights into the mechanism of PA regulation by VqWRKY56 in grapevine and provides a basis for further metabolic engineering of PA biosynthesis to improve PM resistance.

First assessment of leaf lipids and fatty acids as biomarkers of grapevine tolerance/susceptibility to Plasmopara viticola

Grapevine is susceptible to several diseases, being downy mildew, caused by Plasmopara viticola, among the most devastating. Control strategies include the application of pesticides every season through almost all the grapevine developmental stages, jeopardizing viticulture sustainability. Understanding resistance molecular processes is crucial to define alternative strategies and select disease resistance traits. Identifying markers that discriminate between tolerant and susceptible genotypes to P. viticola is an important step. It is shown that lipids and fatty acids (FA) play an important role in grapevine defence against downy mildew and in our previous works we have highlighted that susceptible and resistant grapevine genotypes present contrasting FA profiles even without infection. Here, the constitutive lipid and FA composition of four grapevine genotypes, with contrasting behaviour toward P. viticola was analysed. FA desaturases (FAD) gene expression was also analysed. The results indicate that the susceptible genotypes present higher content of plastidial lipids and unsaturated FA and higher expression levels of FAD genes, whereas tolerant genotypes have higher amounts of neutral lipids and phosphatidic acid as well as saturated FA. In this preliminary study, the concentration of monogalactosyldiacylglycerol and phosphatidylcholine was pinpointed as potential tolerance-associated biomarkers and both linoleic acid and plastidial lipids as susceptibility-associated biomarkers.

Alfin-like transcription factor VqAL4 regulates a stilbene synthase to enhance powdery mildew resistance in grapevine.

Resveratrol is a phytoalexin that is synthesized by stilbene synthase (STS). Resveratrol in the human diet is known to have beneficial effects on health. We previously identified six novel STS (VqNSTS) transcripts from the transcriptome data of Vitis quinquangularis accession Danfeng-2. However, the functions of and defensive mechanisms triggered by these VqNSTS transcripts remain unknown. In the present study, we demonstrate that the expression of five of these six novel members, VqNSTS2-VqNSTS6, can be induced by the powdery mildew-causing fungus Uncinula necator. Additionally, overexpression of VqNSTS4 in the V. vinifera susceptible cultivar Thompson Seedless promoted accumulation of stilbenes and enhanced resistance to U. necator by activating salicylic acid (SA) signalling. Furthermore, our results indicate that the Alfin-like (AL) transcription factor VqAL4 can directly bind to the G-rich element (CACCTC) in the VqNSTS4 promoter and activate gene expression. Moreover, overexpression of VqAL4 in Thompson Seedless enhanced resistance to U. necator by promoting stilbene accumulation and activating SA signalling. Conversely, RNA interference-mediated silencing of VqNSTS4 and VqAL4 resulted in increased susceptibility to U. necator. Collectively, our results reveal that VqNSTS4, regulated by VqAL4, enhances grapevine resistance to powdery mildew by activating SA signalling. Our findings may be useful to improve disease resistance in perennial fruit trees.

Study on the Resistance of ‘Cabernet Sauvignon’ Grapevine with Different Rootstocks to Colomerus vitis

In recent years, Colomerus vitis has caused serious economic losses due to reduced grape production in Xinjiang (northwest China). Several rootstock varieties have been reported to improve the resistance of Cabernet Sauvignon to Colomerus vitis. This study explored the influence of Cabernet Sauvignon with different rootstocks on the resistance to Colomerus vitis. In particular, Cabernet Sauvignon/Cabernet Sauvignon (CS/CS) was selected as the control, and Cabernet Sauvignon grafted with five resistant rootstocks (3309C, 1103P, 140R, SO4, and 5C) was employed as the treatment. The infestation rate and injury index of Colomerus vitis to grapevines was investigated, and insect-resistant types of grapevines with different rootstocks were determined. The resveratrol (Res) content, the gene expression of resveratrol synthase (RS), and the activities of peroxidase (POD), polyphenol oxidase (PPO), catalase (CAT), and superoxide dismutase (SOD) in the leaves of each rootstock grapevine were measured. The activity of the four enzymes and the content of Res were negatively correlated with the injury index. The results revealed the ability of the rootstock to improve the resistance of grapevines to Colomerus vitis by increasing the enzyme activity or Res content. In particular, 140R, SO4, and 5C rootstocks can be employed as rootstocks of the ‘Cabernet Sauvignon’ grapevine with resistance to Colomerus vitis. The contents of Res and the four resistance enzymes studied here can be used as indexes to evaluate the insect resistance of rootstock–scion combinations.

Biostimulation can prime elicitor induced resistance of grapevine leaves to downy mildew.

Using plant defense elicitors to protect crops against diseases is an attractive strategy to reduce chemical pesticide use. However, development of elicitors remains limited because of variable effectiveness in the field. In contrast to fungicides that directly target pathogens, elicitors activate plant immunity, which depends on plant physiological status. Other products, the biostimulants, can improve certain functions of plants. In this study, the objective was to determine whether a biostimulant via effects on grapevine physiology could increase effectiveness of a defense elicitor. A new methodology was developed to study biostimulant activity under controlled conditions using in vitro plantlets. Both biostimulant and defense elicitor used in the study were plant extracts. When added to the culture medium, the biostimulant accelerated the beginning of plantlet growth and affected the shoot and root development. It also modified metabolomes and phytohormone contents of leaves, stems, and roots. When applied on shoots, the defense elicitor changed metabolite and phytohormone contents, but effects were different depending on whether plantlets were biostimulated or controls. Defense responses and protection against Plasmopara viticola (downy mildew agent) were induced only for plantlets previously treated with the biostimulant, Therefore, the biostimulant may act by priming the defense elicitor action. In this study, a new method to screen biostimulants active on grapevine vegetative growth was used to demonstrate that a biostimulant can optimize the efficiency of a plant defense elicitor.

Using ontogenetic resistance of grapevine for fungicide reduction strategies

Resistance of grapevine against powdery mildew (Erysiphe necator) and downy mildew (Plasmopara viticola) increases during ontogenetic development of leaves and berries. The aim of this study was to determine whether the use of fungicides against both pathogens could be reduced during the last treatment before harvest (LTBH) by treating only the susceptible leaves of the vines in the upper half of the canopy. For this purpose, field trials were carried out under experimental or practical conditions in vineyards in southern Germany in eight different years. All plants were sprayed after a standard regimen until LTBH, performed at BBCH 77–79. LTBH was either performed on the whole canopy or limited to the upper half of the canopy excluding the fruit zone. For both pathogens, no differences were observed between the two treatments with regard to the disease severity on the grape clusters. Only in one out of eight years, an increased fruit disease incidence could be observed. As far as the disease severity of leaves is concerned, no differences were observed for downy mildew, and in the case of powdery mildew, significantly higher infestation was observed only in two out of eight years. Furthermore, the efficacy of different fungicides for LTBH was investigated in a 3-year experiment. Sole use of a copper fungicide for LTBH had no disadvantages against E. necator, compared to a treatment with synthetic fungicides. Taken together, LTBH at BBCH 77–79 can be limited to the upper canopy without risk to the health of the fruit.

The Characterization of Pathotypes in Grapevine Downy Mildew Provides Insights into the Breakdown of Rpv3, Rpv10, and Rpv12 Factors in Grapevines.

We describe a standard method for characterizing the virulence profile of Plasmopara viticola, the causal agent of grapevine downy mildew. We used 33 European strains to inoculate six grapevine varieties carrying the principal factors for resistance to downy mildew (Rpv1, Rpv3.1, Rpv3.2, Rpv5, Rpv6, Rpv10, and Rpv12) and the susceptible Vitis vinifera 'Chardonnay'. For each interaction, we characterized the level of sporulation by image analysis and the intensity of the grapevine hypersensitive response by visual score. We propose a definition for the breakdown of grapevine quantitative resistances combining these two traits. Among the 33 strains analyzed, 28 are virulent on at least one resistance factor. We identified five different pathotypes across the 33 strains analyzed: two pathotypes overcoming a single resistance factor (vir3.1 and vir3.2) and three complex pathotypes overcoming multiple resistance factors (vir3.1,3.2; vir3.2,12; vir3.1,3.2,10). Our findings confirm the widespread occurrence of P. viticola strains overcoming the Rpv3 haplotypes (28 strains). We also detected the first breakdown of resistance to the Rpv10 by a strain from Germany and the breakdown of Rpv12 factors by a strain from Hungary. The pathotyping method proposed here and the associated differential host range lay the groundwork for the early detection of resistance breakdown in grapevines. This approach will also facilitate the monitoring of the evolution of P. viticola populations at large spatial scales. This is an essential step forward to promoting durable management of the resistant grapevine varieties currently available.

The transcription factors VaERF16 and VaMYB306 interact to enhance resistance of grapevine to Botrytis cinerea infection.

Botrytis cinerea is a fungus that infects cultivated grape (Vitis vinifera); the identification and characterization of resistance mechanisms in the host is of great importance for the grape industry. Here, we report that a transcription factor in the ethylene‐responsive factor (ERF) family (VaERF16) from Chinese wild grape (Vitis amurensis ‘Shuang You’) is expressed during B. cinerea infection and in response to treatments with the hormones ethylene and methyl jasmonate. Heterologous overexpression of VaERF16 in Arabidopsis thaliana substantially enhanced resistance to B. cinerea and the bacterium Pseudomonas syringae DC3000 via the salicylic acid and jasmonate/ethylene signalling pathways. Yeast two‐hybrid, bimolecular fluorescence complementation, and co‐immunoprecipitation assays indicated that VaERF16 interacts with the MYB family transcription factor VaMYB306. Overexpression of VaERF16 or VaMYB306 in grape leaves increased resistance to B. cinerea and caused an up‐regulation of the defence‐related gene PDF1.2, which encodes a defensin‐like protein. Conversely, silencing of either gene resulted in increased susceptibility to B. cinerea. Yeast one‐hybrid and dual‐luciferase assays indicated that VaERF16 increased the transcript levels of VaPDF1.2 by binding directly to the GCC box in its promoter. Notably, VaMYB306 alone did not bind to the VaPDF1.2 promoter, but the VaERF16–VaMYB306 transcriptional complex resulted in higher transcript levels of VaPDF1.2, suggesting that the proteins function through their mutual interaction. Elucidation of this regulatory module may be of value in enhancing resistance of grapevine to B. cinerea infection.

Uncovering the role of DNA methyltransferases in grapevine – Plasmopara viticola interaction: From genome-wide characterization to global methylation patterns

Epigenetic regulation has recently gained prominence in the field of plant-pathogen interactions, providing a deeper understanding of the molecular mechanisms associated with plant infection. In grapevine interaction with pathogens, epigenetic regulation still remains a black box. In this work, we characterized grapevine DNA methyltransferase gene family and identified nine DNA methyltransferases genes across eight grapevine chromosomes coding for 17 proteins. We also assessed the modulation of global cytosine methylation and gene expression levels of these genes with the aim of establishing a connection between DNA methylation and grapevine resistance towards downy mildew. Our results revealed that, in the incompatible interaction, an early hypomethylation, coupled with downregulation of DNMT and CMT genes occurs very early after pathogen inoculation. Additionally, the compatible interaction is characterized by a hypermethylation at 6hpi. A temporal delay is evident between the shifts in DNA methyltransferases gene expression in both compatible and incompatible interactions which in turn may be reflected in the global methylation percentage. Overall, we present the first evidence of an epigenetic regulation role in grapevine defense against P. viticola.

Analysis of Grapevine's Somatic Embryogenesis Receptor Kinase (SERK) Gene Family: VqSERK3/BAK1 Overexpression Enhances Disease Resistance.

The somatic embryogenesis receptor kinase (SERK) gene family has been intensively studied in several plant species. Here we confirmed the existence of five SERK genes in grapevine (Chinese wild grapevine Vitis quinquangularis) and named them VqSERK1, VqSERK2, VqSERK3, VqSERK4, and VqSERK5. Analysis of the predicted structures of these SERK proteins revealed they include a signal peptide domain, a leucine zipper domain, a Ser-Pro-Pro domain, a single transmembrane domain, different leucine-rich repeats, and an intracellular kinase activity domain. The SERK genes of grapevine showed different gene expression patterns when treated with powdery mildew (Erysiphe necator) and hormones (salicylic acid, jasmonic acid, abscisic acid, and ethylene). Subcellular localization assays confirmed that VqSERK family proteins localized to the cell membrane. Moreover, we cloned the SERK3/BAK1 gene from the Chinese wild grapevine V. quinquangularis clone 'Shang-24'. Heterologous VqSERK3/BAK1 expression in the Arabidopsis bak1-4 mutant lines restored control of cell death, increased resistance to powdery mildew, and strengthened stomatal immunity. Our work may provide the foundation for further studies of SERK genes for pathogen resistance and hormone treatment in grapevine.

Comparative Proteomics Reveals the Difference in Root Cold Resistance between Vitis. riparia × V. labrusca and Cabernet Sauvignon in Response to Freezing Temperature.

Grapevines, bearing fruit containing large amounts of bioactive metabolites that offer health benefits, are widely cultivated around the world. However, the cold damage incurred when grown outside in extremely low temperatures during the overwintering stage limits the expansion of production. Although the morphological, biochemical, and molecular levels in different Vitis species exposed to different temperatures have been investigated, differential expression of proteins in roots is still limited. Here, the roots of cold-resistant (Vitis. riparia × V. labrusca, T1) and cold-sensitive varieties (Cabernet Sauvignon, T3) at −4 °C, and also at −15 °C for the former (T2), were measured by iTRAQ-based proteomic analysis. Expression levels of genes encoding candidate proteins were validated by qRT-PCR, and the root activities during different treatments were determined using a triphenyl tetrazolium chloride method. The results show that the root activity of the cold-resistant variety was greater than that of the cold-sensitive variety, and it declined with the decrease in temperature. A total of 25 proteins were differentially co-expressed in T2 vs. T1 and T1 vs. T3, and these proteins were involved in stress response, bio-signaling, metabolism, energy, and translation. The relative expression levels of the 13 selected genes were consistent with their fold-change values of proteins. The signature translation patterns for the roots during spatio-temporal treatments of different varieties at different temperatures provide insight into the differential mechanisms of cold resistance of grapevine.

Fluid dynamic simulations at the interface of the blue-green sharpshooter functional foregut and grapevine xylem sap with implications for transmission of Xylella fastidiosa.

Xylella fastidiosa is a multi-continental, lethal, plant pathogenic bacterium that is transmitted by sharpshooter leafhoppers (Insecta: Hemiptera: Cicadellidae: Cicadellinae) and adult spittlebugs (Hemiptera: Aphrophoridae). The bacterium forms biofilms in plant xylem and the functional foregut of the insect. These biofilms serve as sources of inoculum for insect acquisition and subsequent inoculation to a healthy plant. In this study, 3D fluid dynamic simulations were performed for bidirectional cibarial propulsion of xylem sap through tube-like grapevine xylem and an anatomically accurate model of the functional foregut of the blue-green sharpshooter, Graphocephala atropunctata. The analysis supports a model of how fluid dynamics influence X. fastidiosa transmission. The model supports the hypothesis that X. fastidiosa inoculation is mostly driven by detachment of bacteria from the foregut due to high-velocity flow during egestion (outward fluid flow from the stylets). Acquisition occurs by fluid dynamics during both egestion and ingestion (fluid uptake through the stylets and swallowing). These simulation results are supported by previously reported X. fastidiosa colonization patterns in the functional foregut and sharpshooter stylet probing behaviors. The model indicates that xylem vessel diameter influences drag forces imposed on xylem wall-adherent bacteria; thus, vessel diameter may be an important component of the complex transmission process. Results from this study are directly applicable to development of novel grapevine resistance traits via electropenetrographic monitoring of vector acquisition and inoculation behaviors.