Assessing the influence of autumnal temperature fluctuations on cold hardiness in different grapevine cultivars: variations across vine age and bud positions.

The dynamic fluctuations in autumn temperatures, particularly the marked diurnal variations and the subsequent precipitous drops are key and a pivotal role in viticulture, as they critically influence the acclimation process of grapevines to cold, thereby directly impacting their survival and productivity in cold-climate regions. In this comprehensive study, we investigated the cold hardiness of four grapevine cultivars: 'Itasca', 'Frontenac', 'La Crescent', and 'Marquette', focusing on how these cultivars and their individual buds (1st, 2nd, 3rd, 4th, 5th, 6th, 7th, 8th, and 9th) respond to fluctuating weather and low temperatures typical of autumn [-1.1°C (30°F) -9.4°C (15°F) and -17.8°C (0°F)]. Our results illuminated the striking variability in cold hardiness that was manifest not only among the different cultivars but also within individual buds on the same vine, underscoring the critical influence of bud position on a vine for cold hardiness. 'Frontenac' showed greater cold hardiness at critical temperatures at which 10%, and 50% of the dormant buds were lethally affected by cold (LT10 and LT50) compared to 'Itasca' and 'La Crescent', with 'Marquette' exhibiting intermediate values. However, in cultivars such as 'Itasca' and 'Marquette', certain buds demonstrated a pronounced hardiness when faced with colder temperatures, while others exhibited a heightened sensitivity, thereby revealing a nuanced interplay between bud position and a vine's ability to withstand cold stress. Our study revealed a notable divergence from traditional viticulture understanding; apical buds demonstrated greater cold hardiness than basal buds and opened new paths for research into grapevine physiology. Our results also indicated a significant trend wherein older vines across all studied cultivars displayed enhanced cold hardiness, particularly pronounced at the critical LT50 and the critical temperature at which 90% of the dormant buds were lethally affected by cold (LT90) thresholds, in comparison to younger vines. Moreover, our findings shed light on the impact of autumn's diurnal temperature variations and the subsequent drop in temperatures on vine cold hardiness, thus highlighted the complex interplay between environmental temperature dynamics and dormant bud hardiness. In conclusion, our study showed that the cold damage observed in grapevines in North Dakota was not a result of extreme temperature fluctuations in the fall. This was confirmed by testing the vines after they had reached various threshold temperatures through differential thermal analysis (DTA) and optical differential nucleation and expansion analysis (ODNEAL) methodologies, particularly before the onset of severe pre-winter cold conditions. These comprehensive findings highlighted the complexity of the vine's response to climatic conditions and viticultural management, pointing to the need for specific strategies in vineyard management and cultivar selection to optimize bud hardiness and productivity in the face of various environmental challenges, especially in cold climate viticulture.

Pre-inoculation water deficit effects on grapevine physiology, Xylella fastidiosa titers, and Pierce's disease progression.

Drought and Pierce's disease are common throughout many grapevine-growing regions such as Mexico and the United States. Yet, how ongoing water deficits affect infections of Xylella fastidiosa, the causal agent of Pierce's disease, is poorly understood. Symptoms were observed to be significantly more severe in water-stressed plants one month after X. fastidiosa inoculation, and, in one experiment, titers were significantly lower in water-stressed than well-watered grapevines. Host chemistry examinations revealed overall amino acid and phenolic levels did not statistically differ due to water deficits, but sugar levels were significantly greater in water stressed than well-watered plants. Results highlight the need to especially manage Pierce's disease spread in grapevines experiencing drought.

Characterization of data observing Meloidogyne incognita, Neofusicoccum parvum, and Xylella fastidiosa infection effects on development of grapevine phenolic compound levels and resistance to subsequent Neofusicoccum parvum infections.

Grapevines encounter many different pathogens throughout their lifespans, including the bacterial pathogen Xylella fastidiosa, which causes Pierce's disease that results in vascular occlusion and eventual plant host death, the fungal pathogen Neofusicoccum parvum, which causes stem cankers that kill individual vines and reduce fruit yields, and the root knot nematode Meloidogyne incognita, which destroys root tissues that impacts host vigour. To date, little research has been conducted to examine how one infection could impact subsequent infections by the same or different pathogens despite this is important to ensure healthy vineyards. Therefore, grapevines initially infected with either X. fastidiosa, N. parvum, or M. incognita were subsequently infected with N. parvum eight weeks later to observe developing lesion lengths, which were assessed to determine grapevine resistance to infections. Collected data shows that when prior infections were present, the N. parvum lesions lengths were smaller. This suggests grapevines had induced resistance to combat infections. Further, defence-associated phenolics were measured by high-performance liquid chromatography to determine roles in observed resistance to the secondary N. parvum infections. Data shows that of the different phenolics examined, only stilbenoids were different due to infections, with lowered levels observed in plants that were infected compared with non-infected controls. These data provide insight into how infections by different pathogens could impact grapevine host resistance to new, subsequent pathogen infections.

Studies on pollen micro-morphology, pollen storage methods, and cross-compatibility among grape (Vitis spp.) genotypes.

The knowledge of pollen morphology, suitable storage condition, and species compatibility is vital for a successful grapevine improvement programme. Ten grape genotypes from three different species, viz., Vitis vinifera L., Vitis parviflora Roxb., and Vitis champini Planc., were studied for their pollen structure and pollen storage with the objective of determining their utilization in grape rootstock improvement programs. Pollen morphology was examined through the use of a scanning electron microscope (SEM). The viability of the pollen was assessed using 2,3,5-triphenyltetrazolium chloride (TTC). In vitro pollen germination was investigated using the semi-solid medium with 10 % sucrose, 100 mg/L boric acid, and 300 mg/L calcium nitrate. The results revealed variations in pollen micro-morphology in 10 genotypes, with distinct pollen dimensions, shapes, and exine ornamentation. However, species-wise, no clear difference was found for these parameters. Pollen of V. parviflora Roxb. and Dogridge was acolporated and did not germinate. The remaining eight genotypes exhibited tricolporated pollen and showed satisfactory in vitro pollen germination. Storage temperature and duration interactions showed that, at room temperature, pollen of most of the grape genotypes can be stored for up to 1 day only with an acceptable pollen germination rate (>30 %). However, storage for up to 7 days was successfully achieved at 4 °C, except for 'Pearl of Csaba'. The most effective storage conditions were found to be at -20 °C and -196 °C (in liquid N2), enabling pollen storage for a period of up to 30 days, and can be used for pollination to overcome the challenge of asynchronous flowering. Four interspecific combinations were studied for their compatibility, among which V. parviflora Roxb. × V. vinifera L. (Pusa Navrang) and V. parviflora Roxb. × V. champini Planc. (Salt Creek) showed high cross-compatibility, offering their potential use for grape rootstock breeding. However, V. parviflora Roxb. × V. vinifera L. (Male Hybrid) recorded the lowest compatibility index among studied crosses. In the case of self-pollinated flowers from V. parviflora Roxb. and V. parviflora Roxb. × V. champini Planc. (Dogridge), pollen failed to germinate on the stigma due to male sterility caused by acolporated pollen. As a result, the flowers of these genotypes functioned as females, which means they are ideal female parents for grape breeding without the need for the tedious process of emasculation.

Grape Pomace-Advances in Its Bioactivity, Health Benefits, and Food Applications.

From a circular economy perspective, the appropriate management and valorization of winery wastes and by-products are crucial for sustainable development. Nowadays, grape pomace (GP) has attracted increasing interest within the food field due to its valuable content, comprising nutritional and bioactive compounds (e.g., polyphenols, organic and fatty acids, vitamins, etc.). Particularly, GP polyphenols have been recognized as exhibiting technological and health-promoting effects in different food and biological systems. Hence, GP valorization is a step toward offering new functional foods and contributing to solving waste management problems in the wine industry. On this basis, the use of GP as a food additive/ingredient in the development of novel products with technological and functional advantages has recently been proposed. In this review, we summarize the current knowledge on the bioactivity and health-promoting effects of polyphenolic-rich extracts from GP samples. Advances in GP incorporation into food formulations (enhancement of physicochemical, sensory, and nutritional quality) and information supporting the intellectual property related to GP potential applications in the food industry are also discussed.

Xylem-dwelling pathogen unaffected by local xylem vessel network properties in grapevines (Vitis spp.).

BACKGROUND AND AIMS: Xylella fastidiosa (Xf) is the xylem-dwelling bacterium associated with Pierce's disease (PD), which causes mortality in agriculturally important species, such as grapevine (Vitis vinifera). The development of PD symptoms in grapevines depends on the ability of Xf to produce cell-wall-degrading enzymes to break up intervessel pit membranes and systematically spread through the xylem vessel network. Our objective here was to investigate whether PD resistance could be mechanistically linked to xylem vessel network local connectivity. METHODS: We used high-resolution X-ray micro-computed tomography (microCT) imaging to identify and describe the type, area and spatial distribution of intervessel connections for six different grapevine genotypes from three genetic backgrounds, with varying resistance to PD (four PD resistant and two PD susceptible). KEY RESULTS: Our results suggest that PD resistance is unlikely to derive from local xylem network connectivity. The intervessel pit area (Ai) varied from 0.07 ±â€…0.01 mm2 mm-3 in Lenoir to 0.17 ±â€…0.03 mm2 mm-3 in Blanc do Bois, both PD resistant. Intervessel contact fraction (Cp) was not statically significant, but the two PD-susceptible genotypes, Syrah (0.056 ±â€…0.015) and Chardonnay (0.041 ±â€…0.013), were among the most highly connected vessel networks. Neither Ai nor Cp explained differences in PD resistance among the six genotypes. Bayesian re-analysis of our data shows moderate evidence against the effects of the traits analysed: Ai (BF01 = 4.88), mean vessel density (4.86), relay diameter (4.30), relay density (3.31) and solitary vessel proportion (3.19). CONCLUSIONS: Our results show that radial and tangential xylem network connectivity is highly conserved within the six different Vitis genotypes we sampled. The way that Xf traverses the vessel network may limit the importance of local network properties to its spread and may confer greater importance on host biochemical responses.

Fungicide Use Patterns in Select United States Wine Grape Production Regions.

Wine grape production (Vitis sp.) in the United States requires fungicide inputs for disease control. Currently, there is limited data available on vineyard fungicide use patterns. This information is important in developing tailored recommendations for disease management and fungicide stewardship. In this paper, we summarize the wine grape vineyard fungicide use patterns from four major regions: Napa and Sonoma valleys (California), Willamette Valley (Oregon), Columbia Valley (Washington), and several smaller regions east of the Mississippi River in years 2009 to 2020. We learned that the average in-season total fungicide applications ranged regionally from 5.6 to 8. The most commonly applied Fungicide Resistance Action Committee (FRAC) codes in spray programs were FRAC 3, 13, and M02 across all regions, with some variation to the top four groups in each region. Most applications were made on 14-day intervals; however, shorter intervals (7-day) were favored early season, and longer intervals (21-day) were favored late season. Tank-mixing multiple active ingredients was common east of the Mississippi River during all stages of grape development; this action was typically favored during the bloom period in other regions. In a subset of records that participated in FRAC 11 fungicide resistance testing, the average number of FRAC 11 applications after testing was reduced to either no applications or one application in Napa and Sonoma valleys. This survey provides regionally specific data related to fungicide stewardship practices that could be a focus for future stewardship messaging and fungicide resistance selection training, including total product use (selection events), spray intervals (selection pressure), and tank mixing (selection management).[Formula: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Tissue-specific stilbene accumulation is an early response to wounding/grafting as revealed by using spatial and temporal metabolomics.

Grafting is widely used in horticulture. Shortly after grafting, callus tissues appear at the graft interface and the vascular tissues of the scion and rootstock connect. The graft interface contains a complex mix of tissues, we hypothesised that each tissue has its own metabolic response to wounding/grafting and accumulates different metabolites at different rates. We made intact and wounded cuttings and grafts of grapevine, and then measured changes in bulk flavonoid, phenolic acid and stilbenoid concentration and used metabolite imaging to study tissue-specific responses. We show that some metabolites rapidly accumulate in specific tissues after grafting, for example, stilbene monomers accumulate in necrotic tissues surrounding mature xylem vessels. Whereas other metabolites, such as complex stilbenes, accumulate in the same tissues at later stages. We also observe that other metabolites accumulate in the newly formed callus tissue and identify genotype-specific responses. In addition, exogenous resveratrol application did not modify grafting success rate, potentially suggesting that the accumulation of resveratrol at the graft interface is not linked to graft union formation. The increasing concentration of complex stilbenes often occurs in response to plant stresses (via unknown mechanisms), and potentially increases antioxidant activity and antifungal capacities.

Exploring potential induction of grapevine (Vitis spp.) root phenolic compounds by ring nematodes, Mesocriconema xenoplax.

OBJECTIVE: Ring nematodes can decrease vineyard productivity when plated in conditions favorable for their survival. Resistant rootstocks are available to combat harm due to ring nematodes, and compounds called phenolics were hypothesized as imparting this resistance. Therefore, this study measured phenolic compound levels in four different rootstocks and attempted to find associations with ring nematode populations. Furthermore, select phenolics called stilbenoids were tested in a bioassay to observe if these compounds affect ring nematode survival. This was part of a larger effort to assess the role of phenolics in protecting grapevines from nematodes and other pathogens or pests. DATA DESCRIPTION: This study was conducted over 2 years, 2018 and 2019, and phenolic levels were much greater in 2019 than 2018 likely due to uncontrolled differences in climatic controls. Ring nematode infected grapevines also did not have different phenolic compound levels than healthy controls. Bioassays of different stilbenoid polymers revealed no significant effects on ring nematode survival. These results suggest that analyzed root phenolic compounds were not involved in resistance or susceptibility to ring nematodes. These data should steer future researchers into analyzing other potential sources of nematode resistance.

Grapevine Plants Management Using Natural Extracts and Phytosynthesized Silver Nanoparticles.

Starting from the well-known antimicrobial properties of silver nanoparticles, the goal of this study is to evaluate the influence of two "green" recipes, namely an alcoholic extract of Dryopteris filix-mas (L.) Schott and a dispersion of silver nanoparticles phytosynthesized using the extract on grapevine pathogens. The influence of some grapevine parameters (pith/wood rapport, soluble sugars, starch, total sugars, total water content, length of young shoots, number of grapes) in field experiments was also studied. The study was conducted on four clones (Feteasca alba 97 St., Feteasca neagra 6 St., Feteasca regala 72 St., and Cabernet Sauvignon 131 St.) located in vegetation pots inside a greenhouse. For the phytosynthesis of the silver nanoparticles (AgNPs) we used a scaled-up technology, allowing us to obtain large quantities of nanoparticles-containing solution. The AgNPs analysis by X-ray diffraction and transmission electron microscopy confirmed the synthesis of spherical and quasi-spherical nanoparticles of 17 nm average diameter and 6.72 nm crystallite size. The field experiments registered different responses of the four clones to the treatment, using both the natural extracts and phytosynthesized nanoparticles solution. Both recipes exhibited a protective effect against the Uncinula necator pathogen. For the treatment using phytosynthesized nanoparticles, significant increases in the pith/wood ratio for white wine clones (Feteasca alba 97 St. and Feteasca regala 72 St.) were observed. The biochemical analyses revealed other significant increases of soluble sugars (red wine clones-Feteasca neagra and Cabernet Sauvignon/second year), starch (Feteasca alba and Cabernet Sauvignon in 2021 for both clones), total sugars (Feteasca alba and Feteasca neagra in 2021 for both clones), and of total water content (Feteasca alba and Feteasca neagra in 2021 for both clones), respectively. The applied treatments also led to an increase of young shoots length and grape numbers for all clones as compared to the control (chemical pesticide), which would suggest a potential biostimulant effect of the recipes.

Inflorescence temperature influences fruit set, phenology, and sink strength of Cabernet Sauvignon grape berries.

The temperature during the bloom period leading up to fruit set is a key determinant of reproductive success in plants and of harvest yield in crop plants. However, it is often unclear whether differences in yield components result from temperature effects on the whole plant or specifically on the flower or fruit sinks. We used a forced-convection, free-air cooling and heating system to manipulate the inflorescence temperature of field-grown Cabernet Sauvignon grapevines during the bloom period. Temperature regimes included cooling (ambient -7.5°C), heating (ambient +7.5°C), an ambient control, and a convective control. Cooling significantly retarded the time to fruit set and subsequent berry development, and heating shortened the time to fruit set and accelerated berry development relative to the two controls. Fruit set was decreased in cooled inflorescences, but although the cooling regime resulted in the lowest berry number per cluster, it also decreased seed and berry weight at harvest while not affecting seed number. Cooling inflorescences slightly decreased fruit soluble solids and pH, and increased titratable acidity, but did not affect color density. The inflorescence temperature did not impact leaf gas exchange and shoot growth, and shoot periderm formation occurred independently of the timing of fruit ripening. These results suggest that the temperature experienced by grape flowers during bloom time impacts fruit set and subsequent seed and berry development. Suboptimal temperatures not only reduce the proportion of flowers that set fruit but also limit the sink strength of the berries that do develop after fruit set. Shoot vigor and maturation, and leaf physiology, on the other hand, may be rather insensitive to temperature-induced changes in reproductive development.

Microplate bioassay to examine the effects of grapevine-isolated stilbenoids on survival of root knot nematodes.

OBJECTIVES: Root knot nematodes can be major pests in vineyards and cause significant yield losses over time. Control involves the use of different resistant grapevine rootstocks, but it remains unclear the mechanisms that such rootstocks possess to limit root knot nematode infections. Defense-associated compounds called stilbenoids, a type of phenolic compound, are present in relatively substantial amounts in grapevine root tissues. Therefore, experiments were performed to assess how different stilbenoid compounds impact nematode survival in microplate assays. Data generated were part of a larger effort to understand potential mechanisms that resistant grapevine rootstocks have to limit root knot infections. DATA DESCRIPTION: The percentage of surviving root knot nematodes was assessed 1, 3, and 5 days after J2 juveniles were placed into microplate wells amended with 0, 1.25, 2.5, 5, or 10 ppm of piceid, ε-viniferin, a resveratrol trimer putatively identified as miyabenol C, or a putative mixture of resveratrol tetramers putatively identified as vitisin B and hopeaphenol. Both ε-viniferin and the resveratrol tetramers significantly reduced root knot survival at the higher concentrations. These data provide insight about one potential mechanism that grapevine rootstocks might possess to combat nematodes.

Insight into Carbohydrate Metabolism and Signaling in Grapevine Buds during Dormancy Progression.

Perennial fruit crops enter dormancy to ensure bud tissue survival during winter. However, a faster phenological advancement caused by global warming exposes bud tissue to a higher risk of spring frost damage. Tissue dehydration and soluble sugars accumulation are connected to freezing tolerance, but non-structural carbohydrates also act as metabolic substrates and signaling molecules. A deepened understanding of sugar metabolism in the context of winter freezing resistance is required to gain insight into adaptive possibilities to cope with climate changes. In this study, the soluble sugar content was measured in a cold-tolerant grapevine hybrid throughout the winter season. Moreover, the expression of drought-responsive hexose transporters VvHT1 and VvHT5, raffinose synthase VvRS and grapevine ABA-, Stress- and Ripening protein VvMSA was analyzed. The general increase in sugars in December and January suggests that they can participate in protecting bud tissues against low temperatures. The modulation of VvHT5, VvINV and VvRS appeared consistent with the availability of the different sugar species; challenging results were obtained for VvHT1 and VvMSA, suggesting interesting hypotheses about their role in the sugar-hormone crosstalk. The multifaceted role of sugars on the intricate phenomenon, which is the response of dormant buds to changing temperature, is discussed.

AMF Community Diversity Promotes Grapevine Growth Parameters under High Black Foot Disease Pressure.

Black foot disease is one of the main grapevine root diseases observed worldwide and is especially problematic in New Zealand. Arbuscular mycorrhizal fungi (AMF) have been shown to reduce infection and mitigate the effect of black foot disease on grapevine rootstocks. In contrast to prior studies, which have limited their focus to the effect of one, two or a combination of only a small number of AMF species, this study used whole AMF communities identified from 101-14, 5C and Schwarzmann rootstocks sampled from New Zealand vineyards. The effect of AMF on black foot disease was investigated in a 'home' and 'away' experiment using three commercial grapevine rootstocks. The study produced some evidence that AMF treatments lowered disease incidence at 5 cm and disease severity in vines by 40% to 50% compared to the vines inoculated with the pathogen only. This work also showed that the presence of high disease incidence may have limited the potential disease protective effect of AMF community. However, despite the high disease incidence and severity, AMF inoculation increased vine growth parameters by 60% to 80% compared to the vines inoculated with the pathogen only. This study is the first to provide an understanding on how young grapevine rootstocks inoculated with their 'home' and 'away' AMF communities would respond to challenge with a black foot pathogen species mixture. Further research is required to understand the mechanistic effect of AMF colonization on the increase of grapevine growth parameters under high black foot disease pressure.

Pest categorisation of Arboridia kakogawana.

The EFSA Panel on Plant Health performed a pest categorisation of the Japanese grape leafhopper, Arboridia kakogawana (Matsumura, 1932) (Hemiptera: Cicadellidae), for the EU territory. This species is not included in the EU Commission Implementing Regulation 2019/2072. Adults of A. kakogawana overwinter in broad-leaved and mixed forests and move to vineyards in the spring where there may be up to four generations, before adults move back to forests during late summer-early autumn to overwinter, possibly under diapause. A. kakogawana has a restricted host range (Vitis spp. and Parthenocissus quinquefolia). It is native to Eastern Asia, from where it moved westwards reaching southern Russia in 1999, and subsequently Ukraine, Romania, Bulgaria and Serbia in 2020. A. kakogawana develops on the abaxial side of the leaves causing chlorotic spots that reduce grape quality. Plants for planting of Vitis L. are banned from entering the EU except from Switzerland, where A. kakogawana is not known to occur. Therefore, this can be considered as a closed entry pathway. However, other plants for planting including the host P. quinquefolia and many broad-leaved trees where overwintering takes place, as well as isolated bark and wood with bark provide potential pathways which are partly regulated but remain open. There are no EU records of interception. Additional introductions and further spread of A. kakogawana into/within the EU, coupled with the ample availability of grapevines and the climatic conditions would most probably allow successful establishment in most EU member states. Should this happen, economic impact in table and wine grapes is anticipated. A. kakogawana satisfies all the criteria that are within the remit of EFSA to assess for it to be regarded as a potential Union quarantine pest (UQP).

Cultivated and Wild Grapevines in Tennessee Possess Overlapping but Distinct Virus Populations.

Viruses and viroids prevalent in a population of 42 wild grapevines (i.e., free-living, uncultivated grapevines; Vitis spp.) were compared with those in a population of 85 cultivated grapevines collected in Tennessee, United States by RNA sequencing analysis of pools of ribosomal RNA-depleted total RNA. The sequences of 10 viruses (grapevine fleck virus, grapevine leafroll-associated virus 2, grapevine rupestris stem pitting-associated virus, grapevine Syrah virus 1, grapevine vein-clearing virus, grapevine virus B, grapevine virus E, tobacco ringspot virus, tomato ringspot virus, and a novel nano-like virus) and two viroids (hop stunt viroid and grapevine yellow speckle viroid 1) were detected in both grapevine populations. Sequences of four viruses (grapevine associated tymo-like virus, grapevine leafroll-associated virus 3, grapevine red blotch virus, and grapevine virus H) were identified only from cultivated grapevines. High, moderate, and low numbers of sequence reads were identified only from wild grapevines for a novel caulimovirus, an enamovirus, and alfalfa mosaic virus, respectively. The presence of most virus sequences and both viroids was verified independently in the original samples by reverse-transcription PCR followed by Sanger sequencing. Comparison of viral sequences shared by both populations showed that cultivated and wild grapevines harbored distinct sequence variants, which suggests that there was limited virus movement between the two populations. Collectively, this study represents the first unbiased survey of viruses and viroids in both cultivated and wild grapevines within a defined geographic region.

Mining Grapevine Downy Mildew Susceptibility Genes: A Resource for Genomics-Based Breeding and Tailored Gene Editing.

Several pathogens continuously threaten viticulture worldwide. Until now, the investigation on resistance loci has been the main trend to understand the interaction between grapevine and the mildew causal agents. Dominantly inherited gene-based resistance has shown to be race-specific in some cases, to confer partial immunity, and to be potentially overcome within a few years since its introgression. Recently, on the footprint of research conducted in Arabidopsis, putative genes associated with downy mildew susceptibility have been discovered also in the grapevine genome. In this work, we deep-sequenced four putative susceptibility genes-namely VvDMR6.1, VvDMR6.2, VvDLO1, VvDLO2-in 190 genetically diverse grapevine genotypes to discover new sources of broad-spectrum and recessively inherited resistance. Identified Single Nucleotide Polymorphisms were screened in a bottleneck analysis from the genetic sequence to their impact on protein structure. Fifty-five genotypes showed at least one impacting mutation in one or more of the scouted genes. Haplotypes were inferred for each gene and two of them at the VvDMR6.2 gene were found significantly more represented in downy mildew resistant genotypes. The current results provide a resource for grapevine and plant genetics and could corroborate genomic-assisted breeding programs as well as tailored gene editing approaches for resistance to biotic stresses.

Identification of grapevine Pinot gris virus in free-living Vitis spp. located in riparian areas adjacent to commercial vineyards.

Grapevine Pinot gris virus (GPGV) is a recently identified pathogen of grapevines in California. To advance our knowledge about the epidemiology of GPGV, we investigated if free-living Vitis spp. can represent a source of virus infection. In 2019 a field survey of GPGV infection was conducted in Napa County. During the inspection 60 free-living vines in riparian habitats near commercial vineyards with GPGV infection were sampled. Samples were tested by real-time reverse transcription PCR (RT-PCR), identifying 23 free-living Vitis spp. positive for GPGV. Later, GPGV infection was confirmed in these plants via end-point RT-PCR and Sanger sequencing. Based on sequence analysis, detected GPGV isolates are more related to the asymptomatic variant of the virus. Vitis species ancestry was determined by DNA fingerprinting. GPGV-infected material included V. californica, V. californica × V. vinifera hybrids and hybrid rootstock cultivars. Here, GPGV is reported for the first time in free-living Vitis spp. The results of this study will support the development of management strategies for GPGV in California and beyond.

Fungicide Resistance Profiles of Botrytis cinerea Isolates From Michigan Vineyards and Development of a TaqMan Assay for Detection of Fenhexamid Resistance.

Botrytis cinerea on grapes causes bunch rot at both pre- and postharvest stages, in which losses can reach up to 100%. Chemical control primarily relies on the prophylactic use of site-specific fungicides. Repeated applications of these products raise the risk of fungicide resistance development in B. cinerea populations, which can result in disease control failures. To determine the extent of resistance, B. cinerea isolates were collected from grape clusters in the northwest and southwest grape growing regions of Michigan in 2014 and 2018 (n = 115 and 125, respectively). These isolates were phenotyped using discriminatory doses of eight fungicides to determine the levels of resistance. Fungicide resistance increased from 2014 to 2018, mostly affecting the active ingredients fenhexamid, fluopyram, and iprodione. B. cinerea isolates resistant to multiple fungicides were detected in 2014 and 2018, with a higher frequency of resistance in 2018. TaqMan real-time PCR has been developed to detect B. cinerea fungicide resistance to fenhexamid and to differentiate the erg27 F412S/I/V alleles. The TaqMan assay was tested for sensitivity, specificity, and reproducibility on purified DNA and infected grape tissue samples. Our data provide essential information to growers about the efficacy for B. cinerea control using the available botryticides. Furthermore, the developed fenhexamid markers will be transferred to diagnostic clinics to assist growers in the management of bunch rot before resistance-related control failures occur.

Grapevine Asteroid Mosaic-Associated Virus is Resident and Prevalent in Wild, Noncultivated Grapevine of New York State.

In North America, uncultivated, free-living grapevines (Vitis spp.) frequently grow alongside their cultivated counterparts, thus increasing the potential for exchange of microbiota. For this study, we used high-throughput sequencing (HTS) of small RNAs to survey for virus populations in free-living grapevines of the Finger Lakes region of New York State. Of 32 grapevines analyzed, 23 were free-living vines, while the remaining 9 were commercially grown Vitis vinifera plants from the same region. In total, 18 (78.3%) of the free-living grapevines tested were positive for grapevine asteroid mosaic-associated virus (GAMaV) infection by HTS, with detection confirmed by seminested reverse-transcription PCR and sequencing of nine isolates. Phylogenetic analyses of an ungapped alignment of the New York GAMaV sequences (length: 2,334 nucleotides) with the five known full-length or close to full-length global sequences showed that the New York isolates were broadly grouped. Of the nine cultivated plants, eight were infected with both hop stunt viroid and grapevine yellow speckle viroid 1, three were singly infected with grapevine leafroll-associated virus 3, and one harbored GAMaV. This limited survey of free-living grapevines, one of the first to use HTS, has highlighted the high incidence of a virus associated with disease in commercial V. vinifera.