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Among the main challenges in current viticulture, there is the increasing demand for sustainability in the protection from fungal diseases, such as downy mildew (DM) and powdery mildew (PM). Breeding disease-resistant grapevine varieties is a key strategy for better managing fungicide inputs. This study explores the diversity of grapevine germplasm (cultivated and wild) from Caucasus and neighboring areas to identify genotypes resistant to DM and PM, based on 13 Simple Sequence Repeat (SSR) loci and phenotypical (artificial pathogen inoculation) analysis, and to identify loci associated with DM and PM resistance, via Genome-Wide Association Analysis (GWAS) on Single Nucleotide Polymorphism (SNP) profiles. SSR analysis revealed resistant alleles for 16 out of 88 genotypes. Phenotypic data identified seven DM and 31 PM resistant genotypes. GWAS identified two new loci associated with DM resistance, located on chromosome 15 and 16 (designated as Rpv36 and Rpv37 ), and two with PM resistance, located on chromosome 6 and 17 (designated as Ren14 and Ren15 ). The four novel loci identified genomic regions rich in genes related to biotic stress response, such as genes involved in pathogen recognition, signal transduction and resistance response. This study highlights potential candidate genes associated with resistance to DM and PM, providing valuable insights for breeding programs for resistant varieties. To optimize their utilization, further functional characterization studies are recommended., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2024 Ricciardi, Crespan, Maddalena, Migliaro, Brancadoro, Maghradze, Failla, Toffolatti and De Lorenzis.)

The plant disease onset is a complex event that occurs when the pathogen and the host encounter in a favorable environment. While the plant-pathogen interaction has been much investigated, little attention has been given to the phenological synchrony of the event, especially when both plant and pathogen overwinter, as in the case of grapevines and the downy mildew agent, the oomycete Plasmopara viticola . Oospores allow this obligate parasite to survive grapevine dormancy and, germinating, produce inoculum for primary infections. During overwintering, environmental factors influence the potential oospore germination. This study aimed at investigating the existence of synchrony between the pathogen and the host by identifying and quantifying the most important factors determining oospore maturation and germination and the relationship existing with grapevine phenology. Generalized linear models (GLM and GLMM) were used to analyze the germination dynamics of the oospores overwintered in controlled and field conditions and incubated in isothermal conditions, and oospore viability tests were carried out at different time points. Results showed that the most indicative parameter to describe the germination dynamics is the time spent by the oospores from the start of overwintering. The oospores overwintered in field showed phenological traits related to grapevine phenology not observed in controlled conditions. In particular, they completed the maturation period by the end of grapevine dormancy and germinated more rapidly at plant sprouting, when grapevine reaches susceptibility. Overall, the oospores proved to be able to modulate their behavior in close relationship with grapevine, showing a great adaptation to the host's phenology., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Maddalena, Russo and Toffolatti.)

Downy mildew, caused by the oomycete Plasmopara viticola , is one of the diseases causing the most severe economic losses to grapevine ( Vitis vinifera ) production. To date, the application of fungicides is the most efficient method to control the pathogen and the implementation of novel and sustainable disease control methods is a major challenge. RNA interference (RNAi) represents a novel biotechnological tool with a great potential for controlling fungal pathogens. Recently, a candidate susceptibility gene ( VviLBDIf7 ) to downy mildew has been identified in V. vinifera . In this work, the efficacy of RNAi triggered by exogenous double-stranded RNA (dsRNA) in controlling P. viticola infections has been assessed in a highly susceptible grapevine cultivar (Pinot noir) by knocking down VviLBDIf7 gene. The effects of dsRNA treatment on this target gene were assessed by evaluating gene expression, disease severity, and development of vegetative and reproductive structures of P. viticola in the leaf tissues. Furthermore, the effects of dsRNA treatment on off-target ( EF1α , GAPDH , PEPC , and PEPCK ) and jasmonic acid metabolism ( COI1 ) genes have been evaluated. Exogenous application of dsRNA led to significant reductions both in VviLBDIf7 gene expression, 5 days after the treatment, and in the disease severity when artificial inoculation was carried out 7 days after dsRNA treatments. The pathogen showed clear alterations to both vegetative (hyphae and haustoria) and reproductive structures (sporangiophores) that resulted in stunted growth and reduced sporulation. Treatment with dsRNA showed signatures of systemic activity and no deleterious off-target effects. These results demonstrated the potential of RNAi for silencing susceptibility factors in grapevine as a sustainable strategy for pathogen control, underlying the possibility to adopt this promising biotechnological tool in disease management strategies., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Marcianò, Ricciardi, Marone Fassolo, Passera, Bianco, Failla, Casati, Maddalena, De Lorenzis and Toffolatti.)

Plasmopara viticola (Berk. et Curt.) Berl. and de Toni, the agent of downy mildew, is one of the most important pathogens of European grapevine ( Vitis vinifera L.). Extensive evaluation of cultivated grapevine germplasm has highlighted the existence of resistant phenotypes in the Georgian (Southern Caucasus) germplasm. Resistance is shown as a reduction in disease severity. Unraveling the genetic architecture of grapevine response to P. viticola infection is crucial to develop resistant varieties and reduce the impact of disease management. The aim of this work was to apply a genome-wide association (GWA) approach to a panel of Georgian-derived accessions phenotyped for P. viticola susceptibility and genotyped with Vitis18kSNP chip array. GWA identified three highly significant novel loci on chromosomes 14 ( Rpv29 ), 3 ( Rpv30 ) and 16 ( Rpv31 ) associated with a low level of pathogen sporulation. Rpv29 , Rpv30 , and Rpv31 loci appeared to be associated with plant defense genes against biotic stresses, such as genes involved in pathogen recognition and signal transduction. This study provides the first evidence of resistant loci against P. viticola in V. vinifera germplasm, and identifies potential target genes for breeding P. viticola resistant grapevine cultivars., (Copyright © 2020 Sargolzaei, Maddalena, Bitsadze, Maghradze, Bianco, Failla, Toffolatti and De Lorenzis.)