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 resistanc

Emergent Ascomycetes in viticulture: an interdisciplinary overview

The reduction of pesticide usage is a current imperative and the implementation of sustainable viticulture is an urgent necessity. A potential solution, which is being increasingly adopted, is offered by the use of grapevine cultivars resistant to its main pathogenic threats. This, however, has contributed to changes in defense strategies resulting in the occurrence of secondary diseases, which were previously controlled. Concomitantly, the ongoing climate crisis is contributing to destabilizing the increasingly dynamic viticultural context. In this review, we explore the available knowledge on three Ascomycetes which are considered emergent and causal agents of powdery mildew, black rot and anthracnose. We also aim to provide a survey on methods for phenotyping disease symptoms in fields, greenhouse and lab conditions, and for disease control underlying the insurgence of pathogen resistance to fungicide. Thus, we discuss fungal genetic variability, highlighting the usage and development of molecular markers and barcoding, coupled with genome sequencing. Moreover, we extensively report on the current knowledge available on grapevine-ascomycete interactions, as well as the mechanisms developed by the host to counteract the attack. Indeed, to better understand these resistance mechanisms, it is relevant to identify pathogen effectors which are involved in the infection process and how grapevine resistance genes function and impact the downstream cascade. Dealing with such a wealth of information on both pathogens and the host, the horizon is now represented by multidisciplinary approaches, combining traditional and innovative methods of cultivation. This will support the translation from theory to practice, in an attempt to understand biology very deeply and manage the spread of these Ascomycetes

Marker-assisted breeding for Downy mildew, Powderey mildew and Phylloxera resistance at FEM

Il programma di miglioramento genetico per le resistenze a stress biotici ha avuto inizio presso la Fondazione Edmund Mach (FEM) nel 2010. Inizialmente è stata condotta una caratterizzazione sia genotipica che fenotipica di materiali acquisiti da altri programmi di breeding e di materiale selvatico raccolto in New Jersey. Sia i genotipi conosciuti nei database internazionali che i genotipi sconosciuti, imparentati e non, sono stati impiegati come linee parentali nel processo di introgressione e di piramidazione di loci di interesse. Una volta pianificati e ottenuti gli incroci, la valutazione delle progenie è avvenuta seguendo un processo di Marker-Assisted Selection: dapprima è avvenuta la selezione fenotipica in serra in base al tipo di malattia e al numero di loci attesi per la medesima malattia; successivamente si è proceduto con lo screening molecolare in base ai loci specifici attesi nei parentali. Cinque sono i loci Run/Ren associati alla resistenza all'oidio presenti nel programma FEM; riguardo ai loci associati alla resistenza alla peronospora, quattro Rpv sono ben rappresentati nel piano di incroci. Ad oggi il 26% delle F1 è piramidizzato per quattro loci di resistenza

Studi sui tratti di qualità negativa nelle nuove viti mediamente resistenti alle malattie fungine

The varieties of vines tolerant to the main fungal diseases are the object of growing interest because they require fewer treatments for the production of quality grapes. These varieties are considered qualitatively unsuitable especially for some characteristics such as the foxy aroma, the level of anthocyanin diglucosides and the level of methyl alcohol in wines. In the three-year period 2016-2018, from a population of 74 medium resistant varieties placed in a single collection, in Marlengo (BZ), young leaves were taken for DNA analysis; annually 2 surveys of the OIV 452 453, 455 and 456 descriptors were carried out, and, at 18° Brix, sombunches were collected for the analysis of different metabolites. Nano-vinifications were carried out for 32 genotypes. Genotypes with different levels of stacked loci associated with resistance to downy and powdery mildew were detected, with different resistance behaviors over the years. Only some genotypes have aromas of foxy and only 20% of the genotypes analyzed have levels of anthocyanin diglucosides higher than the levels required by current legislation. Some vinifications showed methanol levels higher than the legal limit set. High percentages of genotypes respected the legal levels of metabolites required by current regulations and only genotypes with the presence of V. labrusca showed the aroma of foxy

Towards Marker-Assisted Breeding for Black Rot Bunch Resistance: Identification of a Major QTL in the Grapevine Cultivar ‘Merzling’

Black rot (BR), caused by Guignardia bidwellii, is an emergent fungal disease threatening viticulture and affecting several mildew-tolerant varieties. However, its genetic bases are not fully dissected yet. For this purpose, a segregating population derived from the cross ‘Merzling’ (hybrid, resistant) × ‘Teroldego’ (V. vinifera, susceptible) was evaluated for BR resistance at the shoot and bunch level. The progeny was genotyped with the GrapeReSeq Illumina 20K SNPchip, and 7175 SNPs were combined with 194 SSRs to generate a high-density linkage map of 1677 cM. The QTL analysis based on shoot trials confirmed the previously identified Resistance to Guignardia bidwellii (Rgb)1 locus on chromosome 14, which explained up to 29.2% of the phenotypic variance, reducing the genomic interval from 2.4 to 0.7 Mb. Upstream of Rgb1, this study revealed a new QTL explaining up to 79.9% of the variance for bunch resistance, designated Rgb3. The physical region encompassing the two QTLs does not underlie annotated resistance (R)-genes. The Rgb1 locus resulted enriched in genes belonging to phloem dynamics and mitochondrial proton transfer, while Rgb3 presented a cluster of pathogenesis-related Germin-like protein genes, promoters of the programmed cell death. These outcomes suggest a strong involvement of mitochondrial oxidative burst and phloem occlusion in BR resistance mechanisms and provide new molecular tools for grapevine marker-assisted breeding

Magnesium supplementation in Gitelman syndrome

The metabolism of potassium and magnesium are closely linked (in situations where potassium and magnesium depletion coexist, magnesium restoration alone is sufficient to correct hypokalemia). Moreover, magnesium deficiency blunts the interplay between circulating calcium and the calciotropic hormones. Renal magnesium wasting, hypokalemia, alkalosis, hypocalciuria, and a tendency towards hypocalcemia characterize Gitelman syndrome. Plasma or intracellular potassium, circulating calcium, and calciotropic hormones were therefore investigated in eight patients (4 females, 4 males, aged 9-20 years) with Gitelman syndrome before and during oral supplementation with magnesium pyrrolidone carboxylate 30 mmol daily for 4 weeks. Magnesium supplementation significantly increased plasma and intracellular magnesium and plasma calcium, but failed to completely restore magnesium deficiency. In contrast, blood levels of parathyroid hormone and calcitriol and plasma and intracellular potassium were not modified following magnesium administration

Effects of Mg pyrrolidone carboxylate treatment in Gitelman's syndrome

The results of this study seem to indicate the use of magnesium pyrrolidone carboxylate in the treatment of Gitelman' syndrom

Genomic Designing for Biotic Stress Resistant Grapevine

Grapevines are challenged by a range of diseases and pests, causing economic losses and requiring often costly approaches to mitigate damage. Public interest in reducing the use of chemicals is a related challenge, along with climate change. Yet, the Vitis gene pool provides vast resources for the development of genetic resistance in rootstock and scion cultivars. Traditional breeding approaches have made great strides in the development of adaptive traits, and recent access to ‘omic technologies has further facilitated the identification of useful loci along with rapid trait introgression from wild species. Moreover, marker technologies are now used to stack multiple genes for the same trait into a single genotype, a heretofore barely accessible technology. Genomic technologies are also impacting germplasm characterization, and thereby facilitating “Breeding by Design” approaches. Genetic transformation and gene-editing technologies are also applicable for both cultivar improvement as well as functional studies of genes. The landscape for acceptance of new resistant cultivars is complex and with wine grapes, subject to high degrees of regulation especially in the European Union. With rootstocks, as well as table/raisin grapes, gaining acceptance in the marketplace for new cultivars developed through either traditional or marker-assisted approaches is routine. Yet even in the highly regulated EU environment, the adoption of new wine cultivars of interspecific origins is beginning to take place in both traditional wine growing regions as well as non-traditional regions nearby

GENETIC-HETEROGENEITY IN TUBULAR HYPOMAGNESEMIA HYPOKALEMIA WITH HYPOCALCIURIA (GITELMAN'S SYNDROME)

To better clarify the genetic inheritance of primary tubular hypomagnesemia-hypokalemia with hypocalciuria, or Gitelman's syndrome (GS), we studied eight families (10 patients aged 11 to 22 years; 16 parents; 9 siblings) in which at least one offspring had GS (plasma magnesium < 0.65 mmol/liter; plasma potassium < 3.6 mmol/liter; high magnesium and potassium fractional excretions; molar urinary calcium/creatinine < 0.10). Two families each had two offspring of different sex with GS, who all had tetanic episodes and/or marked weakness during childhood or adolescence, whereas in three other families two mothers and three offspring presented GS and one father and two other offspring had hypomagnesemia and hypocalciuria but normal plasma potassium. The mean plasma magnesium and potassium levels of the patients of the first two families were significantly lower (P < 0.05) than those of the other three families. Intralymphocytic but not intraerythrocytic magnesium and potassium were significantly lower (P < 0.05) in patients compared to controls. We hypothesize that there are two different types of genetic transmission of GS, one autosomal recessive and one autosomal dominant with high phenotypic variability. It seems that this genetic heterogeneity is associated with a different clinical expression with frequent tetanic episodes and lower plasma potassium and magnesium levels in the autosomal recessive form