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Background: A key step in domestication of the grapevine was the transition fromseparate sexes (dioecy) in wild Vitis vinifera ssp. sylvestris (V. sylvestris) to hermaphroditismincultivated Vitis vinifera ssp. sativa (V. vinifera). It is known that V. sylvestris has an XY systemand V. vinifera a modified Y haplotype (Yh) and that the sex locus is small, but it has notpreviously been precisely characterized.Results: We generate a high-quality de novo reference genome for V. sylvestris, onto whichwe map whole-genome re-sequencing data of a cross to locate the sex locus. Assembly ofthe full X, Y, and Yh haplotypes of V. sylvestris and V. vinifera sex locus and examining theirgene content and expression profiles during flower development in wild and cultivatedaccessions show that truncation and deletion of tapetum and pollen development geneson the X haplotype likely causes male sterility, while the upregulation of a Y allele of acytokinin regulator (APRT3) may cause female sterility. The downregulation of this cytokininregulator in the Yh haplotype may be sufficient to trigger reversal to hermaphroditism.Molecular dating of X and Y haplotypes is consistent with the sex locus being as old as theVitis genus, but the mechanism by which recombination was suppressed remainsundetermined.Conclusions: We describe the genomic and evolutionary characterization of the sex locusof cultivated and wild grapevine, providing a coherent model of sex determination in thelatter and for transition from dioecy to hermaphroditism during domestication.

WRKY proteins comprise a large family of transcription factors that play important roles in plant defence regulatory networks, including responses to various biotic and abiotic stresses. To date, no large-scale study of WRKY genes has been undertaken in grape (Vitis vinifera L.). In this study, a total of 59 putative grape WRKY genes (VvWRKY) were identified and renamed on the basis of their respective chromosome distribution. A multiple sequence alignment analysis using all predicted grape WRKY genes coding sequences, together with those from Arabidopsis thaliana and tomato (Solanum lycopersicum), indicated that the 59 VvWRKY genes can be classified into three main groups (I–III). An evaluation of the duplication events suggested that several WRKY genes arose before the divergence of the grape and Arabidopsis lineages. Moreover, expression profiles derived from semiquantitative PCR and real-time quantitative PCR analyses showed distinct expression patterns in various tissues and in response to different treatments. Four VvWRKY genes showed a significantly higher expression in roots or leaves, 55 responded to varying degrees to at least one abiotic stress treatment, and the expression of 38 were altered following powdery mildew (Erysiphe necator) infection. Most VvWRKY genes were downregulated in response to abscisic acid or salicylic acid treatments, while the expression of a subset was upregulated by methyl jasmonate or ethylene treatments.

The grape is one of the earliest domesticated fruit crops and, since antiquity, it has been widely cultivated and prized for its fruit and wine. Here, we characterize genome-wide patterns of genetic variation in over 1,000 samples of the domesticated grape, Vitis vinifera subsp. vinifera, and its wild relative, V. vinifera subsp. sylvestris from the US Department of Agriculture grape germplasm collection. We find support for a Near East origin of vinifera and present evidence of introgression from local sylvestris as the grape moved into Europe. High levels of genetic diversity and rapid linkage disequilibrium (LD) decay have been maintained in vinifera, which is consistent with a weak domestication bottleneck followed by thousands of years of widespread vegetative propagation. The considerable genetic diversity within vinifera, however, is contained within a complex network of close pedigree relationships that has been generated by crosses among elite cultivars. We show that first-degree relationships are rare between wine and table grapes and among grapes from geographically distant regions. Our results suggest that although substantial genetic diversity has been maintained in the grape subsequent to domestication, there has been a limited exploration of this diversity. We propose that the adoption of vegetative propagation was a double-edged sword: Although it provided a benefit by ensuring true breeding cultivars, it also discouraged the generation of unique cultivars through crosses. The grape currently faces severe pathogen pressures, and the long-term sustainability of the grape and wine industries will rely on the exploitation of the grape's tremendous natural genetic diversity.

• Grapevine trunk diseases have devastating consequences on vineyards worldwide. European wild grapevines (Vitis vinifera subs. sylvestris) from the last viable population in Germany along the Rhine river showed variable degrees of resistance against Neofusicoccum parvum (strain Bt-67), a fungus associated with Botryosphaeriaceae-related dieback. • Representative genotypes from different subclades of this population were mapped with respect to their ability to induce wood necrosis, as well as their defence responses in a controlled inoculation system. • The difference in colonization patterns could be confirmed by cryo-scanning electron microscopy, while there was no relationship between vessel diameter and infection success. Resistant lines accumulated more stilbenes, that were in addition significantly partitioned to nonglycosylated viniferin trimers. By contrast, the susceptible genotypes accumulated less stilbenes with a significantly higher proportion of glycosylated piceid. • We suggest a model in which in the resistant genotypes phenylpropanoid metabolism is channelled rapidly and specifically to the bioactive stilbenes. Our study specifies a resistant chemotype against grapevines trunk diseases and paves a way to breed for resistance against grapevine Botryosphaeriaceae-related dieback.

Across plants, leaves exhibit profound diversity in shape. As a single leaf expands, its shape is in constant flux. Plants may also produce leaves with different shapes at successive nodes. In addition, leaf shape varies among individuals, populations and species as a result of evolutionary processes and environmental influences. Because leaf shape can vary in many different ways, theoretically, the effects of distinct developmental and evolutionary processes are separable, even within the shape of a single leaf. Here, we measured the shapes of > 3200 leaves representing > 270 vines from wild relatives of domesticated grape (Vitis spp.) to determine whether leaf shapes attributable to genetics and development are separable from each other. We isolated latent shapes (multivariate signatures that vary independently from each other) embedded within the overall shape of leaves. These latent shapes can predict developmental stages independent from species identity and vice versa. Shapes predictive of development were then used to stage leaves from 1200 varieties of domesticated grape (Vitis vinifera), revealing that changes in timing underlie leaf shape diversity. Our results indicate that distinct latent shapes combine to produce a composite morphology in leaves, and that developmental and evolutionary contributions to shape vary independently from each other.

Background Calcium-dependent protein kinases (CDPKs) play vital roles in plant growth and development, biotic and abiotic stress responses, and hormone signaling. Little is known about the CDPK gene family in grapevine. Results In this study, we performed a genome-wide analysis of the 12X grape genome ( Vitis vinifera ) and identified nineteen CDPK genes. Comparison of the structures of grape CDPK genes allowed us to examine their functional conservation and differentiation. Segmentally duplicated grape CDPK genes showed high structural conservation and contributed to gene family expansion. Additional comparisons between grape and Arabidopsis thaliana demonstrated that several grape CDPK genes occured in the corresponding syntenic blocks of Arabidopsis , suggesting that these genes arose before the divergence of grapevine and Arabidopsis . Phylogenetic analysis divided the grape CDPK genes into four groups. Furthermore, we examined the expression of the corresponding nineteen homologous CDPK genes in the Chinese wild grape ( Vitis pseudoreticulata ) under various conditions, including biotic stress, abiotic stress, and hormone treatments. The expression profiles derived from reverse transcription and quantitative PCR suggested that a large number of VpCDPKs responded to various stimuli on the transcriptional level, indicating their versatile roles in the responses to biotic and abiotic stresses. Moreover, we examined the subcellular localization of VpCDPKs by transiently expressing six VpCDPK-GFP fusion proteins in Arabidopsis mesophyll protoplasts; this revealed high variability consistent with potential functional differences. Conclusions Taken as a whole, our data provide significant insights into the evolution and function of grape CDPKs and a framework for future investigation of grape CDPK genes.

Grapevine ( ssp. ) is a perennial crop especially important for wine and fruit production. The species is highly polymorphic with thousands of different varieties selected by farmers and clonally propagated. However, it is still debated whether grapevine domestication from its wild ancestor ( ssp. ) has been a single event or rather it occurred on multiple occasions during the diffusion of its cultivation across the Mediterranean. Located in the center of the Basin, Sicily is its largest island and has served as a hotspot for all civilizations that have crossed the Mediterranean throughout history. Hundreds of unique grapevine cultivars are still cultivated in Sicily and its surrounding minor islands, though most of them are menaced by extinction. Wild grapevine is also present with isolated populations thriving along riverbanks. With the aim to evaluate the phylogenetic relationships among Sicilian varieties, and to assess the possible contribution of indigenous wild populations to the genetic makeup of cultivated grapevine, we analyzed 170 domestic cultivars and 125 wild plants, collected from 10 different populations, with 23 SSR markers. We also compared our data with published dataset from Eurasia. Results show that Sicilian wild populations are related to the cultivated Sicilian and Italian germplasm, suggesting events of introgression and/or domestication of local varieties.

Premise of the Study The stem of Vitis vinifera, a climbing vine of global economic importance, is characterized by both wide and narrow vessels and high specific hydraulic conductivity. While the effect of drought stress has been studied in 1‐ and 2‐yr‐old stems, there are few data documenting effects of drought stress on the anatomical structure of the mature, woody stem near the base of the vine. Here we describe mature wood anatomical responses to two irrigation regimes on wood anatomy and specific hydraulic conductivity in Vitis vinifera Merlot vines. Methods For 4 years, irrigation was applied constantly at low, medium, or high levels, or at alternating levels at two different periods during the growing season, either early spring or late summer, resulting in late season or early spring deficits, respectively. The following variables were measured: trunk diameter, annual ring width and area, vessel diameter, specific hydraulic conductivity and stem water potential. Key Results High water availability early in the season (late deficit) resulted in vigorous vegetative growth (greater trunk diameter, ring width and area), wider vessels and increased specific hydraulic conductivity. High water availability early in the season caused a shift of the vessel population towards the wider frequency classes. These late deficit vines showed more negative water potential values late in the season than vines that received low but relatively constant irrigation. Conclusions We concluded that high water availability during vegetative growth period of Vitis increases vessels diameter and hydraulic conductivity and causes the vines to be more vulnerable to drought stress late in the season.

The genetic diversity and relationship between wild (Vitis vinifera L. subsp. sylvestris (Gmel.) Hegi and cultivated (V. vinifera L. subsp. vinifera) grapevine in the western Balkan region and Central Europe have not been studied together previously, although this area has a rich viticultural past. Here, we studied wild grapevine populations sampled from their natural habitats in several countries of the western Balkan region and Central Europe. Their genetic diversity and structure were compared to cultivars that are traditionally in use in this region. A sample set of 243 accessions was genotyped at 20 nuclear microsatellite loci, including 167 sylvestris and 76 diverse vinifera cultivars. The genetic diversity of the wild grapevines was lower than that of cultivars by all genetic parameters. Both hierarchical and nonhierarchical clustering methods differentiated two main groups, indicating clear separation between wild and cultivated vines but also revealed clear gene flow between the cultivated and wild gene pools through overlaps and admixed ancestry values in the graphs. There was greater affinity to the wild grapes in Central European cultivars than in Balkan cultivars. Fine arrangement of the structure among cultivated grapevines showed differentiation among Central European and Balkan cultivars. These results confirm the divergence of wild grapes from vinifera and highlight the “crossroad” role of the western Balkan peninsula in the broader context of European viticulture.

Soil analysis is used to estimate nutrient availability, but nutrient concentrations are not always related to yield in most fruit plants, including grapevines. Thus, additional multivariate mathematical models, such as the compositional nutrient diagnosis (CND), which takes into account leaves nutrient concentration, and yield, can contribute to estimating critical levels or sufficiency bands of elements, as well as to detect deficiency and/or excess of nutrients. The aim of the present study was to establish CND standards, and the critical level and sufficiency band of nutrients, in the grapevine Vitis vinifera L., grown in a subtropical climate region. Leaves were collected in 81 vineyards in the Campanha Gaúcha do Rio Grande do Sul region, Southern Brazil, and analyzed for macro- and micro-nutrient concentration. The yield of each vineyard was assessed. Grapevine nutritional status was calculated through the CND method. CND-r2 indices were effective in establishing the nutritional status of grapevines for macro- and micro-nutrients as sub-optimal, excessive, or balanced. The CND methodology established the critical level and sufficiency bands of nutrients more accurately than the current recommendations for grapevines. Multi-nutrient associations were more effective than the single nutrient determination in defining the threshold of a given nutrient that can reduce grapevine yield.