Dual domestications and origin of traits in grapevine evolution.

We elucidate grapevine evolution and domestication histories with 3525 cultivated and wild accessions worldwide. In the Pleistocene, harsh climate drove the separation of wild grape ecotypes caused by continuous habitat fragmentation. Then, domestication occurred concurrently about 11,000 years ago in Western Asia and the Caucasus to yield table and wine grapevines. The Western Asia domesticates dispersed into Europe with early farmers, introgressed with ancient wild western ecotypes, and subsequently diversified along human migration trails into muscat and unique western wine grape ancestries by the late Neolithic. Analyses of domestication traits also reveal new insights into selection for berry palatability, hermaphroditism, muscat flavor, and berry skin color. These data demonstrate the role of the grapevines in the early inception of agriculture across Eurasia.

Influence of late pruning practice on two red skin grapevine cultivars in a semi-desert climate.

Continually increasing global temperature could severely affect grape berry metabolite accumulation and ultimately wine polyphenol concentration and color intensity. To explore the effect of late shoot pruning on grape berry and wine metabolite composition, field trials were carried out on Vitis vinifera cv. Malbec and cv. Syrah grafted on 110 Richter rootstock. Fifty-one metabolites were detected and unequivocally annotated employing UPLC-MS based metabolite profiling. Integrating the data using hierarchical clustering showed a significant effect of late pruning treatments on must and wine metabolites. Syrah metabolite profiles were characterized by a general trend of higher metabolite content in the late shoot pruning treatments, while Malbec profiles did not show a consistent trend. In summary, late shoot pruning exerts a significant effect, though varietal specific, on must and wine quality-related metabolites, possibly related to enhanced photosynthetic efficiency, which should be taken into consideration when planning mitigating strategies in warm climates.

A Field Collection of Indigenous Grapevines as a Valuable Repository for Applied Research.

The grapevine is an economically important plant, with a historical connection to the development of human culture. Currently, over 6000 accessions are known as individual grapevine varieties, some of which are important to national heritage, valuable for current viticultural practices, and as genetic resources to maintain plasticity under changing climatic conditions, environmental sustainability, and market demands. Recently, the diversity of cultivated grapevines has declined significantly, due to the increased focus of global wine industries on a few major cultivars. Moreover, due to biotic and abiotic stresses, the wild V. vinifera germplasm's genetic diversity has declined, with some varieties on the verge of extinction. Vitis germplasm conservation can be achieved via either in situ (e.g., protected areas) or Ex situ (e.g., field collections, seed banks, and tissue culture collections) methods. This study aims to highlight the importance of Vitis field bank collections. We demonstrate the research done in the Israeli indigenous Vitis vinifera collection. The multi-layer analysis of the varieties enabled the identification of drought stress-resistant varieties, and suggested a mechanism for this resistance through noting the dramatic phenological differences in foliage development between resistant and sensitive varieties. In addition, we show a general characterization of the varieties via major grape characteristics, including bunch and berry shape, as well as their possible utilization based on their aromatic and phenolic profiles.

Integration of the Connectivity Map and Pathway Analysis to Predict Plant Extract's Medicinal Properties-The Study Case of Sarcopoterium spinosum L.

Medicinal properties of plants are usually identified based on knowledge of traditional medicine or using low-throughput screens for specific pharmacological activities. The former is very biased since it requires prior knowledge of plants' properties, while the latter depends on a specific screening system and will miss medicinal activities not covered by the screen. We sought to enrich our understanding of the biological activities of Sarcopoterium spinosum L. root extract based on transcriptome changes to uncover a plurality of possible pharmacological effects without the need for prior knowledge or functional screening. We integrated Gene Set Enrichment Analysis of the RNAseq data to identify pathways affected by the treatment of cells with the extract and perturbational signatures in the CMAP database to enhance the validity of the results. Activities of signaling pathways were measured using immunoblotting with phospho-specific antibodies. Mitochondrial membrane potential was assessed using JC-1 staining. SARS-CoV-2-induced cell killing was assessed in Vero E6 and A549 cells using an MTT assay. Here, we identified transcriptome changes following exposure of cultured cells to the medicinal plant Sarcopoterium spinosum L. root extract. By integrating algorithms of GSEA and CMAP, we confirmed known anti-cancer activities of the extract and predicted novel biological effects on oxidative phosphorylation and interferon pathways. Experimental validation of these pathways uncovered strong activation of autophagy, including mitophagy, and excellent protection from SARS-CoV-2 infection. Our study shows that gene expression analysis alone is insufficient for predicting biological effects since some of the changes reflect compensatory effects, and additional biochemical tests provide necessary corrections. This study defines the advantages and limitations of transcriptome analysis in predicting the biological and medicinal effects of the Sarcopoterium spinosum L. extract. Such analysis could be used as a general approach for predicting the medicinal properties of plants.

The Effect of Irrigation-Initiation Timing on the Phenolic Composition and Overall Quality of Cabernet Sauvignon Wines Grown in a Semi-Arid Climate.

In semi-arid areas, vineyards grown for winemaking are usually mildly irrigated by drip irrigation systems in a manner maintaining drought stress. This practice ensures the proper development of vegetative and reproductive organs on the one hand, and on the other, the development of high-quality grapes which can be hampered by overly abundant water application. In previous work, we have developed and demonstrated an irrigation model suitable for high-quality grape production in semi-arid areas. Here, we tackle the question of proper irrigation initiation dates-should one wait for vines to develop drought stress before the initiation of irrigation, or rather commence irrigation earlier? Our results show that vines which undergo initial irrigation late in the growing season tend to develop a lower midday stem water potential even after irrigation initiation. In addition, these vines tend to produce a lower number of bunches per vine and smaller berry size, leading to lower yields. The wine produced from the late-irrigated treatments had a higher phenolic content, primarily due to higher levels of catechin and epicatechin. Their levels increased as irrigation initiation dates were delayed, while caffeic acid levels showed an opposite trend. Late irrigation also led to higher color intensities compared to those of irrigation at earlier stages, due to higher levels of most anthocyanins. Finally, we show that the overall wine sensory score, representing its overall quality, was approximately five points higher for wines made from delayed irrigation treatments compared to wines made from early season irrigation treatments.

Ecogeographic Conditions Dramatically Affect Trans-Resveratrol and Other Major Phenolics' Levels in Wine at a Semi-Arid Area.

Grapevines are susceptible and responsive to their surrounding environment. Factors such as climate region and terroir are known to affect polyphenolic compounds in wine and therefore, its quality. The uniqueness of the terroir in Israel is the variety of soil types and the climatic conditions, ranging from Mediterranean to arid climates. Thus, understanding the effects of climate on grapevine performance in Israel may be a test case for the effect of climate change on grapevine at other areas in the future. First, we present a preliminary survey (2012-2014) in different climate zones and terroirs, which showed that trans-resveratrol concentrations in Merlot and Shiraz were high, while those of Cabernet Sauvignon were significantly lower. A further comprehensive countrywide survey (2016) of Merlot wines from 62 vineyards (53 wineries) compared several phenolic compounds' concentrations between five areas of different climate and terroir. Results show a connection between trans-resveratrol concentrations, variety, and terroir properties. Furthermore, we show that trans-resveratrol concentrations are strongly correlated to humidity levels at springtime, precipitation, and soil permeability. This work can be considered a glimpse into the possible alterations of wine composition in currently moderate-climate wine-growing areas.

Antidepressant-like effects of a chlorogenic acid- and cynarine-enriched fraction from Dittrichia viscosa root extract.

Dittrichia viscosa is a perennial Mediterranean plant used in traditional medicine for "calming purposes", pointing at a possible antidepressant activity of the plant. We conducted chromatographic and bioassay-guided fractionation of D. viscosa root extract to isolate a specific fraction (fraction "K") with antidepressant-like characteristics in vivo and strong antioxidant properties in vitro. A single dose of "K" reduced immobility time in the forced swim test with a mouse model possessing a depressive-like phenotype. Neurochemical profiling for 5-hydroxytryptamine (5-HT) and its primary metabolite, 5-hydroxyindoleacetic acid (5-HIAA), in prefrontal cortex and hippocampus of "K"-treated mice showed reduction in 5-HIAA, indicative of either serotonin uptake transporter or monoamine oxidase-A inhibition, as well as slight increases in 5-HT content. These neurochemical alterations, as well as the behavioral changes observed, were comparable to the effects of paroxetine. "K" also protected PC12 cells in a H2O2 cytotoxicity assay, thus demonstrating antioxidant properties, yet paroxetine augmented oxidative damage and cell death. Identification of the main compounds in "K" by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) indicated that chlorogenic acid and cynarine comprised 87% of the total components. D. viscosa root extract appears to produce antidepressant and cytoprotective effects and may serve as an attractive alternative to standard therapies for depression.

Grape pomace reduces the severity of non-alcoholic hepatic steatosis and the development of steatohepatitis by improving insulin sensitivity and reducing ectopic fat deposition in mice.

While non-alcoholic fatty liver disease (NAFLD) represents the common cause of chronic liver disease, specific therapies are currently unavailable. The wine industry produces millions of tons of residue (pomace), which contains high levels of bioactive phytochemicals. The aim of this study was to clarify the potential benefits of grape pomace for the treatment of NAFLD at different levels of severity, and to clarify the mechanism of action. C57Bl/6 mice were given high fat diet (HFD) or western diet (WD) as models of obesity and hepatic steatosis or steatohepatitis, respectively, with or without pomace supplementation (50-250 mg/day). Pomace inhibited food intake, and reduced serum leptin and body weight gain. Ectopic fat deposition was reduced, while white adipose tissue mass was preserved. In addition, pomace improved glucose tolerance and insulin sensitivity, prevented the development of adipose tissue inflammation, and reduced hepatic steatosis. Higher expression of genes involved in fatty acids transport and oxidation was observed in adipose tissue, while lipogenic genes were attenuated in the liver of pomace-treated mice. In WD-fed mice, pomace reduced the severity of hepatic steatosis and inflammation and improved blood lipid profile, but was ineffective in reversing hepatic damage of advanced NASH. In conclusion, pomace improved insulin sensitivity and reduced ectopic fat deposition, leading to a healthier metabolic profile. Pomace may hold the potential as a supplement with beneficial health outcomes for the prevention and treatment of hepatic steatosis and other obesity-related pathologies.

Accurate classification of fresh and charred grape seeds to the varietal level, using machine learning based classification method.

Grapevine (Vitis vinifera L.) currently includes thousands of cultivars. Discrimination between these varieties, historically done by ampelography, is done in recent decades mostly by genetic analysis. However, when aiming to identify archaeobotanical remains, which are mostly charred with extremely low genomic preservation, the application of the genomic approach is rarely successful. As a result, variety-level identification of most grape remains is currently prevented. Because grape pips are highly polymorphic, several attempts were made to utilize their morphological diversity as a classification tool, mostly using 2D image analysis technics. Here, we present a highly accurate varietal classification tool using an innovative and accessible 3D seed scanning approach. The suggested classification methodology is machine-learning-based, applied with the Iterative Closest Point (ICP) registration algorithm and the Linear Discriminant Analysis (LDA) technique. This methodology achieved classification results of 91% to 93% accuracy in average when trained by fresh or charred seeds to test fresh or charred seeds, respectively. We show that when classifying 8 groups, enhanced accuracy levels can be achieved using a "tournament" approach. Future development of this new methodology can lead to an effective seed classification tool, significantly improving the fields of archaeobotany, as well as general taxonomy.

Demographic and ecogeographic factors limit wild grapevine spread at the southern edge of its distribution range.

The spatial distribution of plants is constrained by demographic and ecogeographic factors that determine the range and abundance of the species. Wild grapevine (Vitis vinifera ssp. sylvestris) is distributed from Switzerland in the north to Israel in the south. However, little is known about the ecogeographic constraints of this species and its genetic and phenotypic characteristics, especially at the southern edge of its distribution range in the Levant region. In this study, we explore the population structure of southern Levantine wild grapevines and the correlation between demographic and ecogeographic characteristics. Based on our genetic analysis, the wild grapevine populations in this region can be divided into two major subgroups in accordance with a multivariate spatial and ecogeographical clustering model. The identified subpopulations also differ in morphological traits, mainly leaf hairiness which may imply adaptation to environmental stress. The findings suggest that the Upper Jordan River population was spread to the Sea of Galilee area and that a third smaller subpopulation at the south of the Golan Heights may represent a distinguished gene pool or a recent establishment of a new population. A spatial distribution model indicated that distance to water sources, Normalized difference vegetation index, and precipitation are the main environmental factors constraining V. v. sylvestris distribution at its southern distribution range. These factors in addition to limited gene flow between populations prevent further spread of wild grapevines southwards to semi-arid regions.

Temperature Shift Between Vineyards Modulates Berry Phenology and Primary Metabolism in a Varietal Collection of Wine Grapevine.

Global climate change and the expected increase in temperature are altering the relationship between geography and grapevine (V. vinifera) varietal performance, and the implications of which are yet to be fully understood. We investigated berry phenology and biochemistry of 30 cultivars, 20 red and 10 white, across three seasons (2017-2019) in response to a consistent average temperature difference of 1.5°C during the growing season between two experimental sites. The experiments were conducted at Ramat Negev (RN) and Ramon (MR) vineyards, located in the Negev desert, Israel. A significant interaction between vineyard location, season, and variety affected phenology and berry indices. The warmer RN site was generally associated with an advanced phenological course for the white cultivars, which reached harvest up to 2 weeks earlier than at the MR site. The white cultivars also showed stronger correlation between non-consecutive phenological stages than did the red ones. In contrast, harvest time of red cultivars considerably varied according to seasons and sites. Warmer conditions extended fruit developmental phases, causing berry shriveling and cluster collapse in a few cultivars such as Pinot Noir, Ruby Cabernet, and Tempranillo. Analyses of organic acid content suggested differences between red and white cultivars in the content of malate, tartrate, and citrate in response to the temperature difference between sites. However, generally, cultivars at lower temperatures exhibited lower concentrations of pulp organic acids at véraison, but acid degradation until harvest was reduced, compared to the significant pace of acid decline at the warmer site. Sugars showed the greatest differences between sites in both white and red berries at véraison, but differences were seasonal dependent. At harvest, cultivars of both groups exhibited significant variation in hexose/sucrose ratio, and the averages of which varied from 1.6 to 2.9. Hexose/sucrose ratio was significantly higher among the red cultivars at the warmer RN, while this tendency was very slight among white cultivars. White cultivars seem to harbor a considerable degree of resilience due to a combination of earlier and shorter ripening phase, which avoids most of the summer heat. Taken together, our study demonstrates that the extensive genetic capacity of V. vinifera bears significant potential and plasticity to withstand the temperature increase associated with climate change.

Pre-germination plasma treatment of seeds does not alter cotyledon DNA structure, nor phenotype and phenology of tomato and pepper plants.

The impact of cold radiofrequency plasma on CPDs formation, and the morphological and phenological development of tomato and pepper plants and fruits in greenhouse conditions was studied. Quality characteristics of fruits: total sugars, titratable acidity, pH and total solids were determined. Our results show that plasma treatment in the time ranges used for pre-sowing treatments, did not cause the formation of CPDs in the cotyledons, even when the testa was removed before treatment, as opposed to high UV radiation. In addition, plasma treatment did not have a negative effect on the morphology, phenology and quality parameters of plants and fruits that grew up from treated tomato and pepper seeds in the greenhouse.

Isolation and Characterization of Live Yeast Cells from Ancient Vessels as a Tool in Bio-Archaeology.

Ancient fermented food has been studied based on recipes, residue analysis, and ancient-DNA techniques and reconstructed using modern domesticated yeast. Here, we present a novel approach based on our hypothesis that enriched yeast populations in fermented beverages could have become the dominant species in storage vessels and their descendants could be isolated and studied today. We developed a pipeline of yeast isolation from clay vessels and screened for yeast cells in beverage-related and non-beverage-related ancient vessels and sediments from several archaeological sites. We found that yeast cells could be successfully isolated specifically from clay containers of fermented beverages. The findings that genotypically the isolated yeasts are similar to those found in traditional African beverages and phenotypically they grow similar to modern beer-producing yeast strongly suggest that they are descendants of the original fermenting yeast. These results demonstrate that modern microorganisms can serve as a new tool in bio-archaeology research.IMPORTANCE So far, most of the study of ancient organisms has been based mainly on the analysis of ancient DNA. Here we show that it is possible to isolate and study microorganisms-yeast in this case-from ancient pottery vessels used for fermentation. We demonstrate that it is highly likely that these cells are descendants of the original yeast strains that participated in the fermentation process and were absorbed into the clay matrix of the pottery vessels. Moreover, we characterized the isolated yeast strains, their genomes, and the beer they produced. These results open new and exciting avenues in the study of domesticated microorganisms and contribute significantly to the fields of bio- and experimental archaeology that aim to reconstruct ancient artifacts and products.

Placental glucocorticoid receptor and 11ß-hydroxysteroid dehydrogenase-2 recruitment indicates impact of prenatal adversity upon postnatal development in mice.

Prenatal stress may increase concentrations of maternal glucocorticoids, which restrict fetal growth, with variable impact upon postnatal development. Among key regulators of stress hormone effects are the glucocorticoid receptor (GR) and 11ß-hydroxysteroid dehydrogenase-2 (11ßHSD2), the enzyme that inactivates glucocorticoid. This study utilized mice selectively bred for social dominance (Dom) or submissiveness (Sub), respectively exhibiting resilience or sensitivity to stress, to test whether stress-induced alterations in placental GR and 11ßHSD2 protein expression may mediate divergent effects of prenatal adversity upon postnatal development. Pregnant Dom and Sub dams underwent prenatal restraint stress (PRS) for 45 min on gestational days (GD) 15-17. PRS induced a similar spike in serum corticosterone concentrations of dams from each strain on GD15 (p < .001, n = 8), and impaired fetal growth (p < .01, n = 5 litters), although Dom placentae were larger than Sub placentae (p < .01). Among placentae from Dom dams, PRS elevated protein contents of both GR (p < .05, n = 5 litters) and 11ßHSD2 (p < .01) on GD19. In contrast, GR contents were reduced among placentae from PRS-exposed Sub mice (p < .01), without changes in 11ßHSD2 content. Correspondingly, Dom PRS pup growth recovered by PND14, yet Sub PRS pups remained underweight into adolescence (p < .0001, n = 40 pups). Thus, prenatal stress more strongly increased placental GR and 11ßHSD2 levels among Dom mice than in Subs. Increased GR may improve placental function and up-regulate 11ßHSD2 expression, protecting fetuses from effects of prenatal stress upon postnatal development. Placental recruitment of GR and 11ßHSD2 are potential markers of stress-induced developmental disorders, in accordance with maternal resilience or sensitivity to stress.

Development of a 3D seed morphological tool for grapevine variety identification, and its comparison with SSR analysis.

Grapevine (Vitis vinifera L.) is one of the classical fruits of the Old World. Among the thousands of domesticated grapevine varieties and variable wild sylvestris populations, the range of variation in pip morphology is very wide. In this study we scanned representative samples of grape pip populations, in an attempt to probe the possibility of using the 3D tool for grape variety identification. The scanning was followed by mathematical and statistical analysis using innovative algorithms from the field of computer sciences. Using selected Fourier coefficients, a very clear separation was obtained between most of the varieties, with only very few overlaps. These results show that this method enables the separation between different Vitis vinifera varieties. Interestingly, when using the 3D approach to analyze couples of varieties, considered synonyms by the standard 22 SSR analysis approach, we found that the varieties in two of the considered synonym couples were clearly separated by the morphological analysis. This work, therefore, suggests a new systematic tool for high resolution variety discrimination.

Anti-diabetic activity of aerial parts of Sarcopoterium spinosum.

BACKGROUND: Sarcopoterium spinosum (S. spinosum) is used by Bedouin medicinal practitioners for the treatment of diabetes. While the anti-diabetic activity of S. spinosum root extract was validated in previous studies, the activity of aerial parts of the same plants has not been elucidated yet. The aim of this study was to clarify the glucose lowering properties of the aerial parts of the shrub. METHODS: Anti-diabetic properties were evaluated by measuring the activity of carbohydrate digesting enzymes, glucose uptake into 3 T3-L1 adipocytes, and insulin secretion. Insulin signaling cascade was followed in L6 myotubes using Western blot and PathScan analysis. RESULTS: Activity of α-amylase and α-glucosidase was inhibited by extracts of all S. spinosum organs. Basal and glucose-induced insulin secretion was measured in Min6 cells and found to be enhanced as well. Glucose uptake was induced by all S. spinosum extracts, with roots found to be the most effective and fruits the least. The effect of S. spinosum on Akt phosphorylation was minor compared to insulin effect. However, GSK3ß and PRAS40, which are downstream elements of the insulin cascade, were found to be highly phosphorylated by S. spinosum extracts. Inhibition of PI3K and Akt, but not AMPK and ERK, abrogated the induction of glucose uptake by the aerial parts of the shrub. CONCLUSION: The aerial organs of S. spinosum have anti-diabetic properties and may be used as a basis for the development of dietary supplements or to identify new agents for the treatment of type 2 diabetes.

Collection and characterization of grapevine genetic resources (Vitis vinifera) in the Holy Land, towards the renewal of ancient winemaking practices.

The importance and extent of wine consumption in all life aspects at the Holy Land is well documented. The Muslim influence in this region led to the abandonment of winemaking practices, and possible loss of indigenous wine varieties. Here we present a country wide collection of the local grapevine population including wild and cultivated forms, and its characterization by genetic, ampelographic and enological methods. The ampelographic analysis shows clear differences between Sativa and Sylvestris groups in flower, leaf and cluster parameters, and that most Sativa belong to proles orientalis. Genetic population analysis was conducted by analyzing 22 common SSR markers, determining first the unique genotypes, and internally assessing the population's structure, showing the existence of two distinct Sativa and Sylvestris populations, and a third mixed one. Likewise, the relationship between the Israeli grapevine population and grapevine populations in Europe and parts of Asia was investigated, showing that the Israeli Sativa and Sylvestris populations cluster closely together, suggesting a common genetic source. Lastly, the enological characteristics of selected Sativa and Sylvestris genotypes are presented, demonstrating their potential for quality wine production. This research significantly contributes toward the re-establishment of indigenous and traditional local grapevine varieties into the modern international wine industry.

Interaction of cold radiofrequency plasma with seeds of beans (Phaseolus vulgaris).

The impact of cold radiofrequency air plasma on the wetting properties and water imbibition of beans (Phaseolus vulgaris) was studied. The influence of plasma on wetting of a cotyledon and seed coat (testa) was elucidated. It was established that cold plasma treatment leads to hydrophilization of the cotyledon and tissues constituting the testa when they are separately exposed to plasma. By contrast, when the entire bean is exposed to plasma treatment, only the external surface of the bean is hydrophilized by the cold plasma. Water imbibition by plasma-treated beans was studied. Plasma treatment markedly accelerates the water absorption. The crucial role of a micropyle in the process of water imbibition was established. It was established that the final percentage of germination was almost the same in the cases of plasma-treated, untreated, and vacuum-pumped samples. However, the speed of germination was markedly higher for the plasma-treated samples. The influence of the vacuum pumping involved in the cold plasma treatment on the germination was also clarified.

Cold radiofrequency plasma treatment modifies wettability and germination speed of plant seeds.

We report the possibility to modify the wetting properties of the surfaces of a diversity of seeds including: lentils (Lens culinaris), beans (Phaseolus vulgaris) and wheat (Triticum, species C9) by cold radiofrequency air plasma treatment. Air plasma treatment leads to the dramatic decrease in the apparent contact angle. Moreover, the speed of germination and yield (germination rate) of seeds can be modified by preliminary plasma treatment. The change in the wetting properties of seeds is at least partially due to oxidation of their surface under plasma treatment. Significant growth of the peaks corresponding to the nitrogen containing groups in the mass spectra of air plasma treated seeds was registered by TOF-SIMS spectroscopy.