QTL detection and candidate gene analysis of grape white rot resistance by interspecific grape (Vitis vinifera L. × Vitis davidii Foex.) crossing

Grape white rot,a devastating disease of grapevines caused by Coniella diplodiella(Speg.)Sacc.,leads to significant yield losses in grape.Breeding grape cultivars resistant to white rot is essential to reduce the regular use of chemical treatments.In recent years,Chinese grape species have gained more attention for grape breeding due to their high tolerance to various biotic and abiotic factors along with changing climatic conditions.In this study,we employed whole-genome resequencing(WGR)to genotype the parents of‘Manicure Finger’(Vitis vinifera,female)and‘0940’(Vitis davidii,male),along with 101 F1 mapping population individuals,thereby constructing a linkage genetic map.The linkage map contained 9337 single-nucleotide polymorphism(SNP)markers with an average marker distance of 0.3 cM.After 3 years of phenotypic evaluation of the progeny for white rot resistance,we confirmed one stable quantitative trait locus(QTL)for white rot resistance on chromosome 3,explaining up to 17.9%of the phenotypic variation.For this locus,we used RNA-seq to detect candidate gene expression and identified PR1 as a candidate gene involved in white rot resistance.Finally,we demonstrated that recombinant PR1 protein could inhibit the growth of C.diplodiella and that overexpression of PR1 in susceptible V.vinifera increased grape resistance to the pathogen.

欧亚种葡萄(Vitis vinifera L.)的驯化及分类研究进展

欧亚种葡萄是世界上广泛栽培的具有重要经济价值的果树。一般认为欧亚种葡萄栽培亚种是由其野生亚种驯化而来,这个驯化过程包括对色泽引人的大果穗、含糖量高、两性花类型的选择和无性繁殖技术的发现,欧亚种葡萄的驯化问题主要涉及驯化事件发生的时间、地点和数量。本文综述了欧亚种葡萄的驯化和生态地理起源与分布分类研究进展。

Genome and transcriptome analysis of the grapevine(Vitis vinifera L.) WRKY gene family

The plant WRKY gene family represents an ancient and complex class of zinc-finger transcription factors(TFs)that are involved in the regulation of various physiological processes,such as development and senescence,and in plant response to many biotic and abiotic stresses.Despite the growing number of studies on the genomic organisation of WRKY gene family in different species,little information is available about this family in grapevine(Vitis vinifera L.).In the present study,a total number of 59 putative grapevine WRKY transcription factors(VvWRKYs)were identified based on the analysis of various genomic and proteomic grapevine databases.According to their structural and phylogentic features,the identified grapevine WRKY transcription factors were classified into three main groups.In order to shed light into their regulatory roles in growth and development as well as in response to biotic and abiotic stress in grapevine,the VvWRKYs expression profiles were examined in publicly available microarray data.Bioinformatics analysis of these data revealed distinct temporal and spatial expression patterns of VvWRKYs in various tissues,organs and developmental stages,as well as in response to biotic and abiotic stresses.To also extend our analysis to situations not covered by the arrays and to validate our results,the expression profiles of selected VvWRKYs in response to drought stress,Erysiphe necator(powdery mildew)infection,and hormone treatments(salicilic acid and ethylene),were investigated by quantitative real-time reverse transcription PCR(qRT-PCR).The present study provides a foundation for further comparative genomics and functional studies of this important class of transcriptional regulators in grapevine.

The potential role of grape (Vitis vinifera L.) in prevention of threatened abortion via immunomodulatory and anti-inflammatory abilities: a hypothesis

Threatened abortion is a common problem in early pregnancy. This early vaginal bleeding happens in about 25% of pregnant women. The medications including progesterone, uterine muscle relaxant or human chorionic gonadotropin have essential effects in developing pregnancy, but the clinical data are insufficient to prescribe them. In recent decades, medicinal herbs can help us to present new treatments. Grape (Vitis vinifera L.) can protect the fetus from the perspective of Persian medicine. So, we hypothesize about the salutary effects of grape in miscarriage prevention. We found five standard expected mechanisms of grape to prevent threatened abortion: immunomodulatory and anti-inflammatory, anti-oxidant, anti-contraction, hormonal and anti-stress activities. Grape reduces nitric oxide, prostaglandin E2, expression of nuclear factor κB and other pro-inflammatory cytokines like IL (Interleukin)-1β, IL-6, IL-8, and tumor necrosis factor-α. It also elevates anti-inflammatory mediators and expression of peroxisome proliferator-activated receptor-γ. Grape polyphenols have a crucial role in fetus protection with high antioxidant power and other functions such as prevention of stress-triggered abortion with proanthocyanidins, or hormonal effects and inhibition of uterine contractions with resveratrol. So according to these studies, grape probably has effects on the immune and endocrine factors involved in threatened miscarriage.

Anthocyanin profi les and color properties of red wines made from Vitis davidii and Vitis vinifera grapes

Spine grape(Vitis davidii Foex.)is an important wild grape species native to China.Fifteen red spine grape clones and three red Vitis vinifera grape varieties were used to evaluate the differences in the anthocyanin profiles and color properties of wines made from V.davidii and V.vinifera grapes.Among spine wines,‘Junzi#2’wine had the highest total phenolic and total anthocyanin,‘Xiangzhenzhu’wine had the highest total flavonoids,and‘Junzi#1’wine had the highest total tannin.The anthocyanin compositions of all of the spine wines were dominated by Mv-3,5-diglucoside.The total individual anthocyanin contents in spine wines,except‘Gaoshan#5’,‘Junzi#5055’,‘Junzi#5061’,and‘Junzi#5044’,were signifi cantly higher than in V.vinifera wines.Most of the spine wines had a stronger red intensity and a brighter chroma with a bluer hue than V.vinifera wines.Correlation analysis revealed that the color properties were closely related to the anthocyanin composition.These results suggest that wines made from spine grapes may be useful for the wine industry for their color properties and high individual anthocyanin contents.

Comparative metabolic profiling of Vitis amurensis and Vitis vinifera during cold acclimation

Vitis amurensis is a wild Vitis plant that can withstand extreme cold temperatures.However,the accumulation of metabolites during cold acclimation(CA)in V.amurensis remains largely unknown.In this study,plantlets of V.amurensis and V.vinifera cv.Muscat of Hamburg were treated at 4℃ for 24 and 72 h,and changes of metabolites in leaves were detected by gas chromatography coupled with time-of-flight mass spectrometry.Most of the identified metabolites,including carbohydrates,amino acids,and organic acids,accumulated in the two types of grape after CA.Galactinol,raffinose,fructose,mannose,glycine,and ascorbate were continuously induced by cold in V.amurensis,but not in Muscat of Hamburg.Twelve metabolites,including isoleucine,valine,proline,2-oxoglutarate,and putrescine,increased in V.amurensis during CA.More galactinol,ascorbate,2-oxoglutarate,and putrescine,accumulated in V.amurensis,but not in Muscat of Hamburg,during CA,which may be responsible for the excellent cold tolerance in V.amurensis.The expression levels of the genes encodingβ-amylase(BAMY),galactinol synthase(GolS),and raffinose synthase(RafS)were evaluated by quantitative reverse transcription-PCR.The expression BAMY(VIT_02s0012 g00170)and RafS(VIT_05s0077 g00840)were primarily responsible for the accumulation of maltose and raffinose,respectively.The accumulation of galactinol was attributed to different members of GolS in the two grapes.In conclusion,these results show the inherent differences in metabolites between V.amurensis and V.vinifera under CA.

Divergent molecular and growth responses of young“Cabernet Sauvignon”(Vitis vinifera)plants to simple and mixed infections with Grapevine rupestris stem pitting-associated virus

Grapevine rupestris stem pitting associated virus(GRSPaV)is one of the most widely distributed viruses;even so,little is known about its effect on Vitis vinifera.To provide new insights,the effects of single and mixed GRSPaV infections on the V.vinifera cultivar“Cabernet Sauvignon”were studied by evaluating growth parameters,such as measurements of the total plant length,the number and distance of internodes and the number of leaves per shoot.In addition,parameters relating to gas exchange,i.e.,the stomatal conductance,net photosynthetic rate,internal CO2 concentration and leaf transpiration,were also assessed.All the measurements were performed in one-and two-yearold plants with a single GRSPaV infection or mixed infections of GRSPaV and Grapevine fanleaf virus(GFLV).The results show that the plant phytosanitary status did not significantly alter the growth and gas exchange parameters in oneyear-old plants.However,in two-year-old plants,single GRSPaV infections increased shoot elongation,which was accompanied by the overexpression of genes associated with the gibberellic acid response pathway.The gas exchange parameters of these plants were negatively affected,despite exhibiting higher LHCII gene expression.Plants with mixed infections did not have modified growth parameters,although they presented a greater reduction in the primary photosynthetic parameters evaluated with no change in LHCII expression.The results presented here confirm the co-evolution hypothesis for V.vinifera and GRSPaV during the early stages of plant development,and they provide new evidence about the effects of GRSPaV and GFLV co-infections on the“Cabernet Sauvignon”cultivar.

Not All Shrivels Are Created Equal—Morpho-Anatomical and Compositional Characteristics Differ among Different Shrivel Types That Develop during Ripening of Grape (<i>Vitis vinifera</i>L.) Berries

An understanding of physiological disorders associated with ripening of fruits triggered by abiotic stress relies on anatomical and physico-chemical analyses, as it provide insights into their origin and probable causes. The objective of this study was to analyze different ripening disorders of grape (Vitis vinifera L.) berries by dissecting their morpho-anatomy, shriveling nature, and composition. Four well-defined disorders—sunburn, prolonged dehydration (PD), late-season bunch stem necrosis (LBSN), and berry shrivel (BS) were analyzed in field-grown grapevines of the cultivar Cabernet Sauvignon. Early bunch stem necrosis (EBSN) that occurred before ripening was also included in the study. Unlike healthy spherical berries, the pericarp of disordered berries except for sunburn shriveled causing concomitant reductions in fresh weight and volume. The exocarp of PD berries developed well-ordered indentations as distinct from the wrinkles in LBSN berries, whereas BS berries were flaccid with numerous skin folds. The epicuticular wax occurred as upright platelets in all shrivel forms excluding the sun-exposed hemisphere of sunburned berries. A chlorophyllous inflorescence framework persisted in all shrivel forms but in LBSN, wherein the necrotic regions developed tylosis. Unlike the translucent mesocarp of healthy, sunburned, and PD berries, the mesocarp was collapsed in BS and LBSN berries, nevertheless all had well-developed seeds. The composition of healthy berries was optimal, whereas the disordered berries were compositionally distinct from each other, which as a whole differed from the healthy berries. The BS berries had the lowest sugar content, and although sugar concentration was higher in LBSN, sunburned and PD berries, sugar amount per berry was highest in the healthy berries, the same was true for hexoses. Healthy and BS berries exhibited highest amounts of tartaric acid followed by sunburn and PD berries, whereas the LBSN berries had the lowest values. Conversely, healthy and PD berries had the highest amounts of malic acid followed by LBSN, sunburn and BS berries, which collectively displayed similar amounts. The PD berries exhibited the highest calcium content followed by LBSN, healthy, and finally BS and sunburned berries. A linear relationship existed between potassium (K) and pH of the berries. The PD berries had the highest amounts of K followed by healthy, sunburn, LBSN, and BS berries. Overall, the results reported here provided combined morpho-anatomical and compositional analyses of different shrivel types that occurred during a single growing season. Such analysis is needed to make a progress on understanding these ripening disorders culminating in the development of remedial measures.

Understanding Differential Responses of Grapevine (Vitis vinifera L.) Leaf and Fruit to Water Stress and Recovery Following Re-Watering

Among all fruit crops of horticultural importance, grapevines (Vitis vinifera L.) stand out as the most drought tolerant crop species whose tolerance is credited to their proficiency to recover from water stress in both the natural and vineyard growing conditions. However, information on the recovery responses is relatively scant. Studies were conducted to address this issue using potted vines of the grapevine cultivar, Cabernet Sauvignon, which was subjected to water stress and along with anatomical and ultrastructural characterizations, physiological status was assessed in healthy and water stressed vines, and following recovery via rewatering from the water stressed vines. Water stress induced wilting of leaves, drooping of tendrils, and desiccation followed by abscission of shoot tip leaving behind a brown scar at the shoot apex. The wilted leaves accumulated ABA, which correspondingly reduced stomatal conductance and leaf water potential. Upon re-watering, both these parameters made a recovery with values similar to healthy leaves. Likewise, leaf anatomical features following rewatering resembled to that of healthy leaves. In clusters, water stress caused shriveling of preveraison (unripened) berries, which regained full turgor following water resupply, whereas the postveraison (ripening) berries in the same cluster remained unaffected as evidenced by the presence of viable mesocarp cells and epicuticular wax in the form of platelets. The study revealed that shoot tip with leaf primordia was most sensitive to water stress followed by fully expanded leaves and preveraison berries, whereas the postveraison berries remained unaffected. This information could be valuable to implementing irrigation strategies towards sustaining grape production in existing vineyards experiencing episodic droughts and targeted areas prone to drought.

Genomic signatures of different adaptations to environmental stimuli between wild and cultivated Vitis vinifera L

The application of population genetic methods in combination with gene mapping strategies can help to identify genes and mutations selected during the evolution from wild plants to crops and to explore the considerable genetic variation still maintained in natural populations.We genotyped a grapevine germplasm collection of 44 wild(Vitis vinifera subsp.sylvestris)and 48 cultivated(V.vinifera subsp.sativa)accessions at 54 K single-nucleotide polymorphisms(SNPs)to perform a whole-genome comparison of the main population genetic statistics.The analysis of Wright Fixation Index(FST)along the whole genome allowed us to identify several putative“signatures of selection”spanning over two thousand SNPs significantly differentiated between sativa and sylvestris.Many of these genomic regions included genes involved in the adaptation to environmental changes.An overall reduction of nucleotide diversity was observed across the whole genome within sylvestris,supporting a small effective population size of the wild grapevine.Tajima’s D resulted positive in both wild and cultivated subgroups,which may indicate an ongoing balancing selection.Association mapping for six domestication-related traits was performed in combination with population genetics,providing further evidence of different perception and response to environmental stresses between sativa and sylvestris.

葡萄(Vitis vinifera L.)试管繁殖中芽增殖能力衰减的原因及其防治方法的研究

以20个葡萄品种试管苗作为实验材料,经四年、四十多代的继代培养反复对比实验发现:在葡萄试管繁殖中,连续恒温(25±1℃)条件下培养或只采用固体培养方法,不适期进行继代培养均会导致芽增殖能力的衰减,根据生物统计分析,当继代培养超过16～17代后,芽增殖能力即随着继代培养次数的增加而逐渐下降,采取变温培养、变换培养方法、适期继代培养或在增殖培养基中附加适量的椰子乳汁等方法均可防治芽增殖能力的哀减,保持高的芽增殖能力。

Cloning and Bioinformatics Analysis of Resveratrol Synthase Gene from Vitis vinifera

[Objectives] To obtain a resveratrol synthase gene of Vitis vinifera and make bioinformatics analysis. [Methods] Taking total RNA of V. vinifera as the template,by RT-PCR method,a complete c DNA sequence of resveratrol synthase gene was amplified from V. vinifera,and the resveratrol synthase gene was named as RS. The nucleic acid and protein sequences were analyzed using bioinformatics software.[Results]This sequence was 1179 bp in length,the similarity with reported resveratrol synthase gene reached 94%-99%,and the similarity with amino acid sequence reached 96%-99%; the RS gene encoded 392 amino acids,and amino acid sequence contained complete characteristic sequence GVLFGPGLT and active center sequence GCYAGGTVLR of stilbene synthase family; the predicted molecular weight was42. 78 k Da,the theoretical isoelectric point was 6. 57,the instability parameter was 35. 92,and it belonged to stable protein in the classification; the secondary structure was mainly α-helix,random coil and β-folding,α-helix content was 44. 13%,the random coil content was26. 53%,and β-folding content was 17. 66%. [Conclusions] The isolated RS gene is a resveratrol synthase gene from V. vinifera. This experiment is expected to lay a certain foundation for biosynthesis of resveratrol by the genetic engineering method.

The Influence of Photoperiod on the Regulation of Root and Callus Initiation of Perle Noir (<i>V. vinifera</i>L.): Expression of MADS-Box Gene

To study the influence of photoperiod on roots differentiation in the Tunisian grapevine (Vitis vinifera L.) cultivar Perle noir, roots and callus initiation were analyzed under three different conditions of day length: long day (LD), short day (SD) and darkness (D). The photoperiod influenced the number of callus and roots per cuttings;it has a significant effect on the roots and callus initiation. Expression profile analysis of six MADS-box genes (VTM8, VSEP2, VAG12, VAG17-1, VAG17-2 and VSOC1.3) during root and callus development is in agreement with the above-mentioned observation. The expression of the MADS-box genes during root and callus development fluctuated in a tissue-dependent manner. These data suggest that all genes are expressed in roots under three photoperiods. Total darkness gives the number of the most important root per cutting compared to the other two conditions. This photoperiodic condition gave the most important expression of the studied genes VAG12, VAG17-2, VAG17-1, VTM8 and VSEP2 transcripts were not found in callus grown in the dark or in LD conditions, respectively. VSOC1.3 transcripts were not found in callus grown in the dark or in SD conditions, respectively. Transcript abundance of VTM8 and VSOC1 was highest in LD.

Rooting Abilities in <i>Vitis vinifera</i>L.: Total Lipid and Fatty Acids Evolution in Perle Noir Cutting

Rooting ability in Vitis vinifera L. is the phenomena studied. The evolution of the total lipids contents and fatty acids amount of cuttings during the rooting were determined. Total lipid extraction was carried out by an extraction solvent consisting of a mixture of chloroform and methanol. Methylation of fatty acids was carried out by the boron trifluoride methanol complex. Their analysis is performed by gas chromatography. We have obtained a content of merithallus lipids equivalent with then in bibliography. In other hand 3 fatty acids were obtained: eicosapentaenoic acid, oleic acid and docosanoic acid. Among the studied of rooting abilities, we observed decreased levels of total lipids and in the essential fatty acids: eicosapentaenoic acid C20:5 n-3 and oleic acid C18:1 n-9. Minor fatty acids cuttings experimental have a variable evolution. Lipids and certain fatty acids may be markers of rooting in the vine.

Genome-Wide Identification and Expression Profiling of the Shaker K+ Channel and HAK/KUP/KT Transporter Gene Families in Grape (Vitis vinifera L.)

Potassium(K+)is an essential macronutrient for plants to maintain normal growth and development.Shaker-like K+channels and HAK/KUP/KT transporters are critical components in the K+acquisition and translocation.In this study,we identified 9 Shaker-like K+channel(VvK)and 18 HAK/KUP/KT transporter(VvKUP)genes in grape,which were renamed according to their distributions in the genome and relative linear orders among the distinct chromosomes.Similar structure organizations were found within each group according to the exon/intron structure and protein motif analysis.Chromosomal distribution analysis showed that 9 VvK genes and 18 VvKUP genes were unevenly distributed on 7 or 10 putative grape chromosomes.Three pairs of tandem duplicated genes and one pair of segmental duplicated genes were observed in the expansion of the grape VvKUP genes.Gene expression omnibus(GEO)data analysis showed that VvK and VvKUP genes were expressed differentially in distinct tissues.Various cis-acting regulatory elements pertinent to phytohormone responses and abiotic stresses,including K+deficiency response and drought stress,were detected in the promoter region of VvK and VvKUP genes.This study provides valuable information for further functional studies of VvK and VvKUP genes,and lays a foundation to explore K+uptake and utilization in fruit trees.

Evaluation of antioxidant and anti-hypercholesterolemic potential of Vitis vinifera leaves

Background:Vitis vinifera is universally appreciated herb(especially fruit)for their delicacy,nutrition and conventional as functional food.In the present study,we evaluated anti-hypercholesterolemic potential of Vitis vinifera red leaves extract in experimental wistar male rats.Material and methods:Qualitative and quantitative phytochemical screening of vitis vinifera methanolic extract(VVME)and vitis vinifera aqueous extract aqueous(VVAE)extract was carried out to identify the phytoconstituents.The anti-hypercholesterolemic activity was evaluated by inducing hypercholesterolemia with high cholesterol diet for 21 days in experimental animals.During the experimental periods parameters like lipid profile,liver function test,atherogenic index and histopathological studies were analysed.Results:Total tannins,total flavonoids and total phenolic contents were found in major amount in VVME.The lipid levels were significantly attenuated by different doses of VVME and VVAE.Moreover,VVME was found more effective than VVAE and histopathological results confirmed the effectiveness of VVME.Conclusion:It can be concluded that anti-hypercholesterolemic efficacy of Vitis vinifera might be due to presence of active phytoconstituents and its antioxidant efficacy.

Antimutagenic and DNA Damage Protective Activities of a Grape Extract from Vitis vinifera

Antimutagenic and DNA protective effect of an extract VinOserae from Vitis vinifera grapes on oxidative DNA damage was investigated. The extract’s ability to inhibit mutagenicity induced by tert-butyl hydroperoxide (t-BHP) and hydrogen peroxide (H2O2) was determined with Ames test using Salmonella typhimurium His? TA102 strain. Inhibition values of 44.2% and 67.0% were detected for t-BHP and H2O2, respectively. A protective ability of the extract against DNA strand scission induced by hydroxyl radicals was studied with plasmid pBluescript II SK(-). The analysis of DNA strand breaks in plasmid DNA showed a significant inhibition of DNA damage.

Musa Paradaisica and Vitis vinifera Functionalised Ag-NPs: Electrochemical and Optical Detection of Escherichia coli in Seawater

Herein, we demonstrate a simple and inexpensive one-pot green synthesis of silver nanoparticles (Ag-NPs) functionalised with a combination of banana peel (Musa paradisiaca) and grape (Vitis vinifera) fruit extracts. The reaction mixture of aqueous silver nitrate, banana peel and grapefruit extracts revealed a dark brown colour after a reaction time of 18 minutes, which indicates the presence and the successful synthesis of silver nanoparticles. The optical and structural properties of the green synthesised nanoparticles were analysed using UV-Visible spectroscopy (UV-Vis) which confirmed an absorption band at 440 nm. The polydispersity nature and the AgNPs sizes of 30 nm were revealed using small angle X-ray scattering (SAXS) and high-resolution transmission electron microscopy (HR-TEM) techniques. Fourier transform infrared spectroscopy (FT-IR) studies revealed the structure of these nanoparticles which included carbonyl groups, primary amine groups, OH groups and other stabilizing functional groups characteristic of the properties of combined extracts. A simple, quick, less time-consuming surface plasmon resonance (SPR) and electrochemical method in the form of optical and electrochemical sensors have been developed for the detection of Escherichia coli 0157:H7. The obtained limit of detection (LOD) values for SPR and GBPE-Ag-NPs/GCE-based sensor systems were found to be 1 × 102 CFU/mL and 3.5 × 101 CFU/mL, respectively. The obtained values fall within the range for E. coli 0157:H7 in seawater.

Investigation on Viral Pathogens in <i>Vitis vinifera</i>from Four Production Bases in Hangzhou Vicinity of China by sRNAseq and Molecular Validation

This is the first systematic investigation of viral pathogens in Vitis vinifera from Hangzhou vicinity of China. About 7 viruses and 5 viroids were annotated from four production bases “Dushicun”, “Wangjiayuan”, “Xiajiangcun”, and “Yangducun” covering 15 cultivars through sRNAseq technique. At least 3 viruses—grapevine leaf roll-associated virus 3 (GLRaV-3), grapevine fleck virus (GFkV) and grapevine geminivirus A (GGVA), and 4 viroids—hop stunt viroid (HSVd), citrus viroid II (CVd-II), grapevine yellow speckle viroid 1 (GYSVd-1) and grapevine yellow speckle viroid 2 (GYSVd-2) infected all four bases. “Yangducun” base showed 11, the most infected pathogens. GYSVd-1 showed the highest accumulation in host of Wangjiayuan base. The main infected pathogens were verified by reverse-transcription polymerase chain reaction (RT-PCR) technique, the detected rate reached to 85% - 100%. The results provide an important basis for effective and precise detection of viral diseases in the area and for the virus-free cultivation in future.

葡萄(Vitis Vinifera L.)X-射线能谱元素微区污染分析研究

对葡萄果实不同处理的细胞组织多位点进行X -射线能谱微区元素分析。结果 :葡萄不同处理的细胞组织中共发现Mg、Al、Si、P、S、Cl、K、Ca、Fe、Cu十种元素。且不同处理的组织细胞中各元素含量分布有差异 ,未清洗果实表皮处理的Al、Si、S、Ca、Cu含量均显著高于对照 。