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 regu...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.展开更多
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...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.展开更多
Non-embryogenic calli (NEC) was inevitably and heavily produced when grape embryogenic calli (EC) was induced from explants or during the subculture of EC.A stable and highly efficient NEC transformation platform ...Non-embryogenic calli (NEC) was inevitably and heavily produced when grape embryogenic calli (EC) was induced from explants or during the subculture of EC.A stable and highly efficient NEC transformation platform is required to further sort out and verify key genes which determine/switch the identity of NEC and EC.In this research,a vector pA5 containing a chitinase signal sequence fused to gfp (green fluorescent protein) and an HDEL motive was used to target and immobilize into Agrobacterium strain EHA105 to establish a transformation platform for Vitis vinifera L.cv.Chardonnay NEC.It was determined that NEC 10 d after subculture was the best target tissue;30 min for inoculation followed by 3 d co-cultivation with the addition of 200 μmol L-1 acetosyringone (AS) was optimized as protocol.The use of bacterial densities as 1.0 at OD600 did not result in serious tissue hypersensitive reaction and it had higher efficiency.Kanamycin at 200 mg L-1 was picked for positive expression selection.The stable transformation of NEC was proved by reverse transcription-polymerase chain reaction techniques (RT-PCR) and fluorescent microscopy after three sub-cultures of the selected cell line.Highly efficient genetic transformation protocol of grape NEC was achieved and some of the optimized parameters were different from that reported for EC.This transformation platform could facilitate the verification of candidate somatic embryogenesis (SE) decisive genes,and the successfully transformed NEC with certain genes can also be used as bioreactors for the production of functional products,as NEC not only proliferates fast,but also keeps in a rather stable condition.展开更多
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...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.展开更多
We examined the somatic embryogenesis from and histological studies of zygotic embryos of seeds in European Grape 'Moldova' (Vitis vinifera U 'Moldova'). Primary calli were initiated on Nitsch and Nitsch (NN) ...We examined the somatic embryogenesis from and histological studies of zygotic embryos of seeds in European Grape 'Moldova' (Vitis vinifera U 'Moldova'). Primary calli were initiated on Nitsch and Nitsch (NN) medium supplemented with 1.0 mg·L^-1 2,4-D and 0.5 mg·L^-1 6-BA. Embryogenic calli were produced upon transfer to a NN medium with 0.5 mg·L^-1 6-BA and 2 mg·L^-1 NAA and somatic embryos were obtained on a half strength MS medium without plant growth regulators. During the somatic embryo germination, an addition of 1.0 mg·L^-1 6-BA in the medium could accelerate somatic embryos to develop into normal plants and increase the conversion rate from 0 to 43.3%. Histological studies of embryogenic calli and somatic embryos demonstrated dynamic changes of proteins and starch grains. The developmental processes of somatic embryos were similar to those of zygotic embryos, including typical epiderma, cotyledon primordium and vascular tissue.展开更多
MicroRNAs (miRNAs) are a class of newly identified, small, non-coding RNAs that play vital roles in regulation. Based on miRNAs unique features of expression pattern, evolutionary conservation, secondary structure a...MicroRNAs (miRNAs) are a class of newly identified, small, non-coding RNAs that play vital roles in regulation. Based on miRNAs unique features of expression pattern, evolutionary conservation, secondary structure and genetic requirements for biogenesis, computational predication strategy is adopted to predicate the novel miRNAs. In this research, potential miRNAs and their targets in grapevine (Vitis vinifera) were predicted. We used previously known plant miRNAs against grapevine genome sequence databases to search for potential miRNAs. A total of 81 potential miRNAs were detected following a range of strict filtering criteria. Using these potential miRNA sequences, we could further blast the mRNA database to find the potential targets in this species. Comparative analysis of miRNAs in grapevine and other species reveals that miRNAs exhibit an evolutional conservation, the number and function of miRNAs must have significantly expanded during the evolution of land plants. Furthermore divergence made versatile functions of miRNAs feasible. Cluster of miRNAs likely represents an ancient expression mechanism. Predicted target genes include not only transcription factors but also genes implicated in floral development, signal transduction, diseases and stress response. Till now, little is known about experimental or computational identification of miRNA in grapevine species. Increased knowledge of the biological mechanisms of the grapevine will allow targeted approaches to increase the quality of fruit and reduce the impact of parasites together with stress, which could enable a sustainable, environmentally-sound, farming policv.展开更多
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 p...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.展开更多
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 gonado...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.展开更多
To lay a biological foundation for rootstocks and alternate irrigation (AI) popularization, the effects of partial rootzone drying (PRD) on the growth of the grapevine Malvasia grafted on different rootstocks were...To lay a biological foundation for rootstocks and alternate irrigation (AI) popularization, the effects of partial rootzone drying (PRD) on the growth of the grapevine Malvasia grafted on different rootstocks were investigated. Biological effects of 1/2 divided root irrigation on three combinations, i.e., Malvasia/420A, Malvasia/3309C, and Malvasia/110R, were studied by wood-boxed plants. All the plants were separated into three groups for different irrigation strategies. Mass growth of new root in alternate-irrigated plants was remarkably promoted by about 7.8-22.2% higher than the well- watered ones. However, new shoot growth, especially the internode was reduced by alternate irrigation. The average root-shoot ratio of all the three combinations was increased from 1.1 to 1.46. New root growth and internode length were decreased by fixed partial rootzone irrigation (FI) at different amount, M/3309C at 37.9 and 36.9%, M/110R at 18.4 and 22.5%, respectively. Total biomass of all the three combinations under FI decreased at the rate of 19.2-34.3% compared with well-watered ones. Water stress adaptation of grapevine mainly depends on rootstock. 110R is more efficient than 3309C and 420A in water stress adaptation. PRD-AI benefited root growth, thus improved the drought-resistant ability of grapevine.展开更多
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...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.展开更多
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 caus...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.展开更多
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 bo...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.展开更多
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 considerabl...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.展开更多
[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 resveratro...[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.展开更多
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 b...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.展开更多
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 (...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.展开更多
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. T...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.展开更多
基金This studywas funded by the National Key Research and Development Program of China(2021YFD1200200)the National Natural Science Foundation of China(31872057)+1 种基金the China Agriculture Research System(CARS-29)the Agricultural Science and Technology Innovation Program(CAAS-ASTIP-2021-ZFRI).
文摘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.
基金This workwas supported by the Priority Academic Program Development of Modern Horticultural Science in Jiangsu Province.
文摘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.
基金supported by the National Natural Science Foundation of China (30471212,30500347)the Earmarked Fund for Modern Agro-Industry Technology Research System,Ministry of Agriculture,China (NYCYTX-3-CY-04)
文摘Non-embryogenic calli (NEC) was inevitably and heavily produced when grape embryogenic calli (EC) was induced from explants or during the subculture of EC.A stable and highly efficient NEC transformation platform is required to further sort out and verify key genes which determine/switch the identity of NEC and EC.In this research,a vector pA5 containing a chitinase signal sequence fused to gfp (green fluorescent protein) and an HDEL motive was used to target and immobilize into Agrobacterium strain EHA105 to establish a transformation platform for Vitis vinifera L.cv.Chardonnay NEC.It was determined that NEC 10 d after subculture was the best target tissue;30 min for inoculation followed by 3 d co-cultivation with the addition of 200 μmol L-1 acetosyringone (AS) was optimized as protocol.The use of bacterial densities as 1.0 at OD600 did not result in serious tissue hypersensitive reaction and it had higher efficiency.Kanamycin at 200 mg L-1 was picked for positive expression selection.The stable transformation of NEC was proved by reverse transcription-polymerase chain reaction techniques (RT-PCR) and fluorescent microscopy after three sub-cultures of the selected cell line.Highly efficient genetic transformation protocol of grape NEC was achieved and some of the optimized parameters were different from that reported for EC.This transformation platform could facilitate the verification of candidate somatic embryogenesis (SE) decisive genes,and the successfully transformed NEC with certain genes can also be used as bioreactors for the production of functional products,as NEC not only proliferates fast,but also keeps in a rather stable condition.
基金The Youth Innovation Promotion Association of CAS(2015281)National Natural Science Foundation of China(NSFC Accession No.31471857 and 31672132)+1 种基金Science and Technology Service Network Initiative of CAS(KFJ-STSZDTP-025)Grape Breeding Project of Ningxia(NXNYYZ201502)supported this work.
文摘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.
基金supported by "948" Ad-vanced Forestry Technology Introduction Program (Grant No. 2006-4-73).
文摘We examined the somatic embryogenesis from and histological studies of zygotic embryos of seeds in European Grape 'Moldova' (Vitis vinifera U 'Moldova'). Primary calli were initiated on Nitsch and Nitsch (NN) medium supplemented with 1.0 mg·L^-1 2,4-D and 0.5 mg·L^-1 6-BA. Embryogenic calli were produced upon transfer to a NN medium with 0.5 mg·L^-1 6-BA and 2 mg·L^-1 NAA and somatic embryos were obtained on a half strength MS medium without plant growth regulators. During the somatic embryo germination, an addition of 1.0 mg·L^-1 6-BA in the medium could accelerate somatic embryos to develop into normal plants and increase the conversion rate from 0 to 43.3%. Histological studies of embryogenic calli and somatic embryos demonstrated dynamic changes of proteins and starch grains. The developmental processes of somatic embryos were similar to those of zygotic embryos, including typical epiderma, cotyledon primordium and vascular tissue.
文摘MicroRNAs (miRNAs) are a class of newly identified, small, non-coding RNAs that play vital roles in regulation. Based on miRNAs unique features of expression pattern, evolutionary conservation, secondary structure and genetic requirements for biogenesis, computational predication strategy is adopted to predicate the novel miRNAs. In this research, potential miRNAs and their targets in grapevine (Vitis vinifera) were predicted. We used previously known plant miRNAs against grapevine genome sequence databases to search for potential miRNAs. A total of 81 potential miRNAs were detected following a range of strict filtering criteria. Using these potential miRNA sequences, we could further blast the mRNA database to find the potential targets in this species. Comparative analysis of miRNAs in grapevine and other species reveals that miRNAs exhibit an evolutional conservation, the number and function of miRNAs must have significantly expanded during the evolution of land plants. Furthermore divergence made versatile functions of miRNAs feasible. Cluster of miRNAs likely represents an ancient expression mechanism. Predicted target genes include not only transcription factors but also genes implicated in floral development, signal transduction, diseases and stress response. Till now, little is known about experimental or computational identification of miRNA in grapevine species. Increased knowledge of the biological mechanisms of the grapevine will allow targeted approaches to increase the quality of fruit and reduce the impact of parasites together with stress, which could enable a sustainable, environmentally-sound, farming policv.
基金This work was supported by the Fundamental Research Funds for the Central Universities(Grant No.2452019208)the National Key R&D Program on Monitoring,Early Warning and Prevention of Major National Disasters(Grant No.2017YFC1502806)the China Agriculture Research System for Grapes(Grant No.CARS-29-zp-6).
文摘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.
文摘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.
基金supported by the National Natural Science Foundation of China(30471197).
文摘To lay a biological foundation for rootstocks and alternate irrigation (AI) popularization, the effects of partial rootzone drying (PRD) on the growth of the grapevine Malvasia grafted on different rootstocks were investigated. Biological effects of 1/2 divided root irrigation on three combinations, i.e., Malvasia/420A, Malvasia/3309C, and Malvasia/110R, were studied by wood-boxed plants. All the plants were separated into three groups for different irrigation strategies. Mass growth of new root in alternate-irrigated plants was remarkably promoted by about 7.8-22.2% higher than the well- watered ones. However, new shoot growth, especially the internode was reduced by alternate irrigation. The average root-shoot ratio of all the three combinations was increased from 1.1 to 1.46. New root growth and internode length were decreased by fixed partial rootzone irrigation (FI) at different amount, M/3309C at 37.9 and 36.9%, M/110R at 18.4 and 22.5%, respectively. Total biomass of all the three combinations under FI decreased at the rate of 19.2-34.3% compared with well-watered ones. Water stress adaptation of grapevine mainly depends on rootstock. 110R is more efficient than 3309C and 420A in water stress adaptation. PRD-AI benefited root growth, thus improved the drought-resistant ability of grapevine.
基金funding by Comisión Nacional de Ciencia y Tecnología(CONICYT),folio scholarship 21141168We are grateful to I+D Vinos de Chile(Project 12CTI-16788-01)for provide the plant material.
文摘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.
文摘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.
文摘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.
文摘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.
基金Supported by Construction and Expression of Heterologous Biosynthesis Pathway for Resveratrol(BAK:201502bsh)
文摘[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.
文摘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.
文摘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.
文摘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.