Effects of root restriction on nitrogen and gene expression levels in nitrogen metabolism in Jumeigui grapevines(Vitis vinifera L.×Vitis labrusca L.)

To decipher the relationship between the inhibited shoot growth and expression pattern of key enzymes in nitrogen metabolism under root restriction, the effects of root restriction on diurnal variation of expression of nitrate reductase （NR）, nitrite reductase （NiR）, glutamine synthetase （GSI-1, GS1-2, GS2） and glutamate synthase （Fd-GOGAT, NADH-GOGAT） genes and nitrogen levels were evaluated in two-year-old Jumeigui grapevines （Vitis vinifera L.xVitis labrusca L.） when significant differences in shoot growth were observed between treatments at expansion stage （22 days after anthesis）. Grapevines were planted in root-restricting pits as root restriction and in an unrestricted field as the control. Results showed that root restriction significantly reduced shoot growth, but promoted the growth of white roots and fibrous brown roots and improved the fruit quality. （NO3-＋NO2-）-N concentration in all plant parts, NH4＋-N concentration in white roots and total N concentration in leaves and brown roots were significantly reduced under root restriction. Gene expression analysis revealed that mRNA levels of genes related to the GS1/NADH-GOGAT pathway were lower in root-restricted than in control petioles, whereas genes involved in the GS2/Fd-GOGAT pathway were up-regulated under root restriction. Root restriction also resulted in downregulation of genes involved in nitrogen metabolism in leaves, especially at 10：00, while transcript levels of all these genes were enhanced in root-restricted white and brown roots at most time points. This organ-dependent response contributed to the alteration in NO3- reduction and NH4＋ assimilation under root restriction, leading to less NO3- transported from roots and then assimilated in root-restricted leaves. Therefore, this study implied that shoot growth inhibition in grapevines under root restriction is closely associated with down-regulation of gene expression in nitrogen metabolism in leaves.

Dothiorella vidmadera, a novel species from grapevines in Australia and notes on Spencermartinsia

Strains of a coelomycete isolated from grapevine cankers in southeastern Australia and identified as Dothiorella iberica in previous studies are redescribed in this study as a novel species based on morphological characters and phylogenetic analyses of DNA sequences of the internal transcribed spacer region(ITS1-5.8S-ITS2),and partial sequences of the translation elongation factor 1-αandβ-tubulin genes.Dothiorella vidmadera sp.nov.is most closely related to D.iberica,D.americana and D.sarmentorum,but differs in morphological characters and DNA sequences.All four species are known to be associated with Botryosphaeria dieback of grapevines.Additionally,an unidentified ascomycete isolated from dead wood of grapevines in Western Australia is described.Phylogenetically,strains were most closely related to Spencermartinsia viticola,and bore conidia with morphological characters and dimensions consistent with published descriptions.However,ascospores were shorter and narrower than previously reported and lacked the terminal apiculi that typify the genus.

A Comparative Study on Antiradical and Antimicrobial Properties of Red Grapes Extracts Obtained from Different <i>Vitis vinifera</i>Varieties

The present work is devoted to the study of antiradical and antimicrobial activities of phenolic compounds extracted from different grapevine varieties grown in the Bekaa plane-Lebanon. The amount of phenolic compounds in selected grape extracts was determined by the Folin-Ciocalteau method. Phenolic composition was specified by high performance liquid chromatography. Free radical scavenging activity was examined by using the (2,2’-diphenyl-1-picrylhy-drazyl) DPPH assay. The potential antimicrobial activity was studied using a new quantitative method developed during this work. This activity was tested against several microbial pathogens, including a Gram-positive strain (Listeria monocytogenes), two Gram-negative strains (Escherichia coli and Salmonella arizonae) and a fungal strain (Candida albicans). According to the results of the present screening study, a great variability in the composition of phenolic com-pounds in red grape extracts was detected. All phenolic compounds extracts, demonstrated important scavenging properties and antimicrobial effect against bacterial and fungal strains. Yet, a different response degree was noticed depending on the tested microorganism and the phenolic composition of grape extracts. Antimicrobial activity was more effective against Gram-positive than Gram-negative and yeast strains. Furthermore, our results highlighted a significant role of synergistic effect between various phenolic compounds in the free radical scavenging and antimicrobial activities.

First record of Eutypella vitis causing branch dieback on new host trees in Canada

Eutypella vitis is reported for the first time on several new host trees viz.Fagus grandifolia,Fraxinus pennsylvanica and Syringa reticulata with associated branch dieback symptoms in the Niagara Region of Southern Ontario,Canada.Morphological characteristics and phylogenetic analyses of ITS sequence data confirmed the species identification.Eutypella vitis is known as a woody plant pathogen causing dieback of grapevines and other economically important fruit trees widely cultivated in Ontario.The fungus ability to infect several alternate new hosts can promote its faster and wider spread across vineyards and consequently increase a dieback hazard to viticulture in Canada.

A sensitive SYBR Green RT-qPCR method for grapevine virus E and its application for virus detection in different grapevine sample types

To develop a rapid and high-sensitivity method for detection of grapevine virus E(GVE),a SYBR Green based real-time fluorescence quantitative RT-PCR method(RT-qPCR)was established.This method could be used to detect GVE specifically,and the sensitivity was about 100 times greater than conventional RT-PCR.An excellent linear correlation(R=0.997)and a high amplification efficiency(E=97.5%)were obtained from the standard curve of this method.Reproducibility tests revealed that the coefficients of variation in the intra-and inter-assay results were 0.31-1.03%and 0.82--262%,respectively,indicating a good reproduiblity.The RT-qPCR method could be used to detect GVE in a wide range of grapevine sample types.The detection rates of RT-qPCR for nearly all sample types from different positions and seasons were higher than conventional RT-PCR.The detection rates in spring,summer,autumn and winter increased gradually.Samples in autumn and winter were best for detection,and the detection rates of most samples were 80-100%,which were 10 to 40%higher than conventional RT-PCR.In general,old petioles and branches were the best tissues for GVE detection.The detection rates of these samples in each season were all 100%,which were 20 to 40%higher than conventional RT-PCR.The second highest rates were in the old leaf,with detection rates for RT-qPCR of 80-100%in all seasons,which were 20 to 40%higher than conventional RT-PCR.GVE could be difficultly detected in young leaves by conventional RT-PCR,and the detection rates were only 0-50%,while by RT-qPCR the rates could increase to 0--80%.A total of 33 out of 363 samples(belonging to 68 cultivars)from 20 regions in China were detected to be positive by RT-qPCR(9.1%),which was more than twice the rate of the conventional RT-PCR(3.9%).

Effects of Partial Rootzone Drying on the Growth of Vitis vinifera cv.Malvasia Grafted on Different Rootstocks

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.

Somatic Embryogenesis in Iberian Grapevine (Vitis vinifera) Cultivars Using Carpels as Initial Explants: Protocol Establishment and Histological Evaluation

Trincadeira and Aragonez are two important grapevine cultivars in the Iberian Peninsula, used for high quality red wines production. Both cultivars are strongly affected by fungal diseases, with consequent high loses on plant productivity and fruit quality. A successful protocol for plant regeneration via somatic embryogenesis (SE) was established for both cultivars allowing further plant improvement based on gene transfer technology. Several factors were evaluated during the three different phases which characterize an SE plant regeneration protocol. The culture room temperature during the induction phase, a parameter usually accepted as standard by most researchers, proved in these trials to significantly affect the embryo induction rates. Concerning embryo conversion, it was specially affected by the embryo developmental stage, by the intensity and duration of the chilling treatment and by the supplementation of conversion culture medium with activated charcoal. The responses obtained, both for induction and conversion, proved to be highly genotype dependent. Calli structure, as well as embryo integrity, was histologically observed, allowing to characterize embryonic and non-embryonic masses and to identify abnormalities on embryo development.

Versatile physiological functions of the Nudix hydrolase family in berry development and stress response in grapevine

Nudix hydrolases are widely distributed across all classes of organisms and provide the potential capacity to hydrolyze a wide range of organic pyrophosphates. Although Nudix hydrolases are involved in plant detoxification processes in response to abiotic and biotic stresses, the biological functions of Nudix hydrolases remain largely unclear in grapevine.In the present study, a total of 25 putative grapevine Nudix hydrolases(VvNUDXs) were identified by bioinformatics analysis and classified into eight subfamilies based to their preferred substrates. Both tandem and segmental duplications were responsible for the evolution and expansion of the NUDX gene family in grapevine. To investigate the regulatory roles of VvNUDX genes during growth and development, as well as in response to abiotic and biotic stresses in grapevine, the expression patterns were revealed in publicly available microarray data. The spatial and temporal expression patterns of the VvNUDX genes indicated that they might play important roles in multiple developmental processes. Transcriptome and qRT-PCR analyses showed that ten VvNUDX genes were specifically expressed in grapevine berries, suggesting potential roles in grapevine berry development. Expression and phylogenetic analyses demonstrated that VvNUDX1 and VvNUDX3 might be involved in terpenoid biosynthesis in grapevine. Furthermore,most VvNUDX genes active toward the ADP-ribose/NADH showed different patterns in response to various abiotic and biotic stresses, such as salinity and drought, as well as different types of biotic treatments, such as Erysiphe necator,Bois Noir phytoplasma and leaf-roll-associated virus-3(GLRaV-3). These results indicated that VvNUDX genes were associated with plant detoxification processes in response to abiotic and biotic stresses, and regulate the disease immunity and resistance pathways. The information obtained here may provide good opportunities to explore the physiological functions of VvNUDX genes in berry development and stress response networks in grapevine.

Advancements in plant regeneration and genetic transformation of grapevine(Vitis spp.)

Grapevine(Vitis spp.)is one of the most economically important fruit crops worldwide,and there is considerable interest in improving its major agronomic and enological traits in response to ever-changing agricultural environments and consumer demands.Molecular genetic techniques in particular,associated with rapid technological advancements,provide an attractive alternative to conventional breeding approaches for developing new grapevine varieties with enhanced yield performance,quality,stress tolerance and disease resistance.To date,several grapevine varieties have been transformed with genes associated with diverse functions through biolistic bombardment and/or Agrobacterium-mediated transformation,and transgenic grape lines have been obtained using established regeneration systems.Nevertheless,a wide range of factors,including genotype,explant source and culture medium,have been shown to affect the efficiency of plant regeneration.Moreover,the selection and use of acceptor materials,bacterial strain and cell density,selectable markers and selection methods also influence transformation efficiency.This paper provides an overview of recent advances in grapevine regeneration and genetic transformation and in-depth discussion of the major limiting factors,and discusses promising future strategies to develop robust plant regeneration and genetic transformation in grapevine.

Identification of the DEAD-box RNA helicase family members in grapevine reveals that VviDEADRH25a confers tolerance to drought stress

Grapevine growing areas are increasingly affected by drought,which has greatly limited global wine production and quality.DEAD-box is one of the largest subfamilies of the RNA helicase family,and its members play key roles in the growth and development of plants and their stress responses.Previous studies have shown the potential of DEAD-box genes in the drought stress responses of Arabidopsis and tomato,rice,and other crop species.However,information about DEAD-box genes in grapevine remains limited.In this report,a total of 40 DEAD-box genes were identified in grapevine and their protein sequence characteristics and gene structures were analyzed.By comparing the expression profiles of VviDEADRHs in response to drought stress in different grapevine varieties,nine candidate genes(VviDEADRH10c,-13,-22,-25a,-25b,-33,-34,-36,and-39)were screened based on expression profiling data.Combined with qRTPCR results,Vvi DEADRH25a was selected for functional verification.Heterologous overexpression of Vvi DEADRH25a in Arabidopsis showed the transgenic plants were more sensitive to drought stress than the control.Both electrolyte permeability and malondialdehyde content were significantly increased in transgenic plants,whereas the chlorophyll content and superoxide dismutase(SOD),peroxidase(POD),catalase(CAT),and ascorbate peroxidase(APX)enzyme activities were significantly decreased.Furthermore,VviDEADRH25a-overexpressing plants showed down-regulated expression levels of several drought stress-related marker genes,namely At COR15a,At RD29A,At ERD15,and At P5CS1,which indicated that they participated in the drought stress response.In summary,this study provides new insights into the structure,evolution,and participation of DEAD-box RNA helicase genes in the response to drought stress in grapevines.

Elimination of grapevine fleck virus and grapevine rupestris stem pitting-associated virus from Vitis vinifera 87-1 by ribavirin combined with thermotherapy

Vitis vinifera 87-1 plants infected by grapevine fleck virus(GFkV) and grapevine rupestris stem pitting-associated virus(GRSPaV) were used as the plant materials for virus elimination treatment. This study evaluated the effects of ribavirin at different concentrations(15 and 25 μg mL^(–1);R15 and R25, respectively), thermotherapy(37°C;T), and the combination of ribavirin and thermotherapy(R15+T and R25+T) on eliminating viruses from grapevine plants in vitro. Both R15 and R25 had phytotoxic effects and weakened plant growth. Thermotherapy positively affected the growth of grapevine plants. Plant height was significantly greater in T, R15+T, and R25+T than in CK, R15 and R25. The proportion of dead plants after T, R15+T, and R25+T was 51.4, 11.4, and 8.6%, respectively. The survival rates of regenerated plants after all treatments were >68.0%. Ribavirin concentration and treatment time were related to the regeneration of shoot tips and elimination efficiencies of the two viruses. The survival rates of plants after R15+T for 30, 40, and 50 days were 97.3, 90.7, and 74.4%, respectively. The elimination rates of GRSPaV from plants in the three time quantum were 55.6, 84.6, and 93.8%, respectively. The elimination rate of GFkV was 23.9% higher in R25(35/44) than in R15(25/45), and that of GRSPaV was 7.0% higher in R25 than in R15. The combination of thermotherapy and chemotherapy was found to have a positive effect on the eradication of GFkV and GRSPaV, and R25+T for 50 days was able to completely eliminate the two viruses from in vitro grapevines.

Wild grapevine(Vitis vinifera subsp.sylvestris)in the Hyrcanian relict forests of northern Iran:an overview of current taxonomy,ecology and palaeorecords

Due to severe anthropogenic impacts on lowland and submontane zones of the Hyrcanian forests of northern Iran, wild grapevine(Vitis vinifera subsp. sylvestris), a sporadically distributed woody liana, is currently considered an endangered species. Using data from the literature and 34 studied populations, herbarium assessments and nine palynological sites, we provide an overview of its taxonomy,distribution and ecology in the first part of the investigation.The separation of the two subspecies, namely V. sylvestris subsp. anebophylla and V. sylvestris subsp. trichophylla(sensu Flora Iranica), based on their leaf indumentums, could not be confirmed by our examination of herbarium materials and field observations. Indumentum of the leaves is a result of leaf polymorphism in different Vitis specimens and can be strongly influenced by environmental conditions. Vitis vinifera subsp. sylvestris grows in a wide range of habitats including wetlands, seasonal stream sides in closed forests,alluvial beds of large rivers, sand dune shrublands and forested wetlands(alder forests). Parrotia persica and Carpinus betulus were the most frequent host species in the studied populations. In the Hyrcanian region, no pollen record of Vitis older than the Mid-or Late-Holocene has yet been established. Despite the intrinsic problem in pollen identification by normal(i.e., light) microscopy of wild from cultivated grapevines, the significant values and persistent occurrence of Vitis pollen since the Mid-Holocene(before the Bronze Age) in the Hyrcanian pollen records may imply the onset of viticulture in low-to mid-elevation sites in the region. This represents an argument to consider the Hyrcanian region as a possible domestication center for V. vinifera.However, the question of wild versus cultivated origin of grapevines in the Hyrcanian pollen records and the possible date of its domestication and/or cultivation will remain open until further palynological studies are undertaken.

Neopestalotiopsis eucalypti,a causal agent of grapevine shoot rot in cutting nurseries in China

Grapevine(Vitis vinifera L.)is an economically important fruit crop in the world,and China ranks first in the production of grapes with approximately 15%of the world’s total yield.However,diseases that cause the death of grapevine shoots pose a severe threat to the production of grapes.In this study,the fungus Neopestalotiopsis eucalypti was identified as a causal pathogen of grapevine shoot rot based on the morphology of conidia and a phylogenetic analysis.The phylogenetic analysis was performed with three isolates based on the combined sequence of internal transcribed spacer(ITS)region of ribosomal DNA,part of the translation elongation factor 1-alpha(Tef)and theβ-tubulin(Tub2)genes.The three isolates were all identified as N.eucalypti.Pathogenicity tests of the three fungal isolates were conducted on grapevines shoots in vitro and in vivo.The results showed that all three fungal isolates caused severe rot lesions on the inoculated grapevine shoots,and N.eucalypti was re-isolated from the inoculated grapevine shoots.Therefore,N.eucalypti was confirmed as a causal agent of the grapevine shoot rot.This is the first report of N.eucalypti causing grapevine shoot disease in China.

Characterization of an Early Berry Development Grapevine Somatic Variant (Vitis labrusca L. cv. Isabel Precoce)

Over the past 10 years significant advances have been made towards the description of genetics and molecular mechanisms controlling grapevine berry growth. Regardless of this, many aspects of early fruit morphogenesis and its development control remain to be elucidated. In an attempt to understand gene expression patterns associated with the berry growth development, the contrasting phenotype between the cv. Isabel (Vitis labrusca L.) and its early berry development mutant “Isabel Precoce” has been explored by a candidate gene approach. “Isabel Precoce” (Vitis labrusca L.) was confirmed as an EDV (Essentially Derived Variety) of Isabel, with a 30-35-day reduction in the berry growth phase when compared to the wild type and thus, it constitutes an informative model to investigate many aspects of fruit growth and development. Phenotypic analysis showed that “Isabel Precoce” develops fruits that are smaller in diameter and volume despite of following similar development kinetics. The expression of many genes associated with plant growth and development (MIKCC-type MADS box genes), sugar transport and with the control of flavonoid biosynthetic pathway have been evaluated. The majority of the genes presented a remarkably similar transcription profile. However, a higher induction of transcript accumulation for some genes has been observed in the “Isabel Precoce” genetic background.

Rt-PCR Analysis and Evolutionary Relationship of Some Hungarian <i>Grapevine leafroll associated virus</i>1 and 3 Isolates

Hungarian isolates of Grapevine leafroll associated virus 1 and 3 (GLRaV-1, GLRaV-3) were tested using serological (DAS-ELISA) and molecular (RT-PCR) methods. Five hundred bp long PCR products of the part of HSP70 gene of one serologically positive GLRaV-1 and four GLRaV-3 isolates were sequenced. These sequences were applied for phylogenetic analysis and compared to foreign virus isolates of NCBI GenBank. Phylogenetic analysis of GLRaV-1 HSP70 gene supported the earlier results that it could be divided into two clusters: E and A. The Hungarian isolate 6.4.1 belonged to the group E. This isolate showed the highest homology with the AY754914 isolate from the Czech Republic. GLRaV-3 sequence data could cluster five groups. Hungarian 2.2;3.5 and 4.2 isolates were estimated belonging to the group II. The 1.4 isolate from the same vineyard as 2.2 varied in sequence data so it belonged to the other, IV. variant group with two South African, two Austrian and a Syrah isolate. According to the phylogenetic analysis, two variant groups occurred in Hungary. These isolates related with each other, but showed higher similarity of foreign counties. In some cases, they were similar to isolates of the neighbour countries such as Slovakia and Austria. It could be supposed that mainly the exchange of virus infected propagation materials caused the dissemination of GLRaV isolates.

Characterization of the Fungi Involved with Grapevine Trunk Diseases in Castilla-La Mancha Region,Spain

Grapevine trunk diseases (GTDs) are currently one of the most important problems for grapevine plant and wine industry, causing important economic cost in Castilla-La Mancha, Spain. Traditionally, GTDs were associated with diseases present in mature grapevines, however in recent years the incidence of young vine decline has been increasing. In this work, from 2009 to 2016, 250 young grapevine plants have been analysed by fungal isolation and characterization. In addition, during 2017, 32 mature plants showing decline symptoms were analysed. Both kinds of plants were analysed in order to identify the main fungal trunk pathogens present in Castilla-La Mancha region. As a result, it was possible to isolate species associated with black-foot disease (Cylindrocarpon-like anamorphs), esca and Petri disease (Phaeomoniella chlamydospora and Phaeoacremonium spp.), and Botryosphaeria dieback. All these pathogens were isolated from young and mature plants, this fact could suggest a common origin. The lack of effective control measures for these diseases makes it necessary to implement an integrated disease management strategy that provides an efficient approach to reduce infections in nurseries and get better results in infected vineyards.

Mass production, fermentation, formulation of <i>Pseudomonas putida</i>for controlling of die back and phomopsis diseases on grapevine

The export oriented agricultural and horticultural crops depends on the export of residue free produce and has created a great potential and demand for the incorporation of biopesticides in crop protection. PGPR with wide scope for commercialization includes Pseudomonas putida. Pseudomonas putida was effective in reducing die back caused by a fungus Botryodiplodia theobromae and phomopsis or arm death, caused by a fungus, Phomopsis viticola of grapevine in vitro and in vivo. Pseudomonas putida showed optimum Siderophore pseudobactin production at 72 h, and growth peak at 120 h. glycerol as a source of carbon and yeast as a source of nitrogen Pseudomonas putida was formulated using polysaccharide dextran starch, which led to the elongation of conservation and the effectiveness of the biofungicide. Pseudomonas putida was very effective as biocontrol agents to reduce the die back and phomopsis diseases on grapevine. The obtained results showed that both bacterial treatments significantly increased the growth parameters as well as fruits dry weights and yield of grapevine.

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.

谈谈grapevine

《新英汉词典》grapevine一词的释义为:(1)葡萄藤;(2)谣言,传闻,小道新闻。这两个词义似乎没有什么关系,那么,它们是如何联系在一起的呢? 追根溯源,grapevine本来是“葡萄藤”,其新义的产生恐怕要首先归功于电报发明家塞缪尔·莫尔斯(Samuel Morse)。莫尔斯原是一个美国画家,后来改行,立志从事电报的发明研究。为了完成用电传递信息的伟大使命,

Success Through the Grapevine

How one wine importer has become dedicated to supplying China with superior quality South African wine WINE from Africa? To some Chinese wine drinkers,this is a strange concept.In fact,there are vineyards all over Africa.But it is down south in the Cape region of South Africa where the continent’s, and even the world’s,finest wines are produced in the eyes of Si Yang,a South African