单氰胺对葡萄休眠过程中冬芽水分和碳水化合物的影响

以酿酒葡萄品种'赤霞珠'和'霞多丽'为材料,对其冬芽休眠进程和休眠过程中冬芽水分和碳水化合物含量的变化进行分析.结果表明,(1)在陕西杨陵地区气候条件下,'霞多丽'、'赤霞珠'分别在12月20日和1月9日达到深度休眠,随后开始休眠解除阶段.(2)在芽休眠过程中,两个品种冬芽的总水分含量、自由水含量和淀粉含量均随休眠的加深而降低,随休眠的解除而增加,而冬芽的束缚水含量、束缚水/自由水比值、可溶性糖含量均在休眠加深阶段持续上升,在休眠解除阶段逐渐降低.(3)单氰胺处理后,两品种冬芽中自由水含量显著增加,束缚水的比例同时显著降低,且'赤霞珠'的变化幅度大于'霞多丽',但其总水分、可溶性糖和淀粉等含量在休眠过程中无显著变化.(4)葡萄冬芽中的水分含量及存在状态、可溶性糖含量和淀粉含量的变化与其休眠进程密切相关;单氰胺处理能够增加冬芽自由水含量,降低其束缚水含量,从而有效打破葡萄休眠.     

Rootstock control of scion transpiration and its acclimation to water deficit are controlled by different genes

The stomatal control of transpiration is one of the major strategies by which plants cope with water stress. Here, we investigated the genetic architecture of the rootstock control of scion transpiration-related traits over a period of 3 yr. The rootstocks studied were full sibs from a controlled interspecific cross (Vitis vinifera cv. Cabernet Sauvignon × Vitis riparia cv. Gloire de Montpellier), onto which we grafted a single scion genotype. After 10 d without stress, the water supply was progressively limited over a period of 10 d, and a stable water deficit was then applied for 15 d. Transpiration rate was estimated daily and a mathematical curve was fitted to its response to water deficit intensity. We also determined δ(13) C values in leaves, transpiration efficiency and water extraction capacity. These traits were then analysed in a multienvironment (year and water status) quantitative trait locus (QTL) analysis. Quantitative trait loci, independent of year and water status, were detected for each trait. One genomic region was specifically implicated in the acclimation of scion transpiration induced by the rootstock. The QTLs identified colocalized with genes involved in water deficit responses, such as those relating to ABA and hydraulic regulation. Scion transpiration rate and its acclimation to water deficit are thus controlled genetically by the rootstock, through different genetic architectures.     

四个酿酒葡萄品种组培快繁体系的初建

为了提高酿酒葡（Vitis vinifera）苗木繁殖速度及苗木品质,以‘赤霞珠’、‘西拉’、‘霞多丽’和‘美乐’4／p品种为试材,研究无菌外植体建立、启动培养、增殖培养和驯化移栽环节的关键技术,初步建立酿酒葡萄组培快繁体系。结果表明,以半木质化茎段为外植体接种成活率高,在培养基MS＋IBA0．2mg·L-1＋6．BA1．0mg·L-1＋KT0．5mg·L-1上启动培养外植体单芽萌发率最高,以培养基1／2MS＋IBA0．2mg·L-1＋KT1．0mg·L-1增殖培养兼生根诱导,组培苗生长健壮,繁殖率高。增殖培养6代后,‘赤霞珠’、‘西拉’、‘霞多丽’和‘美乐’分别由12株葡萄苗扩繁为l383株、1095株、744株和100株。组培苗驯化培养3周后移栽至营养钵,4个品种成活率均在72％以上。此组培快繁体系基本适用于4个酿酒葡萄品种,可应用于科学研究及工业化大规模生产。     

Abscisic Acid Sprays Significantly Increase Yield per Plant in Vineyard-Grown Wine Grape (<Emphasis Type="Italic">Vitis vinifera</Emphasis> L.) cv. Cabernet Sauvignon Through Increased Berry Set with No Negative Effects on Anthocyanin Content and Total Polyphenol Index of Both Juice and Wine

In many cultivars of Vitis vinifera periods of mild water stress during ripening are thought to increase grape quality for winemaking, even though yields may be negatively affected. Because abscisic acid (ABA) is involved in the signaling of water stress in plants, we examine the effects of the ABA signal being given without the concomitant water stress. ABA at 250 mg l⁻¹ was sprayed weekly or biweekly from bud-burst until harvest onto the leaves of vineyard-grown plants of cv. Cabernet Sauvignon. For ABA-treated plants berry yield per bunch and per plant was significantly increased (1.5- to 2.0-fold) across three consecutive harvests (2005 through 2007). Number of berries per bunch and per plant was the primary basis for the significant crop increases, although bunches per plant also tended to increase (1.1- to 1.3-fold) across all three harvests. Other parameters assessed included number of internodes, shoot length, leaf area, leaf water potential at midday, photosynthesis, and stomatal conductance. These parameters showed no significant change with ABA treatment, although shoot length tended to be reduced, as was leaf area relative to control plants. The significantly increased fruit yields were thus accomplished without accompanying increases in leaf photosynthesis and leaf areas. Juice at harvest had equal levels of sugars (Brix) and somewhat higher levels of anthocyanins and total polyphenols relative to control values. The two latter trends continued for the resultant wine across two vintage years. In conclusion, three seasons of experimental trials have demonstrated that ABA application can significantly enhance yield per plant in the field-grown grape (cv. Cabernet Sauvignon) by favoring increased berry set without diminishing the quality of the fruit for winemaking use.     

基于温度-伤害度关系分析酿酒葡萄根系及芽抗寒性

采用差热分析系统(DTA)对8个主栽酿酒葡萄品种的芽和根系进行低温放热分析(LTE),建立各品种芽、根系韧皮部及木质部的温度-伤害度(LT-I)回归方程,评估不同品种的根系及芽抗寒性.结果表明： 8个品种的根系韧皮部伤害度50%的温度从高到低为马瑟兰>品丽珠>赤霞珠>小芒森>霞多丽>蛇龙珠>贵人香>熊岳白;不同品种木质部伤害度50%的温度从高到低为马瑟兰>霞多丽>赤霞珠>小芒森>品丽珠>蛇龙珠>贵人香>熊岳白;芽伤害度50%的温度从高到低为赤霞珠>小芒森>蛇龙珠>品丽珠>霞多丽>贵人香>马瑟兰>熊岳白.利用模糊隶属函数值法综合评价根系及芽的抗寒性,马瑟兰根系的抗寒性最差,熊岳白根系的抗寒性最好;赤霞珠、品丽珠、小芒森和蛇龙珠芽的抗寒性最差,贵人香和熊岳白芽的抗寒性最好.     

Performance and diacetyl production of commercial strains of malolactic bacteria in wine

This study compares 11 commercial cultures of Leuconostoc oenos and Lactobacillus plantarum in Cabernet Sauvignon, Pinot Noir and Chardonnay wines. Performance of the cultures was found to be greatly influenced by wine type. Better survival of the bacteria was observed in Cabernet Sauvignon and Pinot Noir wines. The time necessary to complete malolactic fermentation (MLF) was 65 ± 14 d for Chardonnay, 71 ± 3 d for Cabernet Sauvignon, and 25 ± 8 d for Pinot Noir. The maximal rate of malate utilization was 0·4 g d^-1 for Pinot Noir, and 0·2 g d^-1 for the two other wine types. Final diacetyl concentration was lower in Chardonnay wines (highest 0·58 mg l^-1) compared to the other wines (highest 5·8 mg l^-1). Malic and citric acid were co-metabolized by all strains. None of the strains metabolized glycerol. Significant differences in final diacetyl concentration of wine vinified with the different strains were found. Panelists could reliably differentiate MLF wines from non-MLF wines, irrespective of their diacetyl content, indicating that diacetyl is not the only important MLF flavour.     

Genetic Transformation of Major Wine Grape Cultivars of Vitis Vinifera L.

We have developed an Agrobacterium-mediated transformation system for a number of important grapevine cultivars used in wine production. Transgenic plants were obtained for the seven cultivars: Cabernet Sauvignon, Shiraz, Chardonnay, Riesling, Sauvignon Blanc, Chenin Blanc and Muscat Gordo Blanco. Embryogenic callus was initiated from anther filaments and genotypic differences were observed for initiation and subsequent proliferation with Chardonnay responding most favourably to culture conditions. The transformation system allowed the recovery of germinating transgenic embryos 10–12 weeks after Agrobacterium inoculation and plants within 18 weeks. Examination of the expression patterns of the green fluorescent protein gene under the control of the CAMV35S promoter in leaf tissue of transgenic plants showed that for up to 35% of plants the pattern was not uniform. The successful transformation of a genetically diverse group of wine grape cultivars indicates that the transformation system may have general application to an even wider range of Vitis vinifera cultivars.     

2-Methoxy-3-isobutylpyrazine in grape berries and its dependence on genotype

2-Methoxy-3-isobutylpyrazine (MIBP) contributes a bell pepper aroma to many grape cultivars and has a reported aroma threshold of ～2 ng L(-1) in water. The purpose of this study was twofold: (1) develop a procedure using headspace solid phase micro-extraction combined with GC-MS in the selected ion monitoring mode (HS-SPME-GC-MS-SIM) for analysis of MIBP in grape berries, and (2) determine the location of MIBP biosynthesis in grapevines by approach grafting clusters of Vitis vinifera L. cvs Cabernet Sauvignon and Muscat blanc onto each other. The soluble solids and pH of the grape juice/homogenate matrix from different grape berry developmental stages influenced the method precision; therefore, quantification via the method of standard addition was used. Using our developed method, the limit of detection (LOD) and limit of quantitation (LOQ) of MIBP were 0.1 ng L(-1) and 2 ng L(-1), respectively, measured in a model juice and non-MIBP containing Chardonnay juice. Spiked recoveries averaged between 91% and 112% in Cabernet Sauvignon and Pinot noir homogenates and the overall relative standard deviation was less than 10%. The method was used to analyze MIBP in 29 grape cultivars and in fruit from clusters grafted to Cabernet Sauvignon or Muscat vines. Quantifiable levels were found only in Cabernet franc, Cabernet Sauvignon, Merlot, Sauvignon blanc and Semillon, providing information on the genetic connection for the occurrence of MIBP in grapes. No MIBP was detected in the berries of Muscat blanc clusters grafted onto Cabernet Sauvignon vines when sampled at fruit maturity. MIBP was detected in all berries of Cabernet Sauvignon regardless the graft configuration. The data indicate that MIBP or its precursors originate in the berry and its formation depends upon grape genotype.     

Changes in protein abundance during powdery mildew infection of leaf tissues of Cabernet Sauvignon grapevine (Vitis vinifera L.)

A comparative analysis of differentially expressed proteins in a susceptible grapevine (Vitis vinifera ‘Cabernet Sauvignon’) during the infection of Erysiphe necator, the causal pathogen of grapevine powdery mildew (PM), was conducted using iTRAQ. The quantitative labeling analysis revealed 63 proteins that significantly changed in abundance at 24, 36, 48, and 72 h post inoculation with powdery mildew conidiospores. The functional classification of the PM‐responsive proteins showed that they are involved in photosynthesis, metabolism, disease/defense, protein destination, and protein synthesis. A number of the proteins induced in grapevine in response to E. necator are associated with the plant defense response, suggesting that PM‐susceptible Cabernet Sauvignon is able to initiate a basal defense but unable to restrict fungal growth or slow down disease progression.     

Preference and performance responses of California grape phylloxera to different Vitis rootstocks

Laboratory experiments on host preference and performance were simultaneously conducted with Cabernet Sauvignon, AXR#1, and 5C Vitis rootstocks for the California biotypes A and B grape phylloxera. Preference bioassays recorded over a 3-day period indicated that phylloxera exhibit differential host choice. AXR#1 is antixenotic to biotype A, and 5C is antixenotic to both phylloxera biotypes. Preferences observed with biotype B for Cabernet Sauvignon and AXR#1 were not statistically different, whereas biotype A had shown a significant preference for Cabernet Sauvignon. On 5C the preferences scored were significantly lower than on either of the other two rootstocks, and there was no significant difference beetween biotypes. Performance bioassays recorded over a 29-day period indicated that survival, development, and reproduction of both biotypes were affected by Vitis rootstock. Both biotypes were unable to survive or develop on 5C suggesting the presence of antibiotic resistance in 5C against these phylloxera biotypes. The antibiotic effects observed with AXR#1 against biotype A were expressed as significant reductions in survival, development, and reproduction. Performance of biotype B on AXR#1 was similar to its performance on Cabernet Sauvignon. For both AXR#1 and 5C rootstocks and both phylloxera biotypes A and B the antibiotic mechanism was considerably stronger than the antixenotic mechanism. This research indicated that host preference and performance are positively correlated in grape phylloxera with the rootstocks and phylloxera biotypes tested.     

根瘤蚜侵染对不同抗性葡萄品种根系超微结构的影响

【目的】探讨不同抗性葡萄品种对葡萄根瘤蚜Daktulosphaira vitifolia Fitch侵染后的组织结构响应。【方法】以砧木140Ru (Vitis rupestris×V. berlandieri)和栽培品种赤霞珠Vitis vinifera cv. Cabernet Sauvignon离体三级根及140Ru盆栽苗为试材接种根瘤蚜卵, 接种两周后取刺吸位点组织作超薄切片并观察超微结构变化, 取盆栽140Ru不同时期根结测定多酚含量。【结果】超微结构显示, 砧木140Ru根系周皮层最外层细胞壁的厚度（1 031.25 nm）及周皮层细胞层数（6～7层）显著高于赤霞珠品种的周皮层细胞厚度（543.75 nm）及层数（3～4层）, 140Ru根系韧皮部酚类物质含量比赤霞珠高出35％; 侵染后, 砧木140Ru周皮层细胞第3层以内的细胞壁加厚并积聚大量的多酚类物质, 140Ru葡萄新根被根瘤蚜侵染后多酚含量呈升高趋势, 在接种20 d时是对照的2.4倍; 赤霞珠粗根被根瘤蚜侵染后薄壁细胞中细胞质变浓, 出现大量淀粉粒, 线粒体及内质网数量增多。【结论】砧木140Ru的周皮层组织结构较赤霞珠不利于根瘤蚜口针穿刺, 被根瘤蚜侵染后发生了不利于根瘤蚜侵染取食的变化。     

Study of the intra-cultivar variability of grape DNA using RFLP and PCR methods

Evaluation of DNA polymorphism among Vitis vinifera varieties using RFLP and PCR methods has been performed to choose a DNA technology for detection of grape intracultivar variation. DNA polymorphism of clones of the varieties Riparia Gluar, Riparia x Rupestris 101-14, Cabernet Sauvignon, Riestling reinskiy has been studied using Southern hybridization and amplification techniques. It has been shown that grape intracultivar variability of rDNA in Riparia x Rupestris 101-14 and Cabernet Sauvignon clones was caused by the modification in Alul restriction sites of rDNA. DNA variability of the randomly amplified and inter-SSR sequences of the Riparia Gluar, Riparia x Rupestris 101-14, Cabernet Sauvignon clones was also detected. A set of molecular DNA loci which can be used for grape clone identification has been obtained.     

南亚热带酿酒葡萄品种(系)引种研究

1998～1999年,对8804、真芳德(Zinfandel)、梅尔诺(Merlot Noir)、赤霞珠(Cabernet Sauvignon)等4个欧亚种优良酿酒葡萄品种(系)在我国南亚热带季风气候下的栽培适应性做了初步观察.结果表明,8804具有较强的抗病性和早期丰产性;真芳德早期丰产性突出,但抗真菌病害能力较差;梅尔诺和赤霞珠早期丰产性较差,抗病性一般,能获得较好的成熟度.     

Genetic variability of photosynthesis and water use in Balearic grapevine cultivars

A comparison of photosynthetic characteristics of 20 cultivars of grapevine ( Vitis vinifera L. ) from Mallorca (Balearic Islands, Spain) and two widespread cultivars, Cabernet Sauvignon and Chardonnay, was made under irrigation as well as in response to drought. Although these cultivars share a common origin, a high variability was found for several photosynthetic characters under irrigation. Interestingly, these variations were significant for gas-exchange parameters (net CO₂ assimilation, stomatal conductance and intrinsic water use efficiency) but not for chlorophyll fluorescence parameters (maximum photochemical efficiency, electron transport rate and non-photochemical quenching). Since water stress is the most limiting factor for plant production under the Mediterranean climate, it is presumable that these findings reflect specific selection pressures over physiological characteristics related to a balance between net carbon gain and water use. Some cultivars presented high carbon assimilation at the expense of a high water loss, whereas others were water savers, accompanied by low CO₂ assimilation even under irrigation. Escursach was found to be an interesting cultivar, presenting low water consumption at the same time as reasonably high carbon assimilation. These cultivars also showed different responses to drought, which allowed their classification in two main groups: alarmist cultivars, which showed strong reductions of stomatal conductance in response to relatively low decreases of leaf water potential, and luxurious cultivars, showing low reductions of stomatal conductance under water stress.     

Quantitative proteomic analysis of cabernet sauvignon grape cells exposed to thermal stresses reveals alterations in sugar and phenylpropanoid metabolism

Grapes (Vitis vinifera) are a valuable fruit crop and wine production is a major industry. Global warming and expanded range of cultivation will expose grapes to more temperature stresses in future. Our study investigated protein level responses to abiotic stresses, with particular reference to proteomic changes induced by the impact of four different temperature stress regimes, including both hot and cold temperatures, on cultured grape cells. Cabernet Sauvignon cell suspension cultures grown at 26°C were subjected to 14 h of exposure to 34 and 42°C for heat stress, and 18 and 10°C for cold stress. Cells from the five temperatures were harvested in biological triplicates and label‐free quantitative shotgun proteomic analysis was performed. A total of 2042 non‐redundant proteins were identified from the five temperature points. Fifty‐five proteins were only detected in extreme heat stress conditions (42°C) and 53 proteins were only detected at extreme cold stress conditions (10°C). Gene Ontology (GO) annotations of differentially expressed proteins provided insights into the metabolic pathways that are involved in temperature stress in grape cells. Sugar metabolism displayed switching between alternative and classical pathways during temperature stresses. Additionally, nine proteins involved in the phenylpropanoid pathway were greatly increased in abundance at extreme cold stress, and were thus found to be cold‐responsive proteins. All MS data have been deposited in the ProteomeXchange with identifier PXD000977 ( http://proteomecentral.proteomexchange.org/dataset/PXD000977 ).     

Mass spectrometry in the analysis of grape and wine proteins

Wine proteins play an important role in the quality of wine, because they affect taste, clarity and stability of product. The majority of wine proteins are in the range of 20-30 kDa. Different mass spectrometry (MS) techniques have been successfully applied to study the grape and wine proteins. By liquid chromatography (LC) electrospray ionization (ESI) MS and nano-LC/MS, nine dipeptides and 80 peptides were unambiguously identified in Champagne and Sauvignon Blanc wines, respectively. Using matrix-assisted laser desorption/ionization (MALDI) time-of-flight (TOF) and surface-enhanced laser desorption/ionization TOF, the protein and peptide fingerprints in Chardonnay, Sauvignon Blanc and Muscat of Alexandria wines were determined. MALDI-TOF identified the mesocarp proteome of six Vitis grape varieties. Proteins in different grape tissue extracts were also studied. The major grape pathogenic-related proteins are chitinases and thaumatin-like proteins, which both persist through the vinification process and cause hazes and sediments in bottled wines. ESI-MS, LC/ESI-MS and MALDI-TOF analysis of these proteins in grape and wine were also used to characterize different grape varieties.