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

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

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

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

The Efficiency of the Moire Technique for Three-Dimensional Measures of Grape Bunches

This study was carried out in the University of Campinas to study the three-dimensional reconstruction of grape bunches through the application of the Moire technique as an alternative and innovative method, using low-cost equipment for data acquisition. This study proposed the three-dimensional visualization of grapes and topography on Xi axes through the Moire projection technique. The artificial grapes with a 20 cm long bunch and 18.45 mm average diameter for each grape were used. A projector was used to generate a grid with light and dark lines. The grape bunch was placed on a holder, illuminated by a sinusoidal grid and photographed. After processing the images, a three-dimensional map of the fruit and topography on Xi axes were obtained. The three-dimensional fruit observation allows the visualization of the higher and lower regions of the three-dimensional configuration through a color map. This result makes it possible to obtain size and shape of the grape bunch, allowing for the process automation of product selection and classification.

Canopy Management of Table Grapes Cultivar in Tropical Conditions

The production of some seedless table grapes under tropical conditions presents low bud fertility on the main canes and low yield, which leads to the need of pruning in the lateral shoots. The objective of this study was to evaluate the influence of the canopy management for the formation of lateral shoots associated with dellsity of canes on the yield and quality of grapes ＂Sugraone＂ in the Sao Francisco Valley. The experiment was carried out over two growing seasons （2011-2012） in a commercial vineyard of Sugraone in Petrolina, Pernambuco state, Brazil. The treatments consisted of two canopy managements （shoot topping associated to elimination of lateral shoots and shoot topping and formation of lateral shoots） combined with two to three densities of canes after pruning （1.8, 2.8 and 3.8 canes/m^2）. The formation of ＂lateral shoots＂ increased yield and number of clusters per plant during two consecutive growing seasons, besides using 2.8 varas/m^2 resulted in highest yields only in 2011 growing season. The variables mass bunch, mass berry, soluble solids （SS）, soluble sugar, titxatable acidity （TA） and pulp firmness were not affected by treatments, while the attributes related to color, as like brightness and hue angle of the skin, and total extractable polyphenols content were influenced by canopy management, especially in the 2012 growing season. Shoot topping to induce formation of laterals shoots associated with density of 2.8 canes/m2 increased the yield of seedless grapes cultivar Sugraone grown in the Sao Francisco Valley, not significantly affecting the quality of the grape.

葡萄AFLP技术体系建立及其在超藤与藤稔葡萄品种鉴别中的应用

超藤葡萄是藤稔葡萄的一个芽变品种,由于2者亲缘关系十分接近,应用形态学手段以及RAPD分子标记不易区分。针对葡萄叶片富含多糖和多酚类等杂质的特点,从基因组DNA纯化入手,对AFLP流程进行了优化改进,建立了葡萄AFLP体系,并成功地对此2个品种进行了区分。采用引物组合M-CAT+E-ACG进行AFLP扩增,超藤葡萄出现167bp特异带。

Effects of Temperature Acclimation Pretreatment on the Ultrastructure of Mesophyll Cells in Young Grape Plants （Vitis vinifera L. cv. Jingxiu） Under Cross-Temperature Stresses

Leaves from annual young grape plants （Vitis vinifera L. cv. Jingxiu） were used as experimental materials. The ultrastructural characteristics of mesophyll cells in chilling-treated plants after heat acclimation （HA） and in heat-treated plants after cold acclimation （CA） were observed and compared using transmission electron microscopy. The results showed that slight injury appeared in the ultrastructure of mesophyll cells after either HA （38℃ for 10 h） or CA （8℃ for 2.5 d）, but the tolerance to subsequent extreme temperature stress was remarkably improved by HA or CA pretreatment. The increases in membrane permeability and malondialdehyde concentration under chilling （0℃） or heat （45℃） stress were markedly inhibited by HA or CA pretreatment. The mesophyll cells of plants not pretreated with HA were markedly damaged following chilling stress. The chloroplasts appeared irregular in shape, the arrangement of the stroma lamellae was disordered, and no starch granules were present. The cristae of the mitochondria were disrupted and became empty. The nucleus became irregular in shape and the nuclear membrane was digested. In contrast, the mesophyll cells of HA-pretreated plants maintained an intact ultrastructure under chilling stress. The mesophyll cells of control plants were also severely damaged under heat stress. The chloroplast became round in shape, the stroma lamellae became swollen, and the contents of vacuoles formed clumps. In the case of mitochondria of control plants subjected to heat stress, the outer envelope was digested and the cristae were disrupted and became many small vesicles. Compared with cellular organelles in control plants, those in CA plant cells always maintained an integrated state during whole heat stress, except for the chloroplasts, which became round in shape after 10 h heat stress. From these data, we suggest that the stability of mesophyll cells under chilling stress can be increased by HA pretreatment. Similarly, CA pretreatment can protect chloroplasts, mitochondria, and the nucleus against subsequent heat stress; thus, the thermoresistance of grape seedlings was improved. The results obtained in the present study are the first, to our knowledge, to offered cytological evidence of cross-adaptation to temperature stresses in grape plants.

Impact of a Non-<i>Saccharomyces</i>Yeast Isolated in the Equatorial Region in the Acceptance of Wine Aroma

The aim of this work was to isolate and identify the yeasts prevalent in fresh grapes cultivated in the “S&atilde;o Francisco Valley” region (Brazil), as well as evaluating the cell growth of these indigenous yeasts during the fermentation of grape musts and their contribution to the improvement of wine aroma. The Chenin Blanc grape must fermented by H. opuntiae presented higher acceptance means at the three points analyzed (6.74, 6.78 and 7.30) and in the fermentation carried out by the yeasts H. opuntiae and S. cerevisiae, the highest mean acceptance (7.22) was observed after 120 hours, with no statistical difference from the sample fermented by H. opuntiae alone. Since these samples that showed higher acceptance means also receiving higher scores for purchasing intention, corresponding to the concepts of “definitely would buy” and “probably would buy”. The present study suggests that the fermentations of grape musts carried out by the yeast H. opuntiae and mixed cultures of H. opuntiae and Saccharomyces cerevisiae, positively influenced the sensory qualities of the wines and showed greater potential to increase the aroma of the musts and to develop specific wine styles.

基于改进BiSeNet的葡萄黑麻疹病害程度分级预测

为了准确对葡萄(Vitis vinifera L.)黑麻疹病害程度进行分级预测,通过语义分割模型将叶片部分和病斑部分分割出来,以同一叶片上病斑面积与总叶面积的比值作为疾病严重程度分级的依据,对葡萄黑麻疹病害程度进行分级预测。精确标注了PlantVillage公开数据库中的419张葡萄疾病图像,细分为背景、叶片和病斑3个类别,并应用了数据增强技术增加样本多样性。以BiSeNet作为基准模型,引入GhostNet作为上下文路径的主干提取网络,不仅保持了较小的模型参数量,而且在精度上实现了明显提升,满足病害程度分级预测的需求。提出了累加空洞空间金字塔池化(CASPP)模块,用来替换BiSeNet模型中单一的上下文嵌入模块,以增强BiSeNet模型的多尺度上下文信息提取能力,提高了模型的分割精度。经过测试,本研究模型在测试集中的平均交并比为94.11%,在对葡萄黑麻疹病害程度进行分级预测时,准确率达98.21%,能够精确地对葡萄黑麻疹病害程度进行分级预测。

葡萄无核基因定位与作图的研究

以UBC 269484和GSLP1569的序列为支点,设计合成了包括UBC 269和GSLP1在内的9条引物,以葡萄 有核亲本红地球和无核亲本红光无核的DNA为模板,对这9个引物进行筛选,结果GSLP1、39970524 5号引物和 39970524 6号引物在无核亲本红光无核上扩增出了特异标记GSLP1569、39970524 5 564、39970524 6 1538和 39970524 6 1200。用这3个特异引物在红地球、红光无核、无核白和红地球×红光无核杂交组合F1代163株杂 种的DNA样上进行PCR扩增,结果4个特异标记在F1群体中与无核主效基因共分离。4个特异标记也出现于所 用组合中无核基因原始供给者无核白上。这些标记和葡萄无核主效基因相连锁。用QTXb17遗传作图软件,对葡 萄无核主效基因S定位与作图,当P=0.01时,LOD值在32.7～46.4之间,置信界限在0.2～9.9之间。这4个 特异标记和无核主效基因S处于在同一连锁群,位于无核主效基因S的两侧,覆盖基因组12.3cM。特异标记 39970524 5 564、GSLP1569、39970524 6 1538、39970524 6 1200距S基因的遗传距离分别为0.6cM、1.2cM、 4.9cM和11.1cM。

欧亚种葡萄花色素苷的积累及UFGT基因的RT-PCR表达分析

以欧亚种葡萄为试材,研究南方设施栽培条件下果实中花色素苷的积累与着色。结果表明,转熟期花色素苷开始积累,果实颜色由绿转红直至成熟,其积累的花色素苷以单糖苷类为主,不同于欧美杂交种,品种间花色素苷组分也存在差异。设施条件下与离体培养结果均表明外源应用ABA和鼠李糖能够促进欧亚种葡萄红地球果实花色素苷的积累,芍药色素糖苷含量增加显著,蔗糖则起抑制作用。半定量RT-PCR分析显示,葡萄糖类黄酮转移酶RGufgt基因(GenBank:DQ513314)在葡萄幼叶中表达,在绿果时不表达,果实转熟着色后即开始表达,直至果实成熟,同花色素苷积累过程完全一致。

南方欧亚种葡萄引种评价和栽培技术研究

评价了35个欧亚种葡萄品种,认为适合南方栽培的欧亚种葡萄基因型有9个,以其中2个表现优良的主栽品种为试材,研究了南方欧亚种葡萄优质稳产栽培的几个主要技术影响因子。结果表明,矢富罗莎、红地球分别选择SO4、巨峰作砧木,生长结果最优,平均粒重较自根苗分别增加21.6%、10.7%,可溶性固形物含量分别提高14.8%、2.8%;2个品种采用单十字飞鸟型架式较双十字V型架产量分别提高20.7%、15.0%;2个品种采用5-4-3-2-1摘心法较常规方法结果枝率分别提高51.9%、28.6%;使用细胞酶制剂叶面肥粒重增加1.2￣1.6g,可溶性固形物含量提高1%,对降低果实酸度也有一定的影响,明显改善红色品种果实外观。

温度逆境交叉适应过程中葡萄幼苗质膜Ca^(2+)-ATPase的细胞化学定位与活性变化

目的研究植物交叉适应现象及其生理生化机制。方法以京秀葡萄(VitisviniferaL.cv.Jingxiu)为试材,研究了38℃/10h的高温锻炼预处理和8℃/2.5d的低温锻炼预处理分别对随后0℃低温和45℃高温胁迫期间葡萄叶片细胞质膜Ca2+-ATPase的影响。从生化水平测定了定位在质膜、液泡膜上的Ca2+-ATPase的活性,并用氯化铈沉淀的电镜细胞化学方法,定位观察了质膜Ca2+-ATPase的活性。结果高温锻炼预处理提高了微粒体膜上的Ca2+-ATPase酶的活性,并且在随后的低温胁迫条件下,保持了酶活性的稳定性;低温锻炼提高了微粒体在高温胁迫下的ATP酶活性。结论高温锻炼诱导的抗冷性和低温锻炼所诱导的耐热性的提高与微粒体膜上Ca2+-ATPase活性的变化有关,且高温锻炼和低温锻炼具有相同的调控机制。

利用GenBank中大量葡萄EST序列分离有效基因的电子表达分析平台

【目的】充分利用GenBank上的大量葡萄EST序列,建立本地化基因电子表达分析平台,实现基因表达模式的大规模快速分析。【方法】利用生物信息学方法对GenBank上的362 193条葡萄EST序列进行本地化及后续处理,建立电子分析平台,并通过RT-PCR技术对其电子分析准确性进行验证。【结果】建立了包含叶、花等11个组织类别和GA3等5个胁迫类型的基因电子表达分析平台。通过VvAG、VvCHS、VvF3H、VvLDOX等4个葡萄基因的RT-PCR和电子表达分析结果比较发现,平台预测效率较高,在预测EST来源数较多的果实、花序、花组织的表达效果较好,而对叶等EST来源较少的组织的预测效果需结合试验判断。随着dbEST库中源于葡萄各组织EST序列数量的大量增加,平台的预测效果将更加符合基因表达的实际情况。【结论】葡萄基因电子表达分析平台预测效率较高,为葡萄基因大规模快速表达分析以及重要相关信息的挖掘提供了很好的分析工具。

2个葡萄品系外植体愈伤组织诱导和植株再生

进行了无核白、红地球2个葡萄品系外植体愈伤组织诱导和植株再生的研究。通过葡萄品系无核白、红地球叶片和叶柄外植体的愈伤组织的诱导、再分化,以器官发生途径实现植株再生。结果表明:(1)愈伤组织诱导以MS+BA5mg/L+NAA0.5mg/L+蔗糖40g/L培养基最佳,诱导率为35%;同一葡萄品种的叶片与叶柄诱导愈伤组织的频率基本相同,而红地球外植体诱导愈伤组织的频率高于无核白;(2)不定芽的诱导再生以1/2MS+BA0.5mg/L+NAA0.05mg/L+CH500mg/L+蔗糖30g/L培养基最佳,平均诱导率达35.4%;叶柄来源的愈伤组织诱导不定芽的频率高于叶片愈伤组织;(3)根系的诱导以1/2MS+KT0.5mg/L+IBA1.0mg/L+AC2g/L+蔗糖15g/L培养基最佳,无核白和红地球来源的不定芽诱导生根频率基本相同,都在80%以上。

预处理方式对提子类葡萄贮藏期间质地的影响

质地是鲜食型水果重要指标之一,为探讨不同预处理方式对鲜食型提子类葡萄贮藏期间质地的变化,采用5种预处理方式对红白两种不同欧亚种提子类葡萄进行处理,然后在(4±1)℃下冷藏,分别在0,10,20,30,40d利用质构仪质地多面分析法测定红白提子类葡萄果实的质地,得到红白提子类葡萄贮藏期间果实质地参数变化规律。结果表明:果实硬度与胶着性、弹性、断裂性均呈很好正相关(r=0.943~1.00),与内聚性呈负相关(r=-0.722^-0.708);果实断裂性与弹性、胶着性和咀嚼性呈较好的正相关(r=0.846~0.996);弹性与胶着性、咀嚼性呈正相关(r=0.961~0.968),胶着性与咀嚼性呈显著正相关(r=0.954~0.944)。果实的硬度与胶着性和咀嚼性呈正相关(r=0.846~0.996),确定以提子类葡萄果实硬度、胶着性和咀嚼性用于比较提子类葡萄质地变化的参数。5种预处理后冷藏的提子类葡萄各项质地参数变化规律总体呈现下降趋势,但不同的预处理表现出的质地下降趋势不同,其中B处理(抗菌保鲜处理)下降趋势最明显。试验证明:贮藏前预处理技术是影响产品贮藏期间质地的重要因素,不同的预处理改变了提子类葡萄的质地,增大了果实的硬度、胶着性和咀嚼性。

SO_2对葡萄采后呼吸强度及内源激素的影响

采用气相色谱法和改进的高效液相色谱法研究发现：‘无核白’葡萄果粒为非跃变型呼吸类型，其乙烯释放量很低，但释放潜力很高，切伤后乙烯释放速率迅速增高；果梗和穗梗为跃变型呼吸类型，有很高的呼吸高峰和乙烯释放高峰。适量的ＳＯ２保鲜处理对葡萄的呼吸强度、脱落酸（ＡＢＡ）含量及乙烯释放具有明显抑制作用，对生长素（ＩＡＡ）、赤霉素（ＧＡ）有促进作用，既可抑制葡萄生理衰老，又可减轻葡萄的落粒。

福建野生葡萄松散型愈伤组织的诱导及其继代保持

以福建野生葡萄(Vitis amurensis Rupr.)带节茎段为外植体,建立无菌株系,并以此无菌株系小苗的茎段或叶片为外植体,进行愈伤组织诱导、继代增殖和长期保持等方面的研究。结果表明,光照条件下,愈伤组织诱导以MS+BA 2.0mg·L-1+NAA 0.2mg·L-1+蔗糖30g.L-1培养基最佳,诱导率可达100%;愈伤组织的继代增殖以MS+2,4-D 1.0mg·L-1+蔗糖30g.L-1培养基最佳,可以获得生长状态良好的松散型愈伤组织;采用MS+2,4-D 1.0mg·L-1+蔗糖30g.L-1培养基继代培养几代后转到MS+2,4-D 1.0mg·L-1+KT 0.5mg·L-1+AgNO35.0mg·L-1+蔗糖30g.L-1培养基上培养1代,如此交替进行,同时在光照条件下,可长期继代保持野生葡萄愈伤组织,并可保持其旺盛的生长能力。

丛枝菌根真菌接种对葡萄扦插苗生长效应的影响

在温室盆栽条件下,通过在不同接种时期对葡萄扦插苗接种不同剂量的丛枝菌根真菌,研究了丛枝菌根真菌对葡萄扦插苗生长效应的影响。结果表明：在葡萄扦插过程中接种丛枝菌根真菌,能促进扦插苗的生长发育,但其促进作用受到接种时期和接种剂量的影响,一般扦插后14～21 d接种30 spores·unit-1的AM菌剂作用最佳,能显著促进葡萄扦插苗根系对P、N等矿质养分的吸收和积累,提高扦插苗的生长效应。

葡萄不同摘心处理下qRT-PCR内参基因的筛选与验证

摘心处理是保证葡萄产量和品质的关键栽培技术之一,选择稳定可靠的内参基因,有利于葡萄摘心响应分子机制相关基因表达的研究。本研究以不同摘心处理的葡萄叶片和茎杆为材料,利用qRT-PCR分析15个候选内参基因Actin、AP-2、Cyclophilin、EF1-α、GAPDH、VAG、SAND、α-Tubulin、β-Tubulin、UBQ-L40、NAD5、ADH2、60SRP、18S rRNA、UBQ等在所供材料中的转录水平,并用RefFinder软件对候选内参基因稳定性进行综合评价。试验结果表明,葡萄叶片为材料的试验体系的最稳定内参基因为SAND和VAG,而葡萄茎杆为材料的试验体系以AP-2和Actin为最稳定的内参基因,且采用双内参基因组合,可以进一步提高试验结果的精确度;通过目的基因VvSS4转录水平数据标准化的验证,所获得的2组内参基因在各自的试验体系中可靠性高,可用于后续相关目的基因定量表达的研究。

葡萄叶片中提取总RNA的三种方法比较

采用试剂盒法、热酚法和LiCl沉淀法3种方法提取葡萄叶片总RNA,通过琼脂糖凝胶电泳和Agilent 2100计检测总RNA的完整性和提取纯度。对红地球葡萄叶片总RNA提取方法做比较,以得到完整的、高质量的葡萄RNA。结果表明:LiCl沉淀法提取的RNA收率高,完整性好,OD260/OD280均在1.80～2.00,纯度高,28S、18S条带清晰完整。可以满足后续分子生物学的研究。