葡萄初级核心种质资源MybA基因型分析

探究MybA类基因在不同类型葡萄品种中的分布,可为葡萄品种鉴定,以及有色葡萄育种的亲本选择提供依据。本研究以欧亚种、欧美杂种、法美杂种、山欧杂种以及美洲种在内的118个葡萄初级核心种质为材料,对其MybA基因型进行分析。结果表明:欧亚种及其杂种普遍具有VvmybA1基因的等位基因VvmybA1a,仅10个欧亚种及其杂种品种中没有检测到VvmybA1a基因;欧亚种、欧美杂种以及法美杂种中普遍同时具有VvmybA1、VvmybA2和VvmybA3基因,仅少数品种未检测到VvmybA2或VvmybA3基因;山欧杂种中北玫、公酿1号和熊岳白葡萄同时具有VvmybA1、VvmybA2和VvmybA3基因,北醇和北红中仅检测到VvmybA1和VvmybA3基因;仅在具有美洲种血缘的葡萄品种中检测到VlmybA2基因,而5个认为是美洲种的品种未检测到VlmybA2基因,且检测到了欧亚种特有的VvmybA1a等位基因,推测它们为含美洲种血缘较多的欧美杂种,而非纯美洲种。     

中国欧亚种酿酒葡萄种植分布的主要气候影响因子与气候适宜性

开展酿酒葡萄气候适宜性研究对于优化酿酒葡萄布局、气候资源开发利用具有重要意义。基于欧亚种酿酒葡萄(Vitis vinifera L.)分布数据和影响其分布的气候因子,利用最大熵模型(MaxEnt)和地理信息系统(ArcGIS),研究影响欧亚种酿酒葡萄种植分布的主导气候因子及其气候适宜性。结果表明:MaxEnt模型能够很好地模拟我国欧亚种酿酒葡萄的潜在分布,模拟效果达到"非常好"(AUC平均值0.936)的水平。基于气候因子对欧亚种酿酒葡萄地理分布影响的贡献确定了主导气候因子,即无霜期、干燥度、极端最低气温、年降水量、生长季日照时数、≥10℃活动积温。当前,我国欧亚种酿酒葡萄种植分布的气候高适宜区、适宜区、次适宜区分别占次适宜及以上区域总面积的2.9%、20.4%和76.7%。欧亚种酿酒葡萄气候高适宜区主要分布在宁夏、山西、陕西、内蒙古、山东、河北、新疆、甘肃等省,只考虑气候因子,陕西、山西、内蒙古具有较大的发展空间。     

Chloroplast phylogenomics of the New World grape species (Vitis,Vitaceae)

Vitis L. (the grape genus) is the economically most important fruit crop, as the source of grapes and wine. Phylogenetic relationships within the genus have been highly controversial. Herein, we employ sequence data from whole plastomes to attempt to enhance Vitis phylogenetic resolution. The results support the New World Vitis subgenus Vitis as monophyletic. Within the clade, V. californica is sister to the remaining New World Vitis subgenus Vitis. Furthermore, within subgenus Vitis, a Eurasian clade is robustly supported and is sister to the New World clade. The clade of Vitis vinifera ssp. vinifera and V. vinifera ssp. sylvestris is sister to the core Asian clade of Vitis. Several widespread species in North America are found to be non‐monophyletic in the plastome tree, for example, the broadly defined Vitis cinerea and V. aestivalis each needs to be split into several species. The non‐monophyly of some species may also be due to common occurrences of hybridizations in North American Vitis. The classification of North American Vitis by Munson into nine series is discussed based on the phylogenetic results. Analyses of divergence times and lineage diversification support a rapid radiation of Vitis in North America beginning in the Neogene.     

Anthocyanin Profiles in Grape Berry Skins of Different Species of Wine Grapes in Shanxi,China

To understand the anthocyanin characteristics of wine grape varieties, the anthocyanin composition and content of 31 wine grape varieties were analyzed to explore the use of anthocyanins as chemical fingerprints to distinguish varieties. Results showed that a total of 21 anthocyanins were detected in the skins, including cyanidin, delphinidin, petunidin, peonidin and malvidin 3-monoglucosides (or 3,5-diglucosides) along with the corresponding acetyl and p-coumaroyl derivatives. The highest and lowest total amount of anthocyanins were detected in ‘Ruby Cabernet’ and ‘Muscat Rouge’, respectively. In the 21 Vitis vinifera grapes, there were 3~11 monoglucoside anthocyanins detected, however, there were 4 to 9 monoglucoside anthocyanins and 1~7 diglucoside anthocyanins detected in the 10 other species of grapes. Except for ‘Zhesexiang’ ‘Seibel Noir’, ‘44-6-7-1’ and ‘Beibinghong’, the contents of diglucoside anthocyanins in the other six varieties accounted for more than 52% of the total anthocyanins. Except for ‘Zhesexiang’, ‘Muscat Rouge’ and ‘Beibinghong’, the content of methylated anthocyanins accounted for more than 75% of total anthocyanins. There were significant differences in the anthocyanin types and contents in the skins among V. vinifera and other grapes. The results of the principal component analysis and the cluster classification of 31 grape varieties (lines) were nearly consistent, which suggested that anthocyanins can be used as chemical fingerprints to distinguish wine grape varieties.     

Activity of grape extracts from Greek varieties of Vitis vinifera against mutagenicity induced by bleomycin and hydrogen peroxide in Salmonella typhimurium strain TA102

Several in vivo and in vitro studies have shown that grape extracts could prevent certain steps in carcinogenesis and a few mechanisms have been proposed for this activity. In this study, the potential antimutagenic activity of methanolic and aqueous extracts from two Greek grape varieties of Vitis vinifera against DNA damage induced by reactive oxygen species (ROS) was assessed as a potential novel chemopreventive mechanism, using Salmonella typhimurium strain TA102. The two grape varieties were Assyrtiko (white grapes) and Mandilaria (red grapes), while the oxidant mutagens used were bleomycin (BLM) and hydrogen peroxide (H(2)O(2)). Since it has been considered that polyphenols present in grapes are their most potent biologically active compounds, we also tested the effects of polyphenol-rich fractions as well as some of the more common grape polyphenols on the activity of the two test mutagens. These polyphenols were quercetin, (+)-catechin, (-)-epicatechin, trans-resveratrol, gallic acid and protocatechuic acid. Almost all extracts showed inhibitory activity against both mutagens. On the other hand, polyphenol-rich fractions as well as individual polyphenols at concentrations found in the extracts either did not diminish or did enhance the activity of the mutagens. These results suggest that the protection of DNA from mutations induced by ROS may be one of the mechanisms accounting for the chemopreventive activity of grape extracts. However, it seems that this protective activity may not be attributed to polyphenols but rather to a synergism of many compounds in the grapes.     

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.     

Comparison of a retrotransposon-based marker with microsatellite markers for discriminating accessions of Vitis vinifera

Identification and knowledge concerning genetic diversity are fundamental for efficient management and use of grapevine germplasm. Recently, new types of molecular markers have been developed, such as retrotransposon-based markers. Because of their multilocus pattern, retrotransposon-based markers might be able to differentiate grapevine accessions with just one pair of primers. In order to evaluate the efficiency of this type of marker, we compared retrotransposon marker Tvv1 with seven microsatellite markers frequently used for genotyping of the genus Vitis (VVMD7, VVMD25, VVMD5, VVMD27, VVMD31, VVS2, and VZAG62). The reference population that we used consisted of 26 accessions of Vitis, including seven European varieties of Vitis vinifera, four North American varieties and hybrids of Vitis labrusca, and 15 rootstock hybrids obtained from crosses of several Vitis species. Individually, the Tvv1 and the group of seven SSR markers were capable of distinguishing all accessions except 'White Niagara' compared to 'Red Niagara'. Using the Structure software, the retrotransposon marker Tvv1 generated two clusters: one with V. vinifera plus North American varieties and the other comprising rootstocks. The seven SSR markers generated five clusters: V. vinifera, the North American varieties, and three groups of rootstock hybrids. The percentages of variation explained by the first two components in the principal coordinate analysis were 65.21 (Tvv1) and 50.42 (SSR markers) while the Mantel correlation between the distance matrixes generated by the two types of markers was 42.5%. We conclude that the Tvv1 marker is useful for DNA fingerprinting, but it lacks efficiency for discrimination of structured groups.     

Anthocyanin, antioxidant activity and stress-induced gene expression in high CO2-treated table grapes stored at low temperature

A pretreatment with 20kPa CO2+20 kPa O2+60 kPa N2 for 3 days proved effective in maintaining the fruit quality and controlling decay in table grapes (Vitis vinifera cv. Cardinal) stored at 0 degrees C. In the present work, we analyzed whether total anthocyanin content, the molecular mechanism implicated in their biosynthesis and antioxidant activity is related to the beneficial effect of this gaseous treatment. We isolated partial cDNAs that codified for enzymes implicated in the anthocyanin biosynthesis such as l-phenylalanine ammonia-lyase (PAL) and chalcone synthase (CHS), and an antioxidant enzyme such as ascorbate peroxidase (APX). Low temperatures induced an accumulation of total anthocyanin content in the skin of both treated and non-treated grapes, although levels were lower in CO2-treated fruit. By contrast, antioxidant activity decreased during storage at 0 degrees C in non-treated grapes but did not change in CO2-treated grapes. The up-regulation of anthocyanin biosynthesis gene expression and VcAPX mRNA observed in non-treated grape is not enhanced in CO2-treated grapes, which presented low total decay. These results point out the ability of CO2-treated grapes to prevent the generation of reactive oxygen species rather than their inactivation by means of induction of studied defense systems.     

Yeasts associated to Malbec grape berries from Mendoza, Argentina

AIMS: The aims of this work were to evaluate different pre-isolation treatments applied to complete yeast extraction from grapes and to identify the yeast microflora associated to Malbec grapes from two vineyards located in Mendoza, Argentina. METHODS AND RESULTS: The pre-isolation treatments evaluated were shaking, jet streaming with pressurized water and grape blending. The overall results clearly indicated that when a more vigorous and disruptive pre-isolation treatment was applied; larger numbers of yeast species were recovered. The yeast population on healthy and ripe Malbec grapes was in the order of 10(5)-10(6) CFU g(-1). Eight different yeast species were isolated from berries, including Kloeckera apiculata, Metschnikowia pulcherrima, Pichia membranifaciens, Saccharomycodes ludwigii, Candida species (Candida stellata and Candida raghi), Issatchenkia orientalis and Rhodotorula spp. CONCLUSIONS: Grape blending gave the highest yeast counts. Rainfall near grape harvest time quantitatively and qualitatively modifies the yeast microflora. The yeast species identified on ripe grapes from the Mendoza region, partially match those previously documented in different parts of the world related. S. ludwigii has not been previously reported in grapes. SIGNIFICANCE AND IMPACT OF THE STUDY: The report is on yeast microbiota in grapes from Mendoza, Argentina. Saccharomycodes ludwigii was found in high percentage (17%), this species has not been described before on grapes surface. The importance of pre-isolation steps to the recovery of high number of yeasts was shown. Influence of climatic conditions near harvest time on microflora was confirmed.     

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 Diversity in Wild Grapes Native to China Based on Randomly Amplified Polymorphic DNA (RAPD) Analysis

The use of the RAPD technique was investigated on a set of 73 genotypes of 18 wild grape species native to China, and one interspecific hybrid, seven Vitis vinifera L. cultivars, one rootstock cultivar and one strain of V. riparia L. Genetic diversity among these grapes was investigated based on RAPD analysis. The screening of 280 decamer oligonucleotides allowed the selection of 20 primers used for the analysis. A total of 191 RAPD markers were produced from the 20 selected primers. Relationships among the 83 clones or accessions based on their genetic distances were clustered using unweighted pair-group method arithmetic average (UPGMA) analysis in a dendrogram. Twenty-two clusters which fortunately adapted to 22 grape species level were clearly resolved on the dendrogram. The 18 wild grape species native to China were grouped into ten subclusters. The largest distance was found between V. riparia L., V. vinifera L., interspecific hybrid ( V. vinifera L.× V. larbrusca L.) and the wild grapes native to China. Among the wild grapes native to China, the largest distance was found between V. hancockii Hance and the other wild species. V. qinlingensis P.C.He was the second. Large genetic variation occurred among the different flower-type clones in one species.     

Identification of microsatellite sequences in Vitis riparia and their applicability for genotyping of different Vitis species.

A Vitis riparia genomic library was screened for the presence of (GA)n simple sequence repeats (SSR) and 18 primer pairs yielding amplification products of the expected size were designed. Heterologous amplification with the primer pairs in related species (V. rupestris, V. berlandieri, V. labrusca, V. cinerea, V. aestivalis, V. vinifera, and interspecific hybrids) was successful in most primer-species combinations. Therefore, the new markers are applicable to the genotyping of a range of Vitis species. Variations in the SSR flanking sequence were detected between and within the species. The degree of polymorphism and performance of the markers were determined in up to 120 individuals of V. vinifera. Four of fifteen alleles per locus were detected and expected heterozygosity ranged between 0.37 and 0.88. Null alleles were shown to be present at two loci by a lack of heterozygous individuals and by transmission of the null alleles in a controlled cross. Regular Mendelian inheritance is indicated for all but one loci by a preliminary segregation analysis in 36 offspring. Thirteen of the markers were found suitable for the genotyping of grapevines (V. vinifera).     

De novo synthesis of monoterpenes by Saccharomyces cerevisiae wine yeasts

This paper reports the production of monoterpenes, which elicit a floral aroma in wine, by strains of the yeast Saccharomyces cerevisiae. Terpenes, which are typical components of the essential oils of flowers and fruits, are also present as free and glycosylated conjugates amongst the secondary metabolites of certain wine grape varieties of Vitis vinifera. Hence, when these compounds are present in wine they are considered to originate from grape and not fermentation. However, the biosynthesis of monoterpenes by S. cerevisiae in the absence of grape derived precursors is shown here to be of de novo origin in wine yeast strains. Higher concentration of assimilable nitrogen increased accumulation of linalool and citronellol. Microaerobic compared with anaerobic conditions favored terpene accumulation in the ferment. The amount of linalool produced by some strains of S. cerevisiae could be of sensory importance in wine production. These unexpected results are discussed in relation to the known sterol biosynthetic pathway and to an alternative pathway for terpene biosynthesis not previously described in yeast.     

Accumulation of tocopherols and tocotrienols during seed development of grape (Vitis vinifera L. cv. Albert Lavallée)

Tocopherols and tocotrienols are present in mature seeds. Yet, little is known about the physiological role and the metabolism of these compounds during seed development. Here we present data on tocopherol and tocotrienol accumulation during seed development in Vitis vinifera L. cv. Albert Lavallée (Royal). This species was chosen for its ability to synthesize both tocopherols and tocotrienols. It is shown here for the first time that during seed development there are significant differences in localization and accumulation kinetics of tocopherols and tocotrienols. Tocopherols are found homogeneously dispersed throughout all tissues of the seed, in concentrations ranging from 20 to 100 microg tocopherol per g dry weight. Tocopherol levels decrease gradually during seed development. In contrast, tocotrienols are only found in the endosperm of the seeds, accumulating in a sigmoid fashion during the maturation period of seed development. Tocotrienol levels were found to be (54+/-7.4) microg/g dry seed in 90-day-old seeds of V. vinifera L. Furthermore, tocotrienol biosynthesis is demonstrated in these seeds during tocotrienol accumulation and in an endosperm fraction isolated at 75 days after flowering.     

美国酿酒葡萄品种在北京地区的生长和适应性表现

调查研究了首次引进的13个美国酿酒葡萄品种在北京地区栽培后的植物学性状、果实品质和抗寒及抗霜霉病能力,并与传统的欧亚种酿酒葡萄及砧木品种进行了比较与分析,总结了它们之间的植物学性状与抗逆性的差异。本试验选出了在抗寒及抗霜霉病方面表现优良的美国酿酒葡萄品种,为这些品种在我国的实际栽培推广提供科学依据。     

Application and evaluation of denaturing gradient gel electrophoresis to analyse the yeast ecology of wine grapes

The performance of denaturing gradient gel electrophoresis (DGGE) for analysing yeasts associated with wine grapes was compared with cultural isolation on malt extract agar (MEA). After optimisation of PCR and electrophoretic conditions, the lower limit of yeast detection by PCR-DGGE was 10(2) cfuml(-1), although this value was affected by culture age and the relative populations of the species in mixed culture. In mixed yeast populations, PCR-DGGE detected species present at 10-100-fold less than other species but not when the ratio exceeded 100-fold. Aureobasidium pullulans was the main species isolated from immature, mature, and both damaged and undamaged grapes. It was not detected by PCR-DGGE when present at populations less than 10(3) cfug(-1). When approaching maturity, damaged grapes gave a predominance of Metschnikowia and Hanseniaspora species (10(5)-10(7) cfug(-1)), all detectable using PCR-DGGE. However, various species of Rhodotorula, Rhodosporidium and Cryptococcus were not detected by this method, even when populations were as high as 10(4) cfug(-1). PCR -DGGE was less sensitive than culture on MEA for determining the yeast ecology of grapes and could not reliably detect species present at populations less than 10(4) cfug(-1). However, this method detected a greater diversity of species than agar plating.     

A Basic Peroxidase Isoenzyme, Marker of Resistance Against Plasmopara viticola in Grapevines, is Induced by an Elicitor from Trichoderma viride in Susceptible Grapevines

The constitutive expression of basic peroxidase isoenzymes in the Plasmopara viticola -resistant ( Vitis vinifera × Vitis rupestris) × Vitis riparia crossing and in the P. viticola -susceptible V. vinifera parent species was studied. The results illustrate that both leaves and stems of the ( V. vinifera × V. rupestris) × V. riparia crossing showed the differential expression of a basic peroxidase isoenzyme B₃ (pl = 8.9), this being almost completely absent from the P. viticola -susceptible V. vinifera parent species. To test whether the basic peroxidase isoenzyme B₃ may be considered as a molecular marker of disease resistance in Vitis spp., suspension cell cultures derived from the P. viticola -susceptible V. vinifera parent species were treated with an elicitor (cellulase Onoztika R-10) from the soil fungus Trichoderma viride , a specific and well-known elicitor of disease resistance reactions in grapevines. The results showed that treatment with the elicitor induces, simultaneously with the activation of the disease resistance mechanism, the appearance of B₃ in the cell cultures. These results suggest that the basic peroxidase isoenzyme B₃ may be considered as a marker of disease resistance in Vitis species since it is present in the P. viticola -resistant ( V. vinifera × V. rupestris) × V. riparia hybrid and is induced by the elicitor Onozuka R-10 in cell cultures of the P. viticola -susceptible Vitis vinifera parent species. This conclusion is supported by the presence of this isoenzyme in other resistant and its absence in other susceptible Vitis spp.     

Increasement of Heterologous Expression of Recombinant Vit v 1 in Pichia pastoris KM71 by Nonionic Detergents as a Cost-effective Approach

Applied Biochemistry and Microbiology - Vit v 1 as a lipid-transfer protein is a major allergen of grapes (Vitis vinifera) that elicits food allergy in many patients in Iran. Todays, recombinant...