产地与品种对葡萄酒中酚类物质含量的影响

选取同一产地(天水)3 个葡萄品种：“西拉(Shiraz)”、“蛇龙珠 (Carbernet Gernischet)”、“赤霞珠(Cabernet Sauvignon)”所酿制的新鲜干红葡萄酒以及5 个不同产地(天水、和硕、玛纳斯、延庆和怀来)赤霞珠葡萄酒,采用高效液相色谱(HPLC)法分析葡萄酒中11 种酚酸、5 种黄烷-3- 醇、10 种黄酮醇和2 种黄酮的含量。结果表明：同一产地不同品种和同一品种不同产地间的葡萄酒中酚类物质的含量都存在显著差异。其中,赤霞珠葡萄酒是3 个葡萄品种中酚类物质含量最高的品种;在5 个不同产地中,延庆赤霞珠葡萄酒中酚酸含量最高,黄烷-3- 醇含量最高的为和硕赤霞珠葡萄酒,而怀来赤霞珠葡萄酒中所含黄酮醇含量最高。     

几个酿酒葡萄品种及品系的有机酸分析

本研究利用反相高效液相色谱技术,分析同一地区几个酿酒葡萄品种(赤霞珠、美乐和品丽珠等)及其品系的果实中4种有机酸(酒石酸、苹果酸、柠檬酸和草酸)的含量.结果表明:同一品种不同品系间果实有机酸含量存在显著差异;不同定植地点的赤霞珠果实有机酸含量也存在一定差异;不同品种的酿酒葡萄其有机酸含量也有所不同;而有机酸含量差异主要与苹果酸和酒石酸合成代谢中碳的流向改变有关.这些结果为品种及品系的选择及酿造工艺与产品类型的制定提供依据.     

The microbial ecology of wine grape berries

Grapes have a complex microbial ecology including filamentous fungi, yeasts and bacteria with different physiological characteristics and effects upon wine production. Some species are only found in grapes, such as parasitic fungi and environmental bacteria, while others have the ability to survive and grow in wines, constituting the wine microbial consortium. This consortium covers yeast species, lactic acid bacteria and acetic acid bacteria. The proportion of these microorganisms depends on the grape ripening stage and on the availability of nutrients. Grape berries are susceptible to fungal parasites until véraison after which the microbiota of truly intact berries is similar to that of plant leaves, which is dominated by basidiomycetous yeasts (e.g. Cryptococcus spp., Rhodotorula spp. Sporobolomyces spp.) and the yeast-like fungus Aureobasidium pullulans. The cuticle of visually intact berries may bear microfissures and softens with ripening, increasing nutrient availability and explaining the possible dominance by the oxidative or weakly fermentative ascomycetous populations (e.g. Candida spp., Hanseniaspora spp., Metschnikowia spp., Pichia spp.) approaching harvest time. When grape skin is clearly damaged, the availability of high sugar concentrations on the berry surface favours the increase of ascomycetes with higher fermentative activity like Pichia spp. and Zygoascus hellenicus, including dangerous wine spoilage yeasts (e.g. Zygosaccharomyces spp., Torulaspora spp.), and of acetic acid bacteria (e.g. Gluconobacter spp., Acetobacter spp.). The sugar fermenting species Saccharomyces cerevisiae is rarely found on unblemished berries, being favoured by grape damage. Lactic acid bacteria are minor partners of grape microbiota and while being the typical agent of malolactic fermentation, Oenococcus oeni has been seldom isolated from grapes in the vineyard. Environmental ubiquitous bacteria of the genus Enterobacter spp., Enterococcus spp., Bacillus spp., Burkholderia spp., Serratia spp., Staphylococcus spp., among others, have been isolated from grapes but do not have the ability to grow in wines. Saprophytic moulds, like Botrytis cinerea, causing grey rot, or Aspergillus spp., possibly producing ochratoxin, are only active in the vineyard, although their metabolites may affect wine quality during grape processing.The impact of damaged grapes in yeast ecology has been underestimated mostly because of inaccurate grape sampling. Injured berries hidden in apparently sound bunches explain the recovery of a higher number of species when whole bunches are picked. Grape health status is the main factor affecting the microbial ecology of grapes, increasing both microbial numbers and species diversity. Therefore, the influence of abiotic (e.g. climate, rain, hail), biotic (e.g. insects, birds, phytopathogenic and saprophytic moulds) and viticultural (e.g. fungicides) factors is dependent on their primary damaging effect.     

中国野生刺葡萄（Vitis davidii Foex）及其单品种葡萄酒中花旗松素含量分析

利用高效液相色谱-串联质谱联用技术对原产中国的野生刺葡萄果实的果皮、果肉、种子及单品种葡萄酒中的花旗松素含量（以鲜质量计算）进行测定与分析。结果表明：刺葡萄果实果皮中花旗松素含量最高,其含量在刺葡萄黑、白果实中分别为（2.44±0.18）、（2.03±0.14） mg/kg,在种子中分别为（1.66±0.13）、（1.38±0.12） mg/kg,果肉中含量最低,分别为（0.36±0.02）、（0.25±0.02） mg/kg。刺葡萄黑、白果实中各个部位花旗松素含量均无明显差异,但对照品种黑色果实果皮中的花旗松素含量显著高于白色果实,这说明花旗松素含量的高低与葡萄果皮颜色并无直接关系。在单品种酒中,白葡萄酒由于酿造工艺不同,果皮和种子中的花旗松素无法进入酒中,花旗松素含量仅为红葡萄酒的20%;葡萄酒中的花旗松素含量略低于果实是因为提取果实中花旗松素的溶剂是甲醇,而葡萄酒中只是水和乙醇的自然浸渍。     

Effect of freezing grape berries and heating must samples on extraction of components and composition parameters of red wine grape varieties

Juices from grape samples subjected to two storage methods (fresh and frozen) and two post-homogenisation treatments (21^oC and 71^oC) were examined for extraction of grape berry components and compared to a microvinified wine made from the same lot of fresh grapes. Cabernet Franc ( Vitis vinifera ) and Cynthiana ( Vitis aestivalis ) grapes were evaluated in 2003. Cabernet Franc, Cabernet Sauvignon ( Vitis vinifera ), Chambourcin (interspecific hybrid) and Cynthiana grapes were evaluated in 2004. Fresh grapes from each variety were sampled, analysed and prepared for microvinification. Frozen grapes of all varieties were thawed and analysed at the same time. For all varieties in both years, heating the must of fresh or frozen grapes increased the titratable acidity (11–58%), tartaric acid levels (12–110%), potassium content (15–62%), darkness (31–97%), red colour (54–1517%) and total red pigment values (36–1171%) of juice as compared to the juice from unheated must. For all varieties in 2004, heating the must from fresh or frozen grapes increased total phenolics (43–619%) of juice as compared to the juice from unheated must. Compared to fresh grape processing, freezing the grapes and then processing at 21^oC or 71^oC had varying results (increases and decreases) regarding the extraction of components in all varieties and both years but usually provided juice with composition most similar to that of the red wine produced by microvinification. Although the degree of extraction of individual components in the juice is influenced by selection of grape storage and must processing procedures, consistency of handling and procedure provided consistent results with low variability.     

Fungi and mycotoxins in vineyards and grape products

Many fungi may occur on grapes during growth in the vineyard, but the main concern from the viewpoint of mycotoxin contamination is the black Aspergilli, Aspergillus carbonarius and A. niger. These fungi are capable of producing ochratoxin A (OA) which may contaminate grapes and grape products such as wine, grape juice and dried vine fruit. Understanding the ecology and physiology of the black Aspergilli can provide tools for management of OA at all stages of grape production and processing. In the vineyard, careful management of cultivation, irrigation and pruning can assist in minimising the levels of black Aspergilli in the soil, which in turn, can minimise contamination of grapes by these fungi. Minimising damage to grapes on the vine by the use of open vine canopies, grape varieties with resistance to rain damage and by the management of insect pests and fungal diseases (e.g., mildew, Botrytis bunch rot) can reduce the incidence of Aspergillus rot in mature berries. The risk of OA in table grapes can be minimised by careful visual inspection to avoid damaged and discoloured berries. In wine, harvesting grapes with minimal damage, rapid processing and good sanitation practices in the winery assist in minimising OA. During vinification, pressing of grapes, and clarification steps which remove grape solids, grape proteins and spent yeast can also remove a significant proportion of OA. For dried vine fruit production, avoiding berry damage, rapid drying, and final cleaning and sorting to remove dark berries can reduce overall OA levels in finished products.     

谈酿出色泽深红葡萄酒的条件

色泽是影响葡萄酒感官质量的重要指标之一,如何酿出色泽深红葡萄酒是葡萄酒酿造技术关注的焦点。红葡萄酒呈色物质主要是葡萄浆果中的花色苷,花色苷类物质的浓度及组成决定红葡萄酒色泽的深度及稳定性。红葡萄酒的色泽深浅受多方面的影响,如酿造葡萄酒用的原料品种及成熟度、酵母菌、发酵工艺及设备、倒灌、皮渣分离等,从多方面对影响红葡萄酒色泽的因素进行了分析和探索,提出了一些改进的方法和建议。     

Understanding the microbial ecosystem on the grape berry surface through numeration and identification of yeast and bacteria

Microbial species present on the surface of grape berries at harvest play an important role in winemaking, thus counting and identifying them is of great importance. The use of conventional microbial techniques and molecular methods allowed a quantitative and qualitative inventory of the different microbial species present on the grape berries. These experiments were carried out in several areas of the Bordeaux region on the red grape varieties Merlot, Cabernet Sauvignon and Cabernet Franc. Populations and species clearly varied according to berry development stage. The most widespread yeast species at berry set, Aureobasidium pullulans was never detected at harvest. Fermentative yeasts were detected at harvest and not in the first stage of grape growth. Oenoccocus oeni was detected on immature as well as on mature berries. Gluconobacter oxydans was detected mainly at harvest. Detection of Pediococcus parvulus , was dependent on the vineyard. Veraison appeared to be a key stage for yeast colonisation and the increase in population involved a change in the proportion of each species. The number of A. pullulans fell significantly at veraison as it was superseded by fermentative yeasts. Microbial populations peaked at harvest when the berry surface available for adhesion was largest and no agrochemical treatments had been applied for some weeks. Soil, grape variety and grapegrowing practices may also influence this microbial ecosystem. Based on these and published data, we formulated hypotheses to describe this microbial ecosystem, thus enabling us to develop the concept of a microbial biofilm.     

本土酿酒酵母的筛选及其低醇葡萄酒特性研究

以天津宝坻产区的三种葡萄为原料,分别从自然发酵的葡萄汁中筛选出340株酵母菌,经生理生化,产酸、酯、尿酶、H2S,发酵能力测定及26SrRNA测序,确定4株为酿酒酵母,用于贵人香低醇白葡萄酒的酿造。以商用酵母为对照,定量分析了不同酵母所酿酒中的香气成分、氨基甲酸乙酯含量,并进行了理化指标和感官测定,发现酵母C508和G611酿造的低醇酒具有更独特的香气特点,氨基甲酸乙酯的含量较低,微生物稳定性较高,更适合低醇酒的酿造。     

陕西合阳地区酿酒葡萄果实成熟特性研究

该实验研究了合阳地区5个红色和5个白色酿酒葡萄品种在成熟期的可溶性固形物、还原糖、总酸含量和成熟系数变化,分析了成熟葡萄浆果的物理品质、花色苷和单体酚的组成和含量。结果表明,转色期后12 d,各品种葡萄的成熟系数均＞20。成熟葡萄浆果还原糖含量为180～235 g/L,总酸含量＜6 g/L。综合来看,赤霞珠、品丽珠、黑比诺、马瑟兰、爱格丽、小芒森和贵人香适合在合阳地区种植。赤霞珠、品丽珠、黑比诺、马瑟兰和贵人香最适采收期在转色期后24 d左右,爱格丽和小芒森在转色期后44 d左右。马瑟兰的成熟浆果花色苷含量较高,达29.85 mg/g;赤霞珠、马瑟兰和爱格丽成熟浆果单体酚含量较高,分别为0.65 mg/g、0.43 mg/g和0.26 mg/g。研究结果将为陕西合阳地区酿酒葡萄种植与推广提供参考。     

Role of non-Saccharomyces yeasts in Korean wines produced from Campbell Early grapes: Potential use of Hanseniaspora uvarum as a starter culture

Several yeasts were isolated from Campbell Early grapes (Vitis labrusca cultivar Campbell Early), the major grape cultivar in Korea, grown in two different regions. PCR-RFLP analysis of the ITS I-5.8S-ITS II region showed that 34 isolates out of a total of 40 were in the same group. Phylogenetic analysis revealed that the major strain belonged to one species, Hanseniaspora uvarum, although they displayed some nucleotide mismatches between them. During spontaneous alcohol fermentation at 20 °C, the two grape musts containing 24 °Brix sugar exhibited similar fermentation patterns with differences in final alcohol production and yeast viable counts. PCR analysis of the yeasts randomly isolated during the fermentation using an intron splice site primer showed changes in yeast flora between 8 and 10 days of fermentation. We found that the dominant yeasts displaying various PCR patterns using the primer remained the same throughout the early stages of fermentation, as determined by molecular typing of their ITS regions using PCR-RFLP, and these yeasts were identical to those isolated from grape berries. Among the isolates, the strain designated SS6 was selected based on its potassium metabisulfite resistance, alcohol production (distillation method), and flavor (by sniffing test) of grape juice. When Campbell Early grape must was inoculated with H. uvarum SS6 cells, no differences in fermentation pattern were observed compared with that inoculated with cells of Saccharomyces cerevisiae W-3, an industrial wine yeast strain. However, SS6 wine showed higher levels of organic acid (especially lactic acid), aldehydes, and minor alcohols (except n-propyl alcohol), as well as a higher score in sensory evaluation, compared to those of W-3 wine.     

红提葡萄酒发酵酵母菌的分离及耐受性研究

该实验从红提葡萄发酵过程中分离出酵母菌株,并对其耐受性进行研究,旨在提高红提葡萄酒酿造品质。通过培养不同发酵时期菌株,并用肉眼和显微镜分别观察其菌落和细胞形态,再测定其发酵能力、耐糖性、耐酸性、耐酒精度。结果表明,共分离出酵母菌140株,经菌落形状和细胞形态鉴定后,获得典型的三类共6株酵母菌;在发酵过程中,加入分离菌株比自然发酵可多发酵1.6%葡萄糖,多产生0.78% CO2和0.82%vol的酒精,且发酵趋势和安琪酵母、自然发酵过程一致;以安琪酵母为对照,对6株菌进行耐糖性、耐酸性、耐酒精度试验,得出综合耐受性较好的酵母菌株为A3、B7和C1。     

Managing ochratoxin A risk in the grape-wine food chain

The main source of ochratoxin A (OTA) in the wine food chain is the infection of grapes by "black aspergilli" in the field. OTA-producing black aspergilli include principally Aspergillus carbonarius, followed by A. niger and possibly A. tubingensis. They are opportunistic fungi that develop particularly on damaged berries at ripening, although they may occur and form OTA on grapes from veraison to harvest. Climatic conditions (high humidity and temperature) and geographical location are important factors favouring OTA accumulation in grape berries. The severity of aspergillus rot is influenced by excessive irrigation and rainfall prior to harvest, which causes berry splitting. In addition, berry wounds caused by insect attack provide preferential entries for black aspergilli. High OTA levels occur in grapes severely damaged by the grape moth, Lobesia botrana, particularly in Mediterranean areas. Some grape varieties display greater susceptibility to aspergillus rot due to intrinsic genetic characteristics and bunch conformation (i.e. compact>sparse). Control measures for toxigenic mycoflora in the vineyards must consider these critical control points. Proper fungicidal and insecticidal treatments can reduce OTA contamination. Nevertheless, knowledge about the fate of OTA and its distribution in wine and winery by-products is important to manage OTA risk in contaminated stock. In our wine-making experiments, only 4% of the OTA present in grapes remained in the wine--the majority is retained in pressed grape pomaces. OTA concentration remained unchanged in wine after a 1-year aging as well as in all liquid fractions collected during vinification (i.e. must, free run wine, and wine after first and second decantation). Activated carbon can reduce OTA levels in wine but negatively affects wine quality.     

Diversity of Saccharomyces and non-Saccharomyces yeasts in three red grape varieties cultured in the Serranía de Ronda (Spain) vine-growing region

For the first time, an ecological survey of wine yeasts present in grapes growing in two vineyards located in the region of “Serranía de Ronda” (Málaga, southern Spain) has been carried out. During the 2006 and 2007 vintages, grapes from different varieties were aseptically collected and allowed to ferment spontaneously in the laboratory. From a total of 1586 colonies isolated from microvinifications, 1281 were identified according to ITS polymorphisms and their identity confirmed by sequencing of the D1/D2 region of 26S rDNA. Most of the isolates (84%) corresponded to thirteen different non-Saccharomyces species with Kluyveromyces thermotolerans, Hanseniaspora guilliermondii, Hanseniaspora uvarum and Issatchenkia orientalis accounting for 42.7% of the total. Mitochondrial DNA restriction analysis from the Saccharomyces cerevisiae isolates revealed a low diversity since only eleven different profiles were found, nine of them corresponding to local strains and two to commercial ones that had been used in different campaigns and that very likely were disseminated from the winery to the adjacent vineyard. A different distribution of strains was found in the three grape varieties studied.     

Application of molecular methods to demonstrate species and strain evolution of acetic acid bacteria population during wine production

The growth of acetic acid bacteria on grapes or throughout the winemaking process influences the quality of wine, mainly because it increases the volatile acidity. The objective of this study was to analyse how the acetic acid bacteria population evolves in the changing environment of the grape surface and during wine fermentation. We have analysed the influence of yeast inoculation and SO2 addition on acetic acid bacteria populations. These bacteria were analysed at both the species and the strain level by molecular methods such as Restriction Fragment Length Polimorfism (RFLP) of amplified 16S rDNA, and amplification by polymerase chain reaction of Enterobacterial Repetitive Intergenic Consensus (ERIC-PCR) and Repetitive Extragenic Palindromic (REP-PCR). Our results show that the increases in population size are normally accompanied by a proliferation of Acetobacter aceti, which is the main species during fermentation. The diversity of strains is considerable in natural environments such as the grape surface. Changes in the environment during alcoholic fermentation substantially reduce the survival and the diversity of acetic acid bacteria. Few strains are able to survive these conditions and they seem to originate from both the grapes and the winery. To the best of our knowledge this is the first time that acetic acid bacteria are analysed at the strain level in grape surfaces and during winemaking.     

Grape and wine analysis - enhancing the power of spectroscopy with chemometrics.

Spectroscopic techniques offer the potential to simplify and reduce analytical times for a range of grape and wine analytes. It is this aspect, together with the ability to simultaneously measure several analytes, which was the impetus for developing spectroscopic methods. The Australian Wine Research Institute (AWRI) has long used spectroscopic analysis of wines in the ultraviolet (UV) and visible (Vis) wavelengths, and since 1998 has been investigating applications of spectroscopic techniques in the near infrared (NIR) and mid-infrared (MIR) wavelength regions of the electromagnetic spectrum for the rapid analysis and quality control of both grapes and wine by the Australian wine industry. This paper reviews the use of several spectroscopic techniques, including NIR, MIR, and Vis, combined with chemometrics, to assess grape and wine composition in the Australian wine industry. The achievements, current research, and proposed further applications of different spectroscopic techniques are discussed in studies into the assessments of red grape composition and of fungal diseases in grapes, monitoring phenolic compounds during red wine fermentations, quality grading of red, white and fortified wine styles, monitoring wine distillation processes, and yeast strain classification.     

Effect of co-winemaking in phenolic composition, color and antioxidant capacity of young red wines from La Mancha region

The influence of co-winemaking technique on the phenolic profiles, color and antioxidant capacity of wines made from red grape varieties cultivated in La Mancha region (Spain) was investigated. Co-winemaking red wines were obtained by pre-fermentative blend of grapes (1:1, w/w) from the predominant Cencibel (Tempranillo) variety in this region and the minor varieties Bobal, Moravia Agria, Moravia Dulce, Tortosí and Rojal, together with a three varieties blend (1:1:1, w/w/w) of Cencibel, Bobal and Moravia Agria grapes. The phenolic profiles obtained in co-winemaking wines showed more quantity of different compounds than single-variety wine of Cencibel used as reference. The best results were obtained using Bobal, Moravia Agria and Moravia Dulce varieties for co-winemaking, especially an improvement of color characteristics. All co-winemaking wines significantly increased the total resveratrol content as compared to the Cencibel reference wine, although cis-isomers largely predominated in all wines. Moravia Agria and Rojal varieties produced co-winemaking wines with decreased antioxidant capacity, whereas Bobal and Moravia Dulce varieties increased it, even when the co-winemaking wine from Moravia Dulce contained less total polyphenols than Cencibel reference wine. Co-winemaking technique can be suggested as a way to widen the offer of wines to consumers, incorporating characteristic properties of minor grape varieties. In addition, this enological technique has the benefit of avoiding the disappearance of native species, thus improving the biodiversity of La Mancha region.     

Yeast diversity investigation of wine-related samples from two different Slovakian wine-producing areas through a multistep procedure

A multistep procedure was applied in order to investigate the yeasts diversity during the must fermentation of two wine varieties Frankovka Modra (Blaufränkisch) and Veltlinske Zelene (Grüner Veltliner) from two different Slovakian wine-producing areas. The procedure combined a cultivation step with a PCR-based selection step (f-ITS PCR; fluorescence internal transcribed spacer 2 polymerase chain reaction), and a final identification step oriented to the sequencing of ITS1-5.8S rDNA-ITS2 region of the selected yeasts. The strategy demonstrated to be suitable for the rapid analysis of a large panel of isolates. The f-ITS PCR was able to discriminate the isolated yeasts by one PCR run and capillary electrophoresis separation. The investigation showed the presence of non-Saccharomyces strains on grapes and during the initial and middle stages of must fermentation. Members of the genus Saccharomyces appeared only during the middle fermentation stage and they were the predominant species until the final phase of the process. The presented simple and reliable multistep procedure can be useful for the investigation of the yeast microflora from any wine-producing area and also for studying other kinds of food environments.     

烟台贵人香葡萄自然发酵醪中本土酿酒酵母的选育及酿造特性研究

自葡萄原料生产地选育优良酿造特性的本土酵母是开发地方特色葡萄酒品种的重要措施。本研究以烟台地区贵人香葡萄自然发酵醪为材料,筛选酿酒酵母并对其发酵特点和耐受能力进行测定,进而选育本土酿酒酵母用于霞多丽干白和赤霞珠干红葡萄酒的酿造试验。采用孟加拉红选择性培养基筛选本土酵母菌,并经WL鉴定培养基和5.8S ITS序列鉴定获得12株酿酒酵母。通过测定发酵后残糖量和酒精度及耐受能力(酒精度、SO2、酸、高糖),选育4株酵母用于酿造霞多丽干白葡萄酒,对酒样指标、主要挥发物和香气组分分析,显示菌株YGF2、YGF5和YGF10的酿造特性和发酵指标较好,并用于酿造赤霞珠干红葡萄酒。结果表明菌株YGF2酿造的葡萄酒质量较好,香气浓郁,部分指标优于商品酿酒酵母,具备生产区域特色葡萄酒的潜力。     

Ochratoxin A production by Aspergillus carbonarius on some grape varieties grown in Italy

Fungi responsible for the presence of Ochratoxin A (OTA) in grapes have been identified as belonging to the black aspergilli, Aspergillus section Nigri, among which Aspergillus carbonarius is the main producer. Despite the widespread occurrence of OTA in various types of wine, there is no specific information on the ability of black aspergilli to infect berries and to produce OTA in different grape varieties. The aim of this study was to investigate the susceptibility to A carbonarius of twelve grape varieties representative of a wide range of characteristics, including geographical distribution. In all the conditions of berry status (intact and damaged) and temperature (20 and 25 °C) tested, the incidence of colonised berries and their OTA contents were always low in ‘Bianco d'Alessano’, ‘Pampanuto’ and ‘Uva di Troia’. In contrast, ‘Cabernet Sauvignon’, ‘Trebbiano’ and ‘Verdeca’ showed high fungal incidence and OTA content. The information gathered is relevant for defining risk factors for OTA presence in grapes, even if these data need to be confirmed by observations in vineyards. Copyright © 2004 Society of Chemical Industry