Determination of piceid and resveratrol in Spanish wines deriving from Monastrell (Vitis vinifera L.) grape variety

The presence of stilbenes in wine is becoming an important issue due to their claimed relation to a low incidence in coronary diseases and their increasing implication as cancer chemopreventive and neuroprotective agents. Total resveratrol content, quantified as glucoside and aglycone forms of resveratrol, has been determined in a survey of 45 Monastrell monovarietal Spanish red wine types (around 135 wine samples), belonging to Alicante and Bullas appellations. The average between ratio glucoside/aglycone forms of resveratrol in these wines was considerably high, ranging from 82 to 91% of resveratrol in its glycosidic form. This characteristic was observed in a high percentage of the studied wines, which were made under different winemaking procedures, and from different vintages (1995-2002). In addition, wines made using macerative fermentations with double amount of solid parts ("doble pasta") reached the highest levels of total stilbene content expressed as resveratrol equivalent, i.e., 30 mg/L (average of 18.8 mg/L). It can be concluded that high resveratrol glucoside concentration and low free isomer content can be considered characteristics of the Monastrell variety, as it happens to red wines deriving from other varieties grown at warm climates. This fact, also observed for other French and Portuguese red varieties, might play an important role in food habits involving these types of wines.     

硅对黄瓜白粉病抗性的影响及其生理机制

以黄瓜感病品种为材料,研究了施硅和诱导接种白粉菌对抗坏血酸过氧化物酶(AsA-POD)、脱氢抗坏血酸还原酶(DR)、苯丙氨酸解氨酶(PAL)、多酚氧化酶(PPO)活性与酚类物质含量变化的影响。结果表明:诱导接种后,营养液中加硅酸盐显著降低黄瓜白粉病病情指数,显著提高了黄瓜叶片中AsA-POD、PPO的活性,而DR、PAL活性低于不施硅处理,但差异不显著。施硅处理的酚类物质含量较高,并显著降低病情指数。     

Mass spectrometric evidence for the existence of oligomeric anthocyanins in grape skins

The fractionation of a grape skin extract by multilayer countercurrent chromatography coupled with step gradient elution allowed the preparation of a fraction almost devoid of free anthocyanins. This fraction appeared to be almost exclusively polymeric, as judged by liquid chromatographic-mass spectrometric (LC-MS) analysis, color-bleaching tests with sulfur dioxide, and thiolysis. Electrospray mass spectrometric analysis indicated that the pigmented material in this fraction was chiefly composed of direct condensation products of anthocyanin extending up to trimers. With regard to their linkages, the anthocyanin units in the oligomers were possibly linked by either an A-type (by both carbon-carbon and ether bonds) or B-type (by carbon-carbon bond) linkage, like proanthocyanidins. The terminal anthocyanin unit of the oligomers is consistently in the flavylium form but the extension units are in the flavan form for the A-type oligomers and in the flavene form for the B-type oligomers. Although their linkages still need to be defined rigorously, this is the first mass spectrometric evidence confirming the existence of anthocyanin oligomers in the grape skin extract.     

不同施锌量对酿酒葡萄“赤霞珠”锌含量及其品质的影响

以4年生"赤霞珠(Cabernet Sauvignon)"葡萄为研究对象,进行连续2年滴灌[0 kg/hm~2(CK)、24.5 kg/hm~2(T_1)、34.0 kg/hm~2(T_2)、43.5 kg/hm~2(T_3)、53.0 kg/hm~2(T_4)、62.5 kg/hm~2(T_5)]和转色期叶面喷施[0 kg/hm~2(CK)、0.5 kg/hm~2(T_1)、1.0 kg/hm~2(T_2)、1.5 kg/hm~2(T_3)、2.0 kg/hm~2(T_4)、2.5 kg/hm~2(T_5)]两种方式配合施EDTA-Zn的田间试验,探讨强碱性灰钙土(pH8.5)补充锌肥对酿酒葡萄锌含量、生长和品质的影响。结果表明:与不施锌肥(CK)相比,施锌肥处理酿酒葡萄锌含量、粒径、穗长、百粒重和产量分别增加了42.1%～78.7%、2.6%～28.2%、10.1%～22.3%、3.0%～8.8%和17.2%～216.6%。同时,与不施锌(CK)相比,酿酒葡萄的单宁含量降低42.0%～67.9%,花色苷、总酚、可溶性固形物和可溶性糖含量分别提高了0.7%～71.9%、5.0%～80.6%、9.3%～24.0%和10.4%～29.4%,而可滴定酸含量除T_4外没有显著增加。相关分析表明,锌含量与穗长、百粒重、粒径、产量、总酚和可溶性糖均显著相关。综合酿酒葡萄锌含量和品质分析得出T_3处理效果最好。在宁夏贺兰山东麓石灰性土壤条件下,EDTA-Zn施用量45 kg/hm~2左右对于提高酿酒葡萄的锌含量、产量和品质较为适宜。     

Identification of new sources for resistance to powdery mildew in H. spontaneum-derived winter barley lines

Twenty four barley lines derived from the F ₇generation of crosses between two winter barley cultivars and different accessions of Hordeum spontaneum Koch collected in Israel were tested against a set of ten European and five Israeli powdery mildew cultures, possessing virulence genes which completely match the spectrum of known mildew resistance genes. The comparison of reaction patterns justified the conclusion that new genes for resistance have been conferred from H. spontaneum which also differ from genes previously identified in other wild barley accessions from Israel. Participation in the expression of the resistance reaction of one of the two barley cultivars used in the development of the lines is well documented in some cases. The majority of the lines was found to be highly resistant against all or almost all European isolates, while various kinds of reactions were observed against the Israeli cultures. The effective transfer of novel mildew resistance from H. spontaneum into winter barley confirms similar results with spring barley, indicating natural wild barley populations in Israel as a significant gene pool for yet unexploited mildew resistance in barley.     

Cellular and behavioral effects of stilbene resveratrol analogues: implications for reducing the deleterious effects of aging

Research suggests that polyphenolic compounds contained in fruits and vegetables that are rich in color may have potent antioxidant and anti-inflammatory activities. The present studies determined if stilbene (e.g., resveratrol) compounds would be efficacious in reversing the deleterious effects of aging in 19 month old Fischer 344 rats. Experiment I utilized resveratrol and six resveratrol analogues and examined their efficacies in preventing dopamine-induced decrements in calcium clearance following oxotremorine-induced depolarization in COS-7 cells transfected with M1 muscarinic receptors (MAChR) that we have shown previously to be sensitive to oxidative stressors. Experiment II utilized the most efficacious analogue (pterostilbene) from experiment I and fed aged rats a diet with a low (0.004%) or a high (0.016%) concentration of pterostilbene. Results indicated that pterostilbene was effective in reversing cognitive behavioral deficits, as well as dopamine release, and working memory was correlated with pterostilbene levels in the hippocampus.     

Benzothiadiazole induces the accumulation of phenolics and improves resistance to powdery mildew in strawberries

Benzothiadiazole (BTH) enhanced the accumulation of soluble and cell-wall-bound phenolics in strawberry leaves and also improved the resistance to powdery mildew infection under greenhouse conditions. The most pronounced change was seen in the levels of ellagitannins, which increased up to 2- to 6-fold 4 days after the BTH application, but persisted only in the inoculated plants. The induction of phenolic metabolism by BTH was also reflected in the fruits, several compounds being increased in inoculated, BTH-treated plants. Basal salicylic acid (SA) content was high in strawberry leaves, but increased in a similar fashion to other phenolics after the treatments. Several phenolic compounds were identified in strawberries for the first time. For example, ellagic acid deoxyhexose, three agrimoniin-like ellagitannins, sanguiin H-10- and lambertianin C-like ellagitannins in the leaves, ellagic acid, p-coumaric acid, gallic acid, and kaempferol hexose in the cell-wall-bound fraction of the leaves, and kaempferol malonylglucoside in the fruits. The findings show that BTH can enhance the accumulation of phenolics in strawberry plants which may then be involved in the BTH-induced resistance to powdery mildew.     

基于因子分析-BP神经网络的小麦叶片白粉病反演模型

明确白粉病胁迫下小麦叶片受害程度并构建误差小、精度高的反演模型,是实现小麦白粉病遥感监测和精确防控的基础。基于大田小区小麦白粉病人工接种试验,采用高光谱仪测试不同白粉病危害程度下冬小麦叶片光谱反射率,利用常规光谱特征参数、比值指数和归一化指数及因子分析(factor analysis,FA)与BP神经网络(back propagation neural network,BPNN)相结合的方法对小麦叶片白粉病严重度进行模型模拟并对模型拟合精度与适用性比较。结果表明:对小麦白粉病反应敏感的光谱波段为415、485~495、620~640 nm。常规光谱参数中表现较好的光谱植被指数和两波段比值及归一化植被指数的决定系数范围为0.6~0.8,均方根误差范围为8.5~11.5,其中,RI(670,855)、NDVI(680,880)、RGRcn和PSRI对白粉病反演精度及误差控制表现得相对较好。经过FA提取敏感波段的公共因子,进而利用BPNN算法进行模拟,较常规光谱参数有效提高了病情严重度的估算精度,各个测定时期模拟检验决定系数大于0.80,模型的检验均方根误差小于8.09,整个灌浆期反演模型检验的均方根误差和相对误差分别为7.84和7.56%,反演模型对小麦白粉病的整个病症期均具有很好的适用性。由此可得,基于FA-BPNN法所建立的反演模型精度高、误差小,对小麦白粉病病害时期兼容性好,研究结果对植物病害精确防控具有重要意义。     

An LC-MS method for analyzing total resveratrol in grape juice, cranberry juice, and in wine.

Resveratrol is an antioxidant found in grapes, grape products, and some other botanical sources with antiinflammatory and anticancer properties. In grapes and wine, it occurs both as free resveratrol and piceid, the 3beta-glucoside of resveratrol. Here we report a liquid chromatography-mass spectrometry method to analyze total resveratrol (including free resveratrol and resveratrol from piceid) in fruit products and wine. Samples were extracted using methanol, enzymatically hydrolyzed, and analyzed using reversed phase HPLC with positive ion atmospheric pressure chemical ionization (APCI) mass spectrometric detection. Following APCI, the abundance of protonated molecules was recorded using selected ion monitoring (SIM) of m/z 229. An external standard curve was used for quantitation, which showed a linear range of 0.52-2260 pmol of trans-resveratrol injected on-column with a correlation coefficient 0.9999. The coefficient of variance of the response factor over the same concentration range was determined to be 5.8%, and the intra-assay coefficient of variance was determined to be 4.2% (n = 7). The limit of quantitation, defined as signal-to-noise 10:1, was determined to be 0.31 pmol injected on-column. The extraction efficiency of the method was determined to be 92%. The stability of resveratrol under different conditions was also examined. For example, resveratrol was stable for up to 5 days at 4 degrees C in the dark but was not stable at room temperature without protection from light. Resveratrol was detected in grape, cranberry, and wine samples. Concentrations ranged from 1.56 to 1042 nmol/g in Concord grape products, and from 8.63 to 24.84 micromol/L in Italian red wine. The concentrations of resveratrol were silmilar in cranberry and grape juice at 1.07 and 1.56 nmol/g, respectively.     

硅酸钠对裸仁美洲南瓜酚类物质含量的影响与抗白粉病的关系

以裸仁美洲南瓜抗、 感病品种为材料,测定了硅酸钠溶液喷雾处理南瓜幼苗后诱导接种白粉菌对不同抗病品种裸仁美洲南瓜叶片中酚类物质、 绿原酸、 类黄酮3种物质在不同处理时期含量变化的影响。结果表明,硅酸钠诱导处理后不同抗性品种南瓜叶片中的酚类物质、 绿原酸、 类黄酮等物质含量均明显升高,抗病品种中酚类物质含量在第5 d、 7 d、 9 d时分别高于对照和接菌处理的,绿原酸含量在各个处理时间均高于对照和接菌处理的,类黄酮含量在第7 d、 9 d时分别高于对照和接菌处理的; 感病品种中酚类物质含量和绿原酸含量分别在第5 d、 7 d和9 d时高于对照和接菌处理; 类黄酮含量在第3 d、 5 d、 7 d和9 d时均高于其他处理; 硅酸钠诱导处理后,大多数情况下抗病品种的酚类物质（除第7 d外）、 绿原酸（除第7 d、 9 d外）、 类黄酮等含量均高于感病品种,差异显著,且在接种后第10 d和第15 d白粉病病情指数明显下降。     

行间生草对葡萄园土壤肥力和葡萄叶片养分的影响

在赤霞珠葡萄生产园行间播种多年生黑麦草、紫花苜蓿、白三叶草.结果表明与清耕相比,行间生草使土壤全N、全K平均含量升高,但对土壤全P含量影响不大;土壤碱解N、速效P和速效K平均含量均降低,土壤pH值升高.对于土壤中的微量元素,Mn含量升高、Zn含量降低,而且行间生草使叶片N、K、Ca、Mg、Mn、Zn都有升高趋势.豆科植物紫花苜蓿和白三叶草有利于土壤中氮的积累,禾本科多年生黑麦草有利于土壤有机质的提高.     

Phenolic composition of Malbec grape skins and seeds from Valle de Uco (Mendoza, Argentina) during ripening. Effect of cluster thinning

The phenolic composition of Malbec (Vitis vinifera L.) grape skins and seeds during ripening and the effect of cluster thinning (CT) in two consecutive seasons (2008-2009) were evaluated by high-performance liquid chromatography-diode array detection/electrospray ionization-mass spectrometry (HPLC-DAD/ESI-MS). Removal of 50% of clusters was performed at 40 days (T1), 80 days (T2), and 100 days after flowering (T3) in a vineyard located in southern Mendoza (Argentina). Yield components, with the exception of cluster weight, were significantly affected by CT in both seasons, but no statistically significant differences were found among treatments. Cluster thinning and its timing had little or no influence on physical parameters and fruit chemical composition, and the differences with respect to the control were mainly due to the season. At harvest in 2008, T1 encouraged the biosynthesis of individual anthocyanins in skins, generating 44.0, 39.6, and 41.2% more glucosylated, acetylated, and total anthocyanins, respectively, as compared to the control, whereas in seeds, T1 and T2 mainly changed the concentrations of (+)-catechin, epicatechin-3-gallate, procyanidin B4, dimer gallate 1, trimer gallate 2, and tetramer. Conversely in 2009, T1 significantly affected the content of flavanols and flavonols in skins, whereas in seeds, T1 and T2 modified the level of (+)-catechin, procyanidins B4 and B6, and trimer gallate 2. Moreover, in 2008 the grapes had a higher concentration of most phenolic compounds, indicating a greater potential for more complex wines. Finally, dihydroquercetin-3-glucoside was the major compound among all nonanthocyanin phenolics detected in Malbec skins and represented 25.7% (2008) and 39.9% (2009) of the total content of those compounds at harvest. This finding could represent a distinctive feature of this grape variety.     

Evaluation of acibenzolar-S-methyl and other low-toxicity products for control of rose powdery mildew (Podosphaera pannosa) in greenhouses

Abstract

In an attempt to find alternative products to classical fungicides, several products with low toxicity were tested against powdery mildew of roses. These products included resistance inducers (Bion, BABA, and ROS), potassium salts (Resistim, monopotassium phosphate), and seed extracts. The best results were obtained with acibenzolar-S-methyl (Bion). The utilization of Bion as prophylactic treatment, watered at a concentration 0.1–0.2 mg/ml, together with good cultural practices can be enough to effectively control powdery mildew on roses. Treatments with Resistim reduced the disease incidence, but not always significantly compared to the controls. None of the other products had effect on powdery mildew.     

Aging effects and grape variety dependence on the content of sulfur volatiles in wine

Thirteen sulfur compounds (boiling points from 35 to 231 degrees C), usually considered as possible off-flavoring volatiles, were quantified by a concurrent headspace-solid phase microextraction method coupled with gas chromatography-mass spectrometry (HS-SPME/GC-MS) on 80 not off-flavoring wines of four varieties (Merlot, Marzemino, and Teroldego as red wines and Chardonnay as a white one) and of five vintages produced in the North Italian Trentino region. The results of the research, the first Italian data-bank per variety on such volatiles, allow us to make a comparison with the data of other winegrowing areas, to investigate the aging effect on the considered volatiles, and, finally, to try a variety discrimination using statistical procedures. Dimethyl sulfide, 3-(methylthio)-1-propanol, diethyl sulfide, and diethyl disulfide were found to increase with time whereas 2-mercaptoethanol and ethylmercaptan showed a decreasing trend. Furthermore, the concentration of several compounds was found to be dependent on the variety. For instance, sulfide, disulfides, benzothiazole, and thioalcohols are at higher levels in Merlot wines, whereas thiols and thioacetates are more abundant in Marzemino and Teroldego wines. Chardonnay products, well apart from the other wines, are the poorest in 3-(methylthio)-1-propanol and rather rich in dimethyl disulfide and in diethyl disulfide, mostly in the aged wines. Applying the principal component analysis to the data, it was possible to demonstrate that Chardonnay and Merlot wines are well-discriminated from the Italian native varietal wines, which on their turn are only partially distinguishable among them. A contribution of these compounds to the variety characteristics of wine is reasonable.     

Genetic studies on the powdery mildew resistance of winter barley lines derived from Hordeum spontaneum accessions collected in Israel

The genetics of the powdery mildew resistance of 15 Hordeum spontaneum-derived winter barley lines were studied in testcrosses with selected cultivars carrying the MIa12 and MIa13 resistance alleles, in order to identify the inheritance of the resistance and to test for linkage with the MIa locus on chromosome 1H (5). A total of 27 genes were identified. Only one of these genes (in line 1–12 x Dura) could eventually be allelic to a gene previously identified in H. spontaneum Koch. A monofactorial dominant inheritance was identified in the majority of the lines. Three of the tested lines possessed recessively inherited genes and in one line a dominant and a recessive gene were identified. In respect to the reaction of the lines against European mildew cultures, lines with a bifactorially inherited resistance had a broader resistance spectrum than those with a monofactorial inheritance, while lines carrying recessive factors showed a moderately expressed resistance. In three cases a linkage with the MIa locus was found, while in two further lines a possible allelism to this locus was indicated by the absence of recombinant progeny plants. By using more than two isolates for linkage analysis, it was possible to identify additional genes for mildew resistance, reaching a maximum number of four genes in the same line. In some cases, a different inheritance was observed for the resistance of the same line against different isolates. It was assumed that different genes exist, each being effective against a different isolate.     

镁肥对缺镁葡萄叶片活性氧含量和保护酶活性的影响

探讨大田条件下,补镁对缺镁葡萄叶片活性氧含量、保护酶活性和产量的影响,为葡萄科学施肥提供理论依据。采用裂区试验,主区设4个土施硫酸镁梯度,分别为0、112. 5、150、187. 5 kg/hm~2,副区设3个叶面喷施硫酸镁浓度,分别是0%、0. 2%、0. 4%。结果发现:葡萄缺镁时,叶片超氧阴离子产生速率加快,过氧化氢含量增加,超氧化物歧化酶活性升高,但过氧化物酶和过氧化氢酶活性显著降低,叶片脯氨酸和丙二醛含量升高,生物膜受到伤害。通过土施和叶面喷施的方式适量补镁,葡萄叶片活性氧产生速率降低,同时保护酶过氧化物酶和过氧化氢酶活性增加,膜脂过氧化产物丙二醛含量降低,生物膜的受伤程度得到缓解,叶片衰老速率降低,葡萄产量提高,但当施入量过大对生物膜受伤程度的缓解效果降低,葡萄产量降低。土施150 kg/hm~2同时叶面喷施0. 4%硫酸镁肥时,叶片超氧阴离子产生速率低,过氧化氢含量少,膜质过氧化产物丙二醛含量低,葡萄产量高,是该土壤环境下的最佳补镁措施。     

Hop as an interesting source of resveratrol for brewers: optimization of the extraction and quantitative study by liquid chromatography/atmospheric pressure chemical ionization tandem mass spectrometry

Nowadays, hop is used almost exclusively by brewers for bitterness and flavor. Although hop polyphenols have been widely studied in the past decade for their antioxidant activity in the boiling kettle, very little is known about their real impact on health. The discovery of resveratrol in hop pellets highlights the potential health-promoting effect of moderate beer consumption. Here, we have optimized a quantitative extraction procedure for resveratrol in hop pellets. Preliminary removal of hydrophobic bitter compounds with toluene and cyclohexane at room temperature allows 99% trans-resveratrol recovery by ethanol:water (75:25, v/v) solid/liquid extraction at 60 degrees C. Reverse phase liquid chromatography proves an excellent means of separating isomers. In addition, we have compared two mass spectrometry ionization methods-atmospheric pressure chemical ionization (APCI) and electrospray ionization (ESI)-in both the positive and the negative modes. On the basis of standard additions applied with the optimized extraction procedure and reverse phase high-performance liquid chromatography-APCI(+)-tandem mass spectrometry, it appears that Tomahawk hop pellets (T90, harvest 2002) contain 0.5 ppm trans-resveratrol, 2 ppm trans-piceid, no cis-resveratrol, and 0.9 ppm cis-piceid.     

Decomposability of barley straw: Effect of cold-water extraction on dry weight and nutrient content

Mature spring barley (Hordeum vulgare L.) straw, cut to a length of 5 cm, was successively extracted with cold water. The influence of leaching on dry weight, nutrient content, and decomposability was investigated in fresh and in partially-decomposed straw.The loss in dry weight was 6% after one extraction, increasing to 9% after three successive extractions. Incubating the straw between extractions increased losses of dry matter and N, whereas losses of ash, K, P, Mg and Ca were not affected by the intervening incubations. Three successive extractions removed 58% of the ash, 87% of the K, 59% of the P, 34% of the Mg and 25% of the Ca. The loss of N increased from 20% without intermittent incubations to 43% for straw samples incubated at 15°C between the extractions (total incubation period, 66 days).The respiration losses of untreated straw samples incubated for 203 days at 5 and 15°C were 9 and 23%, respectively. Extracting the straw before incubation reduced initial respiration losses. After 203 days, the cumulative respiration loss of leached straw incubated at 15°C was 11% of initial straw dry weight, whereas the loss at 5°C was similar to that of untreated straw.     

Toward metrological traceability for DNA fragment ratios in GM quantification. 1. Effect of DNA extraction methods on the quantitative determination of Bt176 corn by real-time PCR

An international CCQM-P60 pilot study involving eight national metrological institutes was organized to investigate if the quantification of genetically modified (GM) corn powder by real-time PCR was affected by the DNA extraction method applied. Four commonly used extraction methods were compared for the extraction of DNA from a GM Bt176 corn powder. The CTAB-based method yielded the highest DNA template quantity and quality. A difference in the 260 nm/230 nm absorbance ratio was observed among the different extraction methods. Real-time amplification of sequences specific for endogenous genes zein and hmg as well as transgenic sequences within the cryIA(b) gene and a fragment covering the junction between the transformed DNA and the plant genome were used to determine the GM percentage. The detection of the transgenic gene was affected by the quantity and quality of template used for the PCR reaction. The Bt176 percentages measured on diluted or purified templates were statistically different depending on the extraction method applied.