Effects of Latitude and Weather Conditions on Proanthocyanidins in Blackcurrant (Ribes nigrum) of Finnish Commercial Cultivars.

Blackcurrants of three Finnish commercial cultivars 'Mortti', 'Ola', and 'Melalahti' cultivated in southern and northern Finland were compared on the basis of the content and composition of proanthocyanidins (PAs). Seventeen B-type PA oligomers (degree of polymerization 2-5 and 7) were detected by hydrophilic interaction liquid chromatography and electrospray ionization mass spectrometry. Total PAs, dimers, trimers, and tetramers were quantified. Among the three cultivars, 'Ola' had the highest contents of both total PAs and PA oligomers. 'Melalahti' was separated from both 'Mortti' and 'Ola' by PA profiles in the partial least-squares discriminant analysis model. All three cultivars revealed distinct responses to latitude and weather conditions. The content of total PAs showed a positive correlation to latitude in 'Ola' and 'Melalahti'. Among the meteorological variables, high temperature and radiation correlated negatively with total PAs, while only specific variables showed a correlation with PA oligomers.

Application of HPLC-PDA-MS metabolite profiling to investigate the effect of growth temperature and day length on blackcurrant fruit.

INTRODUCTION: Blackcurrant (Ribes nigrum L.) is an excellent example of a "super fruit" with potential health benefits. Both genotype and cultivation environment are known to affect the chemical composition of blackcurrant, especially ascorbic acid and various phenolic compounds. Environmental conditions, like temperature, solar radiation and precipitation can also have significant impact on fruit chemical composition. The relevance of the study is further accentuated by the predicted and ongoing changes in global climate. OBJECTIVES: The aim of the present study was to provide new knowledge and a deeper understanding of the effects of post flowering environmental conditions, namely temperature and day length, on fruit quality and chemical composition of blackcurrant using an untargeted high performance liquid chromatography-photo diode array-mass spectrometry (HPLC-PDA-MS) metabolomics approach. METHODS: A phytotron experiment with cultivation of single-stemmed potted plants of blackcurrant cv. Narve Viking was conducted using constant temperatures of 12, 18 or 24 °C and three different photoperiods (short day, short day with night interruption, and natural summer daylight conditions). Plants were also grown under ambient outdoor conditions. Ripe berries were analysed using an untargeted HPLC-PDA-MS metabolomics approach to detect the presence and concentration of molecules as affected by controlled climatic factors. RESULTS: The untargeted metabolomics dataset contained a total of 7274 deconvolved retention time-m/z pairs across both electrospray ionisation (ESI) positive and negative polarities, from which 549 metabolites were identified or minimally annotated based upon accurate mass MS. Conventional principal component analysis (PCA) in combination with the Friedman significance test were applied to first identify which metabolites responded to temperature in a linear fashion. Multi-block hierarchical PCA in combination with the Friedman significance test was secondly applied to identify metabolites that were responsive to different day length conditions. Temperature had significant effect on a total of 365 metabolites representing a diverse range of chemical classes. It was observed that ripening of the blackcurrant berries under ambient conditions, compared to controlled conditions, resulted in an increased accumulation of 34 annotated metabolites, mainly anthocyanins and flavonoids. 18 metabolites were found to be regulated differentially under the different daylength conditions. Moreover, based upon the most abundant anthocyanins, a comparison between targeted and untargeted analyses, revealed a close convergence of the two analytical methods. Therefore, the study not just illustrates the value of non-targeted metabolomics approaches with respect to the huge diversity and numbers of significantly changed metabolites detected (and which would be missed by conventional targeted analyses), but also shows the validity of the non-targeted approach with respect to its precision compared to targeted analyses. CONCLUSIONS: Blackcurrant maturation under controlled ambient conditions revealed a number of insightful relationships between environment and chemical composition of the fruit. A prominent reduction of the most abundant anthocyanins under the highest temperature treatments indicated that blackcurrant berries in general may accumulate lower total anthocyanins in years with extreme hot summer conditions. HPLC-PDA-MS metabolomics is an excellent method for broad analysis of chemical composition of berries rich in phenolic compounds. Moreover, the experiment in controlled phytotron conditions provided additional knowledge concerning plant interactions with the environment.

Comparison of Phenols Content and Antioxidant Activity of Fruits from Different Maturity Stages of Ribes stenocarpum Maxim.

Differences in the content of nine phenols and the antioxidant capacity of Ribes stenocarpum Maxim (RSM) fruits at different stages of maturity were investigated, and the extraction process of polyphenols from RSM was also optimized using Box-Behnken design method. Results showed that the content of the nine phenols varied considerably at different ripening stages; catechin, chlorogenic acid, coumaric acid, and ferulic acid were abundant in immature fruits but decreased with fruit ripening, whereas the levels of rosemary acid and querctin acid were low in immature fruits and increased with time, reaching the highest value after the fruit was completely mature. The phenols extracted from RSM fruits possessed good antioxidant activities for effective and rapid scavenging of DPPH and ABTS free radicals, as well as intracellular ROS. Analysis of the phenols content at different maturity stages indicated that the unripe fruits had significantly higher polyphenols content than mature fruits. Consequently, unripe fruits possessed higher antioxidant activities. According to the overall results of the extraction process optimization, the selected optimal conditions for extracting polyphenols from RSM were as follows: extraction time, 95 min; solvent concentration, 60%; ratio of sample to solvent, 1:25.

Profiles of Volatile Compounds in Blackcurrant ( Ribes nigrum) Cultivars with a Special Focus on the Influence of Growth Latitude and Weather Conditions.

The volatile profiles of three blackcurrant ( Ribes nigrum L.) cultivars grown in Finland and their responses to growth latitude and weather conditions were studied over an 8 year period by headspace solid-phase microextraction (HS-SPME) followed by gas-chromatographic-mass-spectrometric (GC-MS) analysis. Monoterpene hydrocarbons and oxygenated monoterpenes were the major classes of volatiles. The cultivar 'Melalahti' presented lower contents of volatiles compared with 'Ola' and 'Mortti', which showed very similar compositions. Higher contents of volatiles were found in berries cultivated at the higher latitude (66° 34' N) than in those from the southern location (60° 23' N). Among the meteorological variables, radiation and temperature during the last month before harvest were negatively linked with the volatile content. Storage time had a negative impact on the amount of blackcurrant volatiles.

Metabolic Fingerprinting of Dormant and Active Flower Primordia of Ribes nigrum Using High-Resolution Magic Angle Spinning NMR.

Global warming may modify the timing of dormancy release and spring growth of buds of temperate fruit crops. Environmental regulation of the activity-dormancy cycle in perennial plants remains poorly understood at the metabolic level. Especially, the fine-scale metabolic dynamics in the meristematic zone within buds has received little attention. In this work we performed metabolic profiling of intact floral primordia of Ribes nigrum isolated from buds differing in dormancy status using high-resolution magic angle spinning (HR-MAS) NMR. The technique proved useful in monitoring different groups of metabolites, e.g., carbohydrates and amino acids, in floral primordia and allowed metabolic separation of primordia from endo- and ecodormant buds. In addition, due to its nondestructive character, HR-MAS NMR may provide novel insights into cellular compartmentation of individual biomolecules that cannot be obtained using liquid-state NMR. Out results show that HR-MAS NMR may be an important method for metabolomics of intact plant structures.

Analysis and Sensory Evaluation of Volatile Constituents of Fresh Blackcurrant (Ribes nigrum L.) Fruits.

Volatile constituents of fresh blackcurrant (Ribes nigrum L.) berries were isolated via vacuum-headspace extraction and analyzed by capillary gas chromatography-mass spectrometry. In agreement with previous studies with frozen fruits, short-chain esters and terpenes were major compound classes. However, rather high concentrations of C6-compounds (e.g., (E)-hex-2-enal, (Z)-hex-3-enal) constituted a striking difference to data reported for frozen fruits. Frozen storage of blackcurrant berries was shown to result in drastically reduced concentrations of C6-compounds and a shift of the volatile profile in favor of terpenes. The time-dependent enzymatic formation and isomerization of C6-compounds adds an additional element of variability to the spectrum of fresh blackcurrant volatiles. Nevertheless, blackcurrant cultivars can be classified according to the major classes of the volatiles of the fresh fruits, if prerequisites, such as the same growing location and the same state of ripeness, are met. The sensory contributions of volatiles of blackcurrant berries were assessed by gas chromatography-olfactometry in combination with aroma extract dilution analysis. 4-Methoxy-2-methyl-2-butanethiol, (Z)-3-hexenal, ethyl butanoate, 1,8-cineole, oct-1-en-3-one, and alkyl-substituted 3-methoxypyrazines were among the volatiles showing the highest aroma activity values.

Modeling and optimization of red currants vacuum drying process by response surface methodology (RSM).

Fresh red currants were dried by vacuum drying process under different drying conditions. Box-Behnken experimental design with response surface methodology was used for optimization of drying process in terms of physical (moisture content, water activity, total color change, firmness and rehydratation power) and chemical (total phenols, total flavonoids, monomeric anthocyanins and ascorbic acid content and antioxidant activity) properties of dried samples. Temperature (48-78 °C), pressure (30-330 mbar) and drying time (8-16 h) were investigated as independent variables. Experimental results were fitted to a second-order polynomial model where regression analysis and analysis of variance were used to determine model fitness and optimal drying conditions. The optimal conditions of simultaneously optimized responses were temperature of 70.2 °C, pressure of 39 mbar and drying time of 8 h. It could be concluded that vacuum drying provides samples with good physico-chemical properties, similar to lyophilized sample and better than conventionally dried sample.

Antioxidant capacities and anthocyanin characteristics of the black-red wild berries obtained in Northeast China.

Various edible berries widely accessible in nature in Northeast China are poorly exploited. The compositions and contents of anthocyanins in black (Padus maackii, Padus avium, Lonicera caerulea, and Ribes nigrum) and red (Ribes rubrum, Sambucus williamsii, Rubus idaeus, and Ribes procumbens) wild berries in Northeast China were firstly characterized by HPLC-DAD/ESI-MS(2). Twenty-three anthocyanins were detected and identified. Cyanidin glycosides were dominant in both berries. Six anthocyanins were reported for the first time in P. avium, R. rubrum, and Sambucus. Total anthocyanin content (TAC) ranged from 10mg/100gfreshweight (FW) (R. procumbens) to 1058mg/100gFW (P. maackii) among berries. The TACs and antioxidant activities assessed by DPPH and FRAP assays were much higher in black than in red berries. Black-red berries, especially P. maackii and P. avium, can be used in developing functional foods and in improving breeding programs.

Species identification, host range and diversity of Cecidophyopsis mites (Acari: Trombidiformes) infesting Ribes in Latvia.

Cecidophyopsis mites are important pests in all cultivation regions of Ribes causing bud galls and sterility. Despite their economic importance, the knowledge on Cecidophyopsis species infesting Ribes in various areas of the world is still deficient. The present study was carried out to identify Cecidophyopsis species occurring in Latvia on cultivated and wild Ribes, to assess their host range and gain insight into the genetic diversity of these insufficiently studied pests by use of multiplex PCR, rDNA sequences and morphological characters. Cecidophyopsis alpina, C. aurea, C. spicata and C. selachodon were detected to occur in all surveyed habitats. For the first time, C. alpina was identified on blackcurrants and redcurrants, and C. aurea on redcurrants, blackcurrants and alpine currants. The presence of C. ribis was not confirmed with molecular tools during this study. Phylogenetic analyses confirmed the presence of four Cecidophyopsis species identified by multiplex PCR. A close phylogenetic relatedness was found for C. aurea and C. alpina, and for C. ribis and C. spicata highlighting the necessity for additional studies. Our findings suggest a need to consider also other Cecidophyopsis species besides C. ribis in breeding programs for host resistance to mites.

Influence of Controlled Postflowering Temperature and Daylength on Individual Phenolic Compounds in Four Black Currant Cultivars.

The effects of postflowering temperature and daylength on the concentration of individual phenolic compounds were studied in black currant (Ribes nigrum L.) berries under controlled phytotron conditions. The four cultivars studied varied greatly in their concentrations of individual phenolic compounds and temperature stability for accumulation. The concentrations of a wide range of identified phenolic compounds were strongly influenced by temperature over the 12-24 °C range, often with opposite temperature gradient patterns for compounds within the same subclass. Accumulation of anthocyanins and flavonols increased under natural long day conditions, which provided an increased daily light integral, while under identical light energy conditions, photoperiod had little or no effect on the concentration of phenolic compounds. Furthermore, with the exception of members of the hydroxycinnamic acid subclass, the concentration of most phenolic compounds was higher in berries ripened outdoors than in the phytotron, apparently due to screening of UV-B radiation by the glass cover.

Flavonol Glycosides in Currant Leaves and Variation with Growth Season, Growth Location, and Leaf Position.

Flavonol glycosides (FG) were analyzed in the leaves of six currant cultivars (Ribes spp.) with HPLC-DAD, HPLC-MS/MS, and NMR. The average amounts of the 12 major, identified FG constituted 86-93% (9.6-14.1 mg/g DW) of the total of 27 FG found. Quercetin and kaempferol were the major aglycones with trace amounts of myricetin. Quercetin-3-O-(2,6-α-dirhamnopyranosyl-ß-glucopyranoside), quercetin-3-O-(2-ß-xylopyranosyl-6-α-rhamnopyranosyl-ß-glucopyranoside), and kaempferol-3-O-(3,6-α-dirhamnopyranosyl-ß-glucopyranoside) were identified for the first time in currant leaves and existed in a white currant cultivar 'White Dutch' only. Kaempferol-3-O-ß-(6'-malonyl)glucopyranoside was also a new compound existing in abundance in five cultivars but not in the white one. The results show the primary importance of the genetic background of the cultivars. The content of malonylated FG of special importance in cardiovascular health decreased regularly during summer. Time of collection and leaf position were more prominent factors affecting the composition than were the year of harvest or the growth latitude. Randomly collected leaves differed in their FG profiles from those collected from the middle position of new branches.

Cloning of an anthocyanidin synthase gene homolog from blackcurrant (Ribes nigrum L.) and its expression at different fruit stages.

Anthocyanidin synthase (ANS), a 2-oxoglutarate (2OG) and Fe(II)-dependent oxygenase, catalyzes the penultimate step in anthocyanin biosynthesis, from leucoanthocyanidins to anthocyanidins, the first colored compound in the anthocyanin pathway. In this study, a full-length, 1427-bp long cDNA named RnANS1, which is homologous to the anthocyanidin synthase gene, was cloned from blackcurrant using a homologous cloning strategy. RnANS1 is highly homologous to other plant ANS genes at both the nucleotide and amino acid sequence levels. The deduced protein contains domains conserved in the 2OG and Fe(II)-dependent oxygenase, and is phylogenetically closely related to Paeonia suffruticosa and Paeonia lactiflora. The expression of RnANS1 was upregulated during fruit maturation, and correlated with the accumulation of anthocyanins and soluble carbohydrates in the fruit. Further characterization of the structure and expression patterns of RnANS1 will clarify our understanding of anthocyanin biosynthesis in blackcurrant, and support the development of molecular approaches to manipulate anthocyanin production in this plant.

Beneficial Physiological Effects With Blackcurrant Intake in Endurance Athletes.

Blackcurrant contains anthocyanins, known to influence vasorelaxation and peripheral blood flow. We examined the effects of 7 days intake of Sujon New Zealand blackcurrant powder (6g/day) on the lactate curve, maximum oxygen uptake, and cardiovascular responses at rest and during cycling. Thirteen trained triathletes with >3 yrs experience (8 men, age: 38 ± 8 yrs, body mass: 71 ± 9 kg, BF%: 19 ± 5%, mean ± SD) performed two incremental cycling protocols with recording of physiological and cardiovascular responses (Portapres Model 2). Cardiovascular function was also measured in rest. Experimental design was double-blind, placebo-controlled, randomized and cross-over (wash-out 4 wks). Data were analyzed with two-tailed t tests and 2-way ANOVA and significance accepted at p < .05. Plasma lactate was lower at 40%, 50%, 60% and 70% of maximum power by 27%, 22%, 17% and 13%. Intensity at 4 mmol · La(-1) OBLA was 6% higher with blackcurrant without effect on heart rate and oxygen uptake. Maximum values of oxygen uptake, heart rate and power were not affected by blackcurrant, but obtained with 14% lower lactate. In rest, blackcurrant increased stroke volume and cardiac output by 25% and 26%, and decreased total peripheral resistance by 16%, with no changes in blood pressure and heart rate. Cardiovascular responses during exercise at 40%, 50%, 60%, 70% and 80% intensity were not affected. Sujon New Zealand blackcurrant powder affects lactate production and/or clearance during exercise. Sujon New Zealand blackcurrant powder affects physiological and cardiovascular responses in rest and during exercise that may have implications for exercise performance.

Changes in fruit quality parameters of four Ribes species during ripening.

There are no defined maturity parameters for berry fruit which often results in harvesting and marketing of inedible, immature or over ripe berries. In the present study the changes in quality characteristics of different white-, red-, and black-currant cultivars as well as gooseberry and jostaberry fruit were investigated at three maturity stages. Colour parameters, berry juice pH level and the content of sugars, organic acids and phenolic compounds have been evaluated. A total of 65 different phenolic compounds were detected by HPLC-PDA/MS. The results indicate that, during the last 2-4 weeks of fruit ripening, significant changes occur in colour parameters, primary and secondary metabolites and radical scavenging activity. The results suggest that the average content of specific phenolic groups and sugars/acids ratio can be used as a simple and reliable maturity index for optimisation of harvest time.

Polyphenol extraction optimisation from Ceylon gooseberry (Dovyalis hebecarpa) pulp.

Originally from Asia, Dovyalis hebecarpa is a dark purple/red exotic berry now also produced in Brazil. However, no reports were found in the literature about phenolic extraction or characterisation of this berry. In this study we evaluate the extraction optimisation of anthocyanins and total phenolics in D. hebecarpa berries aiming at the development of a simple and mild analytical technique. Multivariate analysis was used to optimise the extraction variables (ethanol:water:acetone solvent proportions, times, and acid concentrations) at different levels. Acetone/water (20/80 v/v) gave the highest anthocyanin extraction yield, but pure water and different proportions of acetone/water or acetone/ethanol/water (with >50% of water) were also effective. Neither acid concentration nor time had a significant effect on extraction efficiency allowing to fix the recommended parameters at the lowest values tested (0.35% formic acid v/v, and 17.6 min). Under optimised conditions, extraction efficiencies were increased by 31.5% and 11% for anthocyanin and total phenolics, respectively as compared to traditional methods that use more solvent and time. Thus, the optimised methodology increased yields being less hazardous and time consuming than traditional methods. Finally, freeze-dried D. hebecarpa showed high content of target phytochemicals (319 mg/100g and 1,421 mg/100g of total anthocyanin and total phenolic content, respectively).

Flavonol glycosides and other phenolic compounds in buds and leaves of different varieties of black currant (Ribes nigrum L.) and changes during growing season.

Phenolic compounds in buds and leaves of three varieties of black currant in Finland were identified by HPLC-DAD-ESI-MS/MS. Forty-three phenolic compounds of flavonol glycosides, proanthocyanidins and phenolic acids were found in variety "Mikael" whereas only thirty-five in "Mortti" and "Jaloste n:o 15". Glycosides of quercetin and kaempferol were the major phenolics. Rutin, hyperoside, isoquercitrin, kaempferol-3-O-rutinoise, kaempferol-3-O-glucoside, quercetin-3-O-(6″-malonyl)-glucoside and a kaempferol-malonylhexoside were the most abundant flavonol glycosides. The contents of flavonol glycosides ranged from 1 to 7 mg/g fresh weight in leaves showing typically an increasing trend from July to August, reaching the highest values in early October in "Mikael" and the end of August in "Mortti" and "Jaloste n:o 15". This is the first systematic report of the composition and content of phenolic compounds in buds and leaves of black currant.

Berry antioxidants: small fruits providing large benefits.

Small berry fruits are consumed because of their attractive colour and special taste, and are considered one of the richest sources of natural antioxidants. Their consumption has been linked to the prevention of some chronic and degenerative diseases. The term 'berry fruits' encompasses the so-called 'soft fruits', primarily strawberry, currants, gooseberry, blackberry, raspberry, blueberry and cranberry. The objective of this review is to highlight the nutraceutical value of berries and to summarize the factors affecting berry fruit antioxidants. Particular attention is given to postharvest and processing operation factors that may affect fruit phytochemical content. The structure-antioxidant relationships for phenolic compounds - the main group of antioxidants in this fruit group - are presented and major areas for future research are identified.

Phenols and ascorbic acid in black currants (Ribes nigrum L.): variation due to genotype, location, and year.

Black currant berries contain many biochemical compounds with proven or potential human health benefits. We studied the content of total and single polyphenols, ascorbic acid, soluble sugars, and titratable acidity for two advanced selections and three cultivars of black currant at two distant locations in Sweden (south: 56°06'N; north: 65°21'N) over a 3 year period. Regression analyses revealed the effect of genotype to be considerably larger than that of location and year. However, significant effects of location, year, and interactions were also revealed. A principal component analysis nevertheless separated the genotypes. The content of ascorbic acid, total phenols, total anthocyanins, and soluble sugars was highest in berries from the south, whereas the content of phenolic acids and titratable acidity was highest in berries from the north. The results show that selection of cultivars and production sites are important for cultivation of high-quality black currant raw material for health-promoting products.

Analysis and sensory evaluation of gooseberry (Ribes uva crispa L.) volatiles.

Volatiles of gooseberries (Ribes uva cripsa L.) were isolated by means of vacuum-headspace-extraction, and the obtained concentrates were analyzed via capillary gas chromatography-mass spectrometry. To ensure the quantitation of highly volatile compounds, headspace analysis was additionally performed on selected batches. C6-components (e.g., (Z)-hex-3-enal, (E)-hex-2-enal), derived from lipid oxidation, and short-chain esters (e.g., ethyl acetate, methyl butanoate, ethyl butanoate) turned out to be the major compound classes in the fresh fruit. The compositional variability was demonstrated by analyzing several gooseberry varieties at different stages of ripeness. The contributions of volatiles to the gooseberry aroma were assessed by using gas chromatography-olfactometry in combination with aroma extract dilution analysis and calculation of odor activity values. C6-components and esters were shown to be responsible for the green and fruity character of fresh gooseberries.

Effects of latitude and weather conditions on phenolic compounds in currant (Ribes spp.) cultivars.

Effects of growth latitude and weather conditions on phenolic compounds of currants (Ribes spp.) were investigated. The berries of red currant cultivar 'Red Dutch', white currant 'White Dutch', and green currant 'Vertti' were collected in seven consecutive years from two growth sites (south and north) with a latitudinal distance of 690 km. The contents of hydroxycinnamic acid conjugates and flavonol glycosides in 'Vertti' were higher than those in 'White Dutch' by 8 and 5 times, respectively, and by 50 and 3 times than those in 'Red Dutch', respectively. The total content of phenolic compounds was 10-19% higher in the north than in the south (p < 0.05). In 'Red Dutch', anthocyanins were 12% richer in berries from the north compared with those from the south (p < 0.05). The total content of hydroxycinnamic acid conjugates in 'Vertti' and 'White Dutch' from the north was 30% higher than those from the south (p < 0.05). High radiation and temperature were associated with low contents of the major phenolic compounds in all the cultivars studied. High humidity correlated with low levels of hydroxycinnamic acid conjugates in green and white currants.