Bilberry metabolomic and proteomic profiling during fruit ripening reveals key dynamics affecting fruit quality.

Bilberry (Vaccinium myrtillus L.) is a wild berry species that is prevalent in northern Europe. It is renowned and well-documented for its nutritional and bioactive properties, especially due to its anthocyanin content. However, an overview of biological systems governing changes in other crucial quality traits, such as size, firmness, and flavours, has received less attention. In the present study, we investigated detailed metabolomic and proteomic profiles at four different ripening stages of bilberry to provide a comprehensive understanding of overall quality during fruit ripening. By integrating omics datasets, we revealed a novel global regulatory network of plant hormones and physiological processes occurring during bilberry ripening. Key physiological processes, such as energy and primary metabolism, strongly correlate with elevated levels of gibberellic acids, jasmonic acid, and salicylic acid in unripe fruits. In contrast, as the fruit ripened, processes including flavour formation, cell wall modification, seed storage, and secondary metabolism became more prominent, and these were associated with increased abscisic acid levels. An indication of the increase in ethylene biosynthesis was detected during bilberry development, raising questions about the classification of non-climacteric and climacteric fruits. Our findings extend the current knowledge on the physiological and biochemical processes occurring during fruit ripening, which can serve as a baseline for studies on both wild and commercially grown berry species. Furthermore, our data may facilitate the optimization of storage conditions and breeding programs, as well as the future exploration of beneficial compounds in berries for new applications in food, cosmetics, and medicines.

Structure of Ground Vegetation and Natural Regeneration of Tree Species in 12- to 15-Year-Old Bilberry Pine Forest-Clear-cut Complex of Middle Taiga Subzone.

Logging in mature stands where part of the forest is harvested in one or several cuts and part is retained (clearcutting and alternate strip cutting) results in the formation of an ecotone complex (EC), which includes a forest (F) zone, a forest edge (FE) as a transition from the forest to the clear-cut under the canopy, a clear-cut edge (CE) as a transition from the forest to the clear-cut outside of the canopy, and the clear-cut proper (C). The composition and structure of ground vegetation and natural regeneration of woody species (Pinus sylvestris L., Picea abies (L.) H. Karst., Betula sp., Populus tremula L., Sorbus aucuparia L., and Juniperus communis L.) were studied in a bilberry pine forest-clear-cut ecotone complex 12-15 years after stand removal. Specific structural features of ground vegetation and undergrowth (including tree regeneration) were observed in each of the four zones of the ecotone complex formed after logging of the mature forest. A typical forest habitat (zone F) showed a minimum number of young regeneration of Pinus sylvestris, Picea abies, Betula sp., Populus tremula, and Sorbus aucuparia and the highest abundance of the lingonberry V. vitis-idaea L. and bilberry Vaccinium myrtillus L. with a maximum height and a maximum yield of bilberry plants. The amount of tree regeneration in the FE zone was much the same as in the F zone. The projective cover, maximum shoot height, and yield of bilberry and the maximum shoot height of lingonberry in the FE zone were significantly lower than in the F zone. The transitional zone on the clear-cut side (CE) and the clear-cut proper (C) strikingly differed from the forest (F and FE) zones of the ecotone complex by a greater number of deciduous and pine regeneration and a low abundance of dwarf shrubs. The clear-cut proper (C) differed from the CE zone by a higher abundance of grasses and forbs and an established tree regeneration layer composed of pine, birch, and aspen.

Carotenoid metabolism during bilberry (Vaccinium myrtillus L.) fruit development under different light conditions is regulated by biosynthesis and degradation.

BACKGROUND: Carotenoids are important pigments and precursors for central signaling molecules associated in fruit development and ripening. Carotenoid metabolism has been studied especially in the climacteric tomato fruit but the content of carotenoids and the regulation of their metabolism have been shown to be highly variable between fruit species. Non-climacteric berries of the genus Vaccinium are among the best natural sources of health-beneficial flavonoids but not studied previously for carotenoid biosynthesis. RESULTS: In this study, carotenoid biosynthetic genes, PSY, PDS, ZDS, CRTISO, LCYB, LCYE, BCH and CYP450-BCH, as well as a carotenoid cleavage dioxygenase CCD1 were identified from bilberry (V. myrtillus L.) fruit and their expression was studied along with carotenoid composition during fruit development under different photoperiod and light quality conditions. Bilberry was found to be a good source of carotenoids among fruits and berries. The most abundant carotenoids throughout the berry development were lutein and ß-carotene, which were accompanied by lower amounts of 9Z-ß-carotene, violaxanthin, neoxanthin, zeaxanthin, antheraxanthin and ß-cryptoxanthin. The expression patterns of the biosynthetic genes in ripening fruits indicated a metabolic flux towards ß-branch of the carotenoid pathway. However, the carotenoid levels decreased in both the ß-branch and ε,ß-branch towards bilberry fruit ripening along with increased VmCCD1 expression, similarly to VmNCED1, indicating enzymatic carotenoid cleavage and degradation. Intense white light conditions increased the expression of the carotenoid biosynthetic genes but also the expression of the cleavage genes VmCCD1 and VmNCED1, especially in unripe fruits. Instead, mature bilberry fruits responded specifically to red/far-red light wavelengths by inducing the expression of both the carotenoid biosynthetic and the cleavage genes indicating tissue and developmental stage specific regulation of apocarotenoid formation by light quality. CONCLUSIONS: This is the first report of carotenoid biosynthesis in Vaccinium berries. Our results indicate that both transcriptional regulation of the key biosynthetic genes and the enzymatic degradation of the produced carotenoids to apocarotenoids have significant roles in the determination of the carotenoid content and have overall effect on the metabolism during the bilberry fruit ripening.

[Analyzing the fine-scale dynamics of two dominant species in a Polytrichum­Myrtillus pine forest. I. A homogeneous Markov chain and cyclicity indices].

Using long-term direct observations in a Polytrichum-Myrtillus pine forest, we have constructed and verified a homogeneous Markov chain model for two dominant species (Vaccinium myrtillus and V. vitis-idaed) at the late stages of succession. The sampling design features a large sample size (2000 quadrats) on permanent transects, several re-examinations with the interval of 5 years, and the use of species rooted frequency. As a model of the process under concern, the discrete Markov chain accounts for the following four states: both species being absent on the quadrat, one of them being present alone, and the joint presence of the both; the model time step coincides with the time interval between observations. The model is calibrated on the data of two successive examinations and verified on that of one more examination. All possible transitions between the states are revealed to realize in quadrats for one time interval, as well as the absence of transitions at each state, which results in the complete digraph (directed graph) of transitions. Major model results are obtained by the formulae of finite Markov chain theory: the steady-state square distribution, cyclicity characteristics, and the mean durations of stages in the fine-scale dynamics. As a steady-state (stable) outcome of succession, the distribution among quadrats is expected where 30% of quadrats are occupied by V. myrtillus alone, 11% by V. vitis-idaea alone, both species are present on 18% of quadrats, and 41% of quadrats are 'empty'. This demonstrates a possibility for V. myrtillus and V. vitis-idaea to coexist stably at the latest stages of succession, with the clear predominance of V. myrtillus, yet without competitive exclusion. The quantitative characteristics of cyclicity and the durations of stages in the fine-scale dynamics enable us to estimate the total duration of secondary post-fire succession as about 45 years (to reach a distribution of states that differs less than 5% from the steady-state one). Out of the four states specified, the quadrats with V. vitis-idaea alone persist for the least time (8 years) on the average, while 'empty' ones persist for the greatest time (18 years). Forecasting the dynamics for one model time step forward and comparing the forecast with the real square distribution have revealed the measure of difference to be 5.4%. This illustrates the efficiency of the (time-)homogeneous Markov chain as a short-term forecast tool, yet leaves open the question whether the homogeneity hypothesis be true in the longer term.

Dwarf shrub hydraulics: two Vaccinium species (Vaccinium myrtillus, Vaccinium vitis-idaea) of the European Alps compared.

Vaccinium myrtillus and Vaccinium vitis-idaea are two dwarf shrubs widespread in the European Alps. We studied the hydraulics of these species hypothesizing that (1) the hydraulic architecture of dwarf shrubs differs from trees, (2) hydraulic properties reflect the species' ecological amplitude and (3) hydraulic properties vary spatially and seasonally. Key hydraulic parameters (osmotic potential, turgor loss point, xylem hydraulic conductivity, vulnerability to drought-induced embolism, stomata closure, drought-induced cell damage and embolism repair) and related wood anatomical traits (conduit diameter and conduit wall reinforcement) were analyzed at four sites in Tyrol, Austria. Both species exhibited low hydraulic safety as well as low hydraulic efficiency. Fifty percentage embolism accumulated at -2.08 (V. myrtillus) and -1.97 MPa (V. vitis-idaea), 88% stomata closure was at -2.19 and -2.35 MPa, respectively. After drought, both species showed embolism repair on re-watering. Site-specific variation within species was low, while seasonal changes in embolism resistance and turgor loss point were observed. Results indicate that studied Vaccinium species have a high risk for embolism formation. This is balanced by refilling capacities, which are probably based on the small growth height of dwarf shrubs. V. vitis-idaea, which occurs on drier sites, showed more efficient repair and a lower turgor loss point than V. myrtillus.

A comparison of fruit quality parameters of wild bilberry (Vaccinium myrtillus L.) growing at different locations.

BACKGROUND: As a part of the ongoing interest in nutritional and nutraceutical properties of locally produced fruits, the aim of the article was to determine sugars, organic acids, polyphenols and antioxidant capacity of wild bilberries grown in Slovenia. Primary and secondary metabolite composition of bilberry fruit was compared among six phyto-climatic conditions, differing in photosynthetic active radiation and light intensity. RESULTS: Three sugars were quantified in bilberry fruit, the predominant being fructose and glucose, along with five organic acids. Bilberry fruit contained high levels of anthocyanins (cyanidin, delphinidin, malvidin, petunidin and peonidin glycosides), hydroxycinnamic acid derivatives and low levels of flavonol glycosides (mainly quercetin and myricetin glycosides), flavan-3-ols, proanthocyanidins and iridoids. The results of the study indicate that bilberries growing in sites with high photosynthetic active radiation (PAR) contained higher levels of total sugars, anthocyanins, flavonols and hydroxycinnamic acids and lower levels of organic acids compared with bilberry fruit from low-light locations. Consequently, total phenolic content and antioxidative capacity of the fruit from high-PAR locations were significantly increased. CONCLUSION: Bilberries from different natural habitats differ significantly in the quantity of sugars, organic acids and phenolic compounds. Our results contribute to a fuller understanding of the relationships between environmental factors and accumulation of primary and secondary metabolites in bilberry fruits.

Monochromatic light increases anthocyanin content during fruit development in bilberry.

BACKGROUND: Light is one of the most significant environmental factors affecting to the accumulation of flavonoids in fruits. The composition of the light spectrum has been shown to affect the production of phenolic compounds during fruit ripening. However, specific information on the biosynthesis of flavonoids in fruits in response to different wavelengths of light is still scarce. In the present study bilberry (Vaccinium myrtillus L.) fruits, which are known to be rich with anthocyanin compounds, were illuminated with blue, red, far-red or white light during the berry ripening process. Following the illumination, the composition of anthocyanins and other phenolic compounds was analysed at the mature ripening stage of fruits. RESULTS: All the three monochromatic light treatments had significant positive effect on the accumulation of total anthocyanins in ripe fruits compared to treatment with white light or plants kept in darkness. The elevated levels of anthocyanins were mainly due to a significant increase in the accumulation of delphinidin glycosides. A total of 33 anthocyanin compounds were detected in ripe bilberry fruits, of which six are novel in bilberry (cyanidin acetyl-3-O-galactose, malvidin acetyl-3-O-galactose, malvidin coumaroyl-3-O-galactose, malvidin coumaroyl-3-O-glucose, delphinidin coumaroyl-3-O-galactose, delphinidin coumaroyl-3-O-glucose). CONCLUSIONS: Our results indicate that the spectral composition of light during berry development has significant effect on the flavonoid composition of ripe bilberry fruits.

Anthropogenic nitrogen deposition in boreal forests has a minor impact on the global carbon cycle.

It is proposed that increases in anthropogenic reactive nitrogen (Nr ) deposition may cause temperate and boreal forests to sequester a globally significant quantity of carbon (C); however, long-term data from boreal forests describing how C sequestration responds to realistic levels of chronic Nr deposition are scarce. Using a long-term (14-year) stand-scale (0.1 ha) N addition experiment (three levels: 0, 12.5, and 50 kg N ha(-1)  yr(-1) ) in the boreal zone of northern Sweden, we evaluated how chronic N additions altered N uptake and biomass of understory communities, and whether changes in understory communities explained N uptake and C sequestration by trees. We hypothesized that understory communities (i.e. mosses and shrubs) serve as important sinks for low-level N additions, with the strength of these sinks weakening as chronic N addition rates increase, due to shifts in species composition. We further hypothesized that trees would exhibit nonlinear increases in N acquisition, and subsequent C sequestration as N addition rates increased, due to a weakening understory N sink. Our data showed that understory biomass was reduced by 50% in response to the high N addition treatment, mainly due to reduced moss biomass. A (15) N labeling experiment showed that feather mosses acquired the largest fraction of applied label, with this fraction decreasing as the chronic N addition level increased. Contrary to our hypothesis, the proportion of label taken up by trees was equal (ca. 8%) across all three N addition treatments. The relationship between N addition and C sequestration in all vegetation pools combined was linear, and had a slope of 16 kg C kg(-1)  N. While canopy retention of Nr deposition may cause C sequestration rates to be slightly different than this estimate, our data suggest that a minor quantity of annual anthropogenic CO2 emissions are sequestered into boreal forests as a result of Nr deposition.

Impacts of winter icing events on the growth, phenology and physiology of sub-arctic dwarf shrubs.

The Arctic is experiencing the greatest climate change in winter, including increases in freeze-thaw cycles that can result in ice encasement of vegetation. Ice encasement can expose plants to hypoxia and greater temperature extremes, but currently the impacts of icing on plants in the field remain little understood. With this in mind, a unique field manipulation experiment was established in heathland in northern Sweden with ice encasement simulated in early March 2008, 2009 and 2010 until natural thaw each spring. In the following summers we assessed the impacts on flowering, bud phenology, shoot growth and mortality and leaf damage (measured by chlorophyll fluorescence and electrolyte leakage) of the three dominant dwarf shrub species Empetrum nigrum, Vaccinium vitis-idaea (both evergreen) and Vaccinium myrtillus (deciduous). Two consecutive winters of icing decreased V. vitis-idaea flowering by 57%, while flowering of V. myrtillus and E. nigrum remained unaffected. Vaccinium myrtillus showed earlier budburst but shoot growth for all species was unchanged. Shoot mortality of V. myrtillus and V. vitis-idaea increased after the first year (by 70 and 165%, respectively) and again for V. myrtillus following the third year (by 67%), while E. nigrum shoot mortality remained unaffected, as were chlorophyll fluorescence and electrolyte leakage in all species. Overall, the sub-arctic heathland was relatively tolerant to icing, but the considerable shoot mortality of V. myrtillus contrasting with the general tolerance of E. nigrum suggests plant community structure in the longer term could change if winters continue to see a greater frequency of icing events.

Vulnerability of black grouse hens to goshawk predation: result of food supply or predation facilitation?

The plant cycle hypothesis says that poor-quality food affects both herbivorous voles (Microtinae spp.) and grouse (Tetraonidae spp.) in vole decline years, leading to increased foraging effort in female grouse and thus a higher risk of predation by the goshawk Accipiter gentilis. Poor-quality food (mainly the bilberry Vaccinium myrtillus) for these herbivores is induced by seed masting failure in the previous year, when the bilberry is able to allocate resources for chemical defence (the mast depression hypothesis; MDH). The predation facilitation hypothesis (PFH) in turn states that increased searching activity of vole-eating predators during or after the decline year of voles disturbs incubating and brooding grouse females. The behaviours used by grouse to avoid these terrestrial predators make them more vulnerable to predation by goshawks. We tested the main predictions of the MDH and PFH by collecting long-term (21-year) data from black grouse Tetrao tetrix hens and cocks killed by breeding goshawks supplemented with indices of bilberry crop, vole abundance and small carnivores in the vicinity of Oulu, northern Finland. We did not find obvious support for the prediction of the MDH that there is a negative correlation of bilberry crop in year t with vole abundance and with predation index of black grouse hens in year t + 1. We did find obvious support for the prediction of the PFH that there is a positive correlation between predator abundance and predation index of grouse hens, because the stoat Mustela erminea abundance index was positively related to the predation index of black grouse hens. We suggest that changes in vulnerability of grouse hens may mainly be caused by the guild of vole-eating predators, who shift to alternative prey in the decline phase of the vole cycle, and thus chase grouse hens and chicks to the talons of goshawks and other avian predators.

Plant growth shapes the effects of elevation on the content and variability of flavonoids in subalpine bilberry stands.

The study of morphological and physiological responses of shrubs to climate is crucial for the understanding of future scenarios regarding climate change. In this light, studying shrub growth and physiological acclimation along an elevation gradient might be insightful. The phenolic metabolic pathway represents a powerful tool to interpret such processes. In the South-Eastern Alps, we investigated the relationships between elevation, plant traits (i.e. age, xylem ring width, annual shoot length), plant-plant interaction (i.e. shrub cover) and flavonoids in Vaccinium myrtillus L. (leaves, berries) in stands above the treeline. The relationships were parsed within causal networks using a confirmatory path analysis. Elevation was the main driver of V. myrtillus growth, having both direct and indirect effects on the leaf flavonoid content, but this was less evident for berries. In particular, the content of foliar flavonoids showed a peak at mid-elevation and where the growth of xylem rings was intermediate, while it decreased in stands with higher shoot length. Flavonoid content variability of both leaves and berries was affected by elevation and shoot length. In berries, flavonoid variability was further related to all growth traits and shrub cover. These findings evidence that flavonoid content is influenced by both elevation and growth traits of V. myrtillus, often showing non-linear relationships. These results suggest a trait-mediated response of this plant to climate conditions as a result of trade-offs between plant growth, plant defence, environmental stress and nutrient/resource availability.

Investigating the interaction between ungulate grazing and resource effects on Vaccinium myrtillus populations with integral projection models.

Dense ungulate populations in forest accompanied by high grazing intensities have the potential to affect plant population dynamics, and such herbivory effects on populations are hypothesised to differ along environmental gradients. We investigated red deer grazing and resource interaction effects on the performance and dynamics of the functionally important boreal shrub Vaccinium myrtillus using integral projection models (IPMs). We sampled data from 900 V. myrtillus ramets in 30 plots in two consecutive years across the boreo-nemoral pine forest on the island Svanøy, western Norway. The plots spanned two environmental gradients: a red deer grazing intensity gradient (assessed by Cervus elaphus faecal pellets), and a relative resource gradient (DCA-ordination of species composition). The use of IPMs enabled projections of population growth rate (lambda) using continuous plant size instead of forcing stage division upon the demographic data. We used the environmental gradients as continuous variables to explain the dynamics of V. myrtillus populations and found that both increasing grazing intensity and resource levels negatively affected lambda of the V. myrtillus populations. Interestingly, these factors interacted: the negative effects of grazing were strongest in the resource-rich vegetation, and higher resource levels reduced lambda more strongly than at low resource levels when grazing intensities became higher. Populations with lambda > 1 were projected if the grazing intensity was less than or equal to the mean grazing intensity on the island, and indicated that V. myrtillus is relatively tolerant of grazing. Variance decomposing showed that the decrease of lambda along the grazing gradient, both at low and high resource levels, was largely caused by reductions in plant growth. The use of IPMs together with important environmental gradients offered novel possibilities to study the synthesised effect of different factors on plant population dynamics. Here, we show that the population response of an abundant boreal shrub to ungulate grazing depends on resource level.

Phenolic composition and antioxidant capacity of bilberry (Vaccinium myrtillus) leaves in Northern Europe following foliar development and along environmental gradients.

Bilberry is a characteristic field layer species in the boreal forests and is an important forage plant for herbivores of the North European ecosystem. Bilberry leaves contain high levels of phenolic compounds, especially hydroxycinnamic acids, flavonols, catechins, and proanthocyanidins. We investigated the phenolic composition of bilberry leaves in two studies, one following foliar development in forest and open areas, and the other along a wide geographical gradient from south to north boreal forests in Finland. An analysis of bilberry leaves collected in open and forest areas showed that major phenolic changes appeared in the first stages of leaf development, but, most importantly, synthesis and accumulation of flavonoids was delayed in the forest compared to the high light sites. Sampling along a geographical gradient in the boreal zone indicated that leaves from higher latitudes and higher altitudes had greater soluble phenolic and flavonol levels, higher antioxidant capacity, and lower contents of chlorogenic acid derivatives. The ecological significance of the results is discussed.

Influence of light availability and soil productivity on insect herbivory on bilberry (Vaccinium myrtillus L.) leaves following mammalian herbivory.

Vegetative parts of bilberry (Vaccinium myrtillus) are important forage for many boreal forest mammal, bird and insect species. Plant palatability to insects is affected by concentration of nutrients and defense compounds in plants. We expected that palatability of bilberry leaves to insect herbivores is influenced by light availability and soil productivity (both affecting nitrogen concentration and constitutive carbon-based defense compound concentration) and herbivory by mammals (affecting nitrogen concentration and induced carbon-based defense compound concentration). We studied bilberry leaf herbivory under different light availability, soil productivity and mammalian herbivory pressure in small sampling units (1m x 1m) in boreal forest in Norway. We used generalized linear mixed models and generalized additive mixed models to model insect herbivory on bilberry leaves as a function of shade, soil productivity and mammalian herbivory. Observed insect herbivory on bilberry leaves increased with increasing shade levels. Predicted insect herbivory increased with increasing previous mammalian herbivory at high shade levels and this response was magnified at higher soil productivity levels. At low to intermediate shade levels, this response was only present under high soil productivity levels. Our results indicate that light availability is more important for variation in bilberry leaf palatability than soil nutrient conditions.

Effects of a warmer climate on seed germination in the subarctic.

BACKGROUND AND AIMS: In a future warmer subarctic climate, the soil temperatures experienced by dispersed seeds are likely to increase during summer but may decrease during winter due to expected changes in snow depth, duration and quality. Because little is known about the dormancy-breaking and germination requirements of subarctic species, how warming may influence the timing and level of germination in these species was examined. METHODS: Under controlled conditions, how colder winter and warmer summer soil temperatures influenced germination was tested in 23 subarctic species. The cold stratification and warm incubation temperatures were derived from real soil temperature measurements in subarctic tundra and the temperatures were gradually changed over time to simulate different months of the year. KEY RESULTS: Moderate summer warming (+2.5 degrees C) substantially accelerated germination in all but four species but did not affect germination percentages. Optimum germination temperatures (20/10 degrees C) further decreased germination time and increased germination percentages in three species. Colder winter soil temperatures delayed the germination in ten species and decreased the germination percentage in four species, whereas the opposite was found in Silene acaulis. In most species, the combined effect of a reduced snow cover and summer warming resulted in earlier germination and thus a longer first growing season, which improves the chance of seedling survival. In particular the recruitment of (dwarf) shrubs (Vaccinium myrtillus, V. vitis-idaea, Betula nana), trees (Alnus incana, Betula pubescens) and grasses (Calamagrostis lapponica, C. purpurea) is likely to benefit from a warmer subarctic climate. CONCLUSIONS: Seedling establishment is expected to improve in a future warmer subarctic climate, mainly by considerably earlier germination. The magnitudes of the responses are species-specific, which should be taken into account when modelling population growth and migration of subarctic species.

Shrub growth and plant diversity along an elevation gradient: Evidence of indirect effects of climate on alpine ecosystems.

Enhanced shrub growth and expansion are widespread responses to climate warming in many arctic and alpine ecosystems. Warmer temperatures and shrub expansion could cause major changes in plant community structure, affecting both species composition and diversity. To improve our understanding of the ongoing changes in plant communities in alpine tundra, we studied interrelations among climate, shrub growth, shrub cover and plant diversity, using an elevation gradient as a proxy for climate conditions. Specifically, we analyzed growth of bilberry (Vaccinium myrtillus L.) and its associated plant communities along an elevation gradient of ca. 600 vertical meters in the eastern European Alps. We assessed the ramet age, ring width and shoot length of V. myrtillus, and the shrub cover and plant diversity of the community. At higher elevation, ramets of V. myrtillus were younger, with shorter shoots and narrower growth rings. Shoot length was positively related to shrub cover, but shrub cover did not show a direct relationship with elevation. A greater shrub cover had a negative effect on species richness, also affecting species composition (beta-diversity), but these variables were not influenced by elevation. Our findings suggest that changes in plant diversity are driven directly by shrub cover and only indirectly by climate, here represented by changes in elevation.

Polyphenolic profiles and antioxidant and antiradical activity of Italian berries from Vaccinium myrtillus L. and Vaccinium uliginosum L. subsp. gaultherioides (Bigelow) S.B. Young.

Total soluble polyphenols (TSP), total monomeric anthocyanins (TMA), radical scavenging activity (RSA), ferric reducing antioxidant power (FRAP), and a number of anthocyanins, phenolic acids, coumarins, flavanols, dihydrochalcones and flavonols were investigated in Tuscan bilberry (i.e. Vaccinium myrtillus) and "false bilberry" (i.e. Vaccinium uliginosum subsp. gaultherioides Bigelow). V. myrtillus berries showed much higher TSP, TMA, RSA and FRAP values than V. uliginosum subsp. gaultherioides fruits. Moreover, very different profiles of individual phenolics were observed in the two species, being V. myrtillus mainly characterised by delphinidin and cyanidin glycosides, together with chlorogenic acid, and V. uliginosum subsp. gaultherioides dominated by malvidin derivatives and flavonols. Strong differences between the two species regarded also metabolites investigated herein for the first time, such as scopoletin, which was approximately two magnitude orders higher in V. uliginosum subsp. gaultherioides than in V. myrtillus berries. Very different abundances were also highlighted for cryptochlorogenic acid and quercetin-3-rhamnoside that were about ten-fold higher in bilberry than in "false bilberry". When the anthocyanin composition pattern of Tuscan "false bilberry" was compared to those elsewhere reported for V. uliginosum fruits harvested in different world areas, some important differences were observed.

Modification of Sunlight Radiation through Colored Photo-Selective Nets Affects Anthocyanin Profile in Vaccinium spp. Berries.

OBJECTIVES: In recent years, the interest on the effects of the specific wavelengths of the light spectrum on growth and metabolism of plants has been increasing markedly. The present study covers the effect of modified sunlight conditions on the accumulation of anthocyanin pigments in two Vaccinium species: the European wild bilberry (V. myrtillus L.) and the cultivated highbush blueberry (V. corymbosum L.). METHODS: The two Vaccinium species were grown in the same test field in the Alps of Trentino (Northern Italy) under modified light environment. The modification of sunlight radiation was carried out in field, through the use of colored photo-selective nets throughout the berry ripening during two consecutive growing seasons. The anthocyanin profile was then assessed in berries at ripeness. RESULTS: The results indicated that the light responses of the two Vaccinium species studied were different. Although both studied species are shade-adapted plants, 90% shading of sunlight radiation was beneficial only for bilberry plants, which accumulated the highest content of anthocyanins in both seasons. The same condition, instead, was not favorable for blueberries, whose maturation was delayed for at least two weeks, and anthocyanin accumulation was significantly decreased compared to berries grown under sunlight conditions. Moreover, the growing season had strong influence on the anthocyanin accumulation in both species, in relation to temperature flow and sunlight spectra composition during the berry ripening period. CONCLUSIONS: Our results suggest that the use of colored photo-selective nets may be a complementary agricultural practice for cultivation of Vaccinium species. However, further studies are needed to analyze the effect of the light spectra modifications to other nutritional properties, and to elucidate the molecular mechanisms behind the detected differences between the two relative Vaccinium species.

Anthocyanin Profile in Berries of Wild and Cultivated Vaccinium spp. along Altitudinal Gradients in the Alps.

Vaccinium spp. berries provide some of the best natural sources of anthocyanins. In the wild bilberry (Vaccinium myrtillus L.), a clear increasing trend in anthocyanin biosynthesis has been reported toward northern latitudes of Europe, but studies related to altitude have given contradictory results. The present study focused on the anthocyanin composition in wild bilberries and highbush blueberry (Vaccinium corymbosum L. cv. Brigitta Blue) growing along altitudinal gradients in the Alps of northern Italy. Our results indicate an increasing accumulation of anthocyanins in bilberries along an altitudinal gradient of about 650 m. The accumulation was due to a significant increase in delphinidin and malvidin glycosides, whereas the accumulation of cyanidin and peonidin glycosides was not affected by altitude. Seasonal differences, especially temperature, had a major influence on the accumulation of anthocyanins in blueberries.

Anthocyanins and glutathione S-transferase activities in response to low temperature and frost hardening in Vaccinium myrtillus (L.).

Anthocyanin (Acy) contents and GST activities of bilberry (Vaccinium myrtillus L.) were investigated in two experiments conducted in June (Exp. I: active growth) and in August September (Exp. II: beginning of frost hardening) in Northern Finland (65 degrees N). Bilberry plants were subjected to +2 degrees C and +18 degrees C in Exp. I or +5/0 degrees C (day/night) and +18/+13 degrees C (day/night) in Exp. II. GST activities were assessed using either 1-chloro-2,4-dinitrobenzene (CDNB) or trans-cinnamic acid (tCA) as substrates. We found temperature to have no effect on Acy during either active growth or frost hardening. Acy increased several-fold from active growth to the beginning of frost hardening, but no increment was observed during the development of frost hardening. This suggests a role of Acy in photoprotection at low temperatures rather than their direct involvement in the development of freezing tolerance. The lack of response of GST activity to frost hardening and to temperature in autumn may indicate an indirect role of GSTs in frost hardening as protective enzymes. GST activity was the same with the two substrates studied (CDNB, tCA), supporting the assumption that GSTs could catalyze reactions with endogenous phenylpropanoids.