The American cranberry (Vaccinium macrocarpon Ait.) is an iconic North American fruit crop of great cultural and economic importance. Cranberry can be considered a fruit crop model due to its unique fruit nutrient composition, overlapping generations, recent domestication, both sexual and asexual reproduction modes, and the existence of cross-compatible wild species. Development of cranberry molecular resources started very recently; however, further genetic studies are now being limited by the lack of a high-quality genome assembly. Here, we report the first chromosome-scale genome assembly of cranberry, cultivar Stevens, and a draft genome of its close wild relative species Vaccinium microcarpum. More than 92% of the estimated cranberry genome size (492 Mb) was assembled into 12 chromosomes, which enabled gene model prediction and chromosome-level comparative genomics. Our analysis revealed two polyploidization events, the ancient γ-triplication, and a more recent whole genome duplication shared with other members of the Ericaeae, Theaceae and Actinidiaceae families approximately 61 Mya. Furthermore, comparative genomics within the Vaccinium genus suggested cranberry-V. microcarpum divergence occurred 4.5 Mya, following their divergence from blueberry 10.4 Mya, which agrees with morphological differences between these species and previously identified duplication events. Finally, we identified a cluster of subgroup-6 R2R3 MYB transcription factors within a genomic region spanning a large QTL for anthocyanin variation in cranberry fruit. Phylogenetic analysis suggested these genes likely act as anthocyanin biosynthesis regulators in cranberry. Undoubtedly, these new cranberry genomic resources will facilitate the dissection of the genetic mechanisms governing agronomic traits and further breeding efforts at the molecular level.

(1) Background: Two Caucasian blueberries Vaccinium myrtillus L. and Vaccinium arctostaphylos L. are famous berry bushes growing in the Caucasus region and used to treat neurological diseases, but the chemistry and bioactivity of leaf extracts are still poorly studied. (2) Methods: Phenolic compounds of V. myrtillus and V. arctostaphylos leaf extracts were profiled and quantified by HPLC-PDA-ESI-tQ-MS. The neurotropic potential of Vaccinium extracts was studied using the model of middle cerebral artery permanent occlusion to determine cerebral blood flow, the area of the brain tissue necrosis, and antioxidant enzyme activity (including superoxide dismutase, succinate dehydrogenase, and cytochrome C oxidase), as well as the concentration of TBARS. (3) Results: Hydroxycinnamates and flavonoids were identified in the leaves of V. myrtillus and V. arctostaphylos, and the dominant metabolite of both extracts was 5-O-caffeoylquinic acid in the amount of 105-226 mg/g. The studied extracts enhanced the cerebral hemodynamics and decreased the frequency of necrotic and lipooxidative processes in the brain tissue, accompanied by an increase in the activity of antioxidant enzymes. The positive effect of V. arctostaphylos was stronger and exceeded the effectiveness of Ginkgo biloba standardized extract. (4) Conclusion: The leaf extracts of Caucasian blueberries V. myrtillus and V. arctostaphylos as a new source of hydroxycinnamates demonstrated a protective effect of the brain ischemia pathology and can be used as therapeutic agents to treat neurological diseases.

Vaccinium species have great significance as fruit crops due to their economic and food values. Here we report the chloroplast genome of V. oxycoccos. The chloroplast genome of V. oxycoccos was 177,088 bp in length with a GC content of 36.74%. LSC, SSC, and IR regions were 104,139 bp, 3031 bp, and 34,959 bp in length, respectively. The chloroplast genome contained 105 different genes, including 73 protein-coding genes, 4 rRNA genes, and 28 tRNA genes. The phylogenetic analysis indicated that V. oxycoccos was closely related to V. microcarpum in the family Ericaceae. This chloroplast genome not only enriches the genome information of Vaccinium, but also will be useful in the evolution study of the family Ericaceae.

Highbush blueberry is a small berry fruit tree belonging to the family Ericaceae and genus Vaccinium, which fruit has high nutritional value. High-throughput sequencing technology was applied in this study to sequence and assemble the whole chloroplast genome of the southern highbush blueberry variety sharpblue. The results of the study showed that the circular genome of sharpblue is 170,737 bp in length, and the GC content of the genome was 36.8%. The complete chloroplast genome of sharpblue has consisted of two inverted repeat regions (IRs), a large single-copy region (LSC, 31, 076 bp), and a small single-copy region (SSC, 3, 044 bp). The chloroplast genome contained a total of 144 functional genes, including 100 mRNA genes, eight rRNA genes, and 36 tRNA genes. In addition, V. corymbosum and V. oldhamii were clustered into one group in this phylogenetic analysis which indicated that they have a close evolutionary relationship. The findings of this investigation are a significant reference source for the phylogeny and evolutionary origin of the Ericaceae family.

Bioinformatic and molecular characterization of satellite repeats was performed to understand the impact of their diversification on Vaccinium genome evolution. Satellite repeat diversity was evaluated in four cultivated and wild species, including the diploid species Vaccinium myrtillus and Vaccinium uliginosum, as well as the tetraploid species Vaccinium corymbosum and Vaccinium arctostaphylos. We comparatively characterized six satellite repeat families using in total 76 clones with 180 monomers. We observed that the monomer units of VaccSat1, VaccSat2, VaccSat5, and VaccSat6 showed a higher order repeat (HOR) structure, likely originating from the organization of two adjacent subunits with differing similarity, length and size. Moreover, VaccSat1, VaccSat3, VaccSat6, and VaccSat7 were found to have sequence similarity to parts of transposable elements. We detected satellite-typical tandem organization for VaccSat1 and VaccSat2 in long arrays, while VaccSat5 and VaccSat6 distributed in multiple sites over all chromosomes of tetraploid V. corymbosum, presumably in long arrays. In contrast, very short arrays of VaccSat3 and VaccSat7 are dispersedly distributed over all chromosomes in the same species, likely as internal parts of transposable elements. We provide a comprehensive overview on satellite species specificity in Vaccinium, which are potentially useful as molecular markers to address the taxonomic complexity of the genus, and provide information for genome studies of this genus.

Wild berries are interesting research subjects due to their rich sources of health-beneficial phenolic compounds. However, the internal microbial communities, endophytes, associated with the wild berry fruits are currently unknown. Endophytes are bacteria or fungi inhabiting inside plant tissues, and their functions vary depending on the host species and environmental parameters. The present study aimed to examine community composition of fungal and bacterial endophytes in fruits of three wild berry species (bilberry Vaccinium myrtillus L., lingonberry Vaccinium vitis-idaea L. and crowberry Empetrum nigrum L.) and the effects of host plant species and their growth sites on shaping the endophytic communities. We found that the endophytic community structures differed between the berry species, and fungi were predominant over bacteria in the total endophytic taxa. We identified previously unknown endophytic fungal taxa including Angustimassarina, Dothidea, Fellozyma, Pseudohyphozyma, Hannaella coprosmae and Oberwinklerozyma straminea. A role of soluble phenolic compounds, the intracellular components in wild berry fruits, in shaping the endophytic communities is proposed. Overall, our study demonstrates that each berry species harbors a unique endophytic community of microbes.

Cranberry (Vaccinium macrocarpon) dietary supplementation can help prevention of urinary tract infections through the supply of proanthocyanidin-type polyphenols (PAC). The main uropathogenic bacteria are members of the intestinal microbiota. A randomized cross-over experiment was done to investigate whether cranberry dietary supplementation affects concentrations of thermotolerant coliforms, Enterococcus spp. and Lactobacillus spp. in rat faeces. Thirteen rats, housed in individual cages, received successively two diets as pellets during 7 days each: a standard diet without polyphenols and the standard diet supplemented with cranberry powder containing 10.9 mg/100 g of PAC. There was a 7 days wash-out period in between with standard diet without polyphenols. Body weight and feed intake were recorded. Faeces were collected on the last day of treatment, and crushed to count the different bacterial populations using the most probable number method. Thermotolerant coliforms were grown in BGBLB tubes and on MacConkey agar. Enterococcus spp. were grown in Rothe and Litsky broths and on KF Streptococcus agar. Lactobacillus spp. were grown in Man Rogosa Sharpe broth. Body mass gains were not affected by cranberry supplementation. This is consistent with equal food intake, cranberry powder not providing significant energy supplement. Cranberry dietary supplementation was associated with changes in fecal concentrations of thermotolerant coliforms, and Enterococcus spp. in some rats, but did not induce significant changes in bacterial fecal concentrations in a global population of 13 rats. In conclusion, we did not observe any significant effect of dietary cranberry supplementation on the fecal microbiota of Wistars rats for a 7-day diet.

The true blueberries (Vaccinium sect. Cyanococcus; Ericaceae), endemic to North America, have been intensively studied for over a century. However, with species estimates ranging from nine to 24 and much confusion regarding species boundaries, this ecologically and economically valuable group remains inadequately understood at a basic evolutionary and taxonomic level. As a first step toward understanding the evolutionary history and taxonomy of this species complex, we present the first phylogenomic hypothesis of the known diploid blueberries.

Bilberry fruit is regarded as one of the best natural sources of anthocyanins and is widely explored for its health-beneficial compounds. Besides anthocyanins, one of the major attributes that determine the berry quality is the accumulation of sugars that provide sweetness and flavor to ripening fruit. In this study, we have identified 25 sugar metabolism-related genes in bilberry, including invertases (INVs), hexokinases (HKs), fructokinases (FKs), sucrose synthases (SSs), sucrose phosphate synthases (SPSs), and sucrose phosphate phosphatases (SPPs). The results indicate that isoforms of the identified genes are expressed differentially during berry development, suggesting specialized functions. The highest sugar content was found in ripe berries, with fructose and glucose dominating accompanied by low sucrose amount. The related enzyme activities during berry development and ripening were further analyzed to understand the molecular mechanism of sugar accumulation. The activity of INVs in the cell wall and vacuole increased toward ripe berries. Amylase activity involved in starch metabolism was not detected in unripe berries but was found in ripe berries. Sucrose resynthesizing SS enzyme activity was detected upon early ripening and had the highest activity in ripe berries. Interestingly, our transcriptome data showed that supplemental irradiation with red and blue light triggered upregulation of several sugar metabolism-related genes, including α- and β-amylases. Also, differential expression patterns in responses to red and blue light were found across sucrose, galactose, and sugar-alcohol metabolism. Our enzymological and transcriptional data provide new understanding of the bilberry fruit sugar metabolism having major effect on fruit quality.

This study was aimed to investigate the effect of aqueous cranberry extract (ACE) on MK-801-induced psychosis in mice.

Vaccinium myrtillus (Bilberry) extracts (VME) were tested for effects on angiogenesis in vitro and in vivo. VME (0.3-30 µg ml(-1)) and GM6001 (0.1-100 µM; a matrix metalloproteinase inhibitor) concentration-dependently inhibited both tube formation and migration of human umbilical vein endothelial cells (HUVECs) induced by vascular endothelial growth factor-A (VEGF-A). In addition, VME inhibited VEGF-A-induced proliferation of HUVECs. VME inhibited VEGF-A-induced phosphorylations of extracellular signal-regulated kinase 1/2 (ERK 1/2) and serine/threonine protein kinase family protein kinase B (Akt), but not that of phospholipase Cγ (PLCγ). In an in vivo assay, intravitreal administration of VME inhibited the formation of neovascular tufts during oxygen-induced retinopathy in mice. Thus, VME inhibited angiogenesis both in vitro and in vivo, presumably by inhibiting the phosphorylations of ERK 1/2 and Akt. These findings indicate that VME may be effective against retinal diseases involving angiogenesis, providing it can reach the retina after its administration. Further investigations will be needed to clarify the major angiogenesis-modulating constituent(s) of VME.

The demand for blueberry Vaccinium corymbosum L. (and hybrids) plants has significantly increased in the last 30 years due to its market expansion. In vitro propagation of sterile plants are required for commercial purposes but also for research applications such as plant transformation. Thus far, tissue culture characteristics of the tropical-adapted blueberry have been scarcely studied. In this study we present the following findings: (i) zeatin, a hormone used to promote plant growth, should be used in the 1-2 mg/L range to promote plant architecture optimal for transformation experiments; (ii) red-blue LED lights induce more production of meristems and biomass than white LED or fluorescent lights; (iii) levels as high as 1000 mg/L of decontamination agents (the antibiotics timentin and cefotaxime) can be used to eliminate Agrobacterium overgrowth without inhibiting plant growth during plant transformation experiments; (iv) kanamycin, paromomycin, and geneticin, which are widely used antibiotics to select transgene-carrying transformants, cannot be efficiently used in this system; (v) glufosinate, a widely used herbicide, shows potential to be used as an effective selectable marker for transformed plants.

Blueberries are one of the few horticultural crops adapted to grow in acidic soils. Neutral to basic soil pH is detrimental to all commonly cultivated blueberry species, including Vaccinium corymbosum (VC). In contrast, the wild species V. arboreum (VA) is able to tolerate a wider range of soil pH. To assess the molecular mechanisms involved in near neutral pH stress response, plants from pH-sensitive VC (tetraploid) and pH-tolerant VA (diploid) were grown at near neutral pH 6.5 and at the preferred pH of 4.5.

Vaccinium is a large genus of shrubs that includes a handful of economically important berry crops. Given the numerous hybridizations and polyploidization events, the taxonomy of this genus has remained the subject of long debate. In addition, berries and berry-based products are liable to adulteration, either fraudulent or unintentional due to misidentification of species. The availability of more genomic information could help achieve higher phylogenetic resolution for the genus, provide molecular markers for berry crops identification, and a framework for efficient genetic engineering of chloroplasts. Therefore, in this study we assembled five Vaccinium chloroplast sequences representing the economically relevant berry types: northern highbush blueberry (V. corymbosum), southern highbush blueberry (V. corymbosum hybrids), rabbiteye blueberry (V. virgatum), lowbush blueberry (V. angustifolium), and bilberry (V. myrtillus). Comparative analyses showed that the Vaccinium chloroplast genomes exhibited an overall highly conserved synteny and sequence identity among them. Polymorphic regions included the expansion/contraction of inverted repeats, gene copy number variation, simple sequence repeats, indels, and single nucleotide polymorphisms. Based on their in silico discrimination power, we suggested variants that could be developed into molecular markers for berry crops identification. Phylogenetic analysis revealed multiple origins of highbush blueberry plastomes, likely due to the hybridization events that occurred during northern and southern highbush blueberry domestication.

This study investigated ericoid mycorrhizal fungi (EMF) diversity in Vaccinium uliginosum across its main wild distribution range in China. Fungal communities in hair roots of V. uliginosum were analyzed using Illumina MiSeq sequencing. Only 22 OTUs were assigned to putative EMF genera. Rhizoscyphus and Meliniomyces dominated EMF communities, followed by Clavaria, Oidiodendron, Lachnum, Acephala, and Phialocephala. There were more dark septate endophytes (DSE) reads from the Greater Khingan Mountains than from other study areas, similar to the results of the percent colonization of DSE by the magnified intersections method. Overall, high-throughput sequencing data provided a rough community-scale sketch of root-associated fungi of V. uliginosum. Two hundred and eighty slow-growing isolates were isolated from root pieces of V. uliginosum, and the isolates matched 16 fungal genera on the basis of morphological and internal transcribed spacer sequence comparison. The isolates of Cryptosporiopsis ericae, Oidiodendron maius, Lachnum sp., Sordariomycetes sp., and Pleosporales sp., formed ericoid hyphal coils via resynthesis trails. The co-existence between EMF and DSE in hair roots was observed via trypan blue staining. A putative model for the co-existence between EMF and DSE in the hair roots of V. uliginosum was proposed. We suggest that under certain environmental stresses, such as low temperature and poor available nutrients, ericoid plants may favor co-colonization by both DSE and EMF.

Fruits and leaves of Vaccinium spp. are known for their high content of bioactive compounds, but the chemical and biological characteristics of mortiño (Vaccinium floribundum Kunth) have not been fully described. In this study, the levels of polyphenols, antioxidant capacity, anthocyanins, antimicrobial activity, and genetic variability were determined in mortiño plants. The Folin-Ciocalteu's, ABTS scavenging, pH differential, and well diffusion methods were used to evaluate the levels of polyphenols, antioxidant capacity, anthocyanins, and antimicrobial activity, respectively. The genetic variability was evaluated by sequencing of the matK and rbcl DNA regions. Polyphenol content was up to 229.81 mg gallic acid equivalents/100 g, the average antioxidant capacity was 11.01 mmol Trolox equivalents/100 g, and anthocyanin content was up to 1,095.39 mg/100 g. Mortiño extracts significantly inhibited the growth of Gram-negative bacteria including Burkholderia gladioli, Burkholderia cepacia, Salmonella Typhimurium, Vibrio parahaemolyticus, Vibrio harveyi, Vibrio vulnificus, Escherichia coli, and Pseudomona aeruginosa, as well as Gram-positive bacteria such as Probionibacterium propionicum, Staphylococcus aureus, and Enterococcus faecalis showing greater inhibition halos than those produced by the antibiotic ampicillin. A polymorphic nucleotide was found in position 739 of the matK region. This study shows the potential of mortiño for the food and pharmaceutical industries.

Products of lingonberries are widely used in the human diet; they are also promising beauty and health therapeutic candidates in the cosmetic and pharmaceutical industries. It is important to examine the sugar profile of these berries, due to potential deleterious health effects resulting from high sugar consumption. The aim of this study was to determine the composition of sugars in wild clones and cultivars or lower taxa of lingonberries by HPLC-ELSD method of analysis. Acceptable system suitability, linearity, limits of detection and quantification, precision, and accuracy of this analytical method were achieved. The same sugars with moderate amounts of fructose, glucose, and low amounts of sucrose were found in wild and cultivated lingonberries. Cultivar 'Erntekrone' and wild lingonberries collected from full sun, dry pine tree forests with lower altitude and latitude of the location, distinguished themselves with exclusive high contents of sugars. The changes in the sugar levels during the growing season were apparent in lingonberries and the highest amounts accumulated at the end of the vegetation. According to our findings, lingonberries seem to be an appropriate source of dietary sugars.

Wufanshu (Vaccinium bracteatum Thunb.), which is a wild member of the genus Vaccinium, accumulates high concentration of anthocyanin in its berries. In this study, the accumulated anthocyanins and their derivatives in Wufanshu berries were identified through UHPLC-MS/MS analysis. Candidate anthocyanin biosynthetic genes were identified from the transcriptome of Wufanshu berries. qRT-PCR analyses showed that the expression of anthocyanin structural genes correlated with anthocyanin accumulation in berries. The R2R3-MYB, VbMYBA, which is a homolog of anthocyanin promoting R2R3-MYBs from other Vaccinium species, was also identified. Transient expression of VbMYBA in Nicotiana tabacum leaves confirmed its role as an anthocyanin regulator, and produced a higher anthocyanin concentration when compared with blueberry VcMYBA expression. Dual-luciferase assays further showed that VbMYBA can activate the DFR and UFGT promoters from other Vaccinium species. VbMYBA has an additional 23 aa at the N terminus compared with blueberry VcMYBA, but this was shown not to affect the ability to regulate anthocyanins. Taken together, our results provide important information on the molecular mechanisms responsible for the high anthocyanin content in Wufanshu berries.

The beneficial effects of anthocyanins consumption on cardiovascular risk are supported by mechanistic and epidemiologic evidence. In order to explore the effects of Vaccinium berries rich in anthocyanins on serum lipids, we conducted a meta-analysis of relevant randomized controlled trials (RCTs). Sixteen studies with 1109 subjects were included in this meta-analysis. Significant heterogeneity confirmed differential effects between Vaccinium subclasses. The whortleberry group is significantly superior to placebo in lipids improvement. Besides, bilberry groups show significant differences in reducing LDL-C and increasing HDL-C in comparison with other treatments. For many of the other subgroups and comparison arms, there was insufficient evidence to draw conclusions about efficacy.

Vaccinium arctostaphylos has traditionally been employed in Iranian folk medicine to treat diabetes. However, the precise molecular mechanisms underlying its antidiabetic properties remain incompletely understood. The current experiment intended to explore the modulatory effects of V. arctostaphylos fruit ethanolic extract (VAE) on biochemical and molecular events in the livers of diabetic rats.