THE GENETIC ASPECTS OF BERRIES: FROM FIELD TO HEALTH Luca Mazzoni1, Patricia Perez-Lopez2, Francesca Giampieri3,*, Jose M Alvarez-Suarez1,4, Massimiliano Gasparrini1, Tamara Y Forbes-Hernandez1,5, Jose L Quiles2, Bruno Mezzetti3, and Maurizio Battino1,6*
1
Dip. Scienze Cliniche Specialistiche ed Odontostomatologiche (DISCO), Facoltà di
Medicina, Università Politecnica delle Marche, Ancona, Italy 2
Dep. Physiology, Institute of Nutrition and Food Technology ‘‘José Mataix”, Biomedical
Research Center, University of Granada, Granada, Spain 3
Dip. Scienze Agrarie, Alimentari e Ambientali, Università Politecnica delle Marche,
Ancona, Italy 4
Facultad de Ciencias de la Salud, Universidad Nacional de Chimborazo, Riobamba -
Ecuador 5
Area de Nutrición y Salud, Universidad Internacional Iberoamericana (UNINI), Campeche,
C.P.24040, Mexico 6
Director Centre for Nutrition & Health, Universidad Europea del Atlantico (UEA),
Santander 39011, Spain
*Correspondence
to:
Prof.
Maurizio
Battino,
PhD,
DSc,
MS,
MD
([email protected]) or Dr. Francesca Giampieri ([email protected]),
(Hon) DISCO,
Facoltà di Medicina, Università Politecnica delle Marche, Via Ranieri 65, 60131 Ancona, Italy, Telephone +39 071 2204646/2204136; fax +39 071 2204123.
This article has been accepted for publication and undergone full peer review but has not been through the copyediting, typesetting, pagination and proofreading process, which may lead to differences between this version and the Version of Record. Please cite this article as doi: 10.1002/jsfa.7216
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ABSTRACT Berries are a relevant source of micronutrients and nonessential phytochemicals such as polyphenol compounds, that play a synergistic and cumulative role in human health promotion. Several systematic analyses showed that berry phenolics are able to detoxify ROS/RNS blocking their production, to intervene in the cell cycle, participating in the transduction and expression of genes involved in apoptosis, and to repair oxidative DNA damage. As a consequence, the improvement of the nutritional quality of berries has become a new quality target of breeding and biotechnological strategies, to control or to increase the content of specific health-related compounds in fruits. This work reviews, on the basis of the in vitro and in vivo evidences, the main berries phytochemical compounds and their possible mechanisms of action on pathways involved in several type of diseases, with particular attention to cancer, inflammation, neurodegeneration, diabetes and cardiovascular diseases.
KEYWORDS berries, bioactive compounds, genetic improvement, cancer, inflammation, neurological diseases.
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INTRODUCTION The importance of diet in the prevention of several chronic diseases has been widely recognized. The assumption is that consumption of fruit and vegetables is correlated with a high intake of antioxidant and bioactive compounds that play a crucial role in the prevention of a wide range of diseases, such as cancer, cardiovascular, and other age-related degenerative pathologies1-3. The aging process is common for most multicellular organisms, and consists of a gradual impairment of the biological function of all organs in the postreproduction phase, compromising the homeostasis preservation and resulting in an increased risk of death. Research has been directed to the delineation of the most critical factors that may account for a wide range of age-related diseases (ARD), suggesting that one of the major causes of the incidence of age-related illnesses as skin aging, neurodegenerative pathologies including Lou Gehrig’s, Parkinson’s, Alzheimer’s and Huntington’s diseases, cancer and cardiovascular diseases (CVD), is the long-term exposition to oxidative stress (OS)4. OS is caused by an imbalance between the systemic production of reactive oxygen and nitrogen species (ROS and RNS) and the ability of the biological system to detoxify or to repair the ROS/RNS induced damage5. Studies monitoring ROS/RNS production and antioxidant defenses indicate that OS, with the consequent mitochondrial respiration impairment and mitochondrial damage, is related with the pathophysiology of these diseases6. A balanced diet, rich in antioxidant compounds derived from fruits and vegetables, as the Mediterranean one, can deeply influence the susceptibility to OS, counteracting the decrease in antioxidant protection that occurs during illness7,8. Berries are a common and important fruit in the Mediterranean diet and are among the richest fruits in nutritive compounds, including minerals, vitamins, dietary fibers, and especially polyphenolic phytochemicals (flavonoids, phenolic acids, lignans and tannins)9-11, which exhibit strong antioxidant and anti-inflammatory activities that may reduce sensitivity to
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OS12. Therefore, berries may have a role in the prevention of degenerative pathologies, stimulating researchers to implement new biotechnological approaches for the improvement of bioactive compound contents13-14. For example the use of wild germplasm in the genetic improvement programs for the creation of new commercial strawberries with increased antioxidant content is an important option to support a higher intake of healthy compounds even with a lower fruit consumption13, giving that commercial strawberries deriving from wild germplasm breeding programs resulted rich in antioxidant compounds. Two important areas of research are therefore (i) the development of new berry varieties or the improvement of current varieties with improved levels of bioactive compounds production (ii) determination of the mechanisms of bioactive compounds on gene expression in humans to identify which compounds are useful, particularly those with protective effects against ROS/RNS. Both lines of investigation are of critical importance for obtaining consistent results.
BREEDING AND BIOTECHNOLOGICAL PROGRAMS IN BERRIES High throughput technologies for plant genotyping, such as those developed for the detection of Single Nucleotide Polymorphism, accelerated the fine mapping of Quantitative Trait Loci (QTLs) and the positional cloning of the genes responsible for traits involved in berry nutritional quality. The correct integration between genetic resources, genetic improvement and transgenic approaches is essential to obtain new varieties that can help to reach the desired nutritional quality in berries15-17. In the last few years, fruit sensorial and nutritional traits have become the major breeding targets in the commercial octoploid strawberry (Fragaria x ananassa). The first works on the genetic engineering of strawberry were focused on the improvement of agronomical or qualitative (but not nutritional) traits, like size, number, yield, ripening, softening and
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firmness of fruit18,19. Recently efforts have focused on improved nutritional value of strawberry. A first interesting result was the demonstration that the biosynthesis of L-ascorbic acid in ripe strawberry can occur through D-galacturonic acid, encoded by the gene GaiUR20. Another study demonstrate, for the first time, that the rolC gene lead to an improvement of fruit nutritional quality in strawberry, mainly sugar content and total antioxidant capacity13. One of the most important group of compounds with healthy properties are anthocyanidins. A study demonstrated that at least two different enzymes are involved in the glucosylation of anthocyanidins in strawberry: a transferase, which forms the glucosides at early ripening stages, and a glycosyl-transferase (encoded by FaGT1 gene) which catalyzes the glucoside transfer in ripe strawberry fruits21. The anthocyanin accumulation, instead, is regulated by the coordinated expression of genes encoding the flavonoid biosynthetic pathway enzymes, the R2R3MYB and the bHLH transcription factors22. For further improvement projects concerning the octoploid strawberry, the construction of the most comprehensive genetic map for Fragaria x ananassa was a crucial step23. In this study a first reference map for octoploid Fragaria was provided and almost all linkage groups were ranged into the seven expected homoeologous groups (HG). A successive three years study identifies 33 QTLs of 14 studied traits into the seven HGs expected in Fragaria. In addition to QTLs for titratable acidity, pH, plant width, color parameters and other characteristics, in this map the genes and the QTLs for the control of the healthy compounds were indicated: for ascorbic acid, three QTLs (LAAV-M.1, LAAVII-M.1 e LAAIV-F.2), and FaGaiUR gene in the linkage groups (LGs) belonging to HG IV were identified; for anthocyanin, a cluster of QTLs was identified in LG V-M.2, another QTL in LG III-F.1, the anthocyanin biosynthetic gene F3H in HG I and other genes involved in anthocyanin biosynthesis (CHI, ChFaM247, ChFvM234 or FaMYB1) in HG V (Figure 1)24,25. Furthermore, in the last year, an international consortium sequenced the genome of the diploid strawberry (Fragaria vesca), that permitted the access to genomic
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information relevant to Rosaceae, especially for fruit quality (flavor, nutrition and aroma). In particular, for the nutritional quality, some genes have been detected for the proanthocyanidin levels in the seed (FvMyb33)26, for the content of l-ascorbate (FaGaiUR and FaGLHD), for flavonoid metabolism (FaGT6 and FaGT7), for polyphenols anchoring (Fahyprp) and for flavonoid biosynthesis (Fraa1A)27. Genetic transformation in strawberry offers an effective method for creating new varieties that selectively targets a specific interested gene, with particular attention to the nutritional quality of the fruit. Another high-value berry species is raspberry, with a demonstrated beneficial impact on human health28,29. The main characters, objects of genetic improvement, are the visual parameters, in particular color. This parameter, however, not only influenced the appearance and the flavor of the fruit, but is also closely linked to the anthocyanin content (in particular cyanidin and pelargonidin) of the berry30. Since 1980, genes involved in the synthesis of sugars rhamnose, sophorose and xylose (required to give the array of different anthocyanin pigments observed in red raspberry) were detected31. Only recently, genes involved in the anthocyanin
pathway
(bHLH
and
FRUITE4)
and
in
the
flavonol
pathway
(ERubLR_SQ07_2_H02) were found. The bHLH was mapped in the LG1 and the bZIP (FRUITE4) gene on LG4 (Figure 2)32,33. Blueberry, a major berry crop in the United States, has great nutritional and economical value. The bigger problem of its cultivation is the low temperature that reduces crop yields and causes major losses. So a blueberry genomics database was developed to help the researchers to establish new varieties with a better cold acclimation and freeze tolerance34. In the last years, the interest on the nutritional value of blueberry has increased, in particular for the flavonoid biosynthesis. A first study in 2002 detected the genes involved in the flavonoid pathway of bilberry: Phe ammonia-lyase (PAL), chalcone synthase (CHS), flavanone 3hydroxylase (F3H), dihydroflavonol 4-reductase (DFR) and anthocyanidin synthase (ANS)35.
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This latter gene demonstrate a correlation between anthocyanin accumulation and expression of the flavonoid pathway genes during the ripening of berries.
THE GENETIC MECHANISM OF ACTION OF BERRIES BIOACTIVE COMPOUNDS Numerous physiological processes produce reactive oxidant species and free radicals which may endanger the health and favor the onset of several diseases. So often the body needs exogenous sources of antioxidant defenses, mainly supplied by the diet. Berries, as previously discussed, are rich in bioactive compounds36 and their antioxidant activity can act along many mechanisms, such as detoxifying ROS and blocking of ROS production37-39, intervening in the cell cycle and participating in the transduction and expression of genes involving in apoptosis, suppression of tumors, and repair of oxidative DNA damage, that is the first step for the initiation of cancer40. Therefore the ability of berries to reduce oxidative DNA damage may be considered a good therapy for cancer prevention. It has been demonstrated that berry compounds modulate the expression of genes involved in DNA damage41: on the one hand, berries rich diet over-expressed DNA repair genes, such as XPA, DNL3, ERCC5, CYP450, CYP3A442,43, and on the other hand, genes involved in the DNA damage, such as MAPK14 and MAPKK, are down-regulated42. Over the last few years, many researches have demonstrated cancer chemopreventive activities of berries. In studies of the anti-tumoral activity of berries, work has focused on up or down regulation of different genes involved in inhibition of carcinogen activation, stimulation of carcinogen detoxification, scavenging of oxygen reactive species, control of cell-cycle and cell proliferation, induction of apoptosis, inhibition of oncogenes and metastasis44,45. Berry compounds have been shown to regulate several pathways including nuclear factor kB (NF-κB) and cyclooxygenases (COX-2), cell adhesion proteins46,
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phosphoinositide 3-kinase (PI-3K)/protein kinase B (Akt), mitogen-activated protein kinase (MAPK), p38, vascular endothelial growth factor (VEGF-1) for angiogenesis47 and Bcl-2, p21, Bax, cytochrome c and caspase 3, which regulate cellular apoptosis48,49. A study on cranberries confirms the ability of berries to inhibit growth and proliferation of tumor cells reducing ornithine decarboxylase and COX-2 activity, and matrix metalloproteases expression50. COX-2 is implicated also in the anti-inflammatory capacity of berry juice, which inhibits tumor necrosis factor (TNF)-induced expression of COX-2 mediated by the nuclear expression factor NF-kB51. This research group also showed that berry juice inhibits the proliferation of five cancer cell lines by reducing the expression of key regulators of cellcycle (cdk4, cdk6, cyclin D1 and cyclin D3)51. Blackberry extract has the ability to inhibit both VEGF and AP-1 expression mediated by the PI-3K/Akt pathway in presence of benzo(a)pyrene (BaP), a potent cancerogen47. Meanwhile, raspberries and strawberries inhibit BaP-induced Nuclear factor of activated T-cells (NFAT) activation and TNF-alpha (α) expression52. Furthermore, berries stimulate the expression of cytoprotective genes, such as nuclear factor-E2-related factor 2 (Nrf2) which is over-expressed by berry polyphenols and binds to the antioxidant response element in cells53. A range of studies have been carried out to establish the anticancer activity of berries and their bioactive compounds. In colon cancer cells, berry phenolic extracts inhibit cell growth by over-expression of p21WAF1, a member of the cyclin kinase inhibitors54, and stimulates apoptosis through COX-2 expression55. Ellagitannins also showed beneficial effects against colon carcinogenesis by inhibition of Wnt signaling56 and induction of apoptosis by suppression the PI3K/Akt pathway57. Resveratrol has been shown to inhibit proliferation by over expression of sirtuin 1 (SIRT1) and inducing apoptosis by activation of MAPK, which downstream caspases activation lead to cell death58. Recent research has related black raspberry consumption with the decrease in plasma concentrations of interleukine 8 (IL-8)
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and granulocyte macrophage colony stimulating factor (GM-CSF), which suggest beneficial properties of berries treatment59. In relation to breast cancer, several studies indicate the efficacy of berries consumption in the prevention and treatment of mammalian tumors. Berries prevent estradiol-induced mammary tumors by down-regulation the levels of enzymes to estradiol metabolism, such as CYP1A1, P450, 17beta-hydroxysteroid dehydrogenase type 7 (17βHSD7) and estrogen receptor alpha (ERα)60,61, and binding to receptor pathways which participate in breast cancer development, such as ER and tyrosine kinase receptor (TKR), which reduces phosphorylation of several kinases such as PI3K/Akt and extracellular signal-regulated kinase (ERK)/MAPK and inhibits grown factor receptor (GRF) activation, decreasing the tumor development62. With regard to other types of cancer, it was found that the combination of distinct types of berry fruits suppress the growth and invasion capacity of lung cancer cells by downregulation of WNT pathways and different targets of cell cycle progression, such as cyclin D1, cyclin B1, pERK, Matrix metallopeptidase 9 (MMP9) and VEGF63. In addition, berry fruits suppress TNFα-induced NF-κB activation, decreasing cell invasion. Finally, Wang et al.64 have gone a step further using DNA microarray technology for the determination of the gene expression profile on rat esophageal carcinogenesis treated with black raspberries. They have found that the treatment with berries promotes changes in the expression of genes implicated in proliferation, apoptosis, inflammation, angiogenesis and metastasis, among others. The mainly chemoprotective effect of berries on esophageal squamous cell seems to initiate with the inhibition of inducible nitric oxide synthase (iNOS), COX-2, VEGF, NF-κB and AP-1 pathway65. Quercetin, one of the main flavonoids which is a common component of berry fruits, is able to induce apoptosis in leukemia cell lines and B cells of chronic lymphocytic leukemia (BCLL) by down-regulation of myeloid leukemia cell differentiation protein (Mcl-1)
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expression, an antiapoptotic proteins of Bcl-2 family66. Furthermore, berry compounds have been studied as phytochemical suppressors of human T-lymphocytes by decreasing of IL-2 production, which could be used for the treatment of autoimmune and transplant patients67. Exposure to UV-A and UV-B radiation is known to induce DNA damage and reactive oxygen species production which could lead to skin diseases. Giampieri et al.44 showed that strawberry extracts provide photoprotective effects to human fibroblast in vitro against UV-A irradiation damage, according with Bae et al.68, who found that blueberry anthocyanins prevented UV-B-induced skin photoaging by decreasing collagen destruction and inflammatory responses via NF-κB and MAPK signaling. Cyanidin-3-rutinoside, an anthocyanin highly found in black raspberry, is potentially able to protect UV-induced skin damage, by down-regulation of NF-κB and AP-1 activation, inhibition of IL-8 and prevention of pro-caspase-3 cleavage69. Similarly, black raspberry extracts (BRE), topically applied, was effective at reducing the tumor number and the average tumor size in female SKH-1 hairless mice exposed to one minimal erythemal dose of UV-B thrice weekly on nonconsecutive days for 25 weeks and that the reduction was correlated with a significant reduction in tumorinfiltrating CD3+foxp3+ regulatory T-cells70. The same authors found that topical BRE treatment in mice exposed to a single minimal erythemal dose of UV-B significantly reduces edema, p53 protein levels, oxidative DNA damage, and neutrophil activation, highlighting the berry efficacy in the prevention of skin cancers. In addition to the anticancer properties, berry compounds have also shown anti-inflammatory capacity. In general, they are capable of blocking NF-κB (which is involved in decreasing PI3K/Akt signaling), lipopolysaccharide(LPS)-induced pro-inflammatory cytokines, hypoxiainducible factor 1-alpha (HIF-1α) and VEGF expression and TNF-α production. They also can inhibit activation of MAPK (down-regulating COX-2 and IL-6 expression) that participates in the down-regulation of TNF-α production71. Moreover, it has been shown the
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capacity of flavonols to up regulate VCAM-1, ICAM-1 and MCP-1 gene expression and decrease inflammatory markers, such as IL-1alpha (α) and IL-472. Inhibition of COX-2, iNOS and TNF-α have been shown in rats fed with rich-flavonols diet73. The positive effects of antocyanindins were also demonstrated: they reduce prostaglandin E2 (PGE2) levels and platelet-derived growth factor (PDGF)-induced VEGF expression by down-regulation of p38 MAPK and c-Jun N-terminal kinases (JNK) pathways74. All these anti-inflammatory mechanisms of berries have been associated with their beneficial actions in several diseases, including cardiovascular diseases, brain inflammatory stress, obesity and metabolic syndrome. Recent studies demonstrated that bilberry attenuates atherosclerosis lesion in mouse by down-regulation of genes implicated in oxidative stress, inflammation, and angiogenesis, such as AOX1, CYP2E1, TXNIP, JAM-A and VEGRF275,76. Furthermore, berry fruits modulate angiogenesis via PI3K/Akt pathway, improving plasma lipid profile, increasing plasma antioxidant activity and low-density lipoprotein (LDL) resistance to oxidation, and ameliorating endothelial function11. A recent meta-analysis also showed that high anthocyanin intake is associated with a reduction of risk of myocardial infarction (MI) and an inverse association between higher intake of anthocyanins and risk of MI was observed77. At the same time, a strawberry-enriched diet has shown to significantly improve the markers of oxidative stress, by decreasing lipid peroxidation oxidation and ameliorating the serum lipid profile of the subjects, through a reduction of total cholesterol, LDL-C and triglyceride levels, both in healthy78 and in dyslipidemic subjects79.Therefore, the berries polyphenols seem to be the most likely constituents, which exert in vivo effects in the prevention of the CVD risk. In brain, neuroinflammation and neurodegeneration are mediated by berry compounds actuation over NF-κB cascade and MAPK signaling, PI3K/Akt pathway80 and TNFα gene
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expression inhibition81. Berries bioactive compounds may also have a direct effect on signaling to enhance neuronal communication, a buffer capacity against excess of calcium, an enhancement of neuroprotective stress shock proteins and reduction of stress signals. Furthermore, anthocyanins not only have the capacity of scavenge free radicals in the brain, but also to affect intraneural signaling, to inhibit lipid peroxidation and inflammatory mediators COX-1 and COX-282. Flavonoids are also capable to regulate the transcription of antioxidant enzymes related to glutathione via ERK1 and ERK2, which exert an important role in some types of memory83. Some of the deleterious effects of OS in neurodegenerative diseases could be retarded or even reversed by increasing antioxidant levels, and the putative synergistic effects of antioxidants combinations is particularly effective in this regard84. In a two long-term studies on rats, strawberry and blueberry prevented retarded age-related declines in neuronal and cognitive functions: berries supplementation had the greatest effect on GTP-ase activity, and its association with vitamin E showed significant protection against these age-induced deficits also on the other parameters studied, like the motor behavioral and the Morris water maze performances84,85. Moreover, dietary supplementation with blueberry or strawberry extracts in rats, prior the exposition to whole-body irradiation with high-energy and charge particles, showed the ability to retard/reverse age-related deficits in behavior and signal transduction caused by the irradiation. In particular, the strawberry-fed rats showed a better protection against spatial deficits in the maze, and animals were better able to retain place information (a hippocampally mediated behavior) compared to controls, while blueberry diet appeared to improve the reversal learning, a behavior more dependent on intact striatal function86. Recently, human studies suggest that polyphenols present in berries may prevent the occurrence of dementia and improve memory indices83. These findings suggest that berries phytochemicals can be beneficial in retarding age-related functional and
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cognitive-behavioral deficits, as well as in reversing the course of neuronal aging in neurodegenerative diseases, mainly Alzheimer disease87. Finally, because diabetes mellitus is a metabolic disease often associated with aging, the effect of berries on this pathology is also interesting. An in vivo study on rats has demonstrated that a dietary intake of cranberries may protect the pancreatic islets from the morphological and functional decline. In fact, the cranberries consumption inhibited two enzymes required for starch digestion to maltodextrins and thereby conversion into absorbable monosaccharides. The inhibition of these enzymes improved metabolic homeostasis, delaying the age-related decline in basal insulin, ameliorating pancreatic beta (β)-cell glucose responsiveness and augmenting β-cell mass in normal aged rats88.
CONCLUSIONS Several evidences from in vitro and in vivo studies suggest that berry phytochemicals are able to counteract OS, to protect DNA and to regulate the cellular metabolism and apoptosis. Their anti-inflammatory properties can also decrease the risk of developing chronic ARD. The results here presented show the role of berries compounds in regulating cellular processes related to OS, inflammation and cancer proliferation, including the expression of key genes involved in these pathways, providing an insight on the potential of berries as important allies, in the context of a balanced diet such as the Mediterranean one, to protect or reduce the risk and the occurrence of several chronic pathologies. The integration of breeding knowledge with the availability of complete genome will represent in the next future the most efficient approach to improve nutritional and nutraceutical quality for healthier fruits.
ACKNOWLEDGEMENTS
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The authors would like to thank to Ms. Monica Glebocki for extensive editing of the manuscript.
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REFERENCES
1.
Yi-Fang C, Jie S, Xianzhong W, Rui Hai L, Antioxidant and antiproliferative activities of common vegetables. J Agric Food Chem 50: 6010-6916 (2002).
2.
Etminan M, Takkouche B, Caamano-Isorna F, The role of tomato products and licopene in the prevention of prostate cancer: a meta-analysis of observational studies. Cancer Epidemiol Biomarkers Prev 13 (3) (2004).
3.
Tulipani S, Romandini S, Busco F, Bompadre S, Mezzetti B, Battino M, Ascorbate, not urate, modulates the plasma antioxidant capacity after strawberry intake. Food Chem 117: 181-188 (2009).
4.
Joseph JA, Shukitt-Hale B, Denisova NA, Bielinski D, Martin A, McEwen JJ, Bickford PC, Reversals of age-related declines in neuronal signal transduction, cognitive, and motor behavioral deficits with blueberry, spinach, or strawberry dietary supplementation. J Neurosci 15: 8114-8121 (1999).
5.
Patel VP, Chu CT, Nuclear transport, oxidative stress, and neurodegeneration. Int J Clin Exp Pathol 4: 215-229 (2011).
6.
Ramalingam M, Kim SJ, Reactive oxygen/nitrogen species and their functional correlations in neurodegenerative diseases. J Neural Transm 119: 891-910 (2012).
7.
Bach-Faig A, Berry EM, Lairon D, Reguant J, Trichopoulou A, Dernini S, Medina FX, Battino M, Belahsen R, Miranda G, Serra-Majem L; Mediterranean Diet Foundation Expert Group, Mediterranean diet pyramid today. Science and cultural updates. Public Health Nutr 14: 2274-2284 (2011).
8.
Requant-Aleix J, Arbore MR, Bach-Faig A, Serra-Majem L, Mediterranean heritage: an intangible cultural heritage. Public Health Nutr 12: 1591-1594 (2009).
This article is protected by copyright. All rights reserved
9.
Battino M, Beekwilder J, Denoyes-Rothan B, Laimer M, McDougall GJ, Mezzetti B, Bioactive compounds in berries relevant to human health. Nutr Rev 67: Suppl. 1, S145S150 (2009).
10. Giampieri F, Alvarez-Suarez JM, Mazzoni L, Romandini S, Bompadre S, Diamanti J, Capocasa F, Mezzetti B, Quiles JL, Ferreiro MS, Tulipani S, Battino M, The potential impact of strawberry on human health. Nat Prod Res 27: 448-455 (2013). 11. Giampieri F, Tulipani S, Alvarez-Suarez JM, Quiles JM, Mezzetti B, Battino M, The strawberry: composition, nutritional quality, and impact on human health. Nutrition 28: 9-19 (2012). 12. Tulipani S, Mezzetti B, Battino M, Impact of strawberries on human health: insight into marginally discussed bioactive compounds for the Mediterranean diet. Public Health Nutr 12: 1656-1662 (2009). 13. Scalzo J, Battino M, Mezzetti B, Breeding and biotechnology for improving berry nutritional quality. Biofactors 23: 1-8 (2005). 14. Diamanti J, Capocasa F, Balducci F, Battino M, Hancock J, Mezzetti B, Increasing strawberry fruit sensorial and nutritional quality using wild and cultivated germplasm. PLoS ONE 7(10):e46470. doi:10.1371/journal.pone.0046470 (2012). 15. Pérez-de-Castro AM, Vilanova S, Cañizares J, Pascual L, Blanca JM, Díez MJ, Prohens J, Picó B, Application of genomic tools in plant breeding. Curr Genomics 13(3): 179-195 (2012). 16. Mezzetti B, Landi L, Pandolfini T, Spena A, The defH9-iaaM auxin-synthesizing gene increases plant fecundity and fruit production in strawberry and raspberry. BMC Biotechnol 15(4): 4 (2004).
This article is protected by copyright. All rights reserved
17. Matas AJ, Gapper NE, Chung MY, Giovannoni JJ, Rose JK, Biology and genetic engineering of fruit maturation for enhanced quality and shelf-life. Curr Opin Biotechnol 20(2): 197-203 (2009). 18. Qin Y, Teixeira da Silva JA, Zhang L, Zhang S, Transgenic strawberry: state of the art for improved traits. Biotechnol Adv 26: 219-232 (2008). 19. Salentijn EMJ, Aharoni A, Schaart JG, Boone MJ, Krens FA, Differential gene expression analysis of strawberry cultivars that differ in fruit-firmness. Physiol Plant 118: 571-578 (2003). 20. Agius F, Gonzales-Lamothe R, Caballero JL, Munoz-Blanco J, Botella MA, Valpuesta V,
Engineering increased vitamin C levels in plants by overexpression of a D-
galacturonic acid reductase. Nat Biotechnol 21: 177-181 (2003). 21. Griesser M, Hoffman T, Bellido ML, Rosati C, Fink B, Kurtzer R, Aharoni A, MunozBlanco J, Schwab W, Redirection of flavonoid biosynthesis through the down-regulation of an anthocyanidin glucosyltransferase in ripening strawberry fruit. Plant Physiol 146: 1528-1539 (2008). 22. Allan AC, Hellens RP, Laing WA, MYB transcription factors that colour our fruit. Trends Plant Sci 13: 99-102 (2008). 23. Rousseau-Gueutin M, Lerceteau-Kohler E, Barrot L, Sargent DJ, Monfort A, Simpson D, Arus P, Guerin G, Denoyes-Rothan B, Comparative genetic mapping between octoploid and diploid Fragaria species reveals a high level of colinearity between their genomes and the essentially disomic behavior of the cultivated octoploid strawberry. Genetics 179: 2045-2060 (2008). 24. Sargent DJ, Cipriani G, Vilanova S, Gil-Ariza D, Arús P, Simpson DW, Tobutt KR, Monfort A, The development of a bin mapping population and the selective mapping of 103 markers in the diploid Fragaria reference map. Genome 51(2): 120-127 (2008).
This article is protected by copyright. All rights reserved
25. Zorrilla-Fontanesi Y, Cabeza A, Dominguez P, Medina JJ, Valpuesta V, DenoyesRothan B, Sanchez-Sevilla JF, Amaya I, Quantitative trait loci and underlying candidate genes controlling agronomical and fruit quality traits in octoploid strawberry (Fragaria x ananassa). Theor Appl Genet 123: 755-778 (2011). 26. Wei YL, Li JN, Lu J, Tanq ZL, Pu DC, Chai YR, Molecular cloning of Brassica napus TRANSPARENT TESTA 2 gene family encoding potential MYB regulatory proteins of proanthocyanidins biosynthesis. Mol Biol Rep 34(2): 105-120 (2007). 27. Shulaev V, Sargent DJ, Crowhurst RN et al, The genome of woodland strawberry (Fragaria vesca). Nat Genet 43: 109–116 (2011). 28. Stewart D, McDougall GJ, Sungurtas J, Verrall S, Graham J, Martinussen I, Metabolomic approach to identifying bioactive compounds in berries: advances toward fruit nutritional enhancement. Mol Nutr Food Res 51: 645-651 (2007). 29. Zafra-Stone S, Yasmin T, Bagchi M, Chatterjee A, Vinson JA, Bagchi D, Berry anthocyanins as novel antioxidants in human health and disease prevention. Mol Nutr Food Res 51: 756-683 (2007). 30. Wang SY, Chen CT, Wang CY, The influence of light and maturity on fruit quality and flavonoid content of red raspberries. Food Chem 112: 676-684 (2009). 31. Jennings DL, Carmichael E, Anthocyanin variation in the genus Rubus. New Phytol 84: 505-513 (1980). 32. Kassim A, Poette J, Paterson A, Zait D, McCallum S, Woodhead M, Smith K, Hackett CA, Graham J, Environmental and seasonal influences on red raspberry anthocyanins antioxidant contents and identification of quantitative traits loci (QTL). Mol Nutr Food Res 53: 625-634 (2009).
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33. McCallum S, Woodhead M, Hackett CA, Kassim A, Paterson A, Graham J, Genetic and environmental effects influencing fruit colour and QTL analysis in raspberry. Theor Appl Genet 121: 611-627 (2010). 34. Alkharouf NW, Dhanaraj AL, Naik D, Overall C, Matthews BF, Rowland LJ, BBGD: an online database for blueberry genomic data. BMC Plant Biol 7: 5 (2007). 35. Jaakola L, Maatta K, Pirttila AM, Torronen R, Karenlampi S, Hohtola A, Expression of genes involved in anthocyanin biosynthesis in relation to anthocyanin, proanthocyanidin, and flavonol levels during bilberry fruit development. Plant Physiol 130: 729-739 (2002). 36. Szajdek A, Borowska JE, Bioactive compounds and health-promoting properties of berry fruits: a rewiew. Plant Foods Hum Nutr 63: 147-156 (2008). 37. Giampieri F, Alvarez-Suarez JM, Mazzoni L, Forbes-Hernandez TY, Gasparrini M, Gonzàlez-Paramàs AM, Santos-Buelga C, Quiles JL, Bompadre S, Mezzetti B, Battino M, Polyphenol-rich strawberry extract protects human dermal fibroblasts against hydrogen peroxide oxidative damage and improves mitochondrial functionality. Molecules 19: 7798-816 (2014). 38. Giampieri F, Alvarez-Suarez JM, Mazzoni L, Forbes-Hernandez TY, Gasparrini M, Gonzàlez-Paramàs AM, Santos-Buelga C, Quiles JL, Bompadre S, Mezzetti B, Battino M,An anthocyanin-rich strawberry extract protects against oxidative stress damage and improves mitochondrial functionality in human dermal fibroblasts exposed to an oxidizing agent. Food Funct 5: 1939-1948 (2014). 39. Tulipani S, Armeni T, Giampieri F, Alvarez-Suarez JM, Gonzalez-Paramás AM, SantosBuelga C, Busco F, Principato G, Bompadre S, Quiles JL, Mezzetti B, Battino M, Strawberry intake increases blood fluid, erythrocyte and mononuclear cell defenses against oxidative challenge. Food Chem 156: 87-93 (2014).
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40. Giampieri F, Alvarez-Suarez JM, Battino M, Strawberry and Human Health: Effects beyond Antioxidant Activity. J Agric Food Chem 62: 3867–3876 (2014). 41. Paredes-Lopez O, Cervantes-Ceja ML, Vigna-Perez M, Hernandez-Perez T, Berries: improving human health and healthy aging, and promoting quality life – A review. Plant Foods Hum Nutr 65: 299-308 (2010). 42. Aiyer HS, Vadhanam MV, Stoyanova R, Caprio GD, Clapper ML, Gupta RC, Dietary berries and ellagic acid prevent oxidative DNA damage and modulate expression of DNA repair genes. Int J Mol Sci 9: 327-341 (2008). 43. Aiyer HS, Kichambare S, Gupta RC Prevention of oxidative DNA damage by bioactive berry components. Nutr Cancer 60: 36-42 (2008). 44. Giampieri F, Alvarez-Suarez JM, Tulipani S, Gonzales-Paramas AM, Santos-Buelga C, Bompadre S, Quiles JL, Mezzetti B, Battino M, Photoprotective potential of strawberry (Fragaria x ananassa) extract against UV-A irradiation damage on human fibroblasts. J Agric Food Chem 60: 2322-2327 (2012). 45. Schantz M, Mohn C, Baum M, Richling E, Antioxidative efficiency of an anthocyanin rich bilberry extract in the human colon tumor cell lines Caco-2 and HT-29. J Berry Res 1: 25-33 (2010). 46. Stoner GD, Wang LS, Casto BC, Laboratory and clinical studies of cancer chemoprevention by antioxidants in berries. Carcinogenesis 29: 1665-1674 (2008). 47. Huang C, Li J, Song L, Zhang D, Tong Q, Ding M, Bowman L, Aziz R, Stoner GD, Black Raspberry extracts inhibit Benzo(a)Pyrene Diol-Epoxide-induced activator protein 1 activation and VEGF transcription by targeting the phosphotidylinositol 3-kinase/Akt pathway. Cancer Res 66: 581-587 (2006). 48. Neto CC, Cranberries: ripe for more cancer research? J Agric Food Chem 91: 2303-2307 (2011).
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49. Seeram NP, Berry fruits for cancer prevention: currents status and future prospects. J Agric Food Chem 56: 630-635 (2008). 50. Neto CC, Amoroso JW, Liberty AM, Anticancer activities of cranberry phytochemicals: an update. Mol Nutr Food Res 52: 18-27 (2008). 51. Boivin D, Blanchette M, Barrette S, Moghrabi A, Beliveau R, Inhibition of cancer cell proliferation and suppression of TNF-induced activation of NFrB by edible berry juice. Anticancer Res 27: 937-948 (2007). 52. Li J, Zhang D, Stoner GD, Huang C, Differential effects of black raspberry and strawberry extracts on BaPDE-induced activation of transcription factors and their target genes. Mol Carcinog 47: 286-294 (2008). 53. Chiva-Blanch G, Visioli F, Polyphenols and health: moving beyond antioxidants. J Berry Res 2: 63-71 (2012). 54. Wu QK, Koponen JM, Mykkanen HM, Torronen AR, Berry phenolics extracts modulate the expression of p21WAF1 and Bax but not Bcl-2 in HT-29 colon cancer cells. J Agric Food Chem 55: 1156-1163 (2007). 55. Seeram NP, Adams LS, Zhang Y, Lee R, Sand D, Scheuller HS, Heber D, Blackberry, black raspberry, blueberry, cranberry, red raspberry, and strawberry extracts inhibit growth and stimulate apoptosis of human cancer cells in vitro. J Agric Food Chem 54: 9329-9339 (2006). 56. Sharma M, Li L, Celver J, Killian C, Kovoor A, Seeram N, Effects of fruit ellagitannin extracts, ellagic acid, and their colonic metabolite, urolithin A, on Wnt Signaling. J Agric Food Chem 58: 3965-3969 (2010). 57. Umesalma S, Sudhandiran G, Ellagic acid prevents rat colon carcinogenesis induced by 1,2 dimethyl hydrazine through inhibition of AKT-phosphoinositide-3 kinase pathway. Mol Cell Pharmacol 660: 249-258 (2012).
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58. Juan ME, Alfaras I, Planas JM, Colorectal cancer chemoprotection by trans-resveratrol. Pharmacol Res 65: 584-591 (2012). 59. Mentor-Marcel RA, Bobe G, Dietary freeze-dried black raspberries in colorectal cancer patients. Nutr Cancer 64: 820-825 (2012). 60. Ravoori S, Vadhanam MV, Aqil F, Gupta RC, Inhibition of estrogen-mediated mammary tumorigenesis by blueberry and black raspberry. J Agric Food Chem 60: 5547-5555 (2012). 61. Aiyer HS, Gupta RC, Berries and ellagic acid prevent estrogen-induced mammary tumorigenesis by modulating enzymes of estrogen metabolism. Cancer Prev Res 3: 727737 (2010). 62. Aiyer H, Warri AM, Woode DR, Hilakivi-Clarke L, Clarke R, Influence of berrypolyphenols on receptor signaling and cell-death pathways: implication for breast cancer prevention. J Agric Food Chem 60: 5693-5708 (2012). 63. Kausar H, Jeyabalan J, Aqil F, Chabba D, Sidana J, Singh IP, Gupta RC, Berry anthocyanidins synergistically suppress growht and invasive potential of human nonsmall-cell lung cancer cells. Cancer Lett 325: 54-62 (2012). 64. Wang LS, Dombkowski AA, Seguin C, Rocha C, Cukovic D, Mukundan A, Henry C, Stoner GD, Mechanistic basis for the chemopreventive effects of black raspberries at a late stage of rat esophageal carcinogenesis. Mol Carcinog 50: 291-300 (2011). 65. Stoner GD, Wang LS, Chemoprevention of esophageal squamous cell carcinoma with berries. Top Curr Chem 329: 1-20. (2013). 66. Spagnuolo C, Russo M, Bilotto S, Tedesco I, Laratta B, Russo GL, Dietary polyphenols in cancer prevention: the example of the flavonoid quercetin in leukemia. Ann N Y Acad Sci 1259: 95-103 (2012).
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67. Hushmendy S, Jayakumar L, Hahn AB, Bhoiwala D, Bhoiwala DL, Crawfor DR, Select phytochemicals suppress human T-lymphocytes and mouse splenocytes suggesting their use in autoimmunity and transplantion. Nutr Res 29: 568-578 (2009). 68. Bae JY, Lim SS, Kim SJ, Choi JS, Park J, Ju SM, Han S.J, Kang IJ, Kan YH, Bog blueberry anthocyanins alleviate photoaging in ultraviolet-B irradiation-induced human dermal fibroblasts. Mol Nutr Food Res 53: 726-738 (2009). 69. Huang C, Zhang D, Li J, Tong Q, Stoner GD, Differential inhibition of UV-induced activation of NFκB and AP-1 by extracts from black raspberries, strawberries, and blueberries. Nutr Cancer 58: 205-212 (2007). 70. Duncan FJ, Martin JR, Wulff BC, Stoner GD, Tober KL, Oberyszyn TM, Kusewitt DF, Van Buskirk AM. Topical treatment with black raspberry extract reduces cutaneous UVB-induced carcinogenesis and inflammation. Cancer Prev Res (Phila) 2: 665-672 (2009). 71. Pan MH, Lai CS, Ho CT, Anti-inflammatory activity of natural dietary flavonoids. Food Funct 1: 15-31(2010). 72. Tribolo S, Lodi F, Connor C, Suri S, Wilson VG, Taylor MA, Needs PW, Kroon PA, Hughes DA Comparative effects of quercentin and its predominant human metabolites on adhesion molecule expression in activated human vascular endothelial cells. Atherosclerosis 197: 50-56 (2008). 73. Park MJ, Lee EK, Heo HS, Kim MS, Sung B, Kim MK, Lee J, Kim ND, Anton S, Choi JS, Yu BP, Chung HY, The anti-inflammatory effect of kaempferol in aged kidney tissues: the involvement of nuclear factor-kappaB via nuclear factor-inducing kinase/IkappaB kinase and mitogen-activated protein kinase pathways. J Med Food 12: 351-358 (2009).
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74. Oak MH, Bedoui JE, Madeira SV, Chalupsky K, Schini-Kerth VB, Delphinidin and cyanidin inhibit PDGF(AB)-induced VEGF release in vascular smooth muscle cells by preventing activation of p38 MAPK and JNK. Br J Pharmacol 149: 283-290 (2006). 75. Mauray A, Felgines C, Morand C, Mazur A, Scalbert A, Milenkovic D, Nutrigenomic analysis of the protective effects of bilberry anthocyanin-rich extract in apo E-deficient mice. Genes Nutr 4: 343-353 (2010). 76. Mauray A, Felgines C, Morand C, Mazur A, Scalbert A, Milenkovic D, Bilberry anthocyanin-rich extract alters expression of genes related to atherosclerosis development in aorta of apo E-deficient mice. Nutr Metab Cardiovasc Dis 22: 72-80 (2012). 77. Cassidy A, Mukamal KJ, Liu L, Franz M, Eliassen AH, Rimm EB, High anthocyanin intake is associated with a reduced risk of myocardial infarction in young and middleaged women. Circulation 127: 188-196 (2013). 78. Alvarez-Suarez JM, Giampieri F, Tulipani S, Casoli T, Di Stefano G, González-Paramás AM, Santos-Buelga C, Busco F, Quiles JL, Cordero MD, Bompadre S, Mezzetti B, Battino M, One-month strawberry-rich anthocyanin supplementation ameliorates cardiovascular risk, oxidative stress markers and platelet activation in humans. J Nutr Biochem 25: 289-294 (2014). 79. Qin Y, Xia M, Ma J, Hao Y, Liu J, Mou H, Cao L, Ling W, Anthocyanin supplementation improves serum LDL- and HDL-cholesterol concentrations associated with the inhibition of cholesteryl ester transfer protein in dyslipidemic subjects. Am J Clin Nutr 90: 485-492 (2009). 80. Spencer JPE, Vafeiadou K, Williams RJ, Vauzour D, Neuroinflammation: modulation by flavonoids and mechanism of action. Mol Aspects Med 33: 83-97 (2012).
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81. Vauzour D, Vafeiadou K, Rendeiro C, Corona G, Spencer JPE, The inhibitory effects of berry-derived flavonoids against neurodegenerative processes. J Berry Res 1: 45-52 (2010). 82. Joseph JA, Shukitt-Hale B, Willis LM, Grape juice, berries, and walnuts affect brain aging and behavior. J Nutr 139: 1813-1817 (2009). 83. Poulose SM, Carey AN, Shukitt-Hale B, Improving brain signaling in aging: Could berries be the answer? Expert Rev Neurother 12: 887-889 (2012). 84. Joseph JA, Shukitt-Hale B, Denisova NA, Prior RL, Cao G, Martin A, Taglialatela G, Bickford PC, Long-term dietary strawberry, spinach, or vitamin E supplementation retards the onset of age-related neuronal signal-transduction and cognitive behavioral deficits. J Neurosci 18: 8047-8055 (1998). 85. Joseph JA, Shukitt-Hale B, Denisova NA, Bielinski D, Martin A, McEwen JJ, Bickford PC, Reversals of age-related declines in neuronal signal transduction, cognitive, and motor behavioral deficits with blueberry, spinach, or strawberry dietary supplementation. J Neurosci. 15: 8114-21 (1999). 86. Shukitt-Hale B, Cheng V, Bielinski D, Carey AN, Lau FC, Seeram NP, Heber D, Joseph JA, Differential brain regional specificity to blueberry and strawberry polyphenols in improved motor and cognitive function in aged rats. Soc Neurosci Abstr 32: 81.15 (2006). 87. Williams RJ, Spencer JPE, Flavonoids, cognition and dementia: actions, mechanisms, and potential therapeutic utility for Alzheimer disease. Free Radic Biol Med 52: 35-45 (2012). 88. Zhu M, Hu J, Perez E, Phillips D, Kim W, Ghaedian R, Napora JK, Zou S, Effects of long-term cranberry supplementation on endocrine pancreas in aging rats. J Gerontol A Biol Sci Med Sci 66A: 1139-1151 (2011).
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ILLUSTRATIONS
Figure 1: Homoeeology grouups (HGs, named n withh roman nuumerals as the correspponding Linkagee Groups inn the diploidd Fragaria reference r map24) follow m wed by an F (for femalle, LGs) or an M (male Linnkage Groupps). Linkage Groups within w each HG H were nuumbered arrbitrarily althouggh the same number in male and feemale maps indicates thhe homologgous groupss. On the left, thee size of LG Gs are indiccated (in cenntimorgans)). On the riight, the maarker and thhe QTLs are listeed. Source: Zorrilla-Fonntanesi et al. 2011(25) with w some modification m ns.
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Figure 2: Mappedd QTLs in raspberry. r On O the top, LG 1 is reepresented: bHLH, b invoolved in the antthocyanin pathway, p iss the mostt importantt marker. Below, B LG G 4 is shoown and FRUITE E4, also invvolved in thhe anthocyaanin pathwaay, is the most m importaant marker. Source: Kassim m et al. 2009(32) with som me modificaations.
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1
Dip. Scienze Cliniche Specialistiche ed Odontostomatologiche (DISCO), Facoltà di
Medicina, Università Politecnica delle Marche, Ancona, Italy 2
Dep. Physiology, Institute of Nutrition and Food Technology ‘‘José Mataix”, Biomedical
Research Center, University of Granada, Granada, Spain 3
Dip. Scienze Agrarie, Alimentari e Ambientali, Università Politecnica delle Marche,
Ancona, Italy 4
Facultad de Ciencias de la Salud, Universidad Nacional de Chimborazo, Riobamba -
Ecuador 5
Area de Nutrición y Salud, Universidad Internacional Iberoamericana (UNINI), Campeche,
C.P.24040, Mexico 6
Director Centre for Nutrition & Health, Universidad Europea del Atlantico (UEA),
Santander 39011, Spain
*Correspondence
to:
Prof.
Maurizio
Battino,
PhD,
DSc,
MS,
MD
([email protected]) or Dr. Francesca Giampieri ([email protected]),
(Hon) DISCO,
Facoltà di Medicina, Università Politecnica delle Marche, Via Ranieri 65, 60131 Ancona, Italy, Telephone +39 071 2204646/2204136; fax +39 071 2204123.
This article has been accepted for publication and undergone full peer review but has not been through the copyediting, typesetting, pagination and proofreading process, which may lead to differences between this version and the Version of Record. Please cite this article as doi: 10.1002/jsfa.7216
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ABSTRACT Berries are a relevant source of micronutrients and nonessential phytochemicals such as polyphenol compounds, that play a synergistic and cumulative role in human health promotion. Several systematic analyses showed that berry phenolics are able to detoxify ROS/RNS blocking their production, to intervene in the cell cycle, participating in the transduction and expression of genes involved in apoptosis, and to repair oxidative DNA damage. As a consequence, the improvement of the nutritional quality of berries has become a new quality target of breeding and biotechnological strategies, to control or to increase the content of specific health-related compounds in fruits. This work reviews, on the basis of the in vitro and in vivo evidences, the main berries phytochemical compounds and their possible mechanisms of action on pathways involved in several type of diseases, with particular attention to cancer, inflammation, neurodegeneration, diabetes and cardiovascular diseases.
KEYWORDS berries, bioactive compounds, genetic improvement, cancer, inflammation, neurological diseases.
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INTRODUCTION The importance of diet in the prevention of several chronic diseases has been widely recognized. The assumption is that consumption of fruit and vegetables is correlated with a high intake of antioxidant and bioactive compounds that play a crucial role in the prevention of a wide range of diseases, such as cancer, cardiovascular, and other age-related degenerative pathologies1-3. The aging process is common for most multicellular organisms, and consists of a gradual impairment of the biological function of all organs in the postreproduction phase, compromising the homeostasis preservation and resulting in an increased risk of death. Research has been directed to the delineation of the most critical factors that may account for a wide range of age-related diseases (ARD), suggesting that one of the major causes of the incidence of age-related illnesses as skin aging, neurodegenerative pathologies including Lou Gehrig’s, Parkinson’s, Alzheimer’s and Huntington’s diseases, cancer and cardiovascular diseases (CVD), is the long-term exposition to oxidative stress (OS)4. OS is caused by an imbalance between the systemic production of reactive oxygen and nitrogen species (ROS and RNS) and the ability of the biological system to detoxify or to repair the ROS/RNS induced damage5. Studies monitoring ROS/RNS production and antioxidant defenses indicate that OS, with the consequent mitochondrial respiration impairment and mitochondrial damage, is related with the pathophysiology of these diseases6. A balanced diet, rich in antioxidant compounds derived from fruits and vegetables, as the Mediterranean one, can deeply influence the susceptibility to OS, counteracting the decrease in antioxidant protection that occurs during illness7,8. Berries are a common and important fruit in the Mediterranean diet and are among the richest fruits in nutritive compounds, including minerals, vitamins, dietary fibers, and especially polyphenolic phytochemicals (flavonoids, phenolic acids, lignans and tannins)9-11, which exhibit strong antioxidant and anti-inflammatory activities that may reduce sensitivity to
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OS12. Therefore, berries may have a role in the prevention of degenerative pathologies, stimulating researchers to implement new biotechnological approaches for the improvement of bioactive compound contents13-14. For example the use of wild germplasm in the genetic improvement programs for the creation of new commercial strawberries with increased antioxidant content is an important option to support a higher intake of healthy compounds even with a lower fruit consumption13, giving that commercial strawberries deriving from wild germplasm breeding programs resulted rich in antioxidant compounds. Two important areas of research are therefore (i) the development of new berry varieties or the improvement of current varieties with improved levels of bioactive compounds production (ii) determination of the mechanisms of bioactive compounds on gene expression in humans to identify which compounds are useful, particularly those with protective effects against ROS/RNS. Both lines of investigation are of critical importance for obtaining consistent results.
BREEDING AND BIOTECHNOLOGICAL PROGRAMS IN BERRIES High throughput technologies for plant genotyping, such as those developed for the detection of Single Nucleotide Polymorphism, accelerated the fine mapping of Quantitative Trait Loci (QTLs) and the positional cloning of the genes responsible for traits involved in berry nutritional quality. The correct integration between genetic resources, genetic improvement and transgenic approaches is essential to obtain new varieties that can help to reach the desired nutritional quality in berries15-17. In the last few years, fruit sensorial and nutritional traits have become the major breeding targets in the commercial octoploid strawberry (Fragaria x ananassa). The first works on the genetic engineering of strawberry were focused on the improvement of agronomical or qualitative (but not nutritional) traits, like size, number, yield, ripening, softening and
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firmness of fruit18,19. Recently efforts have focused on improved nutritional value of strawberry. A first interesting result was the demonstration that the biosynthesis of L-ascorbic acid in ripe strawberry can occur through D-galacturonic acid, encoded by the gene GaiUR20. Another study demonstrate, for the first time, that the rolC gene lead to an improvement of fruit nutritional quality in strawberry, mainly sugar content and total antioxidant capacity13. One of the most important group of compounds with healthy properties are anthocyanidins. A study demonstrated that at least two different enzymes are involved in the glucosylation of anthocyanidins in strawberry: a transferase, which forms the glucosides at early ripening stages, and a glycosyl-transferase (encoded by FaGT1 gene) which catalyzes the glucoside transfer in ripe strawberry fruits21. The anthocyanin accumulation, instead, is regulated by the coordinated expression of genes encoding the flavonoid biosynthetic pathway enzymes, the R2R3MYB and the bHLH transcription factors22. For further improvement projects concerning the octoploid strawberry, the construction of the most comprehensive genetic map for Fragaria x ananassa was a crucial step23. In this study a first reference map for octoploid Fragaria was provided and almost all linkage groups were ranged into the seven expected homoeologous groups (HG). A successive three years study identifies 33 QTLs of 14 studied traits into the seven HGs expected in Fragaria. In addition to QTLs for titratable acidity, pH, plant width, color parameters and other characteristics, in this map the genes and the QTLs for the control of the healthy compounds were indicated: for ascorbic acid, three QTLs (LAAV-M.1, LAAVII-M.1 e LAAIV-F.2), and FaGaiUR gene in the linkage groups (LGs) belonging to HG IV were identified; for anthocyanin, a cluster of QTLs was identified in LG V-M.2, another QTL in LG III-F.1, the anthocyanin biosynthetic gene F3H in HG I and other genes involved in anthocyanin biosynthesis (CHI, ChFaM247, ChFvM234 or FaMYB1) in HG V (Figure 1)24,25. Furthermore, in the last year, an international consortium sequenced the genome of the diploid strawberry (Fragaria vesca), that permitted the access to genomic
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information relevant to Rosaceae, especially for fruit quality (flavor, nutrition and aroma). In particular, for the nutritional quality, some genes have been detected for the proanthocyanidin levels in the seed (FvMyb33)26, for the content of l-ascorbate (FaGaiUR and FaGLHD), for flavonoid metabolism (FaGT6 and FaGT7), for polyphenols anchoring (Fahyprp) and for flavonoid biosynthesis (Fraa1A)27. Genetic transformation in strawberry offers an effective method for creating new varieties that selectively targets a specific interested gene, with particular attention to the nutritional quality of the fruit. Another high-value berry species is raspberry, with a demonstrated beneficial impact on human health28,29. The main characters, objects of genetic improvement, are the visual parameters, in particular color. This parameter, however, not only influenced the appearance and the flavor of the fruit, but is also closely linked to the anthocyanin content (in particular cyanidin and pelargonidin) of the berry30. Since 1980, genes involved in the synthesis of sugars rhamnose, sophorose and xylose (required to give the array of different anthocyanin pigments observed in red raspberry) were detected31. Only recently, genes involved in the anthocyanin
pathway
(bHLH
and
FRUITE4)
and
in
the
flavonol
pathway
(ERubLR_SQ07_2_H02) were found. The bHLH was mapped in the LG1 and the bZIP (FRUITE4) gene on LG4 (Figure 2)32,33. Blueberry, a major berry crop in the United States, has great nutritional and economical value. The bigger problem of its cultivation is the low temperature that reduces crop yields and causes major losses. So a blueberry genomics database was developed to help the researchers to establish new varieties with a better cold acclimation and freeze tolerance34. In the last years, the interest on the nutritional value of blueberry has increased, in particular for the flavonoid biosynthesis. A first study in 2002 detected the genes involved in the flavonoid pathway of bilberry: Phe ammonia-lyase (PAL), chalcone synthase (CHS), flavanone 3hydroxylase (F3H), dihydroflavonol 4-reductase (DFR) and anthocyanidin synthase (ANS)35.
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This latter gene demonstrate a correlation between anthocyanin accumulation and expression of the flavonoid pathway genes during the ripening of berries.
THE GENETIC MECHANISM OF ACTION OF BERRIES BIOACTIVE COMPOUNDS Numerous physiological processes produce reactive oxidant species and free radicals which may endanger the health and favor the onset of several diseases. So often the body needs exogenous sources of antioxidant defenses, mainly supplied by the diet. Berries, as previously discussed, are rich in bioactive compounds36 and their antioxidant activity can act along many mechanisms, such as detoxifying ROS and blocking of ROS production37-39, intervening in the cell cycle and participating in the transduction and expression of genes involving in apoptosis, suppression of tumors, and repair of oxidative DNA damage, that is the first step for the initiation of cancer40. Therefore the ability of berries to reduce oxidative DNA damage may be considered a good therapy for cancer prevention. It has been demonstrated that berry compounds modulate the expression of genes involved in DNA damage41: on the one hand, berries rich diet over-expressed DNA repair genes, such as XPA, DNL3, ERCC5, CYP450, CYP3A442,43, and on the other hand, genes involved in the DNA damage, such as MAPK14 and MAPKK, are down-regulated42. Over the last few years, many researches have demonstrated cancer chemopreventive activities of berries. In studies of the anti-tumoral activity of berries, work has focused on up or down regulation of different genes involved in inhibition of carcinogen activation, stimulation of carcinogen detoxification, scavenging of oxygen reactive species, control of cell-cycle and cell proliferation, induction of apoptosis, inhibition of oncogenes and metastasis44,45. Berry compounds have been shown to regulate several pathways including nuclear factor kB (NF-κB) and cyclooxygenases (COX-2), cell adhesion proteins46,
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phosphoinositide 3-kinase (PI-3K)/protein kinase B (Akt), mitogen-activated protein kinase (MAPK), p38, vascular endothelial growth factor (VEGF-1) for angiogenesis47 and Bcl-2, p21, Bax, cytochrome c and caspase 3, which regulate cellular apoptosis48,49. A study on cranberries confirms the ability of berries to inhibit growth and proliferation of tumor cells reducing ornithine decarboxylase and COX-2 activity, and matrix metalloproteases expression50. COX-2 is implicated also in the anti-inflammatory capacity of berry juice, which inhibits tumor necrosis factor (TNF)-induced expression of COX-2 mediated by the nuclear expression factor NF-kB51. This research group also showed that berry juice inhibits the proliferation of five cancer cell lines by reducing the expression of key regulators of cellcycle (cdk4, cdk6, cyclin D1 and cyclin D3)51. Blackberry extract has the ability to inhibit both VEGF and AP-1 expression mediated by the PI-3K/Akt pathway in presence of benzo(a)pyrene (BaP), a potent cancerogen47. Meanwhile, raspberries and strawberries inhibit BaP-induced Nuclear factor of activated T-cells (NFAT) activation and TNF-alpha (α) expression52. Furthermore, berries stimulate the expression of cytoprotective genes, such as nuclear factor-E2-related factor 2 (Nrf2) which is over-expressed by berry polyphenols and binds to the antioxidant response element in cells53. A range of studies have been carried out to establish the anticancer activity of berries and their bioactive compounds. In colon cancer cells, berry phenolic extracts inhibit cell growth by over-expression of p21WAF1, a member of the cyclin kinase inhibitors54, and stimulates apoptosis through COX-2 expression55. Ellagitannins also showed beneficial effects against colon carcinogenesis by inhibition of Wnt signaling56 and induction of apoptosis by suppression the PI3K/Akt pathway57. Resveratrol has been shown to inhibit proliferation by over expression of sirtuin 1 (SIRT1) and inducing apoptosis by activation of MAPK, which downstream caspases activation lead to cell death58. Recent research has related black raspberry consumption with the decrease in plasma concentrations of interleukine 8 (IL-8)
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and granulocyte macrophage colony stimulating factor (GM-CSF), which suggest beneficial properties of berries treatment59. In relation to breast cancer, several studies indicate the efficacy of berries consumption in the prevention and treatment of mammalian tumors. Berries prevent estradiol-induced mammary tumors by down-regulation the levels of enzymes to estradiol metabolism, such as CYP1A1, P450, 17beta-hydroxysteroid dehydrogenase type 7 (17βHSD7) and estrogen receptor alpha (ERα)60,61, and binding to receptor pathways which participate in breast cancer development, such as ER and tyrosine kinase receptor (TKR), which reduces phosphorylation of several kinases such as PI3K/Akt and extracellular signal-regulated kinase (ERK)/MAPK and inhibits grown factor receptor (GRF) activation, decreasing the tumor development62. With regard to other types of cancer, it was found that the combination of distinct types of berry fruits suppress the growth and invasion capacity of lung cancer cells by downregulation of WNT pathways and different targets of cell cycle progression, such as cyclin D1, cyclin B1, pERK, Matrix metallopeptidase 9 (MMP9) and VEGF63. In addition, berry fruits suppress TNFα-induced NF-κB activation, decreasing cell invasion. Finally, Wang et al.64 have gone a step further using DNA microarray technology for the determination of the gene expression profile on rat esophageal carcinogenesis treated with black raspberries. They have found that the treatment with berries promotes changes in the expression of genes implicated in proliferation, apoptosis, inflammation, angiogenesis and metastasis, among others. The mainly chemoprotective effect of berries on esophageal squamous cell seems to initiate with the inhibition of inducible nitric oxide synthase (iNOS), COX-2, VEGF, NF-κB and AP-1 pathway65. Quercetin, one of the main flavonoids which is a common component of berry fruits, is able to induce apoptosis in leukemia cell lines and B cells of chronic lymphocytic leukemia (BCLL) by down-regulation of myeloid leukemia cell differentiation protein (Mcl-1)
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expression, an antiapoptotic proteins of Bcl-2 family66. Furthermore, berry compounds have been studied as phytochemical suppressors of human T-lymphocytes by decreasing of IL-2 production, which could be used for the treatment of autoimmune and transplant patients67. Exposure to UV-A and UV-B radiation is known to induce DNA damage and reactive oxygen species production which could lead to skin diseases. Giampieri et al.44 showed that strawberry extracts provide photoprotective effects to human fibroblast in vitro against UV-A irradiation damage, according with Bae et al.68, who found that blueberry anthocyanins prevented UV-B-induced skin photoaging by decreasing collagen destruction and inflammatory responses via NF-κB and MAPK signaling. Cyanidin-3-rutinoside, an anthocyanin highly found in black raspberry, is potentially able to protect UV-induced skin damage, by down-regulation of NF-κB and AP-1 activation, inhibition of IL-8 and prevention of pro-caspase-3 cleavage69. Similarly, black raspberry extracts (BRE), topically applied, was effective at reducing the tumor number and the average tumor size in female SKH-1 hairless mice exposed to one minimal erythemal dose of UV-B thrice weekly on nonconsecutive days for 25 weeks and that the reduction was correlated with a significant reduction in tumorinfiltrating CD3+foxp3+ regulatory T-cells70. The same authors found that topical BRE treatment in mice exposed to a single minimal erythemal dose of UV-B significantly reduces edema, p53 protein levels, oxidative DNA damage, and neutrophil activation, highlighting the berry efficacy in the prevention of skin cancers. In addition to the anticancer properties, berry compounds have also shown anti-inflammatory capacity. In general, they are capable of blocking NF-κB (which is involved in decreasing PI3K/Akt signaling), lipopolysaccharide(LPS)-induced pro-inflammatory cytokines, hypoxiainducible factor 1-alpha (HIF-1α) and VEGF expression and TNF-α production. They also can inhibit activation of MAPK (down-regulating COX-2 and IL-6 expression) that participates in the down-regulation of TNF-α production71. Moreover, it has been shown the
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capacity of flavonols to up regulate VCAM-1, ICAM-1 and MCP-1 gene expression and decrease inflammatory markers, such as IL-1alpha (α) and IL-472. Inhibition of COX-2, iNOS and TNF-α have been shown in rats fed with rich-flavonols diet73. The positive effects of antocyanindins were also demonstrated: they reduce prostaglandin E2 (PGE2) levels and platelet-derived growth factor (PDGF)-induced VEGF expression by down-regulation of p38 MAPK and c-Jun N-terminal kinases (JNK) pathways74. All these anti-inflammatory mechanisms of berries have been associated with their beneficial actions in several diseases, including cardiovascular diseases, brain inflammatory stress, obesity and metabolic syndrome. Recent studies demonstrated that bilberry attenuates atherosclerosis lesion in mouse by down-regulation of genes implicated in oxidative stress, inflammation, and angiogenesis, such as AOX1, CYP2E1, TXNIP, JAM-A and VEGRF275,76. Furthermore, berry fruits modulate angiogenesis via PI3K/Akt pathway, improving plasma lipid profile, increasing plasma antioxidant activity and low-density lipoprotein (LDL) resistance to oxidation, and ameliorating endothelial function11. A recent meta-analysis also showed that high anthocyanin intake is associated with a reduction of risk of myocardial infarction (MI) and an inverse association between higher intake of anthocyanins and risk of MI was observed77. At the same time, a strawberry-enriched diet has shown to significantly improve the markers of oxidative stress, by decreasing lipid peroxidation oxidation and ameliorating the serum lipid profile of the subjects, through a reduction of total cholesterol, LDL-C and triglyceride levels, both in healthy78 and in dyslipidemic subjects79.Therefore, the berries polyphenols seem to be the most likely constituents, which exert in vivo effects in the prevention of the CVD risk. In brain, neuroinflammation and neurodegeneration are mediated by berry compounds actuation over NF-κB cascade and MAPK signaling, PI3K/Akt pathway80 and TNFα gene
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expression inhibition81. Berries bioactive compounds may also have a direct effect on signaling to enhance neuronal communication, a buffer capacity against excess of calcium, an enhancement of neuroprotective stress shock proteins and reduction of stress signals. Furthermore, anthocyanins not only have the capacity of scavenge free radicals in the brain, but also to affect intraneural signaling, to inhibit lipid peroxidation and inflammatory mediators COX-1 and COX-282. Flavonoids are also capable to regulate the transcription of antioxidant enzymes related to glutathione via ERK1 and ERK2, which exert an important role in some types of memory83. Some of the deleterious effects of OS in neurodegenerative diseases could be retarded or even reversed by increasing antioxidant levels, and the putative synergistic effects of antioxidants combinations is particularly effective in this regard84. In a two long-term studies on rats, strawberry and blueberry prevented retarded age-related declines in neuronal and cognitive functions: berries supplementation had the greatest effect on GTP-ase activity, and its association with vitamin E showed significant protection against these age-induced deficits also on the other parameters studied, like the motor behavioral and the Morris water maze performances84,85. Moreover, dietary supplementation with blueberry or strawberry extracts in rats, prior the exposition to whole-body irradiation with high-energy and charge particles, showed the ability to retard/reverse age-related deficits in behavior and signal transduction caused by the irradiation. In particular, the strawberry-fed rats showed a better protection against spatial deficits in the maze, and animals were better able to retain place information (a hippocampally mediated behavior) compared to controls, while blueberry diet appeared to improve the reversal learning, a behavior more dependent on intact striatal function86. Recently, human studies suggest that polyphenols present in berries may prevent the occurrence of dementia and improve memory indices83. These findings suggest that berries phytochemicals can be beneficial in retarding age-related functional and
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cognitive-behavioral deficits, as well as in reversing the course of neuronal aging in neurodegenerative diseases, mainly Alzheimer disease87. Finally, because diabetes mellitus is a metabolic disease often associated with aging, the effect of berries on this pathology is also interesting. An in vivo study on rats has demonstrated that a dietary intake of cranberries may protect the pancreatic islets from the morphological and functional decline. In fact, the cranberries consumption inhibited two enzymes required for starch digestion to maltodextrins and thereby conversion into absorbable monosaccharides. The inhibition of these enzymes improved metabolic homeostasis, delaying the age-related decline in basal insulin, ameliorating pancreatic beta (β)-cell glucose responsiveness and augmenting β-cell mass in normal aged rats88.
CONCLUSIONS Several evidences from in vitro and in vivo studies suggest that berry phytochemicals are able to counteract OS, to protect DNA and to regulate the cellular metabolism and apoptosis. Their anti-inflammatory properties can also decrease the risk of developing chronic ARD. The results here presented show the role of berries compounds in regulating cellular processes related to OS, inflammation and cancer proliferation, including the expression of key genes involved in these pathways, providing an insight on the potential of berries as important allies, in the context of a balanced diet such as the Mediterranean one, to protect or reduce the risk and the occurrence of several chronic pathologies. The integration of breeding knowledge with the availability of complete genome will represent in the next future the most efficient approach to improve nutritional and nutraceutical quality for healthier fruits.
ACKNOWLEDGEMENTS
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The authors would like to thank to Ms. Monica Glebocki for extensive editing of the manuscript.
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REFERENCES
1.
Yi-Fang C, Jie S, Xianzhong W, Rui Hai L, Antioxidant and antiproliferative activities of common vegetables. J Agric Food Chem 50: 6010-6916 (2002).
2.
Etminan M, Takkouche B, Caamano-Isorna F, The role of tomato products and licopene in the prevention of prostate cancer: a meta-analysis of observational studies. Cancer Epidemiol Biomarkers Prev 13 (3) (2004).
3.
Tulipani S, Romandini S, Busco F, Bompadre S, Mezzetti B, Battino M, Ascorbate, not urate, modulates the plasma antioxidant capacity after strawberry intake. Food Chem 117: 181-188 (2009).
4.
Joseph JA, Shukitt-Hale B, Denisova NA, Bielinski D, Martin A, McEwen JJ, Bickford PC, Reversals of age-related declines in neuronal signal transduction, cognitive, and motor behavioral deficits with blueberry, spinach, or strawberry dietary supplementation. J Neurosci 15: 8114-8121 (1999).
5.
Patel VP, Chu CT, Nuclear transport, oxidative stress, and neurodegeneration. Int J Clin Exp Pathol 4: 215-229 (2011).
6.
Ramalingam M, Kim SJ, Reactive oxygen/nitrogen species and their functional correlations in neurodegenerative diseases. J Neural Transm 119: 891-910 (2012).
7.
Bach-Faig A, Berry EM, Lairon D, Reguant J, Trichopoulou A, Dernini S, Medina FX, Battino M, Belahsen R, Miranda G, Serra-Majem L; Mediterranean Diet Foundation Expert Group, Mediterranean diet pyramid today. Science and cultural updates. Public Health Nutr 14: 2274-2284 (2011).
8.
Requant-Aleix J, Arbore MR, Bach-Faig A, Serra-Majem L, Mediterranean heritage: an intangible cultural heritage. Public Health Nutr 12: 1591-1594 (2009).
This article is protected by copyright. All rights reserved
9.
Battino M, Beekwilder J, Denoyes-Rothan B, Laimer M, McDougall GJ, Mezzetti B, Bioactive compounds in berries relevant to human health. Nutr Rev 67: Suppl. 1, S145S150 (2009).
10. Giampieri F, Alvarez-Suarez JM, Mazzoni L, Romandini S, Bompadre S, Diamanti J, Capocasa F, Mezzetti B, Quiles JL, Ferreiro MS, Tulipani S, Battino M, The potential impact of strawberry on human health. Nat Prod Res 27: 448-455 (2013). 11. Giampieri F, Tulipani S, Alvarez-Suarez JM, Quiles JM, Mezzetti B, Battino M, The strawberry: composition, nutritional quality, and impact on human health. Nutrition 28: 9-19 (2012). 12. Tulipani S, Mezzetti B, Battino M, Impact of strawberries on human health: insight into marginally discussed bioactive compounds for the Mediterranean diet. Public Health Nutr 12: 1656-1662 (2009). 13. Scalzo J, Battino M, Mezzetti B, Breeding and biotechnology for improving berry nutritional quality. Biofactors 23: 1-8 (2005). 14. Diamanti J, Capocasa F, Balducci F, Battino M, Hancock J, Mezzetti B, Increasing strawberry fruit sensorial and nutritional quality using wild and cultivated germplasm. PLoS ONE 7(10):e46470. doi:10.1371/journal.pone.0046470 (2012). 15. Pérez-de-Castro AM, Vilanova S, Cañizares J, Pascual L, Blanca JM, Díez MJ, Prohens J, Picó B, Application of genomic tools in plant breeding. Curr Genomics 13(3): 179-195 (2012). 16. Mezzetti B, Landi L, Pandolfini T, Spena A, The defH9-iaaM auxin-synthesizing gene increases plant fecundity and fruit production in strawberry and raspberry. BMC Biotechnol 15(4): 4 (2004).
This article is protected by copyright. All rights reserved
17. Matas AJ, Gapper NE, Chung MY, Giovannoni JJ, Rose JK, Biology and genetic engineering of fruit maturation for enhanced quality and shelf-life. Curr Opin Biotechnol 20(2): 197-203 (2009). 18. Qin Y, Teixeira da Silva JA, Zhang L, Zhang S, Transgenic strawberry: state of the art for improved traits. Biotechnol Adv 26: 219-232 (2008). 19. Salentijn EMJ, Aharoni A, Schaart JG, Boone MJ, Krens FA, Differential gene expression analysis of strawberry cultivars that differ in fruit-firmness. Physiol Plant 118: 571-578 (2003). 20. Agius F, Gonzales-Lamothe R, Caballero JL, Munoz-Blanco J, Botella MA, Valpuesta V,
Engineering increased vitamin C levels in plants by overexpression of a D-
galacturonic acid reductase. Nat Biotechnol 21: 177-181 (2003). 21. Griesser M, Hoffman T, Bellido ML, Rosati C, Fink B, Kurtzer R, Aharoni A, MunozBlanco J, Schwab W, Redirection of flavonoid biosynthesis through the down-regulation of an anthocyanidin glucosyltransferase in ripening strawberry fruit. Plant Physiol 146: 1528-1539 (2008). 22. Allan AC, Hellens RP, Laing WA, MYB transcription factors that colour our fruit. Trends Plant Sci 13: 99-102 (2008). 23. Rousseau-Gueutin M, Lerceteau-Kohler E, Barrot L, Sargent DJ, Monfort A, Simpson D, Arus P, Guerin G, Denoyes-Rothan B, Comparative genetic mapping between octoploid and diploid Fragaria species reveals a high level of colinearity between their genomes and the essentially disomic behavior of the cultivated octoploid strawberry. Genetics 179: 2045-2060 (2008). 24. Sargent DJ, Cipriani G, Vilanova S, Gil-Ariza D, Arús P, Simpson DW, Tobutt KR, Monfort A, The development of a bin mapping population and the selective mapping of 103 markers in the diploid Fragaria reference map. Genome 51(2): 120-127 (2008).
This article is protected by copyright. All rights reserved
25. Zorrilla-Fontanesi Y, Cabeza A, Dominguez P, Medina JJ, Valpuesta V, DenoyesRothan B, Sanchez-Sevilla JF, Amaya I, Quantitative trait loci and underlying candidate genes controlling agronomical and fruit quality traits in octoploid strawberry (Fragaria x ananassa). Theor Appl Genet 123: 755-778 (2011). 26. Wei YL, Li JN, Lu J, Tanq ZL, Pu DC, Chai YR, Molecular cloning of Brassica napus TRANSPARENT TESTA 2 gene family encoding potential MYB regulatory proteins of proanthocyanidins biosynthesis. Mol Biol Rep 34(2): 105-120 (2007). 27. Shulaev V, Sargent DJ, Crowhurst RN et al, The genome of woodland strawberry (Fragaria vesca). Nat Genet 43: 109–116 (2011). 28. Stewart D, McDougall GJ, Sungurtas J, Verrall S, Graham J, Martinussen I, Metabolomic approach to identifying bioactive compounds in berries: advances toward fruit nutritional enhancement. Mol Nutr Food Res 51: 645-651 (2007). 29. Zafra-Stone S, Yasmin T, Bagchi M, Chatterjee A, Vinson JA, Bagchi D, Berry anthocyanins as novel antioxidants in human health and disease prevention. Mol Nutr Food Res 51: 756-683 (2007). 30. Wang SY, Chen CT, Wang CY, The influence of light and maturity on fruit quality and flavonoid content of red raspberries. Food Chem 112: 676-684 (2009). 31. Jennings DL, Carmichael E, Anthocyanin variation in the genus Rubus. New Phytol 84: 505-513 (1980). 32. Kassim A, Poette J, Paterson A, Zait D, McCallum S, Woodhead M, Smith K, Hackett CA, Graham J, Environmental and seasonal influences on red raspberry anthocyanins antioxidant contents and identification of quantitative traits loci (QTL). Mol Nutr Food Res 53: 625-634 (2009).
This article is protected by copyright. All rights reserved
33. McCallum S, Woodhead M, Hackett CA, Kassim A, Paterson A, Graham J, Genetic and environmental effects influencing fruit colour and QTL analysis in raspberry. Theor Appl Genet 121: 611-627 (2010). 34. Alkharouf NW, Dhanaraj AL, Naik D, Overall C, Matthews BF, Rowland LJ, BBGD: an online database for blueberry genomic data. BMC Plant Biol 7: 5 (2007). 35. Jaakola L, Maatta K, Pirttila AM, Torronen R, Karenlampi S, Hohtola A, Expression of genes involved in anthocyanin biosynthesis in relation to anthocyanin, proanthocyanidin, and flavonol levels during bilberry fruit development. Plant Physiol 130: 729-739 (2002). 36. Szajdek A, Borowska JE, Bioactive compounds and health-promoting properties of berry fruits: a rewiew. Plant Foods Hum Nutr 63: 147-156 (2008). 37. Giampieri F, Alvarez-Suarez JM, Mazzoni L, Forbes-Hernandez TY, Gasparrini M, Gonzàlez-Paramàs AM, Santos-Buelga C, Quiles JL, Bompadre S, Mezzetti B, Battino M, Polyphenol-rich strawberry extract protects human dermal fibroblasts against hydrogen peroxide oxidative damage and improves mitochondrial functionality. Molecules 19: 7798-816 (2014). 38. Giampieri F, Alvarez-Suarez JM, Mazzoni L, Forbes-Hernandez TY, Gasparrini M, Gonzàlez-Paramàs AM, Santos-Buelga C, Quiles JL, Bompadre S, Mezzetti B, Battino M,An anthocyanin-rich strawberry extract protects against oxidative stress damage and improves mitochondrial functionality in human dermal fibroblasts exposed to an oxidizing agent. Food Funct 5: 1939-1948 (2014). 39. Tulipani S, Armeni T, Giampieri F, Alvarez-Suarez JM, Gonzalez-Paramás AM, SantosBuelga C, Busco F, Principato G, Bompadre S, Quiles JL, Mezzetti B, Battino M, Strawberry intake increases blood fluid, erythrocyte and mononuclear cell defenses against oxidative challenge. Food Chem 156: 87-93 (2014).
This article is protected by copyright. All rights reserved
40. Giampieri F, Alvarez-Suarez JM, Battino M, Strawberry and Human Health: Effects beyond Antioxidant Activity. J Agric Food Chem 62: 3867–3876 (2014). 41. Paredes-Lopez O, Cervantes-Ceja ML, Vigna-Perez M, Hernandez-Perez T, Berries: improving human health and healthy aging, and promoting quality life – A review. Plant Foods Hum Nutr 65: 299-308 (2010). 42. Aiyer HS, Vadhanam MV, Stoyanova R, Caprio GD, Clapper ML, Gupta RC, Dietary berries and ellagic acid prevent oxidative DNA damage and modulate expression of DNA repair genes. Int J Mol Sci 9: 327-341 (2008). 43. Aiyer HS, Kichambare S, Gupta RC Prevention of oxidative DNA damage by bioactive berry components. Nutr Cancer 60: 36-42 (2008). 44. Giampieri F, Alvarez-Suarez JM, Tulipani S, Gonzales-Paramas AM, Santos-Buelga C, Bompadre S, Quiles JL, Mezzetti B, Battino M, Photoprotective potential of strawberry (Fragaria x ananassa) extract against UV-A irradiation damage on human fibroblasts. J Agric Food Chem 60: 2322-2327 (2012). 45. Schantz M, Mohn C, Baum M, Richling E, Antioxidative efficiency of an anthocyanin rich bilberry extract in the human colon tumor cell lines Caco-2 and HT-29. J Berry Res 1: 25-33 (2010). 46. Stoner GD, Wang LS, Casto BC, Laboratory and clinical studies of cancer chemoprevention by antioxidants in berries. Carcinogenesis 29: 1665-1674 (2008). 47. Huang C, Li J, Song L, Zhang D, Tong Q, Ding M, Bowman L, Aziz R, Stoner GD, Black Raspberry extracts inhibit Benzo(a)Pyrene Diol-Epoxide-induced activator protein 1 activation and VEGF transcription by targeting the phosphotidylinositol 3-kinase/Akt pathway. Cancer Res 66: 581-587 (2006). 48. Neto CC, Cranberries: ripe for more cancer research? J Agric Food Chem 91: 2303-2307 (2011).
This article is protected by copyright. All rights reserved
49. Seeram NP, Berry fruits for cancer prevention: currents status and future prospects. J Agric Food Chem 56: 630-635 (2008). 50. Neto CC, Amoroso JW, Liberty AM, Anticancer activities of cranberry phytochemicals: an update. Mol Nutr Food Res 52: 18-27 (2008). 51. Boivin D, Blanchette M, Barrette S, Moghrabi A, Beliveau R, Inhibition of cancer cell proliferation and suppression of TNF-induced activation of NFrB by edible berry juice. Anticancer Res 27: 937-948 (2007). 52. Li J, Zhang D, Stoner GD, Huang C, Differential effects of black raspberry and strawberry extracts on BaPDE-induced activation of transcription factors and their target genes. Mol Carcinog 47: 286-294 (2008). 53. Chiva-Blanch G, Visioli F, Polyphenols and health: moving beyond antioxidants. J Berry Res 2: 63-71 (2012). 54. Wu QK, Koponen JM, Mykkanen HM, Torronen AR, Berry phenolics extracts modulate the expression of p21WAF1 and Bax but not Bcl-2 in HT-29 colon cancer cells. J Agric Food Chem 55: 1156-1163 (2007). 55. Seeram NP, Adams LS, Zhang Y, Lee R, Sand D, Scheuller HS, Heber D, Blackberry, black raspberry, blueberry, cranberry, red raspberry, and strawberry extracts inhibit growth and stimulate apoptosis of human cancer cells in vitro. J Agric Food Chem 54: 9329-9339 (2006). 56. Sharma M, Li L, Celver J, Killian C, Kovoor A, Seeram N, Effects of fruit ellagitannin extracts, ellagic acid, and their colonic metabolite, urolithin A, on Wnt Signaling. J Agric Food Chem 58: 3965-3969 (2010). 57. Umesalma S, Sudhandiran G, Ellagic acid prevents rat colon carcinogenesis induced by 1,2 dimethyl hydrazine through inhibition of AKT-phosphoinositide-3 kinase pathway. Mol Cell Pharmacol 660: 249-258 (2012).
This article is protected by copyright. All rights reserved
58. Juan ME, Alfaras I, Planas JM, Colorectal cancer chemoprotection by trans-resveratrol. Pharmacol Res 65: 584-591 (2012). 59. Mentor-Marcel RA, Bobe G, Dietary freeze-dried black raspberries in colorectal cancer patients. Nutr Cancer 64: 820-825 (2012). 60. Ravoori S, Vadhanam MV, Aqil F, Gupta RC, Inhibition of estrogen-mediated mammary tumorigenesis by blueberry and black raspberry. J Agric Food Chem 60: 5547-5555 (2012). 61. Aiyer HS, Gupta RC, Berries and ellagic acid prevent estrogen-induced mammary tumorigenesis by modulating enzymes of estrogen metabolism. Cancer Prev Res 3: 727737 (2010). 62. Aiyer H, Warri AM, Woode DR, Hilakivi-Clarke L, Clarke R, Influence of berrypolyphenols on receptor signaling and cell-death pathways: implication for breast cancer prevention. J Agric Food Chem 60: 5693-5708 (2012). 63. Kausar H, Jeyabalan J, Aqil F, Chabba D, Sidana J, Singh IP, Gupta RC, Berry anthocyanidins synergistically suppress growht and invasive potential of human nonsmall-cell lung cancer cells. Cancer Lett 325: 54-62 (2012). 64. Wang LS, Dombkowski AA, Seguin C, Rocha C, Cukovic D, Mukundan A, Henry C, Stoner GD, Mechanistic basis for the chemopreventive effects of black raspberries at a late stage of rat esophageal carcinogenesis. Mol Carcinog 50: 291-300 (2011). 65. Stoner GD, Wang LS, Chemoprevention of esophageal squamous cell carcinoma with berries. Top Curr Chem 329: 1-20. (2013). 66. Spagnuolo C, Russo M, Bilotto S, Tedesco I, Laratta B, Russo GL, Dietary polyphenols in cancer prevention: the example of the flavonoid quercetin in leukemia. Ann N Y Acad Sci 1259: 95-103 (2012).
This article is protected by copyright. All rights reserved
67. Hushmendy S, Jayakumar L, Hahn AB, Bhoiwala D, Bhoiwala DL, Crawfor DR, Select phytochemicals suppress human T-lymphocytes and mouse splenocytes suggesting their use in autoimmunity and transplantion. Nutr Res 29: 568-578 (2009). 68. Bae JY, Lim SS, Kim SJ, Choi JS, Park J, Ju SM, Han S.J, Kang IJ, Kan YH, Bog blueberry anthocyanins alleviate photoaging in ultraviolet-B irradiation-induced human dermal fibroblasts. Mol Nutr Food Res 53: 726-738 (2009). 69. Huang C, Zhang D, Li J, Tong Q, Stoner GD, Differential inhibition of UV-induced activation of NFκB and AP-1 by extracts from black raspberries, strawberries, and blueberries. Nutr Cancer 58: 205-212 (2007). 70. Duncan FJ, Martin JR, Wulff BC, Stoner GD, Tober KL, Oberyszyn TM, Kusewitt DF, Van Buskirk AM. Topical treatment with black raspberry extract reduces cutaneous UVB-induced carcinogenesis and inflammation. Cancer Prev Res (Phila) 2: 665-672 (2009). 71. Pan MH, Lai CS, Ho CT, Anti-inflammatory activity of natural dietary flavonoids. Food Funct 1: 15-31(2010). 72. Tribolo S, Lodi F, Connor C, Suri S, Wilson VG, Taylor MA, Needs PW, Kroon PA, Hughes DA Comparative effects of quercentin and its predominant human metabolites on adhesion molecule expression in activated human vascular endothelial cells. Atherosclerosis 197: 50-56 (2008). 73. Park MJ, Lee EK, Heo HS, Kim MS, Sung B, Kim MK, Lee J, Kim ND, Anton S, Choi JS, Yu BP, Chung HY, The anti-inflammatory effect of kaempferol in aged kidney tissues: the involvement of nuclear factor-kappaB via nuclear factor-inducing kinase/IkappaB kinase and mitogen-activated protein kinase pathways. J Med Food 12: 351-358 (2009).
This article is protected by copyright. All rights reserved
74. Oak MH, Bedoui JE, Madeira SV, Chalupsky K, Schini-Kerth VB, Delphinidin and cyanidin inhibit PDGF(AB)-induced VEGF release in vascular smooth muscle cells by preventing activation of p38 MAPK and JNK. Br J Pharmacol 149: 283-290 (2006). 75. Mauray A, Felgines C, Morand C, Mazur A, Scalbert A, Milenkovic D, Nutrigenomic analysis of the protective effects of bilberry anthocyanin-rich extract in apo E-deficient mice. Genes Nutr 4: 343-353 (2010). 76. Mauray A, Felgines C, Morand C, Mazur A, Scalbert A, Milenkovic D, Bilberry anthocyanin-rich extract alters expression of genes related to atherosclerosis development in aorta of apo E-deficient mice. Nutr Metab Cardiovasc Dis 22: 72-80 (2012). 77. Cassidy A, Mukamal KJ, Liu L, Franz M, Eliassen AH, Rimm EB, High anthocyanin intake is associated with a reduced risk of myocardial infarction in young and middleaged women. Circulation 127: 188-196 (2013). 78. Alvarez-Suarez JM, Giampieri F, Tulipani S, Casoli T, Di Stefano G, González-Paramás AM, Santos-Buelga C, Busco F, Quiles JL, Cordero MD, Bompadre S, Mezzetti B, Battino M, One-month strawberry-rich anthocyanin supplementation ameliorates cardiovascular risk, oxidative stress markers and platelet activation in humans. J Nutr Biochem 25: 289-294 (2014). 79. Qin Y, Xia M, Ma J, Hao Y, Liu J, Mou H, Cao L, Ling W, Anthocyanin supplementation improves serum LDL- and HDL-cholesterol concentrations associated with the inhibition of cholesteryl ester transfer protein in dyslipidemic subjects. Am J Clin Nutr 90: 485-492 (2009). 80. Spencer JPE, Vafeiadou K, Williams RJ, Vauzour D, Neuroinflammation: modulation by flavonoids and mechanism of action. Mol Aspects Med 33: 83-97 (2012).
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81. Vauzour D, Vafeiadou K, Rendeiro C, Corona G, Spencer JPE, The inhibitory effects of berry-derived flavonoids against neurodegenerative processes. J Berry Res 1: 45-52 (2010). 82. Joseph JA, Shukitt-Hale B, Willis LM, Grape juice, berries, and walnuts affect brain aging and behavior. J Nutr 139: 1813-1817 (2009). 83. Poulose SM, Carey AN, Shukitt-Hale B, Improving brain signaling in aging: Could berries be the answer? Expert Rev Neurother 12: 887-889 (2012). 84. Joseph JA, Shukitt-Hale B, Denisova NA, Prior RL, Cao G, Martin A, Taglialatela G, Bickford PC, Long-term dietary strawberry, spinach, or vitamin E supplementation retards the onset of age-related neuronal signal-transduction and cognitive behavioral deficits. J Neurosci 18: 8047-8055 (1998). 85. Joseph JA, Shukitt-Hale B, Denisova NA, Bielinski D, Martin A, McEwen JJ, Bickford PC, Reversals of age-related declines in neuronal signal transduction, cognitive, and motor behavioral deficits with blueberry, spinach, or strawberry dietary supplementation. J Neurosci. 15: 8114-21 (1999). 86. Shukitt-Hale B, Cheng V, Bielinski D, Carey AN, Lau FC, Seeram NP, Heber D, Joseph JA, Differential brain regional specificity to blueberry and strawberry polyphenols in improved motor and cognitive function in aged rats. Soc Neurosci Abstr 32: 81.15 (2006). 87. Williams RJ, Spencer JPE, Flavonoids, cognition and dementia: actions, mechanisms, and potential therapeutic utility for Alzheimer disease. Free Radic Biol Med 52: 35-45 (2012). 88. Zhu M, Hu J, Perez E, Phillips D, Kim W, Ghaedian R, Napora JK, Zou S, Effects of long-term cranberry supplementation on endocrine pancreas in aging rats. J Gerontol A Biol Sci Med Sci 66A: 1139-1151 (2011).
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ILLUSTRATIONS
Figure 1: Homoeeology grouups (HGs, named n withh roman nuumerals as the correspponding Linkagee Groups inn the diploidd Fragaria reference r map24) follow m wed by an F (for femalle, LGs) or an M (male Linnkage Groupps). Linkage Groups within w each HG H were nuumbered arrbitrarily althouggh the same number in male and feemale maps indicates thhe homologgous groupss. On the left, thee size of LG Gs are indiccated (in cenntimorgans)). On the riight, the maarker and thhe QTLs are listeed. Source: Zorrilla-Fonntanesi et al. 2011(25) with w some modification m ns.
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Figure 2: Mappedd QTLs in raspberry. r On O the top, LG 1 is reepresented: bHLH, b invoolved in the antthocyanin pathway, p iss the mostt importantt marker. Below, B LG G 4 is shoown and FRUITE E4, also invvolved in thhe anthocyaanin pathwaay, is the most m importaant marker. Source: Kassim m et al. 2009(32) with som me modificaations.
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