{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,11]],"date-time":"2026-03-11T07:50:44Z","timestamp":1773215444752,"version":"3.50.1"},"reference-count":44,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2020,1,2]],"date-time":"2020-01-02T00:00:00Z","timestamp":1577923200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"PLATAFORMA DE INOVA\u00c7\u00c3O DA VINHA E DO VINHO \u2010 INNOVINE&amp;WINE","award":["NORTE-01-0145-FEDER-000038"],"award-info":[{"award-number":["NORTE-01-0145-FEDER-000038"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Despite the numerous beneficial properties and uses of chitosan in agriculture, the molecular mechanisms behind its elicitation potential are still unclear. This study aimed at understanding the effect of chitosan application in the levels of phenolic compounds of Vitis vinifera L. red grapes berry skin (cv. Tinto C\u00e3o) during veraison. Grapevines were treated with chitosan (0.01% in 0.01% acetic acid) while control grapevines were sprayed with 0.01% acetic acid. Results showed that several monomeric anthocyanins increased significantly in berry skins after treatment with chitosan. Additionally, Catechin, Rutin and Querecetin-3-O-galactoside were also recorded in higher amount upon chitosan treatment. Besides modulating the phenolic content, chitosan treatment also induced modifications in several target genes encoding key enzymes and transporters involved in secondary metabolic pathways. For instance, the genes PAL, CHS, F3H, ANR, UFGT, ABCC1, GST, MATE1 were upregulated in leaves and berry skins at veraison cessation in response to chitosan treatment. Overall, the results demonstrated that chitosan has a stimulatory effect on the accumulation of phenolic compounds, including anthocyanins, mediated by modifications in the transcription of key genes involved in their biosynthesis and transport in grape berries.<\/jats:p>","DOI":"10.3390\/ijms21010306","type":"journal-article","created":{"date-parts":[[2020,1,3]],"date-time":"2020-01-03T04:43:03Z","timestamp":1578026583000},"page":"306","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":41,"title":["Chitosan Application in Vineyards (Vitis vinifera L. cv. Tinto C\u00e3o) Induces Accumulation of Anthocyanins and Other Phenolics in Berries, Mediated by Modifications in the Transcription of Secondary Metabolism Genes"],"prefix":"10.3390","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2536-1967","authenticated-orcid":false,"given":"Rupesh Kumar","family":"Singh","sequence":"first","affiliation":[{"name":"Centro de Qu\u00edmica de Vila Real (CQ-VR), Universidade de Tr\u00e1s-os-Montes e Alto Douro (UTAD), 5000-801 Vila Real, Portugal"},{"name":"Departamento de Agronomia, Universidade de Tr\u00e1s-os-Montes e Alto Douro (UTAD), Quinta de Prados, 5000-801 Vila Real, Portugal"}]},{"given":"Viviana","family":"Martins","sequence":"additional","affiliation":[{"name":"Centro de Biologia Molecular e Ambiental (CBMA), Department of Biology, UMINHO, 4710-057 Braga, Portugal"},{"name":"Centro de Investiga\u00e7\u00e3o e Tecnologias Agro-Ambientais e Biol\u00f3gicas (CITAB), Universidade de Tr\u00e1s-os-Montes e Alto Douro (UTAD), Quinta de Prados, 5000-801 Vila Real, Portugal"}]},{"given":"Bruno","family":"Soares","sequence":"additional","affiliation":[{"name":"Departamento de Agronomia, Universidade de Tr\u00e1s-os-Montes e Alto Douro (UTAD), Quinta de Prados, 5000-801 Vila Real, Portugal"},{"name":"CoLAB Vines&amp;Wines, Associa\u00e7\u00e3o para o Desenvolvimento da Viticultura Duriense (ADVID), R\u00e9gia Douro Park, 5000-033 Vila Real, Portugal"}]},{"given":"Isaura","family":"Castro","sequence":"additional","affiliation":[{"name":"Centro de Investiga\u00e7\u00e3o e Tecnologias Agro-Ambientais e Biol\u00f3gicas (CITAB), Universidade de Tr\u00e1s-os-Montes e Alto Douro (UTAD), Quinta de Prados, 5000-801 Vila Real, Portugal"}]},{"given":"Virg\u00edlio","family":"Falco","sequence":"additional","affiliation":[{"name":"Centro de Qu\u00edmica de Vila Real (CQ-VR), Universidade de Tr\u00e1s-os-Montes e Alto Douro (UTAD), 5000-801 Vila Real, Portugal"},{"name":"Departamento de Agronomia, Universidade de Tr\u00e1s-os-Montes e Alto Douro (UTAD), Quinta de Prados, 5000-801 Vila Real, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2020,1,2]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"603","DOI":"10.1016\/j.progpolymsci.2006.06.001","article-title":"Chitin and chitosan: Properties and application","volume":"31","author":"Rinaudo","year":"2006","journal-title":"Prog. Polym. Sci."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"3019","DOI":"10.3390\/ijms16023019","article-title":"Reactive oxygen and nitrogen species indefense\/stress responses activated by chitosan in sycamore cultured cells","volume":"16","author":"Malerba","year":"2015","journal-title":"Int. J. Mol. Sci."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1016\/j.carbpol.2015.06.008","article-title":"Fluctuation in physicochemical properties of chitins extracted from different body parts of honeybee","volume":"132","author":"Kaya","year":"2015","journal-title":"Carbohydr. Polym."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"285","DOI":"10.1016\/S0147-5975(79)80054-7","article-title":"The fungicidal effect of chitosan on fungi of varying cell wall composition","volume":"3","author":"Allan","year":"1979","journal-title":"Exp. Mycol."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Sharif, R., Mujtaba, M., Rahman, M., Shalmani, A., Ahmad, H., Anwar, T., Tianchan, D., and Wang, X. (2018). The Multifunctional Role of Chitosan in Horticultural Crops; A Review. Molecules, 23.","DOI":"10.3390\/molecules23040872"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"313","DOI":"10.1007\/s12298-018-0633-1","article-title":"Application of chitosan on plant responses with special reference to abiotic stress","volume":"25","author":"Hidangmayum","year":"2018","journal-title":"Physiol. Mol. Biol. Plants"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"263","DOI":"10.1111\/j.1755-0238.2011.00149.x","article-title":"New chitosan formulation prevents grapevine powdery mildew infection and improves polyphenol content and free radical scavenging activity of grape and wine","volume":"17","author":"Iriti","year":"2011","journal-title":"Aust. J. Grape Wine Res."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1862","DOI":"10.1111\/j.1365-2621.2002.tb08737.x","article-title":"Effects of pre-and postharvest chitosan treatments to control storage grey table grapes","volume":"67","author":"Romanazzi","year":"2002","journal-title":"J. Food Sci."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"882","DOI":"10.1111\/j.1365-3059.2010.02312.x","article-title":"Inhibition of Botrytis cinerea growth and suppression of botrytis bunch rot in grapes using chitosan","volume":"59","author":"Reglinski","year":"2010","journal-title":"Plant Pathol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"501","DOI":"10.1016\/j.foodchem.2007.06.012","article-title":"Physiological responses and quality attributes of table grape fruit to chitosan preharvest spray and postharvest coating during storage","volume":"106","author":"Meng","year":"2008","journal-title":"Food Chem."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Singh, R.K., Soares, B., Goufo, P., Castro, I., Cosme, F., Pinto-Sintra, A.L., Ines, A., Olivera, A.A., and Falco, V. (2019). Chitosan upregulates the genes of the ROS pathway, and enhances the antioxidant potential of grape (Vitis vinifera L. \u2018Touriga Francax and \u2019Tinto C\u00e3o\u2019) tissues. Antioxidants, 8.","DOI":"10.3390\/antiox8110525"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1111\/j.1755-0238.2003.tb00228.x","article-title":"Analysis of tannins in seeds and skins of Shiraz grapes throughout berry development","volume":"9","author":"Downey","year":"2003","journal-title":"Aust. J. Grape Wine Res."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"90","DOI":"10.1111\/j.1755-0238.2010.00121.x","article-title":"Effect of pre- and post-veraison water deficit on proanthocyanidin and anthocyanin accumulation during Shiraz berry development","volume":"17","author":"Guiraud","year":"2011","journal-title":"Aust. J. Grape Wine Res."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"10186","DOI":"10.1021\/jf0506737","article-title":"Elicitation of Resveratrol in Peanut Kernels by Application of Abiotic Stresses","volume":"53","author":"Rudolf","year":"2005","journal-title":"J. Agric. Food Chem."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"610","DOI":"10.1002\/pmic.200800386","article-title":"Chitosan treatment induces changes of protein expression profile and stilbene distribution in Vitis vinifera cell suspensions","volume":"9","author":"Ferri","year":"2009","journal-title":"Proteomics"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1016\/j.foodchem.2016.01.086","article-title":"Improvement of grape and wine phenolic content by foliar application to grapevine of three different elicitors: Methyl jasmonate, chitosan, and yeast extract","volume":"15","author":"Portu","year":"2016","journal-title":"Food Chem."},{"key":"ref_17","first-page":"1385","article-title":"Photoregulation of anthocyanin synthesis in apple fruit under UV-B and red light","volume":"29","author":"Arakawa","year":"1988","journal-title":"Plant Cell Physiol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"647","DOI":"10.1007\/s11676-015-0121-1","article-title":"Expression of structural genes related to anthocyanin biosynthesis of Vitis amurensis","volume":"27","author":"Zhao","year":"2016","journal-title":"J. For. Res."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1059","DOI":"10.1104\/pp.111.4.1059","article-title":"Analysis of the expression of anthocyanin pathway genes in developing Vitis vinifera L. cv Shiraz grape berries and the implications for pathway regulation","volume":"111","author":"Boss","year":"1996","journal-title":"Plant Physiol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"924","DOI":"10.1007\/s00425-002-0830-5","article-title":"Myb-related genes of the Kyoho grape (Vitis labruscana) regulate anthocyanin biosynthesis","volume":"215","author":"Kobayashi","year":"2002","journal-title":"Planta"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1016\/j.jplph.2005.07.006","article-title":"Changes and subcellular localizations of the enzymes involved in phenylpropanoid metabolism during grape berry development","volume":"163","author":"Chen","year":"2006","journal-title":"Plant Physiol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1007\/s10142-004-0124-z","article-title":"cDNA microarray analysis of development grape (Vitis vinifera cv Shiraz) berry skin","volume":"5","author":"Waters","year":"2005","journal-title":"Funct. Integr. Genom."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"571","DOI":"10.1186\/1471-2164-7-12","article-title":"Colour variation in red grapevines (Vitis vinifera L.): Genomic organisation, expression of flavonoid 3\u2032-hydroxylase, flavonoid 3\u2032, 5\u2032-hydroxylase genes and related metabolite profiling of red cyanidin-\/blue delphinidin-based anthocyanins in berry skin","volume":"7","author":"Castellarin","year":"2006","journal-title":"BMC Genom."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1016\/0167-7799(94)90097-3","article-title":"Blue roses-a pigment of our imagination?","volume":"12","author":"Holton","year":"1994","journal-title":"Trends Biotechnol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1840","DOI":"10.1105\/tpc.112.102152","article-title":"ABCC1, an ATP binding cassette protein from 695 grape berry, transports anthocyanidin 3-O-Glucosides","volume":"25","author":"Francisco","year":"2013","journal-title":"Plant Cell"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1812","DOI":"10.1105\/tpc.022574","article-title":"A Multidrug Resistance\u2013Associated Protein Involved in 708 Anthocyanin Transport in Zea mays","volume":"16","author":"Goodman","year":"2004","journal-title":"Plant Cell"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1150","DOI":"10.3389\/fpls.2016.01150","article-title":"Kaolin Foliar Application Has a Stimulatory Effect on Phenylpropanoid and Flavonoid Pathways in Grape Berries","volume":"7","author":"Conde","year":"2016","journal-title":"Front. Plant Sci."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Demurtas, O.C., Francisco, R.B., Diretto, G., Ferrante, P., Frusciante, S., Pietrella, M., Aprea, G., Borghi, L., Feeney, M., and Frigerio, L. (2019). ABCC Transporters Mediate the Vacuolar Accumulation of Crocins in Saffron 3 Stigmas. Plant Cell.","DOI":"10.1105\/tpc.19.00193"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"3621","DOI":"10.1093\/jxb\/ern217","article-title":"Purification, molecular cloning, and characterization of glutathione S-transferases (GSTs) from pigmented Vitis vinifera L. cell suspension cultures as putative anthocyanin transport proteins","volume":"59","author":"Conn","year":"2008","journal-title":"J. Exp. Bot."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"501","DOI":"10.1007\/BF00027496","article-title":"Transformed plants with elevated levels of chloroplastic SOD are not more resistant to superoxide toxicity","volume":"14","author":"Tepperman","year":"1990","journal-title":"Plant Mol. Biol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1331","DOI":"10.1093\/jexbot\/53.372.1331","article-title":"Role of superoxide dismutases (SODs) in controlling oxidative stress in plants","volume":"53","author":"Alscher","year":"2002","journal-title":"J. Exp. Bot."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Feng, X., Lai, Z., Lin, Y., Lai, G., and Lian, C. (2015). Genome-wide identification and characterization of the superoxide dismutase gene family in Musa acuminata cv. Tianbaojiao (AAA group). BMC Genom., 16.","DOI":"10.1186\/s12864-015-2046-7"},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Hu, X., Hao, C., Cheng, Z.M., and Zhong, Y. (2019). Genome-Wide Identification, Characterization, and Expression Analysis of the Grapevine Superoxide Dismutase (SOD) Family. Int. J. Genom.","DOI":"10.1155\/2019\/7350414"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"595","DOI":"10.1007\/s11032-013-9892-2","article-title":"Superoxide dismutase multigene family in longan somatic embryos: A comparison of CuZn-SOD, Fe-SOD, and Mn-SOD gene structure, splicing, phylogeny, and expression","volume":"32","author":"Lin","year":"2013","journal-title":"Mol. Breed."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"209","DOI":"10.1139\/g05-102","article-title":"Molecular structure and organization of the wheat genomic manganese superoxide dismutase gene","volume":"49","author":"Baek","year":"2006","journal-title":"Genome"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"637","DOI":"10.1104\/pp.118.2.637","article-title":"Superoxide dismutase in Arabidopsis: An eclectic enzyme family with disparate regulation and protein localization","volume":"2","author":"Kliebenstein","year":"1998","journal-title":"Plant Physiol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"49","DOI":"10.3906\/biy-1403-9","article-title":"Genome-wide distribution of superoxide dismutase (SOD) gene families in Sorghum bicolor","volume":"39","author":"Filiz","year":"2015","journal-title":"Turk. J. Biol."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Molina-Rueda, J.J., Tsai, C.J., and Kirby, E.G. (2013). The Populus superoxide dismutase gene family and its responses to drought stress in transgenic poplar overexpressing a pine cytosolic glutamine synthetase (GS1a). PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0056421"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1016\/j.plgene.2016.02.002","article-title":"Genome-wide analysis of superoxide dismutase gene family in Gossypium raimondii and G. arboretum","volume":"6","author":"Wang","year":"2016","journal-title":"Plant Gene"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1009","DOI":"10.1007\/s00299-016-1934-x","article-title":"Acetic acid acts as an elicitor exerting a chitosan-like effect on xanthone biosynthesis in Hypericum perforatum L. root cultures","volume":"35","author":"Valletta","year":"2016","journal-title":"Plant Cell Rep."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"e12414","DOI":"10.1111\/jfbc.12414","article-title":"Variation of chemical constituents, antioxidant activity, and endogenous plant hormones throughout different ripening stages of highbush blueberry (Vaccinium corymbosum L.) cultivars produced in centre of Portugal","volume":"41","author":"Aires","year":"2017","journal-title":"J. Food Biochem."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"437","DOI":"10.1007\/BF02772687","article-title":"RNA extraction from different apple tissues rich in polyphenols and polysaccharides for cDNA library construction","volume":"22","author":"Gasic","year":"2004","journal-title":"Plant Mol. Biol. Rep."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/1471-2105-9-465","article-title":"QuantPrime\u2014A flexible tool for reliable high-throughput primer design for quantitative PCR","volume":"9","author":"Arvidsson","year":"2008","journal-title":"BMC Bioinform."},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Reid, K.E., Olsson, N., Schlosser, J., Peng, F., and Lund, S.T. (2006). An optimized grapevine RNA isolation procedure and statistical determination of reference genes for real-time RT-PCR during berry development. BMC Plant Biol., 6.","DOI":"10.1186\/1471-2229-6-27"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/21\/1\/306\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,13]],"date-time":"2025-10-13T14:03:31Z","timestamp":1760364211000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/21\/1\/306"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,1,2]]},"references-count":44,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2020,1]]}},"alternative-id":["ijms21010306"],"URL":"https:\/\/doi.org\/10.3390\/ijms21010306","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,1,2]]}}}