Grafting with rootstocks promotes phenolic compound accumulation in grape berry skin during development based on integrative multi-omics analysis

Abstract

In viticulture, grafting has been practiced widely and influences grape development as well as berry and wine quality. However, there is limited understanding of the effects of rootstocks on grape phenolic compounds, which are located primarily in the berry skin and contribute to certain sensory attributes of wine. In this study, scion–rootstock interactions were investigated at the green-berry stage and the veraison stage when grapevines were hetero-grafted with three commonly used rootstock genotypes (5BB, 101-14MG, and SO4). Physiological investigations showed that hetero-grafts, especially CS/5BB, contained higher concentrations of total proanthocyanidins (PAs) and various PA components in berry skins compared with the auto-grafted grapevines. Further metabolomics analysis identified 105 differentially accumulated flavonoid compounds, the majority of which, including anthocyanins, PAs, and flavonols, were significantly increased in the berry skins of hetero-grafted grapevines compared with auto-grafted controls. In addition, transcriptomic analysis of the same samples identified several thousand differentially expressed genes between hetero-grafted and auto-grafted vines. The three rootstocks not only increased the transcript levels of stilbene, anthocyanin, PA, and flavonol synthesis genes but also affected the expression of numerous transcription factor genes. Taken together, our results suggest that hetero-grafting can promote phenolic compound accumulation in grape berry skin during development. These findings provide new insights for improving the application value of grafting by enhancing the accumulation of nutritious phenolic components in grape.

References

[1] Cuadros-Inostroza A, Ruíz-Lara S, González E et al. GC-MS metabolic profiling of cabernet sauvignon and merlot cultivars during grapevine berry development and network analysis reveals a stage- and cultivar-dependent connectivity of primary metabolites. Metabolomics. 2016; 12: 39.

[2] Vivier MA, Pretorius IS . Genetically tailored grapevines for the wine industry. Trends Biotechnol. 2002; 20: 472-8.

[3] Manolescu BN, Oprea E, Mititelu M et al. Dietary anthocyanins and stroke: a review of pharmacokinetic and pharmacodynamic studies. Nutrients. 2019; 11: 1479.

[4] Teixeira A, Eiras-Dias J, Castellarin SD, Geros H . Berry phenolics of grapevine under challenging environments. Int J Mol Sci. 2013; 14: 18711-39.

[5] Kennedy JA, Saucier C, Glories Y . Grape and wine phenolics: history and perspective. Am J Enol Vitic. 2006; 57: 239-48.

[6] Braidot E, Zancani M, Petrussa E et al. Transport and accumulation of flavonoids in grapevine (Vitis vinifera L.) . Plant Signal Behav. 2008; 3: 626-32.

[7] He F, Mu L, Yan G-L et al. Biosynthesis of anthocyanins and their regulation in colored grapes. Molecules. 2010; 15: 9057-91.

[8] Kolb CA, Kopecky J, Riederer M, Pfundel EE . UV screening by phenolics in berries of grapevine (Vitis vinifera) . Funct Plant Biol. 2003; 30: 1177-86.

[9] Basalekou M, Kyraleou M, Pappas C et al. Proanthocyanidin content as an astringency estimation tool and maturation index in red and white winemaking technology. Food Chem. 2019; 299: 125135.

[10] Downey MO, Dokoozlian NK, Krstic MP . Cultural practice and environmental impacts on the flavonoid composition of grapes and wine: a review of recent research. Am J Enol Vitic. 2006; 57: 257-68.

[11] Matus JT, Loyola R, Vega A et al. Post-veraison sunlight exposure induces MYB-mediated transcriptional regulation of anthocyanin and flavonol synthesis in berry skins of Vitis vinifera. J Exp Bot. 2009; 60: 853-67.

[12] Matus JT . Transcriptomic and metabolomic networks in the grape berry illustrate that it takes more than flavonoids to fight against ultraviolet radiation. Front Plant Sci. 2016; 7: 1337.

[13] Matus JT, Cavallini E, Loyola R et al. A group of grapevine MYBA transcription factors located in chromosome 14 control anthocyanin synthesis in vegetative organs with different specificities compared with the berry color locus. Plant J. 2017; 91: 220-36.

[14] Rinaldo AR, Cavallini E, Jia Y et al. A grapevine anthocyanin acyltransferase, transcriptionally regulated by VvMYBA, can produce most acylated anthocyanins present in grape skins. Plant Physiol. 2015; 169: 1897-916.

[15] Cookson SJ, Moreno MJC, Hevin C et al. Graft union formation in grapevine induces transcriptional changes related to cell wall modification, wounding, hormone signalling, and secondary metabolism. J Exp Bot. 2013; 64: 2997-3008.

[16] Corso M, Vannozzi A, Maza E et al. Comprehensive transcript profiling of two grapevine rootstock genotypes contrasting in drought susceptibility links the phenylpropanoid pathway to enhanced tolerance. J Exp Bot. 2015; 66: 5739-52.

[17] Mare C, Mica E, Cattivelli L . Molecular advances in rootstock-scion interaction in grapevine. Acta Hortic. 2016; 1136: 155-60.

[18] Forneck A, Huber L . (a)sexual reproduction - a review of life cycles of grape phylloxera, Daktulosphaira vitifoliae. Entomol Exp Appl. 2009; 131: 1-10.

[19] Anwar SA, McKenry M, Ramming D . A search for more durable grape rootstock resistance to root-knot nematode. Am J Enol Vitic. 2002; 53: 19-23.

[20] Di Filippo M, Vila H . Influence of different rootstocks on the vegetative and reproductive performance of Vitis Vinifera L. Malbec under irrigated conditions. J Int Sci Vigne du Vin. 2011; 45: 75-84.

[21] Warschefsky EJ, Klein LL, Frank MH et al. Rootstocks: diversity, domestication, and impacts on shoot phenotypes. Trends Plant Sci. 2016; 21: 418-37.

[22] Pulko B, Vrsic S, Valdhuber J . Influence of various rootstocks on the yield and grape composition of sauvignon blanc. Czech J Food Sci. 2012; 30: 467-73.

[23] Ali MB, Howard S, Chen S et al. Berry skin development in Norton grape: distinct patterns of transcriptional regulation and flavonoid biosynthesis. BMC Plant Biol. 2011; 11: 7.

[24] Degu A, Hochberg U, Sikron N et al. Metabolite and transcript profiling of berry skin during fruit development elucidates differential regulation between cabernet sauvignon and shiraz cultivars at branching points in the polyphenol pathway. BMC Plant Biol. 2014; 14: 188.

[25] Chitarra W, Perrone I, Avanzato CG et al. Grapevine grafting: scion transcript profiling and defense-related metabolites induced by rootstocks. Front Plant Sci. 2017; 8: 654.

[26] Jin ZX, Sun H, Sun TY et al. Modifications of ’Gold Finger’ grape berry quality as affected by the different rootstocks. J Agr Food Chem. 2016; 64: 4189-97.

[27] Deluc LG, Grimplet J, Wheatley MD et al. Transcriptomic and metabolite analyses of cabernet sauvignon grape berry development. BMC Genomics. 2007; 8: 429.

[28] Kyraleou M, Kotseridis Y, Koundouras S et al. Effect of irrigation regime on perceived astringency and proanthocyanidin composition of skins and seeds of Vitis vinifera L. cv. Syrah grapes under semiarid conditions. Food Chem. 2016; 203: 292-300.

[29] Koyama K, Ikeda H, Poudel PR, Goto-Yamamoto N . Light quality affects flavonoid biosynthesis in young berries of cabernet sauvignon grape. Phytochemistry. 2012; 78: 54-64.

[30] Vidal S, Francis L, Noble A et al. Taste and mouth-feel properties of different types of tannin-like polyphenolic compounds and anthocyanins in wine. Anal Chim Acta. 2004; 513: 57-65.

[31] Cheynier V, Dueñas-Paton M, Salas E et al. Structure and properties of wine pigments and tannins. Am J Enol Vitic. 2006; 57: 298-305.

[32] Snopek L, Mlcek J, Sochorova L et al. Contribution of red wine consumption to human health protection. Molecules. 2018; 23: 1684.

[33] Amor S, Chalons P, Aires V, Delmas D . Polyphenol extracts from red wine and grapevine: potential effects on cancers. Diseases. 2018; 6: 106.

[34] Gatto P, Vrhovsek U, Muth J et al. Ripening and genotype control stilbene accumulation in healthy grapes. J Agr Food Chem. 2008; 56: 11773-85.

[35] Vannozzi A, Wong DCJ, Höll J et al. Combinatorial regulation of stilbene synthase genes by WRKY and MYB transcription factors in grapevine (Vitis vinifera L.) . Plant Cell Physiol. 2018; 59: 1043- 59.

[36] Sun RZ, Cheng G, Li Q et al. Comparative physiological, metabolomic, and transcriptomic analyses reveal developmental stage-dependent effects of cluster bagging on phenolic metabolism in cabernet sauvignon grape berries. BMC Plant Biol. 2019; 19: 583.

[37] Rinaldi A, Villano C, Lanzillo C et al. Metabolic and RNA profiling elucidates proanthocyanidins accumulation in Aglianico grape. Food Chem. 2017; 233: 52-9.

[38] Zombardo A, Crosatti C, Bagnaresi P et al. Transcriptomic and biochemical investigations support the role of rootstock-scion interaction in grapevine berry quality. BMC Genomics. 2020; 21: 468.

[39] Czemmel S, Heppel SC, Bogs J . R2R3 MYB transcription factors: key regulators of the flavonoid biosynthetic pathway in grapevine. Protoplasma. 2012; 249: S109-18.

[40] Wang M, Vannozzi A, Wang G et al. Genome and transcriptome analysis of the grapevine (Vitis vinifera L.) WRKY gene family . Hortic Res. 2014; 1: 14016.

[41] Deluc L, Barrieu F, Marchive C et al. Characterization of a grapevine R2R3-MYB transcription factor that regulates the phenylpropanoid pathway. Plant Physiol. 2006; 140: 499-511.

[42] Gambetta GA, Matthews MA, Shaghasi TH et al. Sugar and abscisic acid signaling orthologs are activated at the onset of ripening in grape. Planta. 2010; 232: 219-34.

[43] Gutierrez-Gamboa G, Gómez-Plaza E, Bautista-Ortín AB et al. Rootstock effects on grape anthocyanins, skin and seed proanthocyanidins and wine color and phenolic compounds from Vitis vinifera L. merlot grapevines. J Sci Food Agr. 2019; 99: 2846-54.

[44] Pertea M, Kim D, Pertea GM et al. Transcript-level expression analysis of RNA-seq experiments with HISAT, StringTie and Ballgown. Nat Protoc. 2016; 11: 1650-67.

[45] Love MI, Huber W, Anders S . Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol. 2014; 15: 550.

[46] Gotz S, García-Gómez JM, Terol J et al. High-throughput functional annotation and data mining with the Blast2GO suite. Nucleic Acids Res. 2008; 36: 3420-35.

[47] Thimm O, Bläsing O, Gibon Y et al. MAPMAN: a user-driven tool to display genomics data sets onto diagrams of metabolic pathways and other biological processes. Plant J. 2004; 37: 914-39.