Skip to main content
SpringerLink
Log in

VvANR silencing promotes expression of VvANS and accumulation of anthocyanin in grape berries

  • Original Article
  • Published:
Protoplasma Aims and scope Submit manuscript

Abstract

Virus-induced gene silencing (VIGS) technology was applied to silence VvANR in cv. Zaoheibao grape berries, and the effects of VvANR silencing on berries phenotype; gene expression level of ANS, LAR1, LAR2, and UFGT; enzyme activity of ANS; and accumulations of anthocyanin and flavan-3-ol were investigated. At the third day after treatment, the VvANR silenced grape berries began to turn red slightly, which was 2 days earlier than that of the control group. And the flavan-3-ol content in VvANR-silenced grape berries had been remarkable within 1 to 5 days, the ANR enzyme activity in VvANR-silenced grapes extremely significantly decreased in 3 days, and LAR enzyme activity also decreased, but the difference was not striking. The ANS enzyme activity of the transformed berries was significantly higher than that of the control after 3 days of infection, and it was exceedingly significantly higher than that of the control after 5 to 10 days. The content of anthocyanin in transformed berries increased of a very marked difference within 3 to 15 days. pTRV2-ANR infection resulted in an extremely significant decrease in the expression of VvANR gene, and the expression of VvLAR1, VvLAR2, VvMYBPA1, VvMYBPA2, and VvDFR were also down-regulated. However, the expression of VvANS and VvUFGT was up-regulated significantly. After VvANR silencing via VIGS, VvANR expression in grape berries was extremely significantly decreased, resulting in decreased ANR enzyme activity and flavan-3-ol content; berries turned red and deeper in advance. In addition, VvANR silencing can induce up-regulation of VvANS and VvUFGT expression, significantly increase ANS enzyme activity, and increase of anthocyanin accumulation.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

References

  • Aharoni A, Vos CHRD, Wein M, Sun Z, O’Connell AP (2010) The strawberry FaMYB1 transcription factor suppresses anthocyanin and flavonol accumulation in transgenic tobacco. Plant J 28:319–332

    Article  Google Scholar 

  • Amr A, Al-Tamimi E (2007) Stability of the crude extracts of Ranunculus asiaticus anthocyanins and their use as food colourants. Int J Food Sci Tech 42:985–991

    Article  CAS  Google Scholar 

  • Becker A, Lange M (2010) VIGS—genomics goes functional. Trends Plant Sci 15:1–4

    Article  CAS  PubMed  Google Scholar 

  • Bogs J, Downey MO, Harvey JS, Ashton AR, Tanner GJ, Robinson SP (2005) Proanthocyanidin synthesis and expression of genes encoding leucoanthocyanidin reductase and anthocyanidin reductase in developing grape berries and grapevine leaves. Plant Physiol 139:652–663

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bogs J, Jaffé FW, Takos AM, Walker AR, Robinson SP (2007) The grapevine transcription factor VvMYBPA1 regulates proanthocyanidin synthesis during fruit development. Plant Physiol 143:1347–1361

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bogs J, Ebadi A, Mcdavid D, Robinson SP (2006) Identification of the flavonoid hydroxylases from grapevine and their regulation during fruit development. Plant Physiol 140:279–291

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Boss PK, Davies C, Robinson SP (1996) Expression of anthocyanin biosynthesis pathway genes in red and white grapes. Plant Mol Biol 32:565–569

    Article  CAS  PubMed  Google Scholar 

  • Bradford MM (1976) A rapid method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  CAS  PubMed  Google Scholar 

  • Chai YM, Jia HF, Li CL, Dong QH, Shen YY (2011) FaPYR1 is involved in strawberry fruit ripening. J Exp Bot 62:5079–5089

    Article  CAS  PubMed  Google Scholar 

  • Cohen SD, Tarara JM, Gambetta GA, Matthews MA, Kennedy JA (2012) Impact of diurnal temperature variation on grape berry development, proanthocyanidin accumulation, and the expression of flavonoid pathway genes. J Exp Bot 63:2655–2665

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Czemmel S, Heppel SC, Bogs J (2012) R2R3 MYB transcription factors: key regulators of the flavonoid biosynthetic pathway in grapevine. Protoplasma 249:109–118

    Article  CAS  Google Scholar 

  • Dellus V, Heller W, Sandermann H, Scalbert A (1997) Dihydroflavonol 4-reductase activity in lignocellulosic tissues. Phytochemistry 45:1415–1418

    Article  CAS  Google Scholar 

  • Ding XS, Mannas SW, Bishop BA, Rao X, Lecoultre M, Kwon S, Nelson RS (2018) An improved brome mosaic virus silencing vector: greater insert stability and more extensive VIGS. Plant Physiol 176:496–510

    Article  CAS  PubMed  Google Scholar 

  • Duan CG, Wang CH, Guo HS (2012) Application of RNA silencing to plant disease resistance. Silence 3:1–8

    Article  CAS  Google Scholar 

  • Fu DQ, Zhu BZ, Zhu HL, Zhang HX, Xie YH, Jiang WB, Luo YB (2006) Enhancement of virus-induced gene silencing in tomato by low temperature and low humidity. Mol Cells 21:153–160

    CAS  PubMed  Google Scholar 

  • Gargouri M, Manigand C, Chloé M, Granier T, d’Estaintot BL, Cala O, Pianet I, Bathany K, Chaudièrea J, Galloisa B (2009) Structure and epimerase activity of anthocyanidin reductase from vitis vinifera. Acta Crystallogr A 65:989–1000

    CAS  Google Scholar 

  • Gowri R, Avendano EE, Chen SY, Wang JQ, Julia M, Bridget G, Novotny JA, Aedín C (2019) Dietary intakes of flavan-3-ols and cardiometabolic health: systematic review and meta-analysis of randomized trials and prospective cohort studies. Am J Clin Nutr 110:1067–1078

    Article  Google Scholar 

  • Huang C, Qian Y, Li Z, Zhou X (2012) Virus-induced gene silencing and its application in plant functional genomics. Sci China Life Sci 55:99–108

    Article  CAS  PubMed  Google Scholar 

  • Huang YF, Vialet S, Guiraud JL, Torregrosa L, Bertrand Y, Cheynier V, This P, Terrier N (2014) A negative myb regulator of proanthocyanidin accumulation, identified through expression quantitative locus mapping in thegrape berry. New Phytol 201:795–809

    Article  CAS  PubMed  Google Scholar 

  • Ido Y, Nakahara KS, Uyeda I (2012) White clover mosaic virus-induced gene silencing in pea. J Gen Plant Pathol 78:127–132

    Article  CAS  Google Scholar 

  • Jarugula S, Willie K, Stewart LR (2018) Barley stripe mosaic virus (BSMV) as a virus-induced gene silencing vector in maize seedlings. Virus Genes 54:616–620

    Article  CAS  PubMed  Google Scholar 

  • Kim J, Park M, Jeong ES, Lee JM, Choi D (2017) Harnessing anthocyanin-rich fruit: a visible reporter for tracing virus-induced gene silencing in pepper fruit. Plant Meth 13:1–10

    Article  CAS  Google Scholar 

  • Kurth EG, Peremyslov VV, Prokhnevsky AI, Kasschau KD, Miller M, Carrington JC, Dolja VV (2012) Virus-derived gene expression and RNA interference vector for grapevine. J Virol 86:6002–6009

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lange M, Yellina AL, Orashakova S, Becker A (2013) Virus-induced gene silencing (VIGS) in plants: An overview of target species and the virus-derived vector systems. Methods Mol Biol 975:1–14

    Article  CAS  PubMed  Google Scholar 

  • Li CH (2015) The accumulation of catechin and epicatechin, expression of anthocyanidin reductase induced by UV-C irradiation in grape berry. Shanxi Agricultural University, 15 (in Chinese)

  • Li G, Zhao J, Qin B, Yin Y, Cao Y (2019) ABA mediates development-dependent anthocyanin biosynthesis and fruit coloration in lycium plants. BMC Plant Biol 19:1–13

    Article  Google Scholar 

  • Li L, Zhu B, Fu D, Luo Y (2011) RIN transcription factor plays an important role in ethylene biosynthesis of tomato fruit ripening. J Sci Food Agr 91:2308–2314

    Article  CAS  Google Scholar 

  • Ma CM, Sato N, Li XY, Nakamura N, Hattori M (2010) Flavan-3-ol contents, anti-oxidative and α-glucosidase inhibitory activities of Cynomorium songaricum. Food Chem 118:116–119

    Article  CAS  Google Scholar 

  • Maged AF, Amin M, Osman H, Nada LAM (2018) Optical absorption spectroscopy of the blackberry dye applied in solar cell sensitizers and gamma radiation effects. Arab J Nucl Sci Appl 51(3):130–138

    Google Scholar 

  • Murugantham M, Moskovitz Y, Haviv S, Horesh T, Fenigstein A, Preez JD, Stephan D, Burger JT, Mawassi M (2009) Grapevine virusa-mediated gene silencing in Nicotiana benthamiana and Vitis vinifera. J Virol Methods 155:167–174

    Article  CAS  Google Scholar 

  • Naing AH, Kyu SY, Pe PPW, Park KI, Lee JM, Lim KB, Kim KC (2019) Silencing of the phytoene desaturase (PDS) gene affects the expression of fruit-ripening genes in tomatoes. Plant Meth 15:1679–1683

    Article  Google Scholar 

  • Ottaviani JI, Carrasquedo F, Keen CL, Lazarus SA, Schmitz HH, Fraga CG (2002) Influence of flavan-3-ols and procyanidins on UVC-mediated formation of 8-oxo-7, 8-dihydro-2′-deoxyguanosine in isolated DNA. Arch Biochem Biophys 406:203–208

    Article  CAS  PubMed  Google Scholar 

  • Padmanabhan MS, Dinesh-Kumar SP (2008) Virus-induced gene silencing (VIGS). Encyclop Virol 3:123–131

    Google Scholar 

  • Qi XL, Li M, Liu CL, Song LL (2018) Construction of TRV-mediated virus induced gene silencing(VIGS) system in sweet cherry fruit. Journal of Fruit Science 35:3–9 (in Chinese)

    Google Scholar 

  • Reddy AM, Reddy VS, Scheffler BE, Wienand U, Reddy AR (2007) Novel transgenic rice overexpressing anthocyanidin synthase accumulates a mixture of flavonoids leading to an increased antioxidant potential. Metab Eng 9:95–111

    Article  CAS  PubMed  Google Scholar 

  • Romero I, Tikunov Y, Bovy A (2011) Virus-induced gene silencing in detached tomatoes and biochemical effects of phytoene desaturase gene silencing. J Plant Physiol 168:1129–1135

    Article  CAS  PubMed  Google Scholar 

  • Saito K, Kobayashi M, Gong ZZ, Tanaka Y, Yamazaki M (1999) Direct evidence for anthocyanidin synthase as a 2-oxoglutarate-dependent oxygenase: molecular cloning and functional expression of cDNA from a red forma of Perilla frutescens. Plant J 17:181–189

    Article  PubMed  Google Scholar 

  • Senthil-Kumar M, Lee HK, Mysore KS (2013) VIGS-mediated forward genetics screening for identification of genesinvolved in nonhost resistance. JoVE 78:1–10.

    Google Scholar 

  • Senthil-Kumar M, Mysore KS (2011) New dimensions for VIGS in plant functional genomics. Trends Plant Sci 16:656–665

    Article  CAS  PubMed  Google Scholar 

  • Simone DC, Gabriele DG, Raffaella M, Alberto N, Enrico P, Sophie P, Anne-Francoise AB, Raffaele T (2006) Colour variation in red grapevines (Vitis vinifera L.): genomic organisation, expression of flavonoid 3’-hydroxylase, flavonoid 3’,5’-hydroxylase genes and related metabolite profiling of red cyanidin-/blue delphinidin-based anthocyanins in berry skin. BMC Genom 7:1–17

    CAS  Google Scholar 

  • Tanner GJ, Francki KT, Abrahams S, Watson JM, Larkin PJ, Ashton AR (2003) Proanthocyanidin biosynthesis in plants-purification of legume leucoanthocyanidin reductase and molecular cloning of its cDNA. J Biol Chem 278:31647–31656

    Article  CAS  PubMed  Google Scholar 

  • Thill J, Regos I, Farag MA, Ahmad AF, Kusek J, Castro A, Schlangen K, Carbonero CH, Gadjev IZ, Smith LMJ, Halbwirth H, Treutter D, Stich K (2012) Polyphenol metabolism provides a screening tool for beneficial effects of Onobrychis viciifolia (sainfoin). Phytochemistry 82:67–80

    Article  CAS  PubMed  Google Scholar 

  • Tian J, Pei HX, Zhang S, Chen JW, Chen W, Yang RY, Meng YL, You J, Gao JP, Ma N (2014) TRV-GFP: a modified tobacco rattle virus vector for efficient and visualizable analysis of gene function. J Exp Bot 65:311–322

    Article  CAS  PubMed  Google Scholar 

  • Vashisth T, Johnson LK, Malladi A (2011) An efficient RNA isolation procedure and identification of reference genes for normalization of gene expression in blueberry. Plant Cell Rep 30:2167–2176

    Article  CAS  PubMed  Google Scholar 

  • Wang H, Chang XX, Lin J, Chang Y, Jiang CZ (2018) Transcriptome profiling reveals regulatory mechanisms underlying corolla senescence in petunia. Hort Res 5:453–480

    Google Scholar 

  • Wen PF, Ji W, Gao MY, Niu TQ, Xing YF, Niu XY (2015) Accumulation of flavanols and expression of leucoanthocyanidin reductase induced by postharvest UV-C irradiation in grape berry. Genet Mol Res 14:7687–7695

    Article  CAS  PubMed  Google Scholar 

  • Wen PF, Xing YF, Niu TQ, Gao MY, Niu XY (2012) Accumulation of flavanols, expression of leucoanthocyanidin reductase induced by UV-C irradiation during grape berry development. Scientia Agricultura Sinica 45:4428–4436 (in Chinese)

    CAS  Google Scholar 

  • Xie DY, Sharma SB, Paiva NL, Ferreira D, Dixon RA (2003) Role of anthocyanidin reductase, encoded by BANYULS in plant flavonoid biosynthesis. Science 299:396–399

    Article  CAS  PubMed  Google Scholar 

  • Xie YH, Zhu BZ, Yang XL, Zhang HX, Fu DQ, Zhu HL, Shao Y, Li YC, Gao HY, Luo YB (2006) Delay of postharvest ripening and senescence of tomato fruit through virus-induced leACS2 gene silencing. Postharvest Biol Tec 42:8–15

    Article  CAS  Google Scholar 

  • Yan HJ, Shi SC, Ma N, Cao XQ, Zhang H, Qiu XQ, Wang QG, Jian HY, Zhou NN, Zhang Z, Tang KX (2018) Graft-accelerated virus-induced gene silencing facilitates functional genomics in rose flowers. J Integr Plant Biol 60:34–44

    Article  CAS  PubMed  Google Scholar 

  • Yoo BC, Kragler F, Varkonyi-Gasic E, Haywood V, Archer-Evans S, Lee YM, Lough TJ, Lucas WJ (2004) A systemic small RNA signaling system in plants. Plant Cell 16:1979–2000

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhang B, Shi JA, Chen JB, Li DD, Li JJ, Guo HL, Zong JQ, Wang Y, Guo AG, Liu JX (2016) Efficient virus-induced gene silencing in Cynodon dactylon and Zoysia japonica using rice tungro bacilliform virus vectors. Sci Hortic 207:97–103

    Article  CAS  Google Scholar 

  • Zhang PF, Dong YM, Wen HY, Liang CM, Niu TQ, Gao Y, Zhang XJ, Wen PF (2019) Knockdown of VvMYBA1 via virus-induced gene silencing decreases anthocyanin biosynthesis in grape berries. Can J Plant Sci 100:175–184

    Article  CAS  Google Scholar 

  • Zhang XL, Liu YJ, Gao KJ, Zhao L, Liu L, Wang YS, Sun ML, Gao LP, Xia T (2012) Characterisation of anthocyanidin reductase from Shuchazao green tea. J Sci Food Agr 92:1533–1539

    Article  CAS  Google Scholar 

  • Zheng XT, Chen YL, Zhang XH, Cai ML, Yu ZC, Peng CL (2019) ANS-deficient Arabidopsis is sensitive to high light due to impaired anthocyanin photoprotection. Funct Plant Biol. https://doi.org/10.1071/FP19042

    Article  PubMed  Google Scholar 

Download references

Acknowledgements

Thanks to the National Grape Germplasm Resources Nursery for the convenience of sample collection.

Funding

This work was supported by the Applied Basic Research Project of Shanxi Province (No. 201901D111222), the National Natural Science Foundation of China (No. 31372013), Key Research and Development Program Key Projects Scientific and Technological Project of Shanxi Province (No. 201803D211016-4).

Author information

Authors and Affiliations

  1. College of Horticulture, Shanxi Agricultural University, Taigu, 030801, Shanxi, China

    Bo Yang, Ying Wei, Jianyong Guo, Tiequan Niu, Pengfei Zhang & Pengfei Wen

  2. College of Information Science and Engineering, Shanxi Agricultural University, Taigu, 030801, Shanxi, China

    Changmei Liang

  3. Shanxi Key Laboratory of Germplasm Improvement and Utilization in Pomology, Shanxi Agricultural University, Taigu, 030801, Shanxi, China

    Tiequan Niu, Pengfei Zhang & Pengfei Wen

Authors
  1. Bo Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ying Wei
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Changmei Liang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jianyong Guo
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Tiequan Niu
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Pengfei Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Pengfei Wen
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Pengfei Wen.

Ethics declarations

Conflict of interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Additional information

Handling Editor: Hanns H. Kassemeyer

Publisher's note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Yang, B., Wei, Y., Liang, C. et al. VvANR silencing promotes expression of VvANS and accumulation of anthocyanin in grape berries. Protoplasma 259, 743–753 (2022). https://doi.org/10.1007/s00709-021-01698-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00709-021-01698-y

Keywords

Search

Navigation