Abstract
The MYB, MYC, and WD40 transcription factors (TFs) are known to regulate the expression of structural biosynthetic genes at different steps depending on the plant species. In this work, we used an agroinfiltration protocol with Tobacco rattle virus (TRV) constructs containing partial sequences from MYB or WD40 for virus-induced gene silencing (VIGS) to demonstrate their participation in the regulation of anthocyanin biosynthesis in chilli pepper (Capsicum eximium) fruits. The accumulation of anthocyanins in chilli pepper fruits of plants transformed with TRV2-MYB and TRV2-WD40 constructs was significantly reduced compared to the control or empty TRV2-transformed plants. A significant reduction in gene expression of both TFs was also detected. The expressions of the chalcone synthase (CHS), chalcone isomerase (CHI), flavonoid 3′,5′-hydroxylase (F3′5′H), dihydroflavonol 4-reductase (DFR), and UDP-glucose:flavonoid 3-O-glucosyltransferase (3GT) genes were decreased in the plants transformed with the TRV2-MYB construct but not the transcription of flavanone 3-hydroxylase (F3H). When chilli pepper plants were infected with the TRV2-WD40 construct, a significant reduction in CHS, F3H, F3′5′H, DFR and 3GT expression, but not in CHI in the fruits was observed.
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Abbreviations
- ANS:
-
anthocyanidin synthase
- CHS:
-
chalcone synthase
- CHI:
-
chalcone isomerase
- DFR:
-
dihydroflavonol 4-reductase
- F3H:
-
flavanone 3-hydroxylase
- F3′H:
-
flavonoid 3′-hydroxylase
- F3′5′H:
-
flavonoid 3′,5′-hydroxylase
- 3GT:
-
UDPglucose: flavonoid 3-O-glucosyltransferase
- GST:
-
glutathione S-transferase
- OMT:
-
O-methyltransferase
- TF:
-
transcription factor
- VIGS:
-
virus-induced gene silencing
References
Aza-González, C., Ochoa-Alejo, N.: Characterization of anthocyanins from fruits of two Mexican chilli peppers (Capsicum annuum L.). — J. mexican chem. Soc. 56: 143–145, 2012.
Aza-González, C., Herrera-Isidrón, L., Núñez-Palenius, H.G., Martínez de la Vega, O., Ochoa-Alejo, N.: Anthocyanin accumulation and expression analysis of biosynthesisrelated genes during chilli pepper fruit development. — Biol. Plant. 57: 49–55, 2013.
Borovsky, Y., Oren-Shamir, M., Ovadia, R., De Jong, W., Paran, I.: The A locus that controls anthocyanin accumulation in pepper encodes a MYB transcription factor homologous to Anthocyanin2 of Petunia. — Theor. appl. Genet. 109: 23–29, 2004.
De Vetten, N., Quattrocchio, F., Mol, J., Koes, R.: The an11 locus controlling flower pigmentation in petunia encodes a novel WD-repeat protein conserved in yeast, plants and animals. — Genes Dev. 11: 1422–1434, 1997.
González, A., Zhao, M., Leavitt, J.M., Lloyd, M.: Regulation of the anthocyanin biosynthetic pathway by the TTG1/bHLH/MYB transcriptional complex in Arabidopsis seedlings. — Plant J. 53: 814–827, 2008.
Gould, B., Kramer, E.M.: Virus-induced gene silencing as a tool for functional analyses in the emerging model plant Aquilegia (columbine, Ranunculaceae). — Plant Meth. 3: 6–18, 2007.
Grotewold, E., Sainz, M.B., Tagliani, L., Hernández, J.M., Bowen, B., Chandler, V.L.: Identification in the residues in the MYB domain of maize C1 that specify the interaction with de bHLH cofactor R. — Proc. nat. Acad. Sci. USA 97: 13579–13584, 2000.
Holton, T.A., Cornish, E.C.: Genetics and biochemistry of anthocyanin biosynthesis. — Plant Cell 7: 1071–1083, 1995.
Koes, R., Verweij, W., Quattrocchio, F.: Flavonoids: a colorful model for the regulation and evolution of biochemical pathways. — Trends Plant Sci. 10: 236–242, 2005.
Lightbourn, G.J., Griesbach, R.J., Novotny, J.A., Clevidence, B.A., Rao, D.D., Stommel, J.R.: Effects of anthocyanin and carotenoid combinations on foliage and immature fruit color of Capsicum annuum L. — J. Hered. 99: 105–111, 2008.
Liu, Y., Schiff, M., Marathe, R., Dinesh-Kumar, S.P.: Tobacco Rar1, EDS1 and NPR1/NIM1 like genes are required for Nmediated resistance to tobacco mosaic virus. — Plant J. 30: 415–429, 2002a.
Liu, Y., Schiff, M., Marathe, R., Dinesh-Kumar, S.P.: Virus induced gene silencing in tomato. — Plant J. 31: 777–786, 2002b.
Ma, H., Pooler, M., Griesbach, R.: Anthocyanin regulatory/structural gene expression in Phalaenopsis. — J. amer. Soc. hort. Sci. 134: 88–96, 2009.
Martin, C., Paz-Ares, J.: MYB transcription factors in plants. — Trends Genet. 13: 67–73, 1997.
Martin, C., Prescott, A., Mackay, S., Bartlett, J., Vrijlandt, E.: Control of anthocyanin biosynthesis in flowers of Anthirrinum majus. — Plant J. 1: 37–49, 1991.
Mehrtens, F., Krnz, H., Bednarek, P., Weishaar, B.: The Arabidopsis transcription factor MYB12 is a flavonolspecific regulator of phenylpropanoid biosynthesis. — Plant Physiol. 138: 1083–1096, 2005.
Payne, C.T., Zhang, F., Lloyd, A.M.: GL3 encodes a bHLH protein that regulates trichome development in Arabidopsis through interaction with GL1 and TTG1. — Genetics 156: 1349–1362, 2000.
Peterson, P.A.: Linkage of fruit shape and color genes in Capsicum. — Genetics 44: 407–419, 1959.
Petroni, K., Tonelli, C.: Recent advances on the regulation of anthocyanin synthesis in reproductive organs. — Plant Sci. 181: 219–229, 2011.
Quattrocchio, F., Wing, J.F., Leppen, T.C., Mol, J.N.M., Koes, R.E.: Regulatory genes controlling anthocyanin pigmentation are functionally conserved among plant species and have distinct sets of target genes. — Plant Cell 5: 1497–1512, 1993.
Quattrocchio, F., Wing, J.F., Van der Woude, K., Mol, J.N.M., Koes, R.: Analysis of bHLH and MYB domain proteins: species-specific regulatory differences are caused by divergent evolution of target anthocyanin genes. Plant J. 13: 475–488, 1998.
Ramsay, N.A., Glover, B.J.: MYB-bHLH-WD40 protein complex and the evolution of cellular diversity. — Trends Plant Sci. 10: 63–70, 2005.
Stommel, J.R., Lightbourn, G.J., Winkel, B.S., Griesbach, R.J.: Transcription factors families regulate the anthocyanin biosynthetic pathway in Capsicum annuum. — J. amer. Soc. hort. Sci. 134: 244–251, 2009.
Stracke, R., Werber, M., Weisshaar, B.: The R2R3-MYB gene family in Arabidopsis thaliana. — Curr. Opin. Plant Biol. 4: 447–456, 2001.
Winkel-Shirley, B.: Flavonoid biosynthesis. A colorful model for genetics, biochemistry, cell biology and biotechnology. — Plant Physiol. 126: 485–493, 2001.
Zhang, F., Gonzalez, A., Zhao, M., Payne, C.T., Lloyd, A.: A network of redundant bHLH proteins functions in all TTG1-dependent pathways in Arabidopsis. — Development 130: 4859–4869, 2003.
Zhao, L., Gao, L., Wang, H., Chen, X., Wang, Y., Yang, H., Wei, C., Wan, X., Xia, T.: The R2R3-MYB, bHLH, WD40, and related transcription factors in flavonoid biosynthesis. — Funct. Integr. Genomics 13: 75–98, 2013.
Zimmermann, I.M., Heim, M.A., Weisshaar, B., Uhrig, J.F.: Comprehensive identification of Arabidopsis thaliana MYB transcription factors interacting with R/B like BHLH proteins. — Plant J. 40: 22–34, 2004.
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Acknowledgements: We thank Dr. César Aza-González for participating in the design of the study and for his help with experiments. Funding of this work came from the Consejo Nacional de Ciencia y Tecnología, Mexico (Conacyt project 55264).
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Aguilar-Barragán, A., Ochoa-Alejo, N. Virus-induced silencing of MYB and WD40 transcription factor genes affects the accumulation of anthocyanins in chilli pepper fruit. Biol Plant 58, 567–574 (2014). https://doi.org/10.1007/s10535-014-0427-4
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DOI: https://doi.org/10.1007/s10535-014-0427-4