Abstract
The accumulation of anthocyanins in plants exposed to salt stress has been largely documented. However, the functional link and regulatory components underlying the biosynthesis of these molecules during exposure to stress are largely unknown. In a screen of second site suppressors of the salt overly sensitive3-1 (sos3-1) mutant, we isolated the anthocyanin-impaired-response-1 (air1) mutant. air1 is unable to accumulate anthocyanins under salt stress, a key phenotype of sos3-1 under high NaCl levels (120 mM). The air1 mutant showed a defect in anthocyanin production in response to salt stress but not to other stresses such as high light, low phosphorous, high temperature or drought stress. This specificity indicated that air1 mutation did not affect anthocyanin biosynthesis but rather its regulation in response to salt stress. Analysis of this mutant revealed a T-DNA insertion at the first exon of an Arabidopsis thaliana gene encoding for a basic region-leucine zipper transcription factor. air1 mutants displayed higher survival rates compared to wild-type in oxidative stress conditions, and presented an altered expression of anthocyanin biosynthetic genes such as F3H, F3′H and LDOX in salt stress conditions. The results presented here indicate that AIR1 is involved in the regulation of various steps of the flavonoid and anthocyanin accumulation pathways and is itself regulated by the salt-stress response signalling machinery. The discovery and characterization of AIR1 opens avenues to dissect the connections between abiotic stress and accumulation of antioxidants in the form of flavonoids and anthocyanins.
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References
Abrahams S, Tanner GJ, Larkin PJ, Ashton AR (2002) Identification and biochemical characterization of mutants in the proanthocyanidin pathway in Arabidopsis. Plant Physiol 130:561–576. doi:10.1104/pp.006189
Abrahams S, Lee E, Walker AR et al (2003) The Arabidopsis TDS4 gene encodes leucoanthocyanidin dioxygenase (LDOX) and is essential for proanthocyanidin synthesis and vacuole development. Plant J 35:624–636
Aeschbacher R, Schrott M, Potrykus I, Saul M (1991) Isolation and molecular characterization of PosF21, an Arabidopsis thaliana gene which shows characteristics of a b-Zip class transcription factor. Plant J 1:303–316. doi:10.1046/j.1365-313X.1991.t01-1-00999.x
Borghesi E, González-Miret ML, Escudero-Gilete ML et al (2011) Effects of salinity stress on carotenoids, anthocyanins, and color of diverse tomato genotypes. J Agric Food Chem 59:11676–11682. doi:10.1021/jf2021623
Brown DE, Rashotte AM, Murphy AS et al (2001) Flavonoids act as negative regulators of auxin transport in vivo in Arabidopsis. Plant Physiol 126:524–535. doi:10.1104/pp.126.2.524
Buer CS, Djordjevic MA (2009) Architectural phenotypes in the transparent testa mutants of Arabidopsis thaliana. J Exp Bot 60:751–763. doi:10.1093/jxb/ern323
Chalker-Scott L (1999) Environmental significance of anthocyanins in plant stress responses. Photochem Photobiol 70:1–9. doi:10.1111/j.1751-1097.1999.tb01944.x
Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16:735–743. doi:10.1046/j.1365-313x.1998.00343.x
Deppmann CD, Acharya A, Rishi V et al (2004) Dimerization specificity of all 67 B-ZIP motifs in Arabidopsis thaliana: a comparison to Homo sapiens B-ZIP motifs. Nucleic Acids Res 32:3435–3445. doi:10.1093/nar/gkh653
Dubos C, Le Gourrierec J, Baudry A et al (2008) MYBL2 is a new regulator of flavonoid biosynthesis in Arabidopsis thaliana. Plant J 55:940–953. doi:10.1111/j.1365-313X.2008.03564.x
Dubrovsky JG, Sauer M, Napsucialy-Mendivil S et al (2008) Auxin acts as a local morphogenetic trigger to specify lateral root founder cells. PNAS 105:8790–8794. doi:10.1073/pnas.0712307105
Edreva A (2005) The importance of non-photosynthetic pigments and cinnamic acid derivatives in photoprotection. Agric Ecosyst Environ 106:135–146. doi:10.1016/j.agee.2004.10.002
Falcone Ferreyra ML, Casas MI, Questa JI et al (2012a) Evolution and expression of tandem duplicated maize flavonol synthase genes. Front Plant Sci. doi:10.3389/fpls.2012.00101
Falcone Ferreyra ML, Rius SP, Casati P (2012b) Flavonoids: biosynthesis, biological functions, and biotechnological applications. Front Plant Sci 3:222. doi:10.3389/fpls.2012.00222
Frary A, Göl D, Keleş D et al (2010) Salt tolerance in Solanum pennellii: antioxidant response and related QTL. BMC Plant Biol 10:58. doi:10.1186/1471-2229-10-58
Fujii H, Zhu J-K (2009) An autophosphorylation site of the protein kinase SOS2 is important for salt tolerance in Arabidopsis. Mol Plant 2:183–190. doi:10.1093/mp/ssn087
Galvan-Ampudia CS, Testerink C (2011) Salt stress signals shape the plant root. Curr Opin Plant Biol 14:296–302. doi:10.1016/j.pbi.2011.03.019
Geisler M, Blakeslee JJ, Bouchard R et al (2005) Cellular efflux of auxin catalyzed by the Arabidopsis MDR/PGP transporter AtPGP1. Plant J 44:179–194. doi:10.1111/j.1365-313X.2005.02519.x
Gong D, Guo Y, Schumaker KS, Zhu J-K (2004) The SOS3 family of calcium sensors and SOS2 family of protein kinases in Arabidopsis. Plant Physiol 134:919–926. doi:10.1104/pp.103.037440
Harborne JB, Williams CA (2000) Advances in flavonoid research since 1992. Phytochemistry 55:481–504
Hernández I, Alegre L, Van Breusegem F, Munné-Bosch S (2009) How relevant are flavonoids as antioxidants in plants? Trends Plant Sci 14:125–132. doi:10.1016/j.tplants.2008.12.003
Hichri I, Barrieu F, Bogs J et al (2011) Recent advances in the transcriptional regulation of the flavonoid biosynthetic pathway. J Exp Bot 62:2465–2483. doi:10.1093/jxb/erq442
Holm M, Hardtke CS, Gaudet R, Deng X-W (2001) Identification of a structural motif that confers specific interaction with the WD40 repeat domain of Arabidopsis COP1. EMBO J 20:118–127. doi:10.1093/emboj/20.1.118
Ishitani M, Liu J, Halfter U, Kim C-S, Shi W, Zhu J-K (2000) SOS3 function in plant salt tolerance requires N-myristoylation and calcium binding. Plant Cell 12:1667–1678. doi:10.1105/tpc.12.9.1667
Keller CP, Volkenburgh EV (1996) Osmoregulation by oat coleoptile protoplasts (effect of auxin). Plant Physiol 110:1007–1016. doi:10.1104/pp.110.3.1007
Ko FN, Chu CC, Lin CN et al (1998) Isoorientin-6″-O-glucoside, a water-soluble antioxidant isolated from Gentiana arisanensis. Biochim Biophys Acta 1389:81–90
Koiwa H, Bressan RA, Hasegawa PM (2006) Identification of plant stress-responsive determinants in arabidopsis by large-scale forward genetic screens. J Exp Bot 57:1119–1128. doi:10.1093/jxb/erj093
Leopoldini M, Russo N, Chiodo S, Toscano M (2006) Iron chelation by the powerful antioxidant flavonoid quercetin. J Agric Food Chem 54:6343–6351. doi:10.1021/jf060986h
Lewis DR, Ramirez MV, Miller ND et al (2011) Auxin and ethylene induce flavonol accumulation through distinct transcriptional networks. Plant Physiol 156:144–164. doi:10.1104/pp.111.172502
Liu J, Zhu J-K (1997) An Arabidopsis mutant that requires increased calcium for potassium nutrition and salt tolerance. PNAS 94:14960–14964
Liu Y-G, Mitsukawa N, Oosumi T, Whittier RF (1995) Efficient isolation and mapping of Arabidopsis thaliana T-DNA insert junctions by thermal asymmetric interlaced PCR. Plant J 8:457–463. doi:10.1046/j.1365-313X.1995.08030457.x
Liu C, Li S, Wang M, Xia G (2012) A transcriptomic analysis reveals the nature of salinity tolerance of a wheat introgression line. Plant Mol Biol 78:159–169. doi:10.1007/s11103-011-9854-1
Ljung K, Hull AK, Kowalczyk M, et al. (2002) Biosynthesis, conjugation, catabolism and homeostasis of indole-3-acetic acid in Arabidopsis thaliana. In: Perrot-Rechenmann C, Hagen G (eds) Auxin molecular biology. Springer Netherlands, pp 249–272
Lois R, Buchanan BB (1994) Severe sensitivity to ultraviolet radiation in an Arabidopsis mutant deficient in flavonoid accumulation. Planta 194:504–509. doi:10.1007/BF00714463
Lukaszewicz M, Matysiak-Kata I, Skala J et al (2004) Antioxidant capacity manipulation in transgenic potato tuber by changes in phenolic compounds content. J Agric Food Chem 52:1526–1533. doi:10.1021/jf034482k
Matsui K, Umemura Y, Ohme-Takagi M (2008) AtMYBL2, a protein with a single MYB domain, acts as a negative regulator of anthocyanin biosynthesis in Arabidopsis. Plant J 55:954–967. doi:10.1111/j.1365-313X.2008.03565.x
Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497. doi:10.1111/j.1399-3054.1962.tb08052.x
Neff M, Chory J (1998) Genetic interactions between phytochrome A, phytochrome B, and cryptochrome 1 during Arabidopsis development. Plant Physiol 118:27–35. doi:10.1104/PP.118.1.27
Oh JE, Kim YH, Kim JH et al (2011) Enhanced level of anthocyanin leads to increased salt tolerance in arabidopsis PAP1-D plants upon sucrose treatment. J Korean Soc Appl Biol Chem 54:79–88. doi:10.3839/jksabc.2011.011
Peer WA, Murphy AS (2007) Flavonoids and auxin transport: modulators or regulators? Trends Plant Sci 12:556–563
Peer WA, Brown DE, Tague BW et al (2001) Flavonoid accumulation patterns of transparent testa mutants of arabidopsis. Plant Physiol 126:536–548
Peer WA, Bandyopadhyay A, Blakeslee JJ et al (2004) Variation in expression and protein localization of the PIN family of auxin efflux facilitator proteins in flavonoid mutants with altered auxin transport in Arabidopsis thaliana. Plant Cell 16:1898–1911. doi:10.1105/tpc.021501
Petroni K, Tonelli C (2011) Recent advances on the regulation of anthocyanin synthesis in reproductive organs. Plant Sci 181:219–229. doi:10.1016/j.plantsci.2011.05.009
Quintero FJ, Martinez-Atienza J, Villalta I et al (2011) Activation of the plasma membrane Na/H antiporter salt-overly-sensitive 1 (SOS1) by phosphorylation of an auto-inhibitory C-terminal domain. PNAS 108:2611–2616. doi:10.1073/pnas.1018921108
Rahman A, Takahashi M, Shibasaki K et al (2010) Gravitropism of Arabidopsis thaliana roots requires the polarization of PIN2 toward the root tip in meristematic cortical cells. Plant Cell 22:1762–1776. doi:10.1105/tpc.110.075317
Ramakrishna A, Ravishankar GA (2011) Influence of abiotic stress signals on secondary metabolites in plants. Plant Signal Behav 6:1720–1731. doi:10.4161/psb.6.11.17613
Re R, Pellegrini N, Proteggente A et al (1999) Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radic Biol Med 26:1231–1237
Reddy AM, Reddy VS, Scheffler BE et al (2007) Novel transgenic rice overexpressing anthocyanidin synthase accumulates a mixture of flavonoids leading to an increased antioxidant potential. Metab Eng 9:95–111. doi:10.1016/j.ymben.2006.09.003
Rice-Evans C, Miller N, Paganga G (1997) Antioxidant properties of phenolic compounds. Trends Plant Sci 2:152–159. doi:10.1016/S1360-1385(97)01018-2
Ringli C, Bigler L, Kuhn BM et al (2008) The modified flavonol glycosylation profile in the Arabidopsis rol1 mutants results in alterations in plant growth and cell shape formation. Plant Cell 20:1470–1481. doi:10.1105/tpc.107.053249
Rowan DD, Cao M, Lin-Wang K et al (2009) Environmental regulation of leaf colour in red 35S:PAP1 Arabidopsis thaliana. New Phytol 182:102–115. doi:10.1111/j.1469-8137.2008.02737.x
Rus A, Yokoi S, Sharkhuu A et al (2001) AtHKT1 is a salt tolerance determinant that controls Na+ entry into plant roots. PNAS 98:14150–14155. doi:10.1073/pnas.241501798
Sanchez-Barrena MJ, Fujii H, Angulo I et al (2007) The structure of the C-terminal domain of the protein kinase AtSOS2 bound to the calcium sensor AtSOS3. Mol Cell 26:427–435. doi:10.1016/j.molcel.2007.04.013
Santelia D, Henrichs S, Vincenzetti V et al (2008) Flavonoids redirect PIN-mediated polar auxin fluxes during root gravitropic responses. J Biol Chem 283:31218–31226. doi:10.1074/jbc.M710122200
Shi H, Ishitani M, Kim C, Zhu J-K (2000) The Arabidopsis thaliana salt tolerance gene SOS1 encodes a putative Na+/H+ antiporter. PNAS 97:6896–6901. doi:10.1073/pnas.120170197
Shi H, Lee B, Wu S-J, Zhu J-K (2003) Overexpression of a plasma membrane Na+/H+ antiporter gene improves salt tolerance in Arabidopsis thaliana. Nat Biotech 21:81–85. doi:10.1038/nbt766
Sun F, Zhang W, Hu H et al (2008) Salt modulates gravity signaling pathway to regulate growth direction of primary roots in Arabidopsis. Plant Physiol 146:178–188. doi:10.1104/pp.107.109413
Vanneste S, Friml J (2009) Auxin: a trigger for change in plant development. Cell 136:1005–1016. doi:10.1016/j.cell.2009.03.001
Wang Y, Zhang W, Li K, Sun F, Han C, Li X (2008) Salt-induced plasticity of root hair development is caused by ion disequilibrium in Arabidopsis thaliana. J Plant Res 121:87–96
Weigel D, Ahn JH, Blázquez MA et al (2000) Activation tagging in Arabidopsis. Plant Physiol 122:1003–1014. doi:10.1104/pp.122.4.1003
Winkel-Shirley B (2001) Flavonoid biosynthesis. A colorful model for genetics, biochemistry, cell biology, and biotechnology. Plant Physiol 126:485–493. doi:10.1104/pp.126.2.485
Wu Y, Zhao Q, Gao L et al (2010) Isolation and characterization of low-sulphur-tolerant mutants of Arabidopsis. J Exp Bot 61:3407–3422. doi:10.1093/jxb/erq161
Yang Q, Chen Z–Z, Zhou X-F et al (2009) Overexpression of SOS (salt overly sensitive) genes increases salt tolerance in transgenic Arabidopsis. Mol Plant 2:22–31. doi:10.1093/mp/ssn058
Zhao Y, Wang T, Zhang W, Li X (2011) SOS3 mediates lateral root development under low salt stress through regulation of auxin redistribution and maxima in Arabidopsis. New Phytol 189:1122–1134. doi:10.1111/j.1469-8137.2010.03545.x
Zhu J-K (2003) Regulation of ion homeostasis under salt stress. Curr Opin Plant Biol 6:441–445. doi:10.1016/S1369-5266(03)00085-2
Zhu H-F, Fitzsimmons K, Khandelwal A, Kranz RG (2009) CPC, a single-repeat R3 MYB, is a negative regulator of anthocyanin biosynthesis in Arabidopsis. Mol Plant 2:790–802. doi:10.1093/mp/ssp030
Acknowledgments
The authors would like to thank Ana Rus for the original screen of sos3-1 suppressor mutants and Becky Fagan for her excellent administrative support. The authors would also like to thank the ABRC stock center for T-DNA mutant lines. A graduate fellowship for M.J.V. was provided by the Ross Fellowship program of Purdue University.
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11103_2013_99_MOESM2_ESM.tif
Fig. S1 air1-1 phenotype under phosphorous deficiency. Three week old plants of gl1, sos3-1 and air1-1 grown on GM media or GM media without phosphorous; Plants were germinated on control GM media for 7 days and then transferred to minus phosphorous plates. Pictures were taken after 2 and 4 weeks as indicated in figure (TIFF 286 kb)
11103_2013_99_MOESM3_ESM.tif
Fig. S2 Quantification of anthocyanins. Three week old plants of gl1, sos3-1, air1-1 and air1-2 sos3-1 grown on GM media or GM media with 120 mM NaCl; the values are reported as A 535—2(A 650) g−1 fresh weight. Asterisks denote significant differences according to Student (P < 0.05) between control and NaCl treated plants (TIFF 61 kb)
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Van Oosten, M.J., Sharkhuu, A., Batelli, G. et al. The Arabidopsis thaliana mutant air1 implicates SOS3 in the regulation of anthocyanins under salt stress. Plant Mol Biol 83, 405–415 (2013). https://doi.org/10.1007/s11103-013-0099-z
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DOI: https://doi.org/10.1007/s11103-013-0099-z