Advertisement

The Arabidopsis thaliana transcription factor MYB59 regulates calcium signalling during plant growth and stress response

  • Elisa Fasani
  • Giovanni DalCorso
  • Alex Costa
  • Sara Zenoni
  • Antonella FuriniEmail author
Article
  • 77 Downloads

Abstract

Key message

Transcription factor MYB59 is involved in plant growth and stress responses by acting as negative regulator of Ca signalling and homeostasis.

Abstract

The Arabidopsis thaliana transcription factor MYB59 is induced by cadmium (Cd) and plays a key role in the regulation of cell cycle progression and root elongation, but its mechanism of action is poorly understood. We investigated the expression of MYB59 and differences between wild-type plants, the myb59 mutant and MYB59-overexpressing lines (obtained by transformation in the mutant genotype) during plant growth and in response to various forms of stress. We also compared the transcriptomes of wild-type and myb59 mutant plants to determine putative MYB59 targets. The myb59 mutant has longer roots, smaller leaves and smaller cells than wild-type plants and responds differently to stress in germination assay. Transcriptomic analysis revealed the upregulation in the myb59 mutant of multiple genes involved in calcium (Ca) homeostasis and signalling, including those encoding calmodulin-like proteins and Ca transporters. Notably, MYB59 was strongly induced by Ca deficiency, and the myb59 mutant was characterized by higher levels of cytosolic Ca in root cells and showed a modest alteration of Ca transient frequency in guard cells, associated with the absence of Ca-induced stomatal closure. These results indicate that MYB59 negatively regulates Ca homeostasis and signalling during Ca deficiency, thus controlling plant growth and stress responses.

Keywords

Arabidopsis thaliana Ca signalling and homeostasis Plant growth MYB transcription factor Stomata Stress response 

Notes

Acknowledgements

The authors are grateful to Dr. Gaétan Glauser of the Neuchâtel Platform of Analytical Chemistry (NPAC, Université de Neuchâtel, Switzerland) for the measurement of IAA content and to Dr. Maria Teresa Scupoli and Dr. Chiara Cavallini of the University Laboratory for Medical Research (LURM, University of Verona, Italy) for the flow cytometry analysis.

Author contributions

EF performed most of the experiments and wrote the article with contribution of AF and GD, AC designed and helped with the Cameleon experiments and provided assistance in understanding the resulting data. SZ provided assistance in microarray analysis and interpretation. AF and GD conceived the project and supervised the experiments. EF and GD equally contributed to this work.

Funding

Funding for E.F.’s PhD and research grant were from Italian Ministry of University and Research (MIUR).

Supplementary material

11103_2019_833_MOESM1_ESM.pdf (272 kb)
Online Resource 1 Semi-quantitative analysis of the expression of the three MYB59 isoforms by real-time RT-PCR under control conditions, Cd treatment, drought, ABA and salt stress. Supplementary material 1 (PDF 272 KB)
11103_2019_833_MOESM2_ESM.pdf (143 kb)
Online Resource 2 Comparison of MYB59 expression in wild-type, myb59 mutant and MYB59- overexpressing lines. Supplementary material 2 (PDF 143 KB)
11103_2019_833_MOESM3_ESM.pdf (146 kb)
Online Resource 3 Distribution of diameter of the protoplast from leaf 4 and 11 of four-week-old plants in wild-type, myb59 mutant and MYB59-overexpressing plants. Supplementary material 3 (PDF 146 KB)
11103_2019_833_MOESM4_ESM.pdf (250 kb)
Online Resource 4 Stress tolerance in wild-type, myb59 mutant and MYB59-overexpressing plants. Supplementary material 4 (PDF 250 KB)
11103_2019_833_MOESM5_ESM.pdf (159 kb)
Online Resource 5 Expression of MYB59 isoforms after 6 and 24 hours and 4 days under low-Ca treatment. Supplementary material 5 (PDF 158 KB)

References

  1. Abe H, Urao T, Ito T, Seki M, Shinozaki K, Yamaguchi-Shinozaki K (2003) Arabidopsis AtMYC2 (bHLH) and AtMYB2 (MYB) function as transcriptional activators in abscisic acid signaling. Plant Cell 15:63–78Google Scholar
  2. Agarwal M, Hao Y, Kapoor A, Dong CH, Fujii H, Zheng X, Zhu JK (2006) A R2R3 type MYB transcription factor is involved in the cold regulation of CBF genes and in acquired freezing tolerance. J Biol Chem 281(49):37636–37645Google Scholar
  3. Allen GJ, Chu SP, Schumacher K, Shimazaki CT, Vafeados D, Kemper A, Hawke SD, Tallman G, Tsien RY, Harper JF, Chory J (2000) Alteration of stimulus-specific guard cell calcium oscillations and stomatal closing in Arabidopsis det3 mutant. Science 289:2338–2342Google Scholar
  4. Allen GJ, Chu SP, Harrington CL, Schumacher K, Hoffmann T, Tang YY, Grill E, Schroeder JI (2001) A defined range of guard cell calcium oscillation parameters encodes stomatal movements. Nature 411(6841):1053–1057Google Scholar
  5. Baliardini C, Meyer CL, Salis P, Saumitou-Laprade P, Verbruggen N (2015) CATION EXCHANGER1 cosegregates with Cadmium tolerance in the metal hyperaccumulator Arabidopsis halleri and plays a role in limiting oxidative stress in Arabidopsis spp. Plant Physiol 169:549–559Google Scholar
  6. Barbez E, Kubeš M, Rolčík J, Béziat C, Pěnčík A, Wang B, Rosquete MR, Zhu J, Dobrev PI, Lee Y, Zažímalovà E (2012) A novel putative auxin carrier family regulates intracellular auxin homeostasis in plants. Nature 485(7396):119–122Google Scholar
  7. Benjamins R, Ampudia CS, Hooykaas PJ, Offringa R (2003) PINOID-mediated signaling involves calcium-binding proteins. Plant Physiol 132(3):1623–1630Google Scholar
  8. Cheng NH, Pittman JK, Barkla BJ, Shigaki T, Hirschi KD (2003) The Arabidopsis cax1 mutant exhibits impaired ion homeostasis, development, and hormonal responses and reveals interplay among vacuolar transporters. Plant Cell 15(2):347–364Google Scholar
  9. Chezem WR, Memon A, Li FS, Weng JK, Clay NK (2017) SG2-type R2R3-MYB transcription factor MYB15 controls defense-induced lignification and basal immunity in Arabidopsis. Plant Cell 29(8):1907–1926Google Scholar
  10. Cho D, Kim SA, Murata Y, Lee S, Jae SK, Nam HG, Kwak JM (2009) De-regulated expression of the plant glutamate receptor homolog AtGLR3.1 impairs long-term Ca2+-programmed stomatal closure. Plant J 58(3):437–449Google Scholar
  11. Cho D, Villiers F, Kroniewicz L, Lee S, Seo YJ, Hirschi KD, Leonhardt N, Kwak JM (2012) Vacuolar CAX1 and CAX3 influence auxin transport in guard cells via regulation of apoplastic pH. Plant Physiol 160(3):1293–1302Google Scholar
  12. Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16:735–743Google Scholar
  13. Cominelli E, Galbiati M, Vavasseur A, Conti L, Sala T, Vuylsteke M, Leonhardt N, Dellaporta SL, Tonelli C (2005) A guard-cell-specific MYB transcription factor regulates stomatal movements and plant drought tolerance. Curr Biol 15(13):1196–1200Google Scholar
  14. Conn SJ, Gilliham M, Athman A, Schreiber AW, Baumann U, Moller I, Cheng NH, Stancombe MA, Hirschi KD, Webb AA, Burton R (2011) Cell-specific vacuolar calcium storage mediated by CAX1 regulates apoplastic calcium concentration, gas exchange, and plant productivity in Arabidopsis. Plant Cell 23(1):240–257Google Scholar
  15. Das M, Harvey I, Chu LL, Sinha M, Pelletier J (2001) Full-length cDNAs: more than just reaching the ends. Physiol Genomics 6:57–80Google Scholar
  16. Dovzhenko A, Dal Bosco C, Meurer J, Koop HU (2003) Efficient regeneration from cotyledon protoplasts in Arabidopsis thaliana. Protoplasma 222(1–2):107–111Google Scholar
  17. Du L, Poovaiah BW (2004) A novel family of Ca2+/calmodulin-binding proteins involved in transcriptional regulation: interaction with fsh/Ring3 class transcription activators. Plant Mol Biol 54(4):549–569Google Scholar
  18. Du H, Zhang L, Liu L, Tang XF, Yang WJ, Wu YM, Huang YB, Tang YX (2009) Biochemical and molecular characterization of plant MYB transcription factor family. Biochemistry 74(1):1–11Google Scholar
  19. Dubos C, Stracke R, Grotewold E, Weisshaar B, Martin C, Lepiniec L (2010) MYB transcription factors in Arabidopsis. Trends Plant Sci 15(10):573–581Google Scholar
  20. Fusco N, Micheletto L, Dal Corso G, Borgato L, Furini A (2005) Identification of cadmium-regulated genes by cDNA-AFLP in the heavy metal accumulator Brassica juncea L. J Exp Bot 56(421):3017–3027Google Scholar
  21. Galbraith DW (2014) Flow cytometry and sorting in Arabidopsis. In: Sanchez-Serrano JJ, Salinas J (eds) Arabidopsis protocols. Humana Press, Totowa, pp 509–537Google Scholar
  22. Glauser G, Vallat A, Balmer D (2014) Hormone profiling. In: Sanchez-Serrano JJ, Salinas J (eds) Arabidopsis protocols. Humana Press, Totowa, pp 597–608Google Scholar
  23. Guo L, Yang H, Zhang X, Yang S (2013) Lipid transfer protein 3 as a target of MYB96 mediates freezing and drought stress in Arabidopsis. J Exp Bot 64:1755–1767Google Scholar
  24. Hickman R, Van Verk MC, Van Dijken AJ, Mendes MP, Vroegop-Vos IA, Caarls L, Steenbergen M, Van der Nagel I, Wesselink GJ, Jironkin A, Talbot A (2017) Architecture and dynamics of the jasmonic acid gene regulatory network. Plant Cell 29:2086–2105Google Scholar
  25. Hinkle PM, Kinsella PA, Osterhoudt KC (1987) Cadmium uptake and toxicity via voltage-sensitive calcium channels. J Biol Chem 262:16333–16337Google Scholar
  26. Hoagland DR, Arnon DI (1950) The water-culture method for growing plants without soil. Circular 347. College of Agriculture, University of California, BerkeleyGoogle Scholar
  27. Huang LQ, Berkelman T, Franklin AE, Hoffman NE (1993) Characterization of a gene encoding a Ca2+-ATPase-like protein in the plastid envelope. Proc Natl Acad Sci USA 90:10066–10070Google Scholar
  28. Huang DW, Sherman BT, Lempicki RA (2009) Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources. Nat Protoc 4(1):44–57Google Scholar
  29. Huang D, Gong X, Liu Y, Zeng G, Lai C, Bashir H, Zhou L, Wang D, Xu P, Cheng M, Wan J (2017) Effects of calcium at toxic concentrations of cadmium in plants. Planta 245:863–873Google Scholar
  30. Journot-Catalino N, Somssich IE, Roby D, Kroj T (2006) The transcription factors WRKY11 and WRKY17 act as negative regulators of basal resistance in Arabidopsis thaliana. Plant Cell 18(11):3289–3302Google Scholar
  31. Katiyar A, Smita S, Lenka SK, Rajwanshi R, Chinnusamy V, Bansal KC (2012) Genome-wide classification and expression analysis of MYB transcription factor families in rice and Arabidopsis. BMC Genom 13:544Google Scholar
  32. Khokon MA, Salam MA, Jammes F, Ye W, Hossain MA, Uraji M, Nakamura Y, Mori IC, Kwak JM, Murata Y (2015) Two guard cell mitogen-activated protein kinases, MPK9 and MPK12, function in methyl jasmonate-induced stomatal closure in Arabidopsis thaliana. Plant Biol 17:946–952Google Scholar
  33. Kim TH, Böhmer M, Hu H, Nishimura N, Schroeder JI (2010) Guard cell signal transduction network: advances in understanding abscisic acid, CO2, and Ca2+ signaling. Annu Rev Plant Biol 61:561–591Google Scholar
  34. Krebs M, Held K, Binder A, Hashimoto K, Den Herder G, Parniske M, Kudla J, Schumacher K (2012) FRET-based genetically encoded sensors allow high-resolution live cell imaging of Ca2+ dynamics. Plant J 69:181–192Google Scholar
  35. Lai AG, Doherty CJ, Mueller-Roeber B, Kay SA, Schippers JHM, Dijkwel PP (2012) CIRCADIAN CLOCK-ASSOCIATED 1 regulates ROS homeostasis and oxidative stress responses. Proc Natl Acad Sci USA 109(42):17129–17134Google Scholar
  36. Li J, Li X, Guo L, Lu F, Feng X, He K, Wei L, Chen Z, Qu LJ, Gu H (2006a) A subgroup of MYB transcription factor genes undergoes highly conserved alternative splicing in Arabidopsis and rice. J Exp Bot 57(6):1263–1273Google Scholar
  37. Li J, Yang X, Wang Y, Li X, Gao Z, Pei M, Chen Z, Qu LJ, Gu H (2006b) Two groups of MYB transcription factors share a motif which enhances trans-activation activity. Biochem Biophys Res Commun 341:1155–1163Google Scholar
  38. Liang YK, Dubos C, Dodd IC, Holroyd GH, Hetherington AM, Campbell MM (2005) AtMYB61, an R2R3-MYB transcription factor controlling stomatal aperture in Arabidopsis thaliana. Curr Biol 15(13):1201–1206Google Scholar
  39. Liao C, Zheng Y, Guo Y (2017) MYB30 transcription factor regulates oxidative and heat stressresponses through ANNEXIN-mediated cytosolic calcium signaling in Arabidopsis. New Phytol 216(1):163–177Google Scholar
  40. Lipsick JS (1996) One billion years of Myb. Oncogene 13:223–235Google Scholar
  41. Liu L, Zhang J, Adrian J, Gissot L, Coupland G, Yu D, Turck F (2014) Elevated levels of MYB30 in the phloem accelerate flowering in Arabidopsis through the regulation of FLOWERING LOCUS T. PLoS ONE 9(2):e89799Google Scholar
  42. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real time quantitative PCR and the 2–∆∆CT method. Methods 25:402–408Google Scholar
  43. Mandadi KK, Misra A, Ren S, McKnight TD (2009) BT2, a BTB protein, mediates multiple responses to nutrients, stresses, and hormones in Arabidopsis. Plant Physiol 150(4):1930–1939Google Scholar
  44. McDonald JH (2014) Handbook of biological statistics, 3rd edn. Sparky House Publishing, BaltimoreGoogle Scholar
  45. Mehrtens F, Kranz H, Bednarek P, Weisshaar B (2005) The Arabidopsis transcription factor MYB12 is a flavonol-specific regulator of phenylpropanoid biosynthesis. Plant Physiol 138(2):1083–1096Google Scholar
  46. Mu RL, Cao YR, Liu YF, Lei G, Zou HF, Liao Y, Wang HW, Zhang WK, Ma B, Du JZ, Yuan M (2009) An R2R3-type transcription factor gene AtMYB59 regulates root growth and cell cycle progression in Arabidopsis. Cell Res 19(11):1291–1304Google Scholar
  47. Munemasa S, Hossain MA, Nakamura Y, Mori IC, Murata Y (2011) The Arabidopsis calcium-dependent protein kinase, CPK6, functions as a positive regulator of methyl jasmonate signaling in guard cells. Plant Physiol 155:553–561Google Scholar
  48. Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497Google Scholar
  49. Nesi N, Jond C, Debeaujon I, Caboche M, Lepiniec L (2001) The Arabidopsis TT2 gene encodes an R2R3 MYB domain protein that acts as a key determinant for proanthocyanidin accumulation in developing seed. Plant Cell 13(9):2099–2114Google Scholar
  50. Nishida S, Kakei Y, Shimada Y, Fujiwara T (2017) Genome-wide analysis of specific alterations in transcript structure and accumulation caused by nutrient deficiencies in Arabidopsis thaliana. Plant J 91:741–753Google Scholar
  51. Novillo F, Alonso JM, Ecker JR, Salinas J (2004) CBF2/DREB1C is a negative regulator of CBF1/DREB1B and CBF3/DREB1A expression and plays a central role in stress tolerance in Arabidopsis. Proc Natl Acad Sci USA 101(11):3985–3990Google Scholar
  52. Ogata K, Morikawa S, Nakamura H, Hojo H, Yoshimura S, Zhang R, Aimoto S, Ametani Y, Hirata Z, Sarai A, Ishii S (1995) Comparison of the free and DNA-complexed forms of the DNA-binding domain from c-Myb. Struct Biol 2:309–320Google Scholar
  53. Oh S, Park S, Han KH (2003) Transcriptional regulation of secondary growth in Arabidopsis thaliana. J Exp Bot 54(393):2709–2722Google Scholar
  54. Paz-Ares J, Ghosal D, Wienand U, Peterson PA, Saedler H (1987) The regulatory c1 locus of Zea mays encodes a protein with homology to myb proto-oncogene products and with structural similarities to transcriptional activators. EMBO J 6(12):3553–3558Google Scholar
  55. Perfus-Barbeoch L, Leonhardt N, Vavaddeur A, Forestier C (2002) Heavy metal toxicity: cadmium permeates through calcium channels and disturbs the plant water status. Plant J 32:539–548Google Scholar
  56. Ramakers C, Ruijter JM, Lekanne Deprez RH, Moorman AFM (2003) Assumption-free analysis of quantitative real-time polymerase chain reaction (PCR) data. Neurosci Lett 339:62–66Google Scholar
  57. Reddy VS, Day IS, Thomas T, Reddy AS (2004) KIC, a novel Ca2+ binding protein with one EF-hand motif, interacts with a microtubule motor protein and regulates trichome morphogenesis. Plant Cell 16(1):185–200Google Scholar
  58. Ren S, Mandadi KK, Boedeker AL, Rathore KS, McKnight TD (2007) Regulation of telomerase in Arabidopsis by BT2, an apparent target of TELOMERASE ACTIVATOR1. Plant Cell 19(1):23–31Google Scholar
  59. Riechmann JL, Heard J, Martin G, Reuber L, Jiang CZ, Keddie J, Adam L, Pineda O, Ratcliffe OJ, Samaha RR, Creelman R (2000) Arabidopsis transcription factors: genome-wide comparative analysis among Eukaryotes. Science 290:2105–2110Google Scholar
  60. Rosinski JA, Atchley WR (1998) Molecular evolution of the Myb family of transcription factors: evidence for polyphyletic origin. J Mol Evol 46:74–83Google Scholar
  61. Ruta LL, Popa VC, Nicolau I, Danet AF, Iordache V, Neagoe AD, Farcasanu IC (2014) Calcium signaling mediates the response to cadmium toxicity in Saccharomyces cerevisiae cells. FEBS Lett 588(17):3202–3212Google Scholar
  62. Schindelin J, Arganda-Carreras I, Frise E, Kaynig V, Longair M, Pietzsch T, Preibisch S, Rueden C, Saalfeld S, Schmid B, Tinevez JY (2012) Fiji: an open-source platform for biological-image analysis. Nat Methods 9(7):676–682Google Scholar
  63. Scholl RL, May ST, Ware DH (2000) Seed and molecular resources for Arabidopsis. Plant Physiol 124:1477–1480Google Scholar
  64. Seo PJ, Lee SB, Suh MC, Park MJ, Go YS, Park CM (2011) The MYB96 transcription factor regulates cuticular wax biosynthesis under drought conditions in Arabidopsis. Plant Cell 23:1138–1152Google Scholar
  65. Stracke R, Werber M, Weisshaar B (2001) The R2R3-MYB gene family in Arabidopsis thaliana. Curr Opin Plant Biol 4:447–456Google Scholar
  66. Tsai YC, Koo Y, Delk NA, Gehl B, Braam J (2013) Calmodulin-related CML24 interacts with ATG4b and affects autophagy progression in Arabidopsis. Plant J 73(2):325–335Google Scholar
  67. Webb AA, Robertson FC (2011) Calcium signals in the control of stomatal movements. In: Luan S (ed) Coding and decoding of calcium signals in plants. Springer, Berlin, pp 63–77Google Scholar
  68. Weston K (1998) Myb proteins in life, death and differentiation. Curr Opin Genetics Dev 8:76–81Google Scholar
  69. Yanhui C, Xiaoyuan Y, Kun H, Meihua L, Jigang L, Zhaofeng G, Zhiqiang L, Yunfei Z, Xiaoxiao W, Xiaoming Q, Yunping S (2006) The MYB transcription factor superfamily of Arabidopsis: expression analysis and phylogenetic comparison with the rice MYB family. Plant Mol Biol 60(1):107–124Google Scholar
  70. Zhou J, Lee C, Zhong R, Ye ZH (2009) MYB58 and MYB63 are transcriptional activators of the lignin biosynthetic pathway during secondary cell wall formation in Arabidopsis. Plant Cell 21(1):248–266Google Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Department of BiotechnologyUniversity of VeronaVeronaItaly
  2. 2.Department of Life SciencesUniversity of MilanoMilanItaly

Personalised recommendations