Characterization of PP2A-A3 mRNA expression and growth patterns in Arabidopsis thaliana under drought stress and abscisic acid

Research Article
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Abstract

Phosphoprotein phosphatase 2A (PP2A) plays a crucial role in cellular processes via reversible dephosphorylation of proteins. The activity of this enzyme depends on its subunits. There is little information about mRNA expression of each subunit and the relationship between these gene expressions and the growth patterns under stress conditions and hormones. Here, mRNA expression of subunit A3 of PP2A and its relationship with growth patterns under different levels of drought stress and abscisic acid (ABA) concentration were analyzed in Arabidopsis thaliana. The mRNA expression profiles showed different levels of the up- and down-regulation of PP2AA3 in roots and shoots of A. thaliana under drought conditions and ABA treatments. The results demonstrated that the regulation of PP2AA3 expression under the mentioned conditions could indirectly modulate growth patterns such that seedlings grown under severe drought stress and those grown under 4 µM ABA had the maximum number of lateral roots and the shortest primary roots. In contrast, the minimum number of lateral roots and the longest primary roots were observed under mild drought stress and 0.5 µM ABA. Differences in PP2AA3 mRNA expression showed that mechanisms involved in the regulation of this gene under drought conditions would probably be different from those that regulate the PP2AA3 expression under ABA. Co-expression of PP2AA3 with each of PIN1-4,7 (PP2A activity targets) depends on the organ type and different levels of drought stress and ABA concentration. Furthermore, fluctuations in the PP2AA3 expression proved that this gene cannot be suitable as a reference gene although PP2AA3 is widely used as a reference gene.

Keywords

Abscisic acid Arabidopsis thaliana In vitro drought stress mRNA expression Subunit A3 of phosphoprotein phosphatase 2A 

Abbreviations

ABA

Abscisic acid

MS medium

Murashige and Skoog nutrient medium

PP2A

Phosphoprotein phosphatase 2A

qRT-PCR

Quantitative reverse transcription polymerase chain reaction

References

  1. Almeida-de-Macedo MM, Ransom N, Feng Y, Hurst J, Wurtele ES (2013) Comprehensive analysis of correlation coefficients estimated from pooling heterogeneous microarray data. BMC Bioinform 14:214CrossRefGoogle Scholar
  2. Biddington NL, Dearman AS (1982) The effect of abscisic acid on root and shoot growth of cauliflower plants. Plant Growth Regul 1:15–24CrossRefGoogle Scholar
  3. Biswas J, Chowdhury B, Bhattacharya A, Mandal AB (2002) In vitro screening for increased drought tolerance in rice. In Vitro Cell Dev Biol Plant 38:525–530CrossRefGoogle Scholar
  4. Blakeslee JJ, Zhou HW, Heath JT, Skottke KR, Barrios JA, Liu SY, De Long A (2008) Specificity of RCN1-mediated protein phosphatase 2A regulation in meristem organization and stress response in roots. Plant Physiol 146:539–553CrossRefPubMedPubMedCentralGoogle Scholar
  5. Bray EA (2004) Genes commonly regulated by water-deficit stress in Arabidopsis thaliana. J Exp Bot 55:2331–2341CrossRefPubMedGoogle Scholar
  6. Cheng WH, Endo A, Zhou L, Penney J, Chen HC, Arroyo A, Leon P, Nambara E, Asami T, Seo M, Koshiba T, Sheen J (2002) A unique short-chain dehydrogenase/reductase in Arabidopsis glucose signaling and abscisic acid biosynthesis and functions. Plant Cell 14:2723–2743CrossRefPubMedPubMedCentralGoogle Scholar
  7. Claeys H, Inzé D (2013) The agony of choice: how plants balance growth and survival under water-limiting conditions. Plant Physiol 162:1768–1779CrossRefPubMedPubMedCentralGoogle Scholar
  8. Cutler SR, Rodriguez PL, Finkelstein RR, Abrams SR (2010) Abscisic acid: emergence of a core signaling network. Annu Rev Plant Biol 61(1):651–679CrossRefPubMedGoogle Scholar
  9. Czechowski T, Stitt M, Altmann T, Udvardi MK, Scheible WR (2005) Genome-wide identification and testing of superior reference genes for transcript normalization in Arabidopsis. Plant Physiol 139:5–17CrossRefPubMedPubMedCentralGoogle Scholar
  10. DeLong A (2006) Switching the flip: protein phosphatase roles in signaling pathways. Curr Opin Plant Biol 9:470–477CrossRefPubMedGoogle Scholar
  11. Deruère J, Jackson K, Garbers C, Söll D, DeLong A (1999) The RCN1-encoded A subunit of protein phosphatase 2A increases phosphatase activity in vivo. Plant J 20:389–399CrossRefPubMedGoogle Scholar
  12. Dubrovsky JG, Gómez-Lomelí LF (2003) Water deficit accelerates determinate developmental program of the primary root and does not affect lateral root initiation in a Sonoran Desert cactus (Pachycereuspringlei, Cactaceae). Am J Bot 90:823–831CrossRefPubMedGoogle Scholar
  13. Farooqa M, Kobayashia N, Ito O, Wahid A, Serraj R (2010) Broader leaves result in better performance of indica rice under drought stress. J Plant Physiol 167:1066–1075CrossRefGoogle Scholar
  14. Finkelstein R (2013) Abscisic acid synthesis and response. Arabidopsis Book 11:e0166CrossRefPubMedPubMedCentralGoogle Scholar
  15. Garbers C, DeLong A, Deruère J, Bernasconi P, Söll D (1996) A mutation in protein phosphatase 2A regulatory subunit A affects auxin transport in Arabidopsis. EMBO J 15:2115–2124PubMedPubMedCentralGoogle Scholar
  16. Geisler-Lee J, O’Toole N, Ammar R, Provart NJ, Millar AH, Geisler M (2007) A predicted interactome for Arabidopsis. Plant Physiol 145:317–329CrossRefPubMedPubMedCentralGoogle Scholar
  17. Gopal J, Iwama K, Jitsuyama Y (2008) Effect of water stress mediated through agar on in vitro growth of potato. In Vitro Cell Dev Biol Plant 44:221–228CrossRefGoogle Scholar
  18. Horan K, Jang C, Bailey-Serres J, Mittler R, Shelton C, Harper JF, Zhu JK, Cushman JC, Gollery M, Girke T (2008) Annotating genes of known and unknown function by large-scale coexpression analysis. Plant Physiol 147:41–57CrossRefPubMedPubMedCentralGoogle Scholar
  19. Hummel I, Pantin F, Sulpice R, Piques M, Rolland G, Dauzat M, Christophe A, Pervent M, Bouteillé M, Stitt M, Gibon Y, Muller B (2010) Arabidopsis plants acclimate to water deficit at low cost through changes of carbon usage: an integrated perspective using growth, metabolite, enzyme, and gene expression analysis. Plant Physiol 154:357–372CrossRefPubMedPubMedCentralGoogle Scholar
  20. Hunter T (1995) Protein kinases and phosphatases: the yin and yang of protein phosphorylation and signaling. Cell 80:225–236CrossRefPubMedGoogle Scholar
  21. Kalefetoglu T, Ekmekci Y (2005) The effects of drought on plants and tolerance mechanisms. GUJ Sci 18:723–740Google Scholar
  22. Kwak JM, Moon JH, Murata Y, Kuchitsu K, Leonhardt N, DeLong A, Schroeder JI (2002) Disruption of a guard cell expressed protein phosphatase 2A regulatory subunit, RCN1, confers abscisic acid insensitivity in Arabidopsis. Plant Cell 14:2849–2861CrossRefPubMedPubMedCentralGoogle Scholar
  23. Leung J, Giraudat J (1998) Abscisic acid signal transduction. Annu Rev Plant Physiol Plant Mol Biol 49(1):199–222CrossRefPubMedGoogle Scholar
  24. Luan S (1998) Protein phosphatases and signaling cascades in higher plants. Acta Bot Sin 40:883–889Google Scholar
  25. Mahajan S, Tuteja N (2005) Cold, salinity and drought stresses: an overview. Arch Biochem Biophys 444:139–158CrossRefPubMedGoogle Scholar
  26. Michniewicz M, Zago MK, Abas L, Weijers D, Schweighofer A, Meskiene I, Heisler MG, Ohno C, Zhang J, Huang F, Schwab R, Weigel D, Meyerowitz EM, Luschnig C, Offringa R, Friml J (2007) Antagonistic regulation of PIN phosphorylation by PP2A and PINOID directs auxin flux. Cell 130:1044–1056CrossRefPubMedGoogle Scholar
  27. Murashige T, Skoog F (1962) A revised medium for rapid growth and bio-assays with tobacco tissue cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar
  28. País SM, Téllez-Iñón MT, Capiati DA (2009) Serine/threonine protein phosphatases type 2A and their roles in stress signaling. Plant Signal Behav 4:1013–1015CrossRefPubMedPubMedCentralGoogle Scholar
  29. Pernas M, García-Casado G, Rojo E, Solano R, Sánchez-Serrano JJ (2007) A protein phosphatase 2A catalytic subunit is a negative regulator of abscisic acid signaling. Plant J 51:763–778CrossRefPubMedGoogle Scholar
  30. Pfaffl MW (2001) A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 29:2002–2007CrossRefGoogle Scholar
  31. Raghavendra AS, Gonugunta VK, Christmann A, Grill E (2010) ABA perception and signalling. Trends Plant Sci 15:395–401CrossRefPubMedGoogle Scholar
  32. Sacks MM, Silk WK, Burman P (1997) Effect of water stress on cortical cell division rates within the apical meristem of primary roots of Maize. Plant Physiol 114:519–527CrossRefPubMedPubMedCentralGoogle Scholar
  33. Saito N, Munemasa S, Nakamura Y, Shimoishi Y, Mori IC, Murata Y (2008) Roles of RCN1, regulatory A subunit of protein phosphatase 2A, in methyl jasmonate signaling and signal crosstalk between methyl jasmonate and abscisic acid. Plant Cell Physiol 49:1396–1401CrossRefPubMedGoogle Scholar
  34. Schuppler U, He PH, John PC, Munns R (1998) Effect of water stress on cell division and cell division-cycle 2-like cell-cycle kinase activity in wheat leaves. Plant Physiol 117:667–678CrossRefPubMedPubMedCentralGoogle Scholar
  35. Sharp RE, Poroyko V, Hejlek LG, Spollen WG, Springer GK, Bohnert HJ, Nguyen HT (2004) Root growth maintenance during water deficits: physiology to functional genomics. J Exp Bot 55:2343–2351CrossRefPubMedGoogle Scholar
  36. Shojaie B, Mostajeran A, Esmaeili A (2015) Different drought conditions could modulate growth responses of Arabidopsis thaliana through regulation of mRNA expression of genes encoding plasma membrane PIN proteins. Int J Adv Res Biol Sci 2:241–254Google Scholar
  37. Smith RD, Walker JC (1996) Plant protein phosphatases. Annu Rev Plant Physiol Plant Mol Biol 47:101–125CrossRefPubMedGoogle Scholar
  38. Stone JM, Walker JC (1995) Plant protein kinase families and signal transduction. Plant Physiol 108:451–457CrossRefPubMedPubMedCentralGoogle Scholar
  39. Taylor SS, Kornev AP (2011) Protein kinases: evolution of dynamic regulatory proteins. Trends Biochem Sci 36:65–77CrossRefPubMedGoogle Scholar
  40. Terol J, Bargues M, Carrasco P, Pérez-Alonso M, Paricio N (2002) Molecular characterization and evolution of the protein phosphatase 2A B regulatory subunit family in plants. Plant Physiol 129:808–822CrossRefPubMedPubMedCentralGoogle Scholar
  41. Uhrig RG, Labandera AM, Moorhead GB (2013) Arabidopsis PPP family of serine/threonine protein phosphatases: many targets but few engines. Trends Plant Sci 18:505–513CrossRefPubMedGoogle Scholar
  42. Usadel B, Obayashi T, Mutwil M, Giorgi FM, Bassel GW, Tanimoto M, Chow A, Steinhauser D, Persson S, Provart NJ (2009) Co-expression tools for plant biology: opportunities for hypothesis generation and caveats. Plant Cell Environ 32:1633–1651CrossRefPubMedGoogle Scholar
  43. Vartanian N, Marcotte L, Giraudat J (1994) Drought rhizogenesis in Arabidopsis thaliana. Plant Physiol 104:761–767CrossRefPubMedPubMedCentralGoogle Scholar
  44. Wang H, Qi Q, Schorr P, Cutler AJ, Crosby WL, Fowke LC (1998) ICK1, a cyclin-dependent protein kinase inhibitor from Arabidopsis thaliana interacts with both Cdc2a and CycD3, and its expression is induced by abscisic acid. Plant J 15:501–510CrossRefPubMedGoogle Scholar
  45. Xu C, Jing R, Mao X, Jia X, Chang X (2007) A wheat (Triticum aestivum) protein phosphatase 2A catalytic subunit gene provides enhanced drought tolerance in tobacco. Ann Bot 99:439–450CrossRefPubMedPubMedCentralGoogle Scholar
  46. Xu J, Gao G, Du J, Guo Y, Yang C (2010) Cell cycle modulation in response of the primary root of Arabidopsis to ABA. Pak J Bot 42:2703–2710Google Scholar
  47. Zhang J, Jia W, Yang J, Ismail AM (2006) Role of ABA in integrating plant responses to drought and salt stresses. Field Crops Res 97:111–119CrossRefGoogle Scholar
  48. Zhang W, Li C, Qian C, Cao L (2009) Studies on the responses of root, shoot and drought resistance in the seedlings of forage Triticale to water stress. J Agr Sci 1:50–57Google Scholar
  49. Zhou HW, Nussbaumer C, Chao Y, DeLong A (2004) Disparate roles for the regulatory A subunit isoforms in Arabidopsis protein phosphatase 2A. Plant Cell 16:709–722CrossRefPubMedPubMedCentralGoogle Scholar
  50. Zokaee-Khosroshahi M, Esna-Ashari M, Ershadi A, Imani A (2014) Morphological changes in response to drought stress in cultivated and wild Almond species. Int J Hortic Sci Technol 1:79–92Google Scholar

Copyright information

© Prof. H.S. Srivastava Foundation for Science and Society 2018

Authors and Affiliations

  1. 1.Department of BiologyPayame Noor UniversityTehranIran
  2. 2.Department of Biology, Faculty of ScienceUniversity of IsfahanIsfahanIran
  3. 3.Faculty of Life and Environmental SciencesUniversity of TsukubaTsukubaJapan

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