Abstract
The alteration of growth patterns, through the adjustment of cell division and expansion, is a characteristic response of plants to environmental stress. In order to study this response in more depth, the effect of heat stress on growth was investigated in tobacco BY-2 cells. The results indicate that heat stress inhibited cell division, by slowing cell cycle progression. Cells were stopped in the pre-mitotic phases, as shown by the increased expression of CycD3-1 and by the decrease in the NtCycA13, NtCyc29 and CDKB1-1 transcripts. The decrease in cell length and the reduced expression of Nt-EXPA5 indicated that cell expansion was also inhibited. Since DNA methylation plays a key role in controlling gene expression, the possibility that the altered expression of genes involved in the control of cell growth, observed during heat stress, could be due to changes in the methylation state of their promoters was investigated. The results show that the altered expression of CycD3-1 and Nt-EXPA5 was consistent with changes in the methylation state of the upstream region of these genes. These results suggest that DNA methylation, controlling the expression of genes involved in plant development, contributes to growth alteration occurring in response to environmental changes.
Similar content being viewed by others
References
Ahmad A, Zhang Y, Cao XF (2010) Decoding the epigenetic language of plant development. Mol Plant 3:719–728
Aquea F, Federici F, Moscoso C, Vega A, Jullian P, Haseloff J, Arce-Johnson P (2012) A molecular framework for the inhibition of Arabidopsis root growth in response to boron toxicity. Plant Cell Environ 35:719–734
Baccarelli A, Wright RO, Bollati V, Tarantini L, Litonjua AA, Suh HH, Zanobetti A, Sparrow D, Vokonas PS, Schwartz J (2009) Rapid DNA methylation changes after exposure to traffic particles. Am J Respir Crit Care Med 179:572–578
Bita CE, Gerats T (2013) Plant tolerance to high temperature in a changing environment: scientific fundamentals and production of heat stress-tolerant crops. Front Plant Sci 4:273
Boyko A, Kathiria P, Zemp FJ, Yao YL, Pogribny I, Kovalchuk I (2007) Transgenerational changes in the genome stability and methylation in pathogen-infected plants (virus-induced plant genome instability). Nucleic Acids Res 35:1714–1725
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
Candaele J, Demuynck K, Mosoti D, Beemster GTS, Inze D, Nelissen H (2014) Differential methylation during maize leaf growth targets developmentally regulated genes. Plant Physiol 164:1350–1364
Causevic A, Gentil MV, Delaunay A, El-Soud WA, Garcia Z, Pannetier C, Brignolas F, Hagege D, Maury S (2006) Relationship between DNA methylation and histone acetylation levels, cell redox and cell differentiation states in sugarbeet lines. Planta 224:812–827
Choi CS, Sano H (2007) Abiotic-stress induces demethylation and transcriptional activation of a gene encoding a glycerophosphodiesterase-like protein in tobacco plants. Mol Genet Genomics 277:589–600
Christensen JH, Christensen OB (2007) A summary of the PRUDENCE model projections of changes in European climate by the end of this century. Clim Chang 81:7–30
de Pinto MC, Tommasi F, De Gara L (2000) Enzymes of the ascorbate biosynthesis and ascorbate-glutathione cycle in cultured cells of tobacco Bright Yellow 2. Plant Physiol Biochem 38:541–550
de Pinto MC, Tommasi F, De Gara L (2002) Changes in the antioxidant systems as part of the signaling pathway responsible for the programmed cell death activated by nitric oxide and reactive oxygen species in tobacco Bright-Yellow 2 cells. Plant Physiol 130:698–708
Evron E, Umbricht CB, Korz D, Raman V, Loeb DM, Niranjan B, Buluwela L, Weitzman SA, Marks J, Sukumar S (2001) Loss of cyclin D2 expression in the majority of breast cancers is associated with promoter hypermethylation. Cancer Res 61:2782–2787
Geilfus CM, Zorb C, Muhling KH (2010) Salt stress differentially affects growth-mediating beta-expansins in resistant and sensitive maize (Zea mays L.). Plant Physiol Biochem 48:993–998
Granier C, Tardieu F (1999) Leaf expansion and cell division are affected by reducing absorbed light before but not after the decline in cell division rate in the sunflower leaf. Plant Cell Environ 22:1365–1376
Hsu A, Wong CP, Yu Z, Williams DE, Dashwood RH, Ho E (2011) Promoter de-methylation of cyclin D2 by sulforaphane in prostate cancer cells. Clin Epigenetics 3:3
Huang NC, Li CH, Lee JY, Yen HE (2010) Cytosine methylation changes in the ice plant Ppc1 promoter during transition from C-3 to Crassulacean acid metabolism. Plant Sci 178:41–46
Jang SJ, Shin SH, Yee ST, Hwang B, Im KH, Park KY (2005) Effects of abiotic stresses on cell cycle progression in tobacco BY-2 cells. Mol Cells 20:136–141
Kawamura K, Murray JAH, Shinmyo A, Sekine M (2006) Cell cycle regulated D3-type cyclins form active complexes with plant-specific B-type cyclin-dependent kinase in vitro. Plant Mol Biol 61:311–327
Kende H, Bradford KJ, Brummell DA, Cho HT, Cosgrove DJ, Fleming AJ, Gehring C, Lee Y, McQueen-Mason S, Rose JKC, Voesenek LACJ (2004) Nomenclature for members of the expansin superfamily of genes and proteins. Plant Mol Biol 55:311–314
Komaki S, Sugimoto K (2012) Control of the plant cell cycle by developmental and environmental cues. Plant Cell Physiol 53:953–964
Kuluev BR, Safiullina MG, Knyazev AV, Chemeris AV (2013) Effect of ectopic expression of NtEXPA5 gene on cell size and growth of organs of transgenic tobacco plants. Russ J Dev Biol 44:28–34
Kumar A, Kaur J (2014) Primer based approach for PCR amplification of high GC content gene: mycobacterium gene as a model. Mol Biol Int 2014:937308
Lecoeur J, Wery J, Turc O, Tardieu F (1995) Expansion of pea leaves subjected to short water-deficit—cell number and cell-size are sensitive to stress at different periods of leaf development. J Exp Bot 46:1093–1101
Li LC, Dahiya R (2002) MethPrimer: designing primers for methylation PCRs. Bioinformatics 18:1427–1431
Link BM, Cosgrove DJ (1998) Acid-growth response and alpha-expansins in suspension cultures of Bright Yellow 2 tobacco. Plant Physiol 118:907–916
Madlung A, Comai L (2004) The effect of stress on genome regulation and structure. Ann Bot London 94:481–495
Menges M, Samland AK, Planchais S, Murray JAH (2006) The D-type cyclin CYCD3;1 is limiting for the G1-to-S-phase transition in Arabidopsis. Plant Cell 18:893–906
Mirouze M, Paszkowski J (2011) Epigenetic contribution to stress adaptation in plants. Curr Opin Plant Biol 14:267–274
Muller B, Bourdais G, Reidy B, Bencivenni C, Massonneau A, Condamine P, Rolland G, Conejero G, Rogowsky P, Tardieu F (2007) Association of specific expansins with growth in maize leaves is maintained under environmental, genetic, and developmental sources of variation. Plant Physiol 143:278–290
Nagata T, Nemoto Y, Hasezawa S (1992) Tobacco BY-2 cell-line as the Hela-cell in the cell biology of higher-plants. Int Rev Cytol 132:1–30
Nakagami H, Kawamura K, Sugisaka K, Sekine M, Shinmyo A (2002) Phosphorylation of retinoblastoma-related protein by the cyclin D/cyclin-dependent kinase complex is activated at the G1/S-phase transition in tobacco. Plant Cell 14:1847–1857
Ortega L, Taleisnik E (2003) Elongation growth in leaf blades of Chloris gayana under saline conditions. J Plant Physiol 160:517–522
Pasternak T, Potters G, Caubergs R, Jansen MAK (2005a) Complementary interactions between oxidative stress and auxins control plant growth responses at plant, organ, and cellular level. J Exp Bot 56:1991–2001
Pasternak T, Rudas V, Potters G, Jansen MAK (2005b) Morphogenic effects of abiotic stress: reorientation of growth in Arabidopsis thaliana seedlings. Environ Exp Bot 53:299–314
Potters G, Pasternak TP, Guisez Y, Palme KJ, Jansen MAK (2007) Stress-induced morphogenic responses: growing out of trouble? Trends Plant Sci 12:98–105
Potters G, Pasternak TP, Guisez Y, Jansen MAK (2009) Different stresses, similar morphogenic responses: integrating a plethora of pathways. Plant Cell Environ 32:158–169
Reisen D, Marty F, Leborgne-Castel N (2005) New insights into the tonoplast architecture of plant vacuoles and vacuolar dynamics during osmotic stress. BMC Plant Biol 5:13
Rymen B, Sugimoto K (2012) Tuning growth to the environmental demands. Curr Opin Plant Biol 15:683–690
Rymen B, Fiorani F, Kartal F, Vandepoele K, Inze D, Beemster GTS (2007) Cold nights impair leaf growth and cell cycle progression in maize through transcriptional changes of cell cycle genes. Plant Physiol 143:1429–1438
Saidi Y, Finka A, Goloubinoff P (2011) Heat perception and signalling in plants: a tortuous path to thermotolerance. New Phytol 190:556–565
Sampedro J, Cosgrove DJ (2005) The expansin superfamily. Genome Biol 6:242
Sgobba A, Paradiso A, Dipierro S, De Gara L, de Pinto MC (2015) Changes in antioxidants are critical in determining cell responses to short- and long-term heat stress. Physiol Plant 153:68–78
Skirycz A, Inze D (2010) More from less: plant growth under limited water. Curr Opin Biotechnol 21:197–203
Smertenko A, Draber P, Viklicky V, Opatrny Z (1997) Heat stress affects the organization of microtubules and cell division in Nicotiana tabacum cells. Plant Cell Environ 20:1534–1542
Sorrell DA, Combettes B, Chaubet-Gigot N, Gigot C, Murray JAH (1999) Distinct cyclin D genes show mitotic accumulation or constant levels of transcripts in tobacco bright yellow-2 cells. Plant Physiol 119:343–351
Sorrell DA, Menges M, Healy JMS, Deveaux Y, Amano C, Su Y, Nakagami H, Shinmyo A, Doonan JH, Sekine M, Murray JAH (2001) Cell cycle regulation of cyclin-dependent kinases in tobacco cultivar Bright Yellow-2 cells. Plant Physiol 126:1214–1223
Suzuki MM, Bird A (2008) DNA methylation landscapes: provocative insights from epigenomics. Nat Rev Genet 9:465–476
Taya Y (1997) RB kinases and RB-binding proteins: new points of view. Trends Biochem Sci 22:14–17
Urano K, Kurihara Y, Seki M, Shinozaki K (2010) ‘Omics’ analyses of regulatory networks in plant abiotic stress responses. Curr Opin Plant Biol 13:132–138
Vanhees K, Coort S, Ruijters EJB, Godschalk RWL, van Schooten FJ, van Doorn-Khosrovani SBV (2011) Epigenetics: prenatal exposure to genistein leaves a permanent signature on the hematopoietic lineage. FASEB J 25:797–807
Verma S, Mishra SN (2005) Putrescine alleviation of growth in salt stressed Brassica juncea by inducing antioxidative defense system. J Plant Physiol 162:669–677
Wada Y, Miyamoto K, Kusano T, Sano H (2004) Association between up-regulation of stress-responsive genes and hypomethylation of genomic DNA in tobacco plants. Mol Genet Genomics 271:658–666
West G, Inze D, Beemster GTS (2004) Cell cycle modulation in the response of the primary root of Arabidopsis to salt stress. Plant Physiol 135:1050–1058
Xiao WY, Custard KD, Brown RC, Lemmon BE, Harada JJ, Goldberg RB, Fischer RL (2006) DNA methylation is critical for Arabidopsis embryogenesis and seed viability. Plant Cell 18:805–814
Conflict of interest
The authors declare that they have no conflict of interest.
Author information
Authors and Affiliations
Corresponding author
Additional information
Handling Editor: Peter Nick
Electronic supplementary material
Below is the link to the electronic supplementary material.
ESM 1
(PDF 285 kb)
Rights and permissions
About this article
Cite this article
Centomani, I., Sgobba, A., D’Addabbo, P. et al. Involvement of DNA methylation in the control of cell growth during heat stress in tobacco BY-2 cells. Protoplasma 252, 1451–1459 (2015). https://doi.org/10.1007/s00709-015-0772-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00709-015-0772-y