Plant Growth Regulation

, Volume 32, Issue 2–3, pp 307–313 | Cite as

Cytokinin-regulated transcripts in tobacco cell culture

  • S. Schäfer
  • S. Krolzik
  • G.A. Romanov
  • T. Schmülling


To study early responses to the hormone cytokinin we identified transcripts in cell culture of Nicotiana tabacum L. Representational difference analysis (RDA) was used to amplify partial cDNAs of genes that were induced or repressed 45 after addition of 5 × 10−7 to the culture medium. Ten different mRNAs whose abundance changed rapidly after cytokinin treatment were identified. Transcripts coding for osmotin, a peroxidase, pectin methyl esterase, a subunit of NADH:ubiquinone oxidoreductase, a chloroplast nucleoid DNA binding protein homologue and a protein of unknown function were upregulated. Increases of transcript abundance occurred as early as 10 following hormone treatment. The transcripts of a homologue of the TA20 gene and two hitherto unknown transcripts without significant homology to known genes were strongly repressed by cytokinin. The data indicate that the regulation of genes that play a role in energy production, defence, cell wall modification and chloroplast development is part of the early response to cytokinin.

Cytokinin Gene regulation Nicotiana tabacum Plant hormones 


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  1. Altschul S.F., Gish W., Miller W., Myers E. and Lipman D.J. 1990. Basic local alignment search tool. J. Mol. Biol. 215: 403-410.Google Scholar
  2. Brandstatter I. and Kieber J.J. 1998. Two genes with similarity to bacterial response regulators are rapidly and specifically induced by cytokinin in Arabidopsis. Plant Cell 10: 1009-1019.Google Scholar
  3. Cohn N.S., Zhang L., Mitchell J.P. and Vierheller C.Z.J. 1994. Gibberellin-stimulated changes in abundance of two mRNAs in the developing shoot of dwarf peas (Pisum sativum) L. Int. J. Plant Sci. 155: 498-505.Google Scholar
  4. Crowell D.N. 1994. Cytokinin regulation of a soybean pollen allergen gene. Plant Mol. Biol. 25: 829-835.Google Scholar
  5. Crowell D.N., Kadelecek A.T., John M.C. and Amasino R.M. 1990. Cytokinin-induced mRNAs in cultured soybean cells. Proc. Natl. Acad. Sci. USA 87: 8815-8819.Google Scholar
  6. Deikman J. and Hammer P.E. 1995. Induction of anthocyanin accumulation by cytokinins in Arabidopsis thaliana. Plant Physiol. 108: 47-57.Google Scholar
  7. Diaz de Leon F., Klotz K.L. and Lagrimini L.M. 1993. Nucleotide sequence of the tobacco (Nicotiana tabacum) anionic peroxidase gene. Plant Physiol. 101: 1117-1118.Google Scholar
  8. Dominov J.A., Stenzler L., Lee S., Schwarz J.J., Leisner S. and Howell S.H. 1992. Cytokinins and auxins control the expression of a gene in Nicotiana plumbaginifolia cells by feedback regulation. Plant Cell 4: 451-461.Google Scholar
  9. Flores S. and Tobin E.M. 1988. Cytokinin modulation of LHCP mRNA levels: the involvement of post-transcriptional regulation. Plant Mol. Biol. 11: 409-415.Google Scholar
  10. Gazaryan I.G. and Lagrimini L.M. 1996. Tobacco anionic peroxidase overexpressed in transgenic plants: Aerobic oxidation of indole-3-acetic acid. Phytochem. 42: 1271-1278.Google Scholar
  11. Grießbach H. 1981. Lignins. In: Conn E.E. (ed.), The Biochemistry of Plants. Academic Press, New York, pp. 457-478.Google Scholar
  12. Heiser V., Brennicke A. and Grohmann L. 1996. The plant mitochondrial 22 kDa (PSST) subunit of respiratory chain complex I is encoded by a nuclear gene with enhanced transcript levels in flowers. Plant Mol. Biol. 31: 1195-1204.Google Scholar
  13. Hemerly A.S., Ferreira P., de Almeida Engler J., Van Montagu M., Engler G. and Inze D. 1993. cdc2a expression in Arabidopsis is linked with competence for cell division. Plant Cell 5: 1711-1723.Google Scholar
  14. Hubank M. and Schatz D.G. 1994. Identifying differences in mRNA expression by representational difference analysis of cDNA. Nucl. Acids Res. 22: 5640-5648.Google Scholar
  15. Imamura A., Hanaki N., Umeda H., Nakamura A., Suzuki T., Ueguchi C. et al. 1998. Response regulators implicated in His-to-Asp phosphotransfer signaling in Arabidopsis. Proc. Natl. Acad. Sci. USA 95: 2691-2696.Google Scholar
  16. Klotz K.L. and Lagrimini L.M. 1996. Phytohormone control of the tobacco anionic peroxidase promoter. Plant Mol. Biol. 31: 565-571.Google Scholar
  17. Klotz K.L., Liu Tin Ting Y., Liu L. and Lagrimini L.M. 1998. Expression of the tobacco anionic peroxidase gene is tissue-specific and developmentally regulated. Plant Mol. Biol. 36: 509-520.Google Scholar
  18. Koltunow A.M., Truettner J., Cox K.H., Wallroth M. and Goldberg R.B. 1990. Different temporal and spatial gene expression patterns occur during anther development. Plant Cell 2: 1201-1224.Google Scholar
  19. Kusnetsov V.V., Oelmüller R., Sarwat M.I., Porfirova S.A, Cherepneva G.N., Herrmann R.G. et al. 1994. Cytokinins, abscisic acid and light affect accumulation of chloroplast proteins in Lupinus luteus cotyledons without notable effect on steadystate mRNA levels. Planta 194: 318-327.Google Scholar
  20. Lagrimini L.M., Burkhart W., Moyer M. and Rothstein S. 1987. Molecular cloning of complementary cDNA encoding the lignin-forming peroxidase from tobacco: molecular analysis and tissue-specific expression. Proc. Natl. Acad. Sci. USA 84: 7542-7546.Google Scholar
  21. Lamport D.T.A. 1986. In: Greppin H., Penel C. and Gaspar C. (eds), Molecular and Physiological Aspects of Plant Peroxidase. University of Geneva Press, Geneva, pp. 199-208.Google Scholar
  22. Martineau B., Houck C.M., Sheehy R.E. and Hiatt W.R. 1994. Fruit-specific expression of the A. tumefaciens isopentenyl transferase gene in tomato: effects on fruit ripening and defense-related gene expression in leaves. Plant J. 5: 11-19.Google Scholar
  23. Memelink J., Hoge J.H.C. and Schilperoort R.A. 1987. Cytokinin stress changes the developmental regulation of several defencerelated genes in tobacco. EMBO J. 6: 3579-3583.Google Scholar
  24. Murashige T. and Skoog F. 1962. A revised medium for plant growth and bioassays with tobacco tissue culture. Plant Physiol. 15: 473-497.Google Scholar
  25. Nakano T., Murakami S., Shoji T., Yoshida S., Yamada Y. and Sato F. 1997. A novel protein with DNA binding activity from tobacco chloroplast nucleoids. Plant Cell 9: 1673-1682.Google Scholar
  26. Neale A.D., Wahleithner J.A., Lund M., Bonnett H.T., Kelly A., Meeks-Wagner D.R. et al. 1990. Chitinase, b-1,3-glucanase, osmotin and extensin are expressed in tobacco explants during flower formation. Plant Cell 2: 673-684.Google Scholar
  27. Puissant C. and Houdebine L.M. 1990. An improvement of the single-step method of the RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. BioTechniques 8: 148-149.Google Scholar
  28. Raghothama K.G., Maggio A., Narasimhan M.L., Kononowicz A.K., Wang G., Paino D'urzo M. et al. 1997. Tissue-specific activation of the osmotin gene by ABA, C2H4 and NaCl involves the same promoter region. Plant Mol. Biol. 34: 393-402.Google Scholar
  29. Riou-Khamlichi C., Huntley R., Jacqmard A. and Murray J.A.H. 1999. Cytokinin activation of Arabidopsis cell division through a D-type cyclin. Science 283: 1541-1544.Google Scholar
  30. Sakakibara H., Suzuki M., Takei K., Deji A A., Taniguchi M. and Sugiyama T. 1998. A response-regulator homologue possibly involved in nitrogen signal transduction mediated by cytokinin in maize. Plant J. 14: 337-344.Google Scholar
  31. Samuelson M.E., Campbell W.H. and Larsson C.M. 1995. The in-fluence of cytokinins in nitrate regulation of nitrate reductase activity and expression in barley. Physiol. Plant 93: 533-539.Google Scholar
  32. Sambrook J., Fritsch E.F. and Maniatis T. (eds) 1989. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.Google Scholar
  33. Sano H. and Youssefian S. 1994. Light and nutritional regulation of transcripts encoding a wheat protein kinase homolog is mediated by cytokinins. Proc. Natl. Acad. Sci. USA 91: 2582-2586.Google Scholar
  34. Schmülling T., Schäfer S. and Romanov G. 1997. Cytokinins as regulators of gene expression. Physiol. Plant 100: 505-519.Google Scholar
  35. Schmülling T., Rupp H.M., Frank M. and Schäfer S. 1999. Recent advances in cytokinin research: Receptor candidates, primary response genes, mutants and transgenic plants. In: Strnad M., Pec P. and Beck E. (eds), Advances in Regulation of Plant Growth and Development. Peres Company, Prague, pp. 95-96.Google Scholar
  36. Silver D.L., Pinaev A., Chen R. and de Bruijn F.J. 1996. Postranscriptional regulation of the Sesbania rostrata early nodulin gene SrEnod2 by cytokinin. Plant Physiol. 112: 559-567.Google Scholar
  37. Suty L., Moureaux T., Leydecker M. and Teyssendier de la Serve B. 1993. Cytokinin affects nitrate reductase expression through the modulation of polyadenylation of the nitrate reductase mRNA transcript. Plant Science 90: 11-19.Google Scholar
  38. Taniguchi M., Kiba T., Sakakibara H., Ueguchi C., Mizuno T. and Sugiyama T. 1998. Expression of Arabidopsis response regulator homologs is induced by cytokinins and nitrate. FEBS Lett. 429: 259-262.Google Scholar
  39. Teramoto H., Momotani E., Takeba G. and Tsuji H. 1994. Isolation of a cDNA clone for a cytokinin-repressed gene in excised cucumber cotyledons. Planta 193: 573-579.Google Scholar
  40. Vögeli-Lange R., Fründt C., Hart C.M., Nagy F. and Meins F. 1994. Developmental, hormonal and pathogenesis-related regulation of the tobacco class I b-1,3-glucanase B promotor. Plant Mol. Biol. 25: 299-311.Google Scholar
  41. Westneat D.F., Noon W.A., Reeve H. and Aquadro C.F. 1988. Improved hybridization conditions for DNA “fingerprints” probed with M13. Nucl. Acids Res. 16: 4161.Google Scholar
  42. Yanisch-Perron C., Vieira J. and Messing J. 1985. Improved M13 phage cloning vectors and host strains: nucleotide sequences of M13mp18 and pUC19 vectors. Gene. 33: 103-119.Google Scholar
  43. Ye Z.H. and Varner J.E. 1994. Expression of an auxin-and cytokinin-regulated gene in cambial region in Zinnia. Proc. Natl. Acad. Sci. USA 91: 6539-6543.Google Scholar
  44. Zabaleta E., Heiser V., Grohmann L. and Brennicke A. 1998. Promoters of nuclear-encoded respiratory chain Complex I genes from Arabidopsis thaliana contain a region essential for anther/ pollen-specific expression. Plant J. 15: 49-59.Google Scholar

Copyright information

© Kluwer Academic Publishers 2000

Authors and Affiliations

  • S. Schäfer
    • 1
  • S. Krolzik
    • 1
  • G.A. Romanov
    • 2
  • T. Schmülling
    • 1
  1. 1.Zentrum für Molekularbiologie der Pflanzen (ZMBP), Allgemeine GenetikUniversität TübingenGermany
  2. 2.Institute of Plant PhysiologyRussian Academy of SciencesMoscowRussia

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