Skip to main content
SpringerLink
Log in

Functional characterisation of a cytokinin oxidase gene DSCKX1 in Dendrobium orchid

  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

Abstract

Cytokinin oxidase plays an important role in the cytokinin regulatory processes. We have cloned a novel putative cytokinin oxidase, DSCKX1 (D endrobium Sonia cytokinin oxidase), by mRNA differential display from shoot apices of Dendrobium Sonia cultured in the presence of BA. The DSCKX1 gene appears to have three alternative splicing forms and its expression of DSCKX1 was induced in a tissue-specific manner by cytokinins. In transgenic orchid plants overexpressing DSCKX1, the elevated level of cytokinin oxidase activity was accompanied by a reduction of cytokinin content. These plants exhibited slow shoot growth with numerous and long roots in vitro. Their calli also showed decreased capability of shoot formation. Conversly, antisense transgenic plants showed rapid proliferation of shoots and inhibition of root growth combined with a higher endogenous cytokinin content than wild-type plants. Thus DSCKX1 appears to play an important role on cytokinin metabolism and the related developmental programmes in orchid.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Auer, C.A. and Cohen, J.D. 1993. Identification of a benzyladenine disaccharide conjugate produced during shoot organogenesis in Petunia leaf explants. Plant Physiol. 102: 541-545.

    Google Scholar 

  • Banowetz, G.M. 1992. The effects of endogenous cytokinin content on benzyladenine-enhanced nitrate reductase induction. Physiol. Plant 86: 341-348.

    Google Scholar 

  • Belarmino, M.M. and Mii, M. 2000. Agrobacterium-mediated genetic transformation of a phalaenopsis orchid. Plant Cell Rep. 19: 435-442.

    Google Scholar 

  • Bilyeu, K.D., Cole, J.L., Laskey, J.G., Riekhof, W.R., Esparza, T.J., Kramer, M.D. and Morris, R.O. 2001. Molecular and biochem-ical characterization of a cytokinin oxidase from maize. Plant Physiol. 125: 378-386.

    Google Scholar 

  • Bradford, M.M. 1976. A rapid and sensitive method for the quanti-tation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72: 248-254.

    Google Scholar 

  • Brzobohatý, B., Moore, I. and Palme, K. 1994. Cytokinin metabolism: Implications for regulation of plant growth and development. Plant Mol. Biol. 26: 1483-1497.

    Google Scholar 

  • Carlson, L.E., Tulsieram, L.K., Glaubitz, J.C., Luk, V.W.K., Kauf-feldt, C. and Rutledge, R. 1991. Segregation of random amplified DNA markers in F1 progeny of conifers. Theor. Appl. Genet. 83: 194-200.

    Google Scholar 

  • Centeno, M.L., Rodríguez, A., Feito, I., Fernández, B. 1996. Re-lationship between endogenous auxin and cytokinin levels and the morphogenic responses in Actinidia deliciosa tissue cultures. Plant Cell Rep. 16: 58-62.

    Google Scholar 

  • Chatfield, J.M. and Armstrong, D.J. 1986. Regulation of cytokinin oxidase activity in callus tissues of Phaseolus vulgaris L. cv Great Northern. Plant Physiol. 80: 493-499.

    Google Scholar 

  • Chaudhury, A.M., Letham, D.S., Craig, S. and Dennis, E.S. 1993. am p1-a mutant with high cytokinin levels and altered embryonic pattern, faster vegetative growth, constitutive photo-morphogenesis and precocious flowering. Plant J. 4: 907-916.

    Google Scholar 

  • Eklöf, S., Ästot, C., Blackwell, J., Moritz, T., Olsson, O. and Sandberg, G. 1997. Auxin-cytokinin interactions in wild-type and transgenic tobacco. Plant Cell Physiol. 38: 225-235.

    Google Scholar 

  • Galuszka, P., Frébort, I., Šbela, M., Sauer, P., Jacobsen, S., Pec, P. 2001. Cytokinin oxidase or dehydrogenase? Eur. J. Biochem. 268: 450-461.

    Google Scholar 

  • Hare, P.D. and van Staden, J. 1994. Cytokinin oxidase: biochemical features and physiological significance. Physiol. Plant. 91: 128-136.

    Google Scholar 

  • Houba-Hérin, N., Pethe, C., d'Alayer, J. and Laloue, M. 1999. Cytokinin oxidase from Zea mays: purification, cDNA cloning and expression in moss protoplasts. Plant J. 17: 615-626.

    Google Scholar 

  • Jones, R.J. and Schreiber, B.M.N. 1997. Role and function of cytokinin oxidase in plants. Plant Growth Regul. 23: 123-134.

    Google Scholar 

  • Kakimoto, T. 2001. Identification of plant cytokinin biosynthetic enzymes as dimethylallyl diphosphate: ATP/ADP isopentenyl-transferases. Plant Cell Physiol. 42: 677-685.

    Google Scholar 

  • Kamínek, M. and Armstrong, D.J. 1990. Genotypic variation in cytokinin oxidase from Phaseolus callus cultures. Plant Physiol. 93: 1530-1538.

    Google Scholar 

  • Knudson, L. 1946. A new nutrient solution for germination of orchid seed. Am. Orchid Soc. Bull. 15: 214-217.

    Google Scholar 

  • Lakshmanan, P., Loh, C.S., Goh, C.J. 1995. An in vitro method for rapid regeneration of a monopodial orchid hybrid Aranda Deborah using thin section culture. Plant Cell Rep. 14: 510-514.

    Google Scholar 

  • Letham, D.S. and Palni, L.M.S. 1983. The biosynthesis and metabolism of cytokinins. Annu. Rev. Plant Physiol. 34: 163-197.

    Google Scholar 

  • Liang, P. and Pardee, A.B. 1992. Differetial display of eukaryotic messenger RNA by means of the polymerase chain reaction. Science 257: 967-971.

    Google Scholar 

  • Libreros-Minotta, C.A. and Tipton, P.A. 1995. A colorimetric assay for cytokinin oxidase. Anal. Biochem. 231: 339-341.

    Google Scholar 

  • Lorkovic, Z.J., Wieczorek Kirk, D.A., Lambermon, M.H.L. and Fil-ipowicz, W. 2000. Pre-mRNA splicing in higher plants. Trends Plant Sci. 5: 160-167.

    Google Scholar 

  • Lou, H., McCullough, A.J. and Schuler, M.A. 1993. 3 splice site selection in dicot plant nuclei is position dependent. Mol. Cell. Biol. 13: 4485-4493.

    Google Scholar 

  • Martin, R.C., Mok, M.C. and Mok, D.W.S. 1999a. A gene encoding the cytokinin enzyme zeatin O-xylosyltransferase of Phaseolus vulgaris. Plant Physiol. 120: 553-558.

    Google Scholar 

  • Martin, R.C., Mok, M.C. and Mok, D.W.S. 1999b. Isolation of a cy-tokinin gene, ZOG1, encoding zeatin O-glucosyltransferase from Phaseolus lunatus. Proc. Natl. Acad. Sci. USA 96: 284-289.

    Google Scholar 

  • Martin, R.C., Mok, M.C., Habben, J.E., and Mok, D.W.S. 2001. Amaize cytokinin gene encoding an O-glucosyltransferase specific to ciszeatin. Proc. Natl. Acad. Sci. USA 98: 5922-5926.

    Google Scholar 

  • Medford, J.I., Horgan, R., El-Sawi, Z. and Klee, H.J. 1989. Al-terations of endogenous cytokinins in transgenic plants using a chimeric isopentenyl transferase gene. Plant Cell 1: 403-413.

    Google Scholar 

  • Moffatt, B.A., McWhinnie, E.A., Burkhart, W.E., Pasternak, J.J. and Rothstein, S.J. 1992. A complete cDNA for adenine phos-phoribosyltransferase from Arabidopsis thaliana. Plant Mol. Biol. 18: 653-662.

    Google Scholar 

  • Mok, M.C. 1994. Cytokinins and plant development-An overview. In: D.W.S. Mok and M.C. Mok (eds.), Cytokinins: Chemistry, Activity and Function, Boca Raton, CRC Press, pp. 155-166.

    Google Scholar 

  • Morris, R.O., Bilyeu, K.D., Laskey, J.G. and Cheikh, N.N. 1999. Isolation of a gene encoding a glycosylated cytokinin oxidase from maize. Biochem. Biophys. Res. Commun. 255: 328-333.

    Google Scholar 

  • Motyka, V. and Kamínek, M. 1990. Regulation of cytokinin catabolism in tobacco callus cultures. In: H.J.J. Nijkamp, L.H.W. Van der Plas and J. Van Aartrijk (eds), Progress in Plant Cellular and Molecular Biology, Dordrecht, Kluwer academic publishers, pp. 492-497.

    Google Scholar 

  • Motyka, V., Faiss, M., Strnad, M., Kamínek, M. and Schülling, T. 1996. Changes in cytokinin content and cytokinin oxidase activity in response to derepression of IPT gene transcrip-tion in transgenic tobacco calli and plants. Plant Physiol. 112: 1035-1043.

    Google Scholar 

  • Mushegian, A.R. and Koonin, E.V. 1995. A putative FAD-binding domain in a distinct group of oxidases including a protein involved in plant development. Prot. Sci. 4: 1243-1244.

    Google Scholar 

  • Palni, L.M.S., Burch, L. and Horgan, R. 1988. The effect of auxin concentration on cytokinin stability and metabolism. Planta 174: 231-234.

    Google Scholar 

  • Redig, P., Shaul, O., Inzé, D., Van Montagu, M. and Van Onckelen, H. 1996. Levels of endogenous cytokinins, indole-3-acetic acid and abscisic acid during cell cycle of synchronized tobacco BY-2 cells. FEBS Lett. 391: 175-180.

    Google Scholar 

  • Schnorr, K.M., Gaillard, C., Biget, E., Nygaard, P. and Laloue, M. 1996. A second form of adenine phosphoribosyltransferase from Arabidopsis thaliana with relative specificity towards cytokinins. Plant J. 9: 891-898.

    Google Scholar 

  • Simpson, G.G. and Filipowicz, W. 1996. Splicing of precursors to mRNA in higher plants: mechanism, regulation and sub-nuclear organisation of the spliceosomal machinery. Plant Mol. Biol. 32: 1-41.

    Google Scholar 

  • Skoog, F. and Miller, C.O. 1957. Chemical regulation of growth and organ formation in plant tissues cultured in vitro. Soc. Exp. Biol. Symp. 11: 118-131.

    Google Scholar 

  • Skoog, F. and Armstrong, D.J. 1970. Cytokinins. Annu. Rev. Plant Physiol. 21: 359-384.

    Google Scholar 

  • Takei, K., Sakakibara, H. and Sugiyama, T. 2001. Identification of genes encoding adenylate isopentenyltransferase, a cytokinin biosynthesis enzyme, in Arabidopsis thaliana. J. Biol. Chem. 276: 26405-26410.

    Google Scholar 

  • Tantikanjana, T., Yong, J.W.H., Letham, D.S., Griffith, M., Hussain, M., Ljung, K., Sandberg, G. and Sundaresan, V. 2001. Control of axillary bud initiation and shoot architecture in Arabidopsis through the SUPERSHOOT gene. Genes Dev. 15: 1577-1588.

    Google Scholar 

  • Trione, E.J., Banowetz, G.M., Krygier, B.B., Kathrein, J.M., Sayavedra-Soto, L.A. 1987. A quantitative fluorescence enzyme immunoassay for plant cytokinins. Anal. Biochem. 22: 301-308.

    Google Scholar 

  • von Schwartzenberg, K., Kruse, S., Reski, R., Moffatt, B. and Laloue, M. 1998. Cloning and characterization of an adenosine kinase from Physcomitrella involved in cytokinin metabolism. Plant J. 13: 249-257.

    Google Scholar 

  • Werner, T., Motyka, V., Strnad, M. and Schmüling, T. 2001. Regu-lation of plant growth by cytokinin. Proc Natl Acad Sci USA 98: 10487-10492.

    Google Scholar 

  • Yu, H. and Goh, C.J. 2000a. Differential gene expression during floral transition in an orchid hybrid Dendrobium Madame Thong-In. Plant Cell Rep. 19: 926-931.

    Google Scholar 

  • Yu, H. and Goh, C.J. 2000b. Identification and charaterization of three orchid MADS-Box genes of the AP1/AGL9 subfamily during floral transition. Plant Physiol. 123: 1325-1336.

    Google Scholar 

  • Yu, H., Yang, S.H. and Goh, C.J. 2000. DOH1, a class 1 knox gene, is required for maintainence of the basic plant architecture and floral transition in orchid. Plant Cell, 12: 2143-2159.

    Google Scholar 

  • Yu, H., Yang, S.H. and Goh, C.J. 2001. Agrobacterium-mediated transformation of a Dendrobium orchid with the class 1 knox gene DOH1.Plant Cell Rep. 20: 301-305.

    Google Scholar 

  • Zazímalová, E., Brezimová, A., Holík, J. and Opatrný, Z. 1996. Par-tial auxin deprivation affects endogenous cytokinins in an auxin-dependent, cytokinin-independent tobacco cell strain. Plant Cell Rep. 16: 76-79.

    Google Scholar 

  • Zazímalová, E., Kamínek, M., Brezinová, A. and Motyka, V. 1999. Control of cytokinin biosynthesis and metabolism. In: P.J.J. Hooykaas, M.A. Hall and K.R. Libbenga (eds), Biochemistry and Molecular Biology of Plant Hormones, Amsterdam, Elsevier Science B.V., pp. 141-160.

  • Zhang, R., Zhang, X., Wang, J., Letham, D.S., McKinney, S.A. and Higgins, T.J.V. 1995. The effect of auxin on cytokinin levels and metabolism in transgenic tobacco tissue expressing an IPT gene. Planta 196: 84-94.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Plant Growth and Development Laboratory, Department of Biological Sciences, Faculty of Science, National University of Singapore, 10 Kent Ridge Crescent, Singapore, 119260, Republic of Singapore

    Shu Hua Yang, Hao Yu & Chong Jin Goh

Authors
  1. Shu Hua Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hao Yu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Chong Jin Goh
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Chong Jin Goh.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Yang, S.H., Yu, H. & Goh, C.J. Functional characterisation of a cytokinin oxidase gene DSCKX1 in Dendrobium orchid. Plant Mol Biol 51, 237–248 (2003). https://doi.org/10.1023/A:1021115816540

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1021115816540

Search

Navigation