Abstract
A polyclonal antibody raised against cellulase (EC 3.2.1.4.) from callus of Nicotiana tabacum L. cv. Petit Havana SR1 reduced cellulase activity and induced shoot formation in tobacco callus in the presence of callus maintaining concentrations of auxin and cytokinin. Shoot induction as well as reduction of the cellulase activity was also obtained by withdrawing auxin from the callus medium. The effect of the two hormones on cellulase activity in the tobacco tissue was examined by varying the concentration of one of the hormones α-naphthylacetic acid (NAA) or benzylaminopurine (BAP) at a time while the other was kept at a level sufficient for either callus growth or shoot induction. While NAA stimulated the enzyme activity increasingly with concentration in the range 5 × 10−7 M to 5 × 10−5 M at both levels of BAP, BAP only stimulated the cellulase activity at an optimum concentration of 5 × 10−6 M when NAA was present at a level sufficient to induce callus growth. The results point to a pivotal role of the downward regulation of cellulase in the initiation of shoot induction. A series of events leading to oriented cell divisions as a result of the lowered cellulase level during the initial phase of the morphogenetic process is discussed.
Similar content being viewed by others
Abbreviations
- Ab:
-
Purified cellulase antibody
- BAP:
-
benzylaminopurine
- MS:
-
Murashige and Skoog medium
- NAA:
-
α-naphthylacetic acid
- PS:
-
Purified preimmune serum
References
Bradford, M.M. (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
Carpita, N.C., Gibeaut, D.M. (1993) Structural models of primary cell walls in flowering plants: consistency of molecular structure with the physical properties of the walls during growth. Plant J. 3, 1–30
Cosgrove, D.J. (1993) How do plant cells extend? Plant Physiol. 102, 1–6
Davies, E., Özbay, O. (1980) A potential role for cellulase in hormone-controlled elongation. Z. Pflanzenphysiol. 99, 461–469
Hoson, T., Nevins, D.J. (1989) Effect of anti-wall protein antibodies on auxin-induced elongation, cell wall loosening, and β-d-glucan degradation in maize coleoptile segments. Physiol. Plant. 77, 208–215
Hoson, T., Masuda, Y., Sone, Y., Misaki, A. (1991) Xyloglucan antibodies inhibit auxin-induced elongation and cell wall loosening of azuki bean epicotyls but not of oat coleoptiles. Plant Physiol. 96, 551–557
Hoson, T., Masuda, Y., Nevins, D.J. (1992) Comparison of outer and inner epidermis. Inhibition of auxin-induced elongation of maize coleoptiles by glucan antibodies. Plant Physiol. 98, 1298–1303
Hush, J.M., Overall, R.L. (1991) Electrical and mechanical fields orient cortical microtubules in higher plant tissue. Cell Biol. Int. Rep. 15, 551–560
Kirkham, M.B., Gardner, W.R., Gerloff, G.C. (1972) Regulation of cell division and cell enlargement by turgor pressure. Plant Physiol. 49, 961–962
Knox, J.P. (1992) Cell adhesion, cell separation and plant morphogenesis. Plant J. 2, 137–141
Linthilhac, P.M., Vesecky, T.B. (1984) Stress-induced alignment of divisions in plant tissues grown in vitro. Nature 307, 363–364
Maliga, P., Breznovits, A.Sz.-, Marton, L. (1973) Streptomycinresistant plants from callus culture of haploid tobacco. Nature New Biology 244, 29–30
Murashige, T., Skoog, F. (1962) A revised medium for rapid growth and bio-assays with tobacco tissue cultures. Physiol. Plant. 15, 473–497
Nishitani, K., Tominaga, R. (1992) Endo-xyloglucan transferase, a novel class of glycosyltransferase that catalyzes transfer of a segment of xyloglucan molecule to another xyloglucan molecule. J. Biol. Chem. 267, 21058–21064
Potter, I., Fry S.C. (1993) Xyloglucan endotransglycosylase activity in pea internodes. Plant Physiol. 103, 235–241
Ruyssen, R., Lauwers, A. (1978) Cellulases. In: Pharmaceutical enzymes. Properties and assay methods, pp. 154–166, Ruyssen, R., Lauwers, A., eds. E. Story-Scientia, Gent
Sexton, R., Durbin, M.L., Lewis, L.N., Thomson, W.W. (1980) Use of cellulase antibodies to study leaf abscission. Nature 283, 873–874
Skoog, F., Miller, C.O. (1965) Chemical regulation of growth and organ formation in plant tissues cultured in vitro. In: Molecular and cellular aspects of development, pp. 481–494, Bell, E. ed. Harper & Row, New York
Truelsen, T.A., Wyndaele, R. (1991) Cellulase in tobacco callus: Regulation and purification. J. Plant Physiol. 139, 129–134
Verma, D.P., MacLachlan, G.A., Byrne, H., Ewings, D. (1975) Regulation and in vitro translation of messenger ribonucleic acid for cellulase from auxin-treated pea epicotyls. J. Biol. Chem. 250, 1019–1026
Author information
Authors and Affiliations
Corresponding author
Additional information
We thank Mr. Poul Fabech for constructing the automatic viscosimetric equipment and Mr. Hans Hjorth for making the computer programme. This work was supported by The Danish Veterinary and Agricultural Research Council.
Rights and permissions
About this article
Cite this article
Truelsen, T.A., Ulvskov, P. The role of cellulase in hormonal regulation of shoot morphogenesis in tobacco callus. Planta 196, 727–731 (1995). https://doi.org/10.1007/BF00197338
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00197338