Abstract
Previously, it was reported that plant protoplasts isolated from Betula platyphylla (white birch) callus secreted bundles of hollow callose fibrils in acidic culture medium containing a high concentration of calcium ions (Ca2+). Here, the callose synthase was characterized from in situ and in vitro perspectives. Localization of callose synthases at the secreting site of callose fiber was indicated from in situ immunostaining observation of protoplasts. For in vitro analyses, membrane proteins were extracted from membrane fraction of protoplasts with a 3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulfonate (CHAPS) treatment. The CHAPS extract aggregated in the presence of a high concentration of Ca2+, suggesting that Ca2+ may promote the arrangement of callose synthases in the plasma membrane. The callose synthase activity was dependent on pH and Ca2+, similar to the callose synthase of Arabidopsis thaliana. However, the synthesized fibril products were longer than those produced by callose synthases of herbaceous plants. This is the first insight into the specific properties of callose synthases of woody plants that secrete of callose hollow fibers.
Acknowledgments
We thank Prof. R. Malcolm Brown Jr. for providing the β-1,3-glucan synthase antibody. We are grateful to the Research Support Center, Research Center for Human Disease Modeling, Kyushu University Graduate School of Medical Sciences, for assistance with analyses using the multimode plate reader, and the Center for Advanced Instrumental and Educational Supports, Faculty of Agriculture, Kyushu University, for assistance with confocal laser scanning microscopy.
Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.
Research funding: None declared.
Employment or leadership: None declared.
Honorarium: None declared.
References
Amor, Y., Haigler, C.H., Johnson, S., Wainscott, M., Delmer, D.P. (1995) A membrane-associated form of sucrose synthase and its potential role in synthesis of cellulose and callose in plants. Proc. Natl. Acad. Sci. 92:9353–9357.10.1073/pnas.92.20.9353Search in Google Scholar PubMed PubMed Central
Barratt, D.H., Kölling, K., Graf, A., Pike, M., Calder, G., Findlay, K., Zeeman, S.C., Smith, A.M. (2011) Callose synthase GSL7 is necessary for normal phloem transport and inflorescence growth in Arabidopsis. Plant Physiol. 155:328–341.10.1104/pp.110.166330Search in Google Scholar PubMed PubMed Central
Bulone, V., Fincher, G.B., Stone, S.A. (1995) In vitro synthesis of a microfibrillar (1→3)-β-glucan by a ryegrass (Lolium multiflorum) endosperm (1→3)-β-glucan synthase enriched by product entrapment. Plant J. 8:213–225.10.1046/j.1365-313X.1995.08020213.xSearch in Google Scholar
Cifuentes, C., Bulone, V., Emons, A.C. (2010) Biosynthesis of callose and cellulose by detergent extracts of tobacco cell membranes and quantification of the polymers synthesized in vitro. J. Integrat. Plant Biol. 52:221–233.10.1111/j.1744-7909.2010.00919.xSearch in Google Scholar PubMed
Cui, X., Shin, H., Song, C., Laosinchai, W., Amano, Y., Brown, M.R. (2001) A putative plant homolog of the yeast β-1,3-glucan synthase subunit FKS1 from cotton (Gossypium hirsutum L.) fibers. Planta 213:223–230.10.1007/s004250000496Search in Google Scholar PubMed
Evans, N.A., Hoyne, P.A. (1982) A fluorochrome from aniline blue: structure, synthesis and fluorescence properties. Aus. J. Chem. 35:2571–2575.10.1071/CH9822571Search in Google Scholar
Fincher, G.B., Bacic, A., Stone, B.Chemistry, Biochemistry, and Biology of 1–3 Beta Glucans and Related Polysaccharides. Elsevier Science Publishing Co Inc, MA, USA, 2009.Search in Google Scholar
Fujii, S., Hayashi, T., Mizuno, K. (2010) Sucrose synthase is an integral component of the cellulose synthesis machinery. Plant Cell Physiol. 51:294–301.10.1093/pcp/pcp190Search in Google Scholar PubMed
Fukumoto, T., Hayashi, N., Sasamoto, H. (2005) Atomic force microscopy and laser confocal scanning microscopy analysis of callose fibers developed from protoplasts of embryogenic cells of a conifer. Planta 223:40–45.10.1007/s00425-005-0065-3Search in Google Scholar PubMed
Hayashi, T., Read, S.M., Bussell, J., Thelen, M., Lin, F.C., Brown, R.M., Delmer, D.P. (1987) UDP-glucose: (1→3)-β-glucan synthases from mung bean and cotton: differential effects of Ca2+ and Mg2+ on enzyme properties and on macromolecular structure of the glucan product. Plant Physiol. 83:1054–1062.10.1104/pp.83.4.1054Search in Google Scholar PubMed PubMed Central
Him, J., Pelosi, L., Chanzy, H., Putaux, J.L., Bulone, V. (2001) Biosynthesis of (1→3)-β-d-glucan (callose) by detergent extracts of a microsomal fraction from Arabidopsis thaliana. Eur. J. Biochem. 268:4628–4638.Search in Google Scholar
Hong, Z., Delauney, A.J., Verma, D.P. (2001) A cell plate-specific callose synthase and its interaction with phragmoplastin. Plant Cell 13:755–768.Search in Google Scholar
Jacobs, A.K., Lipka, V., Burton, R.A., Panstruga, R., Strizhov, N., Lefert, P.S., Fincher, G.B. (2003) An Arabidopsis callose synthase, GSL5, is required for wound and papillary callose formation. Plant Cell 15:2503–2513.10.1105/tpc.016097Search in Google Scholar PubMed PubMed Central
Kondo, T., Magoshi, J., Abe, H., Sasamoto, H. (2000) Method for producing of non cellulosic callose fibers from protoplasts and the callose fiber. Japan Patent No. 3936522 (Application 2000-220419).Search in Google Scholar
Lai-Kee-Him, J., Chanzy, H., Müller, M., Putaux, J.-L., Imai, T., Bulone, V. (2002) In vitro versus in vivo cellulose microfibrils from plant primary wall synthases: structural differences. J. Biol. Chem. 277:36931–36939.10.1074/jbc.M203530200Search in Google Scholar PubMed
Matsuo, S., Takenaga, A., Seyama, T., Kondo, T. (2014) Secretion of a bundle of (1→3)-β-glucan hollow fibrils from protoplasts of callus suspension under a Ca2+-rich and acidic stressed condition. Holzforschung 68:69–73.10.1515/hf-2013-0010Search in Google Scholar
Murashige, T., Skoog, F. (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol. Plant. 15:473–497.10.1111/j.1399-3054.1962.tb08052.xSearch in Google Scholar
Nedukha, O.M. (2015) Callose: localization, functions, and synthesis in plant cells. Cytol. Genet. 49:49–57.10.3103/S0095452715010090Search in Google Scholar
Pelosi, L., Imai, T., Chanzy, H., Heux, L., Buhler, E., Bulone, V. (2003) Structural and morphological diversity of (1→3)-β-d-glucans synthesized in vitro by enzymes from Saprolegnia monoica. Comparison with a corresponding in vitro product from blackberry (Rubus fruticosus). Biochemistry 42:6264–6274.10.1021/bi0340550Search in Google Scholar PubMed
Salnikov, V.V., Grimson, M.J., Seagull, R.W., Haigler, C.H. (2003) Localization of sucrose synthase and callose in freeze-substituted secondary-wall-stage cotton fibers. Protoplasma 221:175–184.10.1007/s00709-002-0079-7Search in Google Scholar PubMed
Sasamoto, H., Ogita, S., Hayashi, N., Wakita, Y., Yokota, S.,Yoshizawa, N. (2003) Development of novel elongated fiber-structure in protoplast cultures of Betula platyphylia and Larix leptolepis. In Vitro Cell. Dev. Biol. Plant 39: 223–228.10.1079/IVP2002388Search in Google Scholar
Seyama, T., Kondo, T. (2012) Morphological responses of Betula protoplasts in fiber spinning. Holzforschung 66:407–411.10.1515/hf.2011.158Search in Google Scholar
Seyama, T., Kimura, S., Sasamoto, H., Abe, H., Kondo, T. (2008) Spinning of a gigantic bundle of hollow fibrils by a spirally moving higher plant protoplast. Planta 227:1187–1197.10.1007/s00425-008-0689-1Search in Google Scholar PubMed
Shedletzky, E., Unger, C., Delmer, D.P. (1997) A microtiter-based fluorescence assay for (1,3)-β-glucan synthases. Analyt. Biochem. 249:88–93.10.1006/abio.1997.2162Search in Google Scholar PubMed
Smith, P.K., Krohn, I.R., Hermanson, G.T., Mallia, A.K., Gartner, F.H., Provenzano, M., Fujimoto, E.K., Goeke, N.M., Olson, B.J., Klenk, D.C. (1985) Measurement of protein using bicinchoninic acid. Analyt. Biochem. 150:76–85.10.1016/0003-2697(85)90442-7Search in Google Scholar
Tagawa, S., Kondo, T. (2018) Secretion of a callose hollow fiber from herbaceous plant protoplasts induced by inhibition of cell wall formation. J. Wood Sci. 64:467–476.10.1007/s10086-018-1726-8Search in Google Scholar
Tagawa, S., Yamagishi, Y., Watanabe, U., Funada, R., Kondo, T. (2019) Dynamics of structural polysaccharides deposition on the plasmamembrane surface of plant protoplasts during cell wall regeneration. J. Wood Sci. 65:47.10.1186/s10086-019-1826-0Search in Google Scholar
Zilly, F.E., Halemani, N.D., Walrafen, D., Spitta, L., Schreiber, A., Jahn, R., Lang, T. (2011) Ca2+ induces clustering of membrane proteins in the plasma membrane via electrostatic interactions. EMBO J. 30:1209–1220.10.1038/emboj.2011.53Search in Google Scholar PubMed PubMed Central
©2020 Walter de Gruyter GmbH, Berlin/Boston
