Summary
When suspensions of sycamore cells are cultured in a synthetic medium containing 1.0 mg/l 2,4-D and 0.25 mg/l kinetin, maximum cell yield is obtained with an initial concentration of 6 per cent sucrose. There is a progressive increase in dry weight per cell, decline in extractive-free weight as a percentage of cell dry weight and increase in lignin content per cell as the initial sucrose concentration is increased from 1 per cent to 15 per cent. The percentage of lignin in the extractive-free cell residue is further enhanced by increasing the level of 2,4-D to 10 mg/l or by growing the cells in an auxin-free medium containing 10 mg/l kinetin.
When the cell suspensions are treated with phloroglucinol/HCl it is found that only a proportion of the cells contain lignin, that this lignin occurs in the protoplasts and in plates between the cells, and that lignin is present in the culture medium.
Electron micrographs confirmed the absence of any secondary wall such as is characteristic of tracheary elements. Cells cultured in the presence of 6 per cent sucrose or higher levels showed numerous amyloplasts and frequently the presence of electron opaque material. This occurs in the irregular but not frequent wall thickenings, as droplets in the vacuoles and as amorphous sheets between the cells. Pictures showing such electron opaque droplets clustered on the inner face of the tonoplasts suggest that this material is formed in the cytoplasm and released into the vacuoles. In addition these cells are characterised by the presence of fine electron opaque granular material in their vacuoles and external to their protoplasts. Cultures richest in lignin showed the highest content of electron opaque globules in, and amorphous sheets between, the cells and it is suggested that these correspond to lignin or a lignin-hemicellulose complex. In the presence of 15 per cent sucrose many cells showing breakdown of organised structure were observed; they were characterised by the persistence of mitochondria and particularly of the amyloplasts and by their high content of the electron opaque material equated with lignin. This material was also present in the dead cells.
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References
Bergmann, L., 1964: Der Einfluß von Kinetin auf die Ligninbildung und Differenzierung und Gewebekulturen vonNicotiana tabacum. Planta (Berl.)62, 221–254.
Bland, D. E., 1961: Eugenol lignin: its chemical properties and significance. Biochem. J.18, 23–28.
Butcher, D. N., andH. E. Street, 1960a: Effects of kinetin on the growth of excised tomato roots. Physiologia Plantarum13, 46–55.
— —, 1960b: The effect of gibberellins on the growth of excised tomato roots. J. exp. Bot.11, 202–216.
Davey, M. R., andH. E. Street, 1971: Studies on the growth in culture of plant cells. IX. Additional features of the fine structure ofAcer pseudoplatanus cells culture in suspension. J. exp. Bot.22, 90–95.
Drummond, D. G., 1950: The preparation of formvar films. J. roy. micr. Soc.70, 45.
Fosket, D. E., andJ. G. Torrey, 1969: Proliferation and xylem differentiation in cultured tissues ofGlycine max var. Biloxi. Plant Physiol. (Lancaster)44, 871–880.
Henshaw, G. G., K. K. Jha, A. R. Mehta, D. J. Shakeshaft, andH. E. Street, 1966: Studies on the growth in culture of plant cells. I. Growth patterns in batch propagated suspension cultu Res. J. exp. Bot.17, 362–377.
Hughes, E. W. D., andH. E. Street, 1960: The effects of inhibitory concentrations of 3-indolylacetic acid and 3-indolylacetonitrile on cell division and tissue differentiation in excised tomato roots. J. exp. Bot.11, 198–205.
Jeffs, R. A., andD. H. Northcote, 1966: Experimental induction of vascular tissue in an undifferentiated plant callus. Biochem. J.101, 146–152.
— —, 1967: Influence of indolyl-3-acetic acid and sugar in the pattern of induced differentiation in plant tissue culture. J. Cell Sci.2, 77–88.
Koblitz, H., 1962a: Die Chemie der Zellwandin vitro kultivierter pflanzlicher Gewebe. II. Kinetininduzierte Ligninbildung, eine neue Eigenschaft des Kinetins. Faserforsch. Textiltech.13, 231–234.
—, 1962b: Die Chemie der Zellwandin vitro kultivierter pflanzlicher Gewebe. III. Die Wirkung von Cocosmilch, Kinetin und Auxinen auf die Zellwand von Karotten-Geweben. Faserforsch. Textiltech.13, 270–275.
Mohr, W. P., andE. C. Cocking, 1967: A method of preparing high vacuolated, senescent, or damaged plant tissue for ultrastructural study. J. Ultrastruct. Res.21, 171–181.
Nelson, N., 1944: A photometric adaptation of the Somogyi method for the determination of glucose. J. biol. Chem.153, 375–380.
Nevins, D. J., P. D. English, andP. Albersheim, 1966: The specific nature of plant cell wall polysaccharides. Plant Physiol. (Lancaster)42, 900–906.
Öpik, H., 1968: Structure, function and developmental changes in mitochondria of higher plants. In: Plant Cell Organelles. pp. 47–88 (J. B. Pridham, ed.). London: Academic Press.
Palade, G. E., 1952: A study of fixation for electron microscopy. J. exp. Med.95, 285–298.
Reynolds, E. S., 1963: Use of lead citrate at high pH as an electron opaque stain in electron microscopy. J. Cell Biol.17, 208–213.
Siegel, S., P. Frost, andF. Porto, 1960: Effects of indoleacetic acid and other oxidation regulators onin vitro peroxidation and experimental conversion of eugenol to lignin. Plant Physiol. (Lancaster)35, 163–167.
Simpkins, I., H. A. Collin, andH. E. Street, 1970: The growth ofAcer pseudoplatanus cells in a synthetic liquid medium: Response to the carbohydrate, nitrogenous and growth hormone constituents. Physiologia Plantarum23, 385–396.
Somogyi, M., 1952: Notes on sugar determination. J. Biol. Chem.195, 19–23.
Stafford, H. A., 1960: Differences between lignin-like polymers formed by peroxidation of eugenol and ferulic acid in leaf sections. Plant Physiol. (Lancaster)35, 108–114.
Street, H. E., H. A. Collin, K. Short, andI. Simpkins, 1968: Hormonal control of cell division and expansion in suspension cultures ofAcer pseudoplatanus L.—the action of kinetin. In: Biochemistry and Physiology of Plant Growth Substances, pp. 484–504 (F. Wightman andG. Setterfield, eds.). Ottawa: Runge Press Ltd.
—,P. J. King, andK. J. Mansfield, 1971: Growth control in plant cell suspension cultu Res. Proc. 2nd Intern. Conf. Plant Tissue Culture (L. Hirth, ed.), C.N.R.S., Paris. In press.
— andS. M. McGregor, 1952: The carbohydrate nutrition of tomato roots. III. The effect of external sucrose concentration on the growth and anatomy of excised roots. Ann. Bot. (Lond.)16, 185–205.
Stuart, R., andH. E. Street, 1969: Studies on the growth in culture of plant cells. IV. The initiation of division in suspensions of stationary phase cells ofAcer pseudoplatanus L. J. exp. Bot.20, 556–571.
Sutton-Jones, B., andH. E. Street, 1968: Studies on the growth in culture of plant cells. III. Changes in fine structure during the growth ofAcer pseudoplatanus L. cells in suspension culture. J. exp. Bot.19, 114–118.
Thornber, J. P., andD. H. Northcote, 1961: Changes in the chemical composition of a cambial cell during its differentiation to xylem and phloem tissue in trees. I. Main components. Biochem. J.81, 449–455.
Torrey, J. G., 1968: Hormonal control of cytodifferentiation in agar and cell suspension cultu Res. In: Biochemistry and Physiology of Plant Growth Substances. pp. 843–855 (F. Wightman andG. Setterfield, eds.). Ottawa: Runge Press Ltd.
Venverloo, C. J., 1969: The lignin ofPopulus nigra L. cv.italica. A comparative study of the lignified structures in tissue cultures and the tissues of the tree. Acta Bot. Neerl.18, 241–314.
Weston, G. D., andH. E. Street, 1968: The effects of 1-napthaleneacetic acid on the growth of excised tomato roots. J. exp. Bot.19, 628–635.
Wetmore, R. H., andJ. P. Rier, 1963: Experimental induction of vascular tissues in callus in Angiosperms. Amer. J. Bot.50, 418–420.
Wilson, S. B., P. J. King, andH. E. Street, 1971: Studies on the growth in culture of plant cells. XII. A versatile system for the large scale bath or continuous culture of plant cell suspensions. J. exp. Bot.22, 177–207.
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Carceller, M., Davey, M.R., Fowler, M.W. et al. The influence of sucrose, 2,4-D, and kinetin on the growth, fine structure, and lignin content of cultured sycamore cells. Protoplasma 73, 367–385 (1971). https://doi.org/10.1007/BF01273940
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DOI: https://doi.org/10.1007/BF01273940