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. 1972 Oct;50(4):438–445. doi: 10.1104/pp.50.4.438

Cell Wall Regeneration around Protoplasts Isolated from Convolvulus Tissue Culture 1

Randall K Horine a,2, Albert W Ruesink a
PMCID: PMC366163  PMID: 16658192

Abstract

Protoplasts of Convolvulus arvensis L. tissue culture regenerated a wall-like structure within 3 days in culture. Although unusually electron dense and atypically amorphous in the electron microscope, this structure could be digested with Myrothecium cellulase but was resistant to protease, a Rohm and Haas pectinase, and a β-1, 3-exoglucanase just like the original wall. A cytochemical test for callose was negative. Wall regeneration required a readily metabolized external carbon source and was not inhibited by a high concentration of cycloheximide, puromycin, or actinomycin D. Protoplast budding was correlated with the wall regeneration, and the latter was related quantitatively to the sucrose concentration in the medium. Although a concentration of 1 μm 2,4-dichlorophenoxy acetic acid is used normally for both general culture of the tissue and for wall regeneration, concentrations of 0 and 0.1 mm, which are highly deleterious to growth, have no appreciable effect on the incidence of the wall-like structure regenerated around protoplasts. The ability of protoplasts to undergo cell wall regeneration was decreased when they were cultured in the presence of proteolytic enzymes.

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Selected References

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  1. Abdul-Baki A. A., Ray P. M. Regulation by auxin of carbohydrate metabolism involved in cell wall synthesis by pea stem tissue. Plant Physiol. 1971 Apr;47(4):537–544. doi: 10.1104/pp.47.4.537. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. BACHMANN B. J., BONNER D. M. Protoplasts from Neurospora crassa. J Bacteriol. 1959 Oct;78:550–556. doi: 10.1128/jb.78.4.550-556.1959. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bailey R. W., Hassid W. Z. Xylan synthesis from uridine-diphosphate-d-xylose by particulate preparations from immature corncobs. Proc Natl Acad Sci U S A. 1966 Nov;56(5):1586–1593. doi: 10.1073/pnas.56.5.1586. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Baker D. B., Ray P. M. Direct and Indirect Effects of Auxin on Cell Wall Synthesis in Oat Coleoptile Tissue. Plant Physiol. 1965 Mar;40(2):345–352. doi: 10.1104/pp.40.2.345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. COCKING E. C. Action of growth substances, chelating agents and antibiotics on isolated root protoplasts. Nature. 1962 Mar 10;193:998–999. doi: 10.1038/193998a0. [DOI] [PubMed] [Google Scholar]
  6. Cocking E. C. Virus uptake, cell wall regeneration, and virus multiplication in isolated plant protoplasts. Int Rev Cytol. 1970;28:89–124. doi: 10.1016/s0074-7696(08)62541-3. [DOI] [PubMed] [Google Scholar]
  7. EDDY A. A., WILLIAMSON D. H. Formation of aberrant cell walls and of spores by the growing yeast protoplast. Nature. 1959 Apr 18;183(4668):1101–1104. doi: 10.1038/1831101a0. [DOI] [PubMed] [Google Scholar]
  8. Earle E. D., Torrey J. G. Colony Formation by Isolated Convolvulus Cells Plated on Defined Media. Plant Physiol. 1965 May;40(3):520–528. doi: 10.1104/pp.40.3.520. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gabriel M. Formation and regeneration of protoplasts in the mold Rhizopus nigricans. Folia Microbiol (Praha) 1968;13(3):231–234. doi: 10.1007/BF02871039. [DOI] [PubMed] [Google Scholar]
  10. Hassid W. Z. Biosynthesis of oligosaccharides and polysaccharides in plants. Science. 1969 Jul 11;165(3889):137–144. doi: 10.1126/science.165.3889.137. [DOI] [PubMed] [Google Scholar]
  11. Holtzer H., Abbott J., Lash J., Holtzer S. THE LOSS OF PHENOTYPIC TRAITS BY DIFFERENTIATED CELLS IN VITRO, I. DEDIFFERENTIATION OF CARTILAGE CELLS. Proc Natl Acad Sci U S A. 1960 Dec;46(12):1533–1542. doi: 10.1073/pnas.46.12.1533. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kao K. N., Keller W. A., Miller R. A. Cell division in newly formed cells from protoplasts of soybean. Exp Cell Res. 1970 Oct;62(2):338–340. doi: 10.1016/0014-4827(70)90563-x. [DOI] [PubMed] [Google Scholar]
  13. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  14. Motoyoshi F. Protoplasts isolated from callus cells of maize endosperm. Exp Cell Res. 1971 Oct;68(2):452–456. doi: 10.1016/0014-4827(71)90173-x. [DOI] [PubMed] [Google Scholar]
  15. Necas O., Svoboda A., Kopecká M. The effect of cycloheximide (actidione) on cell wall synthesis in yeast protoplasts. Exp Cell Res. 1968 Oct;53(1):291–293. doi: 10.1016/0014-4827(68)90378-9. [DOI] [PubMed] [Google Scholar]
  16. PHILLIPS H. J., TERRYBERRY J. E. Counting actively metabolizing tissue cultured cells. Exp Cell Res. 1957 Oct;13(2):341–347. doi: 10.1016/0014-4827(57)90013-7. [DOI] [PubMed] [Google Scholar]
  17. Poste G. Tissue dissociation with proteolytic enzymes. Adsorption and activity of enzymes at the cell surface. Exp Cell Res. 1971 Apr;65(2):359–367. doi: 10.1016/0014-4827(71)90014-0. [DOI] [PubMed] [Google Scholar]
  18. Power J. B., Cummins S. E., Cocking E. C. Fusion of isolated plant protoplasts. Nature. 1970 Mar 14;225(5237):1016–1018. doi: 10.1038/2251016a0. [DOI] [PubMed] [Google Scholar]
  19. Ruesink A. W., Thimann K. V. Protoplasts from the Avena coleoptile. Proc Natl Acad Sci U S A. 1965 Jul;54(1):56–64. doi: 10.1073/pnas.54.1.56. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Scaife J. F., Brohée H. A quantitative measurement of cell damage by means of 51Cr-binding. Exp Cell Res. 1967 Jun;46(3):612–615. doi: 10.1016/0014-4827(67)90391-6. [DOI] [PubMed] [Google Scholar]
  21. Svoboda A., Necas O. Mechanism of regeneration of yeast protoplasts. VI. An experimental blocking of regeneration of protoplasts. Folia Biol (Praha) 1968;14(5):390–397. [PubMed] [Google Scholar]
  22. Villemez C. L., Lin T. Y., Hassid W. Z. Biosynthesis of the polygalacturonic acid chain of pectin by a particulate enzyme preparation from Phaseolus aureus seedlings. Proc Natl Acad Sci U S A. 1965 Dec;54(6):1626–1632. doi: 10.1073/pnas.54.6.1626. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Villemez C. L., Swanson A. L., Hassid W. Z. Properties of a polygalacturonic acid-synthesizing enzyme system from Phaseolus aureus seedlings. Arch Biochem Biophys. 1966 Sep 26;116(1):446–452. doi: 10.1016/0003-9861(66)90051-8. [DOI] [PubMed] [Google Scholar]
  24. Werz G. Differenzierung und Zellwandibildung in isoliertem Cytoplasma aus Acetabularia. Protoplasma. 1968;65(3):349–357. doi: 10.1007/BF01682538. [DOI] [PubMed] [Google Scholar]
  25. Witham F. H. Effect of 2,4-dichlorophenoxyacetic Acid on the cytokinin requirement of soybean cotyledon and tobacco stem pith callus tissues. Plant Physiol. 1968 Sep;43(9):1455–1457. doi: 10.1104/pp.43.9.1455. [DOI] [PMC free article] [PubMed] [Google Scholar]

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