Skip to main content
SpringerLink
Log in

Unexpected features of exponentially growing Tobacco Bright Yellow-2 cell suspension culture in relation to excreted extracellular polysaccharides and cell wall composition

  • Short Communication
  • Published:
Glycoconjugate Journal Aims and scope Submit manuscript

Abstract

This article presents a new insight about TBY-2 cells; from extracellular polysaccharides secretion to cell wall composition during cell suspension culture. In the medium of cells taken 2 days after dilution (end of lag phase), a two unit pH decrease from 5.38 to 3.45 was observed and linked to a high uronic acid (UA) amount secretion (47.8%) while, in 4 and 7 day-old spent media, pH increased and UA amounts decreased 35.6 and 42.3% UA, respectively. To attain deeper knowledge of the putative link between extracellular polysaccharide excretion and cell wall composition, we determined cell wall UA and neutral sugar composition of cells from D2 to D12 cultures. While cell walls from D2 and D3 cells contained a large amount of uronic acid (twice as much as the other analysed cell walls), similar amounts of neutral sugar were detected in cells from lag to end of exponential phase cells suggesting an enriched pectin network in young cultures. Indeed, monosaccharide composition analysis leads to an estimated percentage of pectins of 56% for D3 cell wall against 45% D7 cell walls indicating that the cells at the mid-exponential growth phase re-organized their cell wall linked to a decrease in secreted UA that finally led to a stabilization of the spent medium pH to 5.4. In conclusion, TBY-2 cell suspension from lag to stationary phase showed cell wall remodeling that could be of interest in drug interaction and internalization study.

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

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

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

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Abbreviations

CW:

cell wall

TBY-2:

tobacco bright yellow-2 cells

UA:

uronic acid

References

  1. Kato, K., Matsumoto, T., Koiwai, S., Mizusaki, S., Nishida, K., Nogushi, M., Tamaki, E.: Liquid suspension culture of tobaco cells. in Fermentation Technology Today. (ed) Terui G (Society of Fermentation Technology, Osaka) 689–695 (1972)

  2. Nagata, T., Nemoto, Y., Hasezawa, S.: Tobacco BY-2 cell line as the “Hela” cell in the cell biology of higher plants. Int. Rev. Cytol. 132, 1–30 (1992)

    Article  CAS  Google Scholar 

  3. Kobayashi, I., Hakuno, H.: Actin-related defense mechanism to reject penetration attempt by a non-pathogen is maintained in tobacco BY-2 cells. Planta. 217, 340–345 (2003)

    CAS  PubMed  Google Scholar 

  4. de Pinto, M.C., Locato, V., Sgobba, A., Romero-Puertas, M.d.C., Gadaleta, C., Delledonne, M., De Gara, L.: S-nitrosylation of ascorbate peroxidase is part of programmed cell death signaling in tobacco bright yellow-2 cells. Plant Physiol. 163, 1766–1775 (2013)

    Article  PubMed  PubMed Central  Google Scholar 

  5. Takahashi, S., Kojo, K.H., Kutsuna, N., Endo, M., Toki, S., Isoda, H., Hasezawa, S.: Differential responses to high- and low-dose ultraviolet-B stress in tobacco bright yellow-2 cells. Front. Plant Sci. 6, 254–264 (2015)

    PubMed  PubMed Central  Google Scholar 

  6. Moscatiello, R., Baldan, B., Navazio, L.: Plant cell suspension cultures. In: Maathuis, F.J.M. (ed.) Plant Mineral Nutrients, pp. 77–93. Humana Press, Totowa, NJ (2013)

    Chapter  Google Scholar 

  7. Srba, M., Černíková, A., Opatrný, Z., Fischer, L.: Practical guidelines for the characterization of tobacco BY-2 cell lines. Biol. Plant. 60, 13–24 (2016)

    Article  CAS  Google Scholar 

  8. Bratic, A., Majic, D., Samardzic, J., Kragl, M.W., Maksimovic, V.: Transient expression in tobacco bright yellow 2 cells and pollen grains: a fast, efficient and reliable system for functional promoter analysis of plant genes. Arch. Biol. Sci. 62, 57–62 (2010)

    Article  Google Scholar 

  9. De Marco, A., Guzzardi, P., Jamet, É.: Isolation of tobacco isoperoxidases accumulated in cell-suspension culture medium and characterization of activities related to cell wall metabolism. Plant Physiol. 120, 371–382 (1999)

    Article  PubMed  PubMed Central  Google Scholar 

  10. Nebenführ, A., Frohlick, J.A., Staehelin, L.A.: Redistribution of Golgi stacks and other organelles during mitosis and cytokinesis in plant cells. Plant Physiol. 124, 135–152 (2000)

    Article  PubMed  PubMed Central  Google Scholar 

  11. Hong, Z., Delauney, A.J., Verma, D.P.S.: A cell plate–specific callose synthase and its interaction with phragmoplastin. Plant Cell. 13, 755–768 (2001)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Yokoyama, R., Tanaka, D., Fujino, T., Itoh, T., Nishitani, K.: Cell wall dynamics in tobacco BY-2 cells. Tobacco BY-2 Cells. 53, 217–230 (2004)

    Article  CAS  Google Scholar 

  13. Kobayashi, M., Ohno, K., Matoh, T.: Boron nutrition of cultured tobacco BY-2 cells. II. Characterization of the boron-polysaccharide complex. Plant Cell Physiol. 38, 676–683 (1997)

    Article  CAS  Google Scholar 

  14. Matoh, T., Takasaki, M., Kobayashi, M., Takabe, K.: Boron nutrition of cultured tobacco BY-2 cells. III. Characterization of the boron-rhamnogalacturonan II complex in cells acclimated to low levels of boron. Plant Cell Physiol. 41, 363–366 (2000)

    Article  CAS  PubMed  Google Scholar 

  15. Goué, N., Mortimer, J.C., Nakano, Y., Zhang, Z., Josserand, M., Ohtani, M., Dupree, P., Kakegawa, K., Demura, T.: Secondary cell wall characterization in a BY-2 inductive system. PCTOC. 115, 223–232 (2013)

    Article  Google Scholar 

  16. Wink, M.: The cell culture medium—a functional extracellular compartment of suspension-cultured cells. PCTOC. 38, 307-319 (1994)

  17. Jamet, E., Canut, H., Boudart, G., Pont-Lezica, R.F.: Cell wall proteins: a new insight through proteomics. Trends Plant Sci. 11, 33–39 (2006)

    Article  CAS  PubMed  Google Scholar 

  18. Webster, J.M., Oxley, D., Pettolino, F.A., Bacic, A.: Characterisation of secreted polysaccharides and glycoproteins from suspension cultures of Pyrus communis. Phytochemistry. 69, 873–881 (2008)

    Article  CAS  PubMed  Google Scholar 

  19. Becker, G.E., Hui, P.A., Albersheim, P.: Synthesis of extracellular polysaccharide by suspensions of Acer pseudoplatanus cells. Plant Physiol. 39, 913 (1964)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Olson, A.C., Evans, J.J., Frederick, D.P., Jansen, E.F.: Plant suspension culture media macromolecules—pectic substances, protein, and peroxidase. Plant Physiol. 44, 1594–1600 (1969)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Liau, D.-F., Boll, W.G.: Extracellular polysaccharide from cell suspension cultures of bush bean (Phaseolus vulgaris cv. Contender). Can. J. Bot. 50, 2031–2037 (1972)

    Article  CAS  Google Scholar 

  22. Moore, T.S.: An extracellular macromolecular complex from the surface of soybean suspension cultures. Plant Physiol. 51, 529–536 (1973)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Oliveira, C.J.F., Cavalari, A.A., Carpita, N.C., Buckeridge, M.S., Braga, M.R.: Cell wall polysaccharides from cell suspension cultures of the Atlantic Forest tree Rudgea jasminoides (Rubiaceae). Trees. 24, 713–722 (2010)

    Article  CAS  Google Scholar 

  24. Moser, J.R, Gonçalves Garcia, M., Maria Viana, A.: Establishment and growth of embryogenic suspension cultures of Ocotea catharinensis Mez.(Lauraceae). PCTOC. 78, 37–42 (2004)

  25. Dubois, M., Gilles, K.A., Hamilton, J.K., Rebers P.A, Smith, F.: Colorimetric method for determination of sugars and related substances. Anal. Chem. 28, 350–356 (1956)

  26. Blumenkrantz, N., Asboe-Hansen, G.: New method for quantitative determination of uronic acids. Anal. Biochem. 54, 484–489 (1973)

    Article  CAS  PubMed  Google Scholar 

  27. Montreuil, J., Bouquelet, S., Debray, H., Fournet, B., Spik, G., Strecker, G.: In: Chaplin MF, Kennedy JF. (Eds), Carbohydrate analysis, a practical approach, IRL Press, Oxford, Washington D.C 143–204. (1986)

  28. Kamerling, J.P., Gerwig, G.J., Vliegenthart, J.F.G., Clamp, J.R.: Characterization by gas-liquid chromatography-mass spectrometry and proton-magnetic-resonance spectroscopy of pertrimethylsilyl methyl glycosides obtained in the methanolysis of glycoproteins and glycopeptides. Biochem. J. 151, 491–495 (1975)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Marga, F., Freyssac, V., Morvan, H.: Rapid gas liquid chromatography microanalysis of carbohydrates in woody plant tissues. J. Trace Microprobe Techn. 13, 473–478 (1995)

    CAS  Google Scholar 

  30. Douchiche, O., Driouich, A., Morvan, C.: Impact of cadmium on early stages of flax fibre differentiation: Ultrastructural aspects and pectic features of cell walls. Plant Physiol. Biochem. 49, 592–599 (2011)

    Article  CAS  PubMed  Google Scholar 

  31. Muschitz, A., Faugeron, C., Morvan, H.: Response of cultured tomato cells subjected to excess zinc: role of cell wall in zinc compartmentation. Acta Physiol. Plant. 31, 1197–1204 (2009)

    Article  CAS  Google Scholar 

  32. Riou, C., Calliste, C.A., Da Silva, A., Guillaumot, D., Rezazgui, O., Sol, V., Leroy-Lhez, S.: Anionic porphyrin as a new powerful cell death inducer of tobacco bright yellow-2 cells. Photochem. Photobiol. Sci. 13, 621–625 (2014)

    Article  CAS  PubMed  Google Scholar 

  33. Yamanaka, A., Hashimoto, A., Matsuo, T., Kanou, M., Suehara, K.-I., Kameoka, T.: Analysis of kinetic uptake phenomena of monosaccharide and disaccharide by suspension TBY-2 cells using an FT-IR/ATR method. Bioproc Biosystems Eng. 30, 457–468 (2007)

    Article  CAS  Google Scholar 

  34. Morvan, H., Priem, B., Verdus, M.C., Kwan, J.S., Morvan, C.: Relationship between the acidity of the pectic acid polymers from suspension cultured cells of white campion and their promoting effects on the growth. Food Hydrocoll. 1, 501–503 (1987)

    Article  CAS  Google Scholar 

  35. Ding, Y., Robinson, D.G., Jiang, L.: Unconventional protein secretion (UPS) pathways in plants. Curr. Op. Cell Biol. 29, 107–115 (2014)

    Article  CAS  PubMed  Google Scholar 

  36. Kim, S.-J., Brandizzi, F.: The plant secretory pathway for the trafficking of cell wall polysaccharides and glycoproteins. Glycobiology. 26, 940–949 (2016)

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

Mohammad Issawi was supported by a Grant from the municipality of Sharkieh (Lebanon). Authors greatly thank Pr M. Guilloton for reading this manuscript.

Author information

Authors and Affiliations

  1. Laboratoire de Chimie des Substances Naturelles, Université de Limoges, EA 1069, 123 avenue Albert Thomas, 87060, Limoges, France

    Mohammad Issawi, Mohammad Muhieddine, Celine Girard, Vincent Sol & Catherine Riou

Authors
  1. Mohammad Issawi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Mohammad Muhieddine
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Celine Girard
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Vincent Sol
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Catherine Riou
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Catherine Riou.

Ethics declarations

Conflicts of interest

The authors declare that they have no conflicts of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Issawi, M., Muhieddine, M., Girard, C. et al. Unexpected features of exponentially growing Tobacco Bright Yellow-2 cell suspension culture in relation to excreted extracellular polysaccharides and cell wall composition. Glycoconj J 34, 585–590 (2017). https://doi.org/10.1007/s10719-017-9782-7

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10719-017-9782-7

Keywords

Search

Navigation