Plant Cell, Tissue and Organ Culture

, Volume 33, Issue 1, pp 81–89 | Cite as

Effect of cytokinins on plastid development and photosynthetic polypeptides during organogenesis ofPinus ponderosa Dougl. cotyledons culturedin vitro

  • Alicia Mazari
  • Edith L. Camm
Original Papers


InPinus ponderosa Dougl., application of the cytokinins, benzyladenine and 2-isopentenyl adenine, to excised cotyledons, promoted thein vitro formation of meristematic centers which led to bud and shoot production. Meristematic cells showed plastids with poorly developed thylakoid membranes and rudimentary grana, whereas cells in non-meristematic tissues and in growth regulator free medium, had chloroplasts with well developed inner membranes, and more thylakoid membranes and grana than plastids of meristematic cells. Chlorophyll and six polypeptides associated with photosynthesis were present in lower concentrations in cytokinin-treated cotyledons than in those cultured in growth regulator free medium. Both benzyladenine and 2-isopentenyl adenine are effective in inhibiting the accumulation of at least two photosynthetic polypeptides in the first 24 h in culture. The ability of cotyledons to respond in this way to cytokinins is lost after three days in culture in growth regulator free medium prior to treatment with cytokinin.

Key words

cytokinins in vitro organogenesis photosynthetic polypeptides Pinus ponderosa plastid 





coupling factor




33kD extrinsic polypeptide


growth regulator free


2-isopentenyl adenine


large subunit of Ribulose-1,5-bisphosphate carboxylase


light harvesting complex of photosystem I


light harvesting complex of photosystem II


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Bonga JM (1982) Vegetative propagation in relation to juvenility, maturity and rejuvenation. In: Bonga JM & Durzan DJ (Eds) Tissue Culture in Forestry (pp 387–412). Martinus Nijhoff, The HagueGoogle Scholar
  2. Camm EL, Green BR, Allred DR & Staehelin LA (1987) Association of the 33 kDa extrinsic polypeptide (watersplitting) with PSII particles: immunochemical quantification of residual polypeptide after membrane extraction. Photos. Res. 13: 69–80CrossRefGoogle Scholar
  3. Chua NH (1980) Electrophoretical analysis of chloroplast proteins. In: Pietro AS (Ed) Methods in Enzymology, Vol 69 (pp 434–446). Academic Press, New YorkGoogle Scholar
  4. Colijn CM, Sijmons P, Mol JNM, Kool AJ & Nijkamp HJJ (1982) Light and benzylaminopurine induce changes in ultrastructure and gene expression in plastids ofPetunia hybrida cell cultures. Curr. Gen. 6: 129–135CrossRefGoogle Scholar
  5. Ellis DD & Judd RC (1987) SDS-PAGE analysis of budforming cotyledons ofPinus ponderosa. Plant Cell Tiss. Org. Cult. 11(1): 57–65CrossRefGoogle Scholar
  6. Flinn BS, Webb DT & Newcomb W (1988) The role of cell clusters and promeristemoids in determination and competence for caulogenesis byPinus strobus cotyledonsin vitro. Can. J. Bot. 66: 1556–1565Google Scholar
  7. Ghosh S, Gepstein S, Heikkila JJ & Dumbroff EB (1988) Use of a scanning densitometer or an ELISA plate reader for measurement of nanogram amounts of protein in crude extracts from biological tissues. Anal. Biochem. 169: 227–233CrossRefPubMedGoogle Scholar
  8. Harvey BMR, Lu BC & Fletcher RA (1974) Benzyladenine accelerates chloroplast differentiation and stimulates photosynthetic enzyme activity in cucumber cotyledons. Can. J. Bot. 52: 2581–2586Google Scholar
  9. Hiscox JD & Israelstam GF (1979) A method for the extraction of chlorophyll from leaf tissue without maceration. Can. J. Bot. 57: 1332–1334Google Scholar
  10. Kumar PP, Bender L & Thorpe TA (1988) Activities of ribulose bisphosphate carboxylase and phosphoenolpyruvate carboxylase and 14-C-bicarbonate fixation duringin vitro culture ofPinus radiata cotyledons. Plant Physiol. 87: 675–679Google Scholar
  11. Kumar PP, Reid DM & Thorpe TA (1987) The role of ethylene and carbon dioxide in differentiation of shoot buds in excised cotyledons ofPinus radiata in vitro. Physiol. Plant. 69: 244–252Google Scholar
  12. Lerbs S, Lerbs W, Klyachko NL, Romanko EG, Kulaeva ON, Wollgiehn R & Parthier B (1984) Gene expression in cytokinin-and light-mediated plastogenesis ofCucurbita cotyledons: ribulose-1,5-bisphosphate carboxylase/oxygenase. Planta 162: 289–298CrossRefGoogle Scholar
  13. Lescure AM & Seyer P (1981) Effect of cytokinin on plastid differentiation in tobacco cell suspensions. In: Guern J & Peaud-Lenoël C (Eds) Metabolism and Molecular Activities of Cytokinins (pp 298–307). Springer-Verlag, BerlinGoogle Scholar
  14. Lichtenthaler H & Buschmann C (1978) Control of chloroplast development by red light, blue light and phytohormones. In: Akoyunoglou G et al. (Eds) Chloroplast development (pp 801–816). Elsevier, N. HollandGoogle Scholar
  15. Mazari-Hiriart A (1991) Effect of cytokinins on tissue structure, plastid development and photosynthetic proteins in tissue culture ofPinus ponderosa Dougl. cotyledons during organogenesis. Ph.D. thesis, The University of British Columbia, Vancouver, B.C., CanadaGoogle Scholar
  16. Mikulovich TP, Romanko ER, Yu-Selivankina S, Kukina IM & Wollgiehn R (1981) Cytokinin action on RNA synthesis in chloroplasts. In: Guern J & Peaud-Lenoël C (Eds) Metabolism and Molecular Activities of Cytokinins (pp 287–297). Springer-Verlag, BerlinGoogle Scholar
  17. Ohya T & Suzuki H (1990) Benzyladenine- and light-stimulated plastid protein synthesis in excised cucumber cotyledons. Plant Physiol. Biochem. 28(1): 27–35Google Scholar
  18. Parthier B, Lerbs S, Lehmann J & Wollgiehn R (1985) Cytokinin-controlled ribulose-1,5-bisphosphate carboxylase gene expression in pumpkin cotyledons. Biol. Plant. 27(2–3): 131–138Google Scholar
  19. Patel KR & Thorpe TA (1984) Histochemical examination of shoot initiation in cultured cotyledon explants of radiata pine. Bot. Gaz. 145(3): 312–322CrossRefGoogle Scholar
  20. Spurr A (1969) A low viscosity epoxy resin embedding medium for electron microscopy. J. Ultrastructure Res. 26: 31–43CrossRefGoogle Scholar
  21. Stabel P, Eriksson T & Engstrom P (1990) Changes in protein synthesis upon cytokinin-mediated adventitious bud induction and during seedling development in Norway spruce,Picea abies. Plant Physiol. 92: 1174–1183Google Scholar
  22. Stabel P, Sundas A & Engstrom P (1991) Cytokinin treatment of embryos inhibits the synthesis of chloroplast proteins in Norway spruce. Planta 183: 520–527CrossRefGoogle Scholar
  23. Tranvan H, Troton D & Calvayrac R (1988) Morphological, histological and lipid changes during adventitious budding inPinus pinaster cultured cotyledons. J. Exp. Bot. 39(204): 907–915Google Scholar
  24. Villalobos VM, Yeung EC & Thorpe TA (1985) Origin of adventitious shoots in excised radiata pine cotyledons culturedin vitro. Can. J. Bot. 63: 2172–2176Google Scholar
  25. Von Arnold S & Eriksson T (1981)In vitro studies of adventitious shoot formation inPinus contorta. Can. J. Bot. 59: 870–874Google Scholar
  26. Von Arnold S & Eriksson T (1985) Initial stages in the adventitious shoot formation inPinus contorta. Can. J. Bot. 59: 870–874Google Scholar
  27. Von Arnold S & Eriksson T (1985) Initial stages in the course of adventitious bud formation on embryos ofPicea abies. Physiol. Plant. 64: 41–47Google Scholar
  28. White MJ & Green BR (1987a) Antibodies to the photosystem I chlorophyll a + b antenna cross-react with polypeptides of CP29 and LHCII. Eur. J. Biochem. 163: 545–551CrossRefPubMedGoogle Scholar
  29. White MJ & Green BR (1987b) Polypeptides belonging to each of the three major chlorophyll a + b protein complexes are present in a chlorophyll-b-less barley mutant. Eur. J. Biochem. 165: 531–535CrossRefPubMedGoogle Scholar
  30. White MJ & Green BR (1988) Intermittent-light chloroplasts are not developmentally equivalent to chlorina f2 chloroplasts in barley. Photos. Res. 15: 195–203CrossRefGoogle Scholar
  31. Zimmermann KH, Chudy M, Preusser E & Goring H (1987) Relationships between light and hormone action on ribulose-1,5-bisphosphate carboxylase in cucumber cotyledons. Biochem. Physiol. Pflanzen 182: 407–415Google Scholar

Copyright information

© Kluwer Academic Publishers 1993

Authors and Affiliations

  • Alicia Mazari
    • 1
  • Edith L. Camm
    • 1
  1. 1.Department of BotanyThe University of British ColumbiaVancouverCanada

Personalised recommendations