, Volume 219, Issue 1, pp 141–146 | Cite as

Regulation of developmental pathways in cultured microspores of tobacco and snapdragon by medium pH

  • Ioulia Barinova
  • Christophe Clément
  • Laurent Martiny
  • Fabienne Baillieul
  • Hana Soukupova
  • Erwin Heberle-Bors
  • Alisher Touraev
Original Article


The regulation of developmental pathways in cultured microspores of tobacco (Nicotiana tabacum L) and snapdragon (Antirrhinum majus L) by medium pH is described for the first time. Unicellular tobacco and snapdragon microspores developed into normal, fertile pollen when cultured in media T1 and AT3 at pH 7.0 and 25°C for 6 and 8 days, respectively. First, pollen mitosis was asymmetric and mature pollen grains were filled with starch granules and germinated upon transfer to a germination medium. However, when tobacco and snapdragon microspores were cultured in media T1 and AT3, respectively, at pH 8.0–8.5 for 4–6 days at 25 °C, the frequency of symmetric division increased significantly with the formation two nuclei of equal size, and the gametophytic pathway was blocked, as seen by the lack of starch accumulation and the inhibition of pollen germination. The transfer of these microspores to embryogenesis medium AT3 at pH 6.5 resulted in the formation of multicellular structures in both species and, in tobacco, in the formation of embryos and plants. In order to understand the possible mechanisms of the action of high pH, sucrose metabolism was analysed in isolated microspores of tobacco cultured at various pH values. Invertase (EC activity in microspores was maximal at pH 5.0 and strongly decreased at higher pH, leading to a slow-down of sucrose cleavage. At the same time the incorporation of 14C-labelled sucrose from the medium into microspores was drastically reduced at high pH. These data suggest that isolated microspores are not able to metabolise carbohydrates at high pH and thus undergo starvation stress, which was shown earlier to block the gametophytic pathway and trigger sporophytic development.


Antirrhinum Nicotiana Microspore Pollen pH Invertase 



The authors specially thank Dr. V. Zarsky (Institute of Experimental Botany, Prague, Czech Republic) for critical reading the manuscript and very useful advice and Dr. Z. Schwarz-Sommer (MPI, Cologne, Germany) for providing A. majus seeds. I. Barinova and Ch. Clément contributed equally to this work.


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Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  • Ioulia Barinova
    • 1
  • Christophe Clément
    • 2
  • Laurent Martiny
    • 3
  • Fabienne Baillieul
    • 2
  • Hana Soukupova
    • 4
  • Erwin Heberle-Bors
    • 1
  • Alisher Touraev
    • 1
  1. 1.Max F. Perutz Laboratories, University Departments at the Vienna BiocenterInstitute of Microbiology and GeneticsViennaAustria
  2. 2.UFR Sciences, URVVC UPRES EA 2069, Biologie et Physiologie VégétalesUniversité de Reims Champagne Ardenne Reims Cédex 2France
  3. 3.UFR Sciences, BiochimieUniversité de Reims Champagne ArdenneReims Cédex 2France
  4. 4.Institute of Experimental BotanyAcademy of Sciences of the Czech RepublicPrague 6 - LysolajeCzech Republic

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