Skip to main content
SpringerLink
Log in

Microspore Embryogenesis and In Vitro Pollen Maturation in Tobacco

  • Protocol
Plant Cell Culture Protocols

Part of the book series: Methods In Molecular Biology™ ((MIMB,volume 111))

Abstract

Microspores have two developmental options when they are isolated from anthers and cultured in vitro. In a rich medium, they can develop into mature pollen grains that are fertile on pollination in vivo (1,2). Their development closely resembles pollen formed in vivo, whereas the in vitro culture conditions simulate the changing environment a microspore/pollen grain experiences inside the anther. The microspore divides by an asymmetrical division, and the daughter cells differentiate into mature pollen grains, with their characteristic cell-cycle arrest (vegetative cell in Gl, generative cells in G2). This simple pathway makes in vitro pollen maturation an excellent system to study developmental processes, such as cell fate determination and cellular differentiation (3). In addition, in vitro pollen maturation has also been used for pollen selection (4) and plant transformation (5). Another potential application is the rescue of sterile or self-incompatible pollen for “self”-pollinations.

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

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 109.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Benito Moreno, R. M., Macke, F, Alwen, A., and Heberle-Bors, E. (1988) In-situ seed production after pollination with in-vitro-matured, isolated pollen Planta 176, 145–148

    Google Scholar 

  2. Stauffer, C, Benito Moreno, R. M., and Heberle-Bors, E (1991) In situ pollination with in vitro matured pollen of Triticum aestivum Theor Appl Genet 81, 576–580.

    Google Scholar 

  3. McCormick, S. (1993) Male gametophyte development Plant Cell 5, 1265–1275

    Article  PubMed  Google Scholar 

  4. Touraev, A., Fink, Ch., Stoger, E., and Heberle-Bors, E (1995). Pollen selection: a transgenic reconstruction approach Proc Natl Acad Set USA 92, 12,165–12,169

    Article  CAS  Google Scholar 

  5. Touraev, A., Stoger, E., Voronin, V., and Heberle-Bors, E. (1998) Plant male germ line transformation. Plant J 12(4), 949–956

    Article  Google Scholar 

  6. Touraev, A, Ilham, A, Vicente, O, and Heberle-Bors, E (1996) Stress induced microspore embryogenesis from tobacco microspores: an optimized system for molecular studies Plant Cell Rep 15, 561–565.

    Article  CAS  Google Scholar 

  7. Touraev, A, Vicente, O, and Heberle-Bors, E. (1997) Initiation of microspore embryogenesis by stress. Trends Plant Sci 2, 297–302

    Article  Google Scholar 

  8. Touraev, A., Indnanto, A., Wratscko, I., Vicente, O., and Heberle-Bors, E. (1996) Efficient microspore embryogenesis in wheat (Triticurn aestivum L ) induced by starvation at high temperatures. Sex Plant Reprod 9, 209–215

    Article  Google Scholar 

  9. Hoekstra, S., van Zijderveld, M H, Louwerse, J. D, Heidekamp, F, and van der Mark, F. (1992) Anther and microspore culture of Hordeum vulgare L cv Ign. Plant Set 86, 89–96

    Article  CAS  Google Scholar 

  10. Ogawa T., Fukuoka, H, and Ohkawa, Y (1994) Induction of cell division of isolated pollen grains by sugar starvation in rice Breeding Sci (Jpn) 44, 75–77

    Google Scholar 

  11. Pechan, P. M. and Keller, W A (1988) Identification of potentially embryogenic microspores in Brassica napus Physiol Plant 74, 377–384.

    Google Scholar 

  12. Morrison, R A and Evans, D A (1988) Haploid plants from tissue culture: new plant varieties in a shortened time frame. Bio/Technology 6, 684–690

    Article  Google Scholar 

  13. Feme, A M. R, Palmer, C E., and Keller, W A (1995). Haploid embryogenesis, in In Vitro Plant Embryogenesis (Thorpe, T A, ed.), Kluwer, Dordrecht, pp 309–344

    Google Scholar 

  14. Jahne, A., Becker, D., Brettschneider, R, and Lorz, A. (1994) Regeneration of transgenic, microspore-derived, fertile barley Theor Appl Genet 89, 525–533

    Article  Google Scholar 

  15. Stoger, E, Fink, C, Pfosser, M., and Heberle-Bors, E. (1995) Plant transformation by particle bombardment of embryogenic pollen Plant Cell Rep 14, 273–278.

    Article  Google Scholar 

  16. Kyo, M. and Harada, H. (1986) Control of the developmental pathway of tobacco pollen in vitro Planta 168, 427–432.

    Article  CAS  Google Scholar 

  17. Tupy, J, R’hová, L., and Zársky, V (1991) Production of fertile tobacco pollen from microspores in suspension cultures and its storage for in situ pollination Sex Plant Reprod 4, 284–287

    Article  Google Scholar 

  18. Read, S. M., Clarke, A. E., and Bacic, A (1993) Stimulation of growth of cultured Nicotiana tabacum W38 pollen tubes by poly (ethylene glycol) and Cu(II) salts Protoplasma 177, 1–14

    Article  CAS  Google Scholar 

  19. Ylstra, B., Touraev, A., Benito Moreno, R. M, Stoger, E., van Tunen, A J., Vicente, O., Mol, J N. M, et al (1992). Flavonols stimulate development, germination, and tube growth of tobacco pollen. Plant Physiol. 100, 902–907

    Article  PubMed  CAS  Google Scholar 

  20. Murashige, T. and Skoog, F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15, 473–497

    Article  CAS  Google Scholar 

  21. Mahga, P, Breznovits, A, and Marton, L (1973) Streptomycin-resistant plants from callus culture of haploid tobacco Nature 244, 29–30.

    Google Scholar 

  22. Vergne, P., Delvallée, I., and Dumas, C. (1987) Rapid assessment of microspore and pollen development stage in wheat and maize using DAPI and membrane permeabilization. Stain Technol. 62, 299–304.

    PubMed  CAS  Google Scholar 

  23. Benito Moreno, R. M., Macke, F,, Hauser, M.-T, Alwen, A., and Heberle-Bors, E (1988) Sporophytes and male gametophytes from in vitro cultured, immature tobacco pollen, in Sexual Reproduction in Higher Plants (Cresti, M, Gon, P and Pacini, E., eds.), Springer, Heidelberg, NY, pp 137–142

    Google Scholar 

  24. Swanson, E B, Coumans, M. P., Wu, S. C, Barsby, T L., and Beversdorf, W. D (1987) Efficient isolation of microspores and the production of microspore-derived embryos from Brassica napus. Plant Cell Rep 6, 94–97

    Google Scholar 

  25. Garrido, D., Charvat, B, Benito Moreno, R. M, Alwen, A, Vicente, O, and Heberle-Bors, E. (1991) Pollen culture for haploid plant production in tobacco, in A Laboratory Guide for Cellular and Molecular Plant Biology (Negrutiu, I. and Gharti-Chhetn, G, eds), Birkhauser, Basel, pp 59–69.

    Google Scholar 

  26. Vogt, T., Pollak, P., Tarlyn, N., and Taylor, L. P. (1994) Pollination-or wound-induced kaempferol accumulation in petunia stigmas enhances seed production Plant Cell 6, 11–23

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Vienna Bwcentre, Institute of Microbiology and Genetics, Vienna University, Vienna, Austria

    Alisher Touraev

  2. Institute of Microbiology and Genetics, Vienna Biocenter, University of Vienna, Vienna, Austria

    Erwin Heberle-Bors

Authors
  1. Alisher Touraev
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Erwin Heberle-Bors
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. CPRO-DLO, Wageningen, The Netherlands

    Robert D. Hall

Rights and permissions

Reprints and permissions

Copyright information

© 1999 Humana Press Inc., Totowa, NJ

About this protocol

Cite this protocol

Touraev, A., Heberle-Bors, E. (1999). Microspore Embryogenesis and In Vitro Pollen Maturation in Tobacco. In: Hall, R.D. (eds) Plant Cell Culture Protocols. Methods In Molecular Biology™, vol 111. Humana Press. https://doi.org/10.1385/1-59259-583-9:281

Download citation

  • DOI: https://doi.org/10.1385/1-59259-583-9:281

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-549-2

  • Online ISBN: 978-1-59259-583-9

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation