Skip to main content
SpringerLink
Log in

Separation of precultured pollen from Nicotiana tabacum by aqueous polymer two-phase partition

  • Published:
Plant Cell, Tissue and Organ Culture Aims and scope Submit manuscript

Abstract

Pollen isolated from cold treated and precultivated anthers of tobacco (Nicotiana tabacum L. var. Wisconsin 38) were separated into different fractions with counter-current distribution using an aqueous Dextran-polyethylene glycol two-phase system. It was possible to distinguish among eight pollen classes differing in developmental stage and in partitioning. A part of each fraction was cultivated for analysis of embryo formation. This was highest in a fraction with an intermediate to high partition in the phase system. Enriched in this fraction were also pollen that were fairly well stained with acetocarmine, contained several nuclei and had a relatively low mitochondrial activity. The enrichment of embryogenic pollen offers several advantages especially to physiological studies on embryogenesis.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Albertsson P-Å, Andersson B, Larsson C, Akerlund H-E (1982) Phase Partition: A Method for purification and analysis of cell organelles and membrane vesicles. In: Glick D (ed) Methods of Biochemical analysis, 28 London, New York: An Interscience Publication, John Wiley & sons, pp 115–150

    Google Scholar 

  2. Appelmans F, Wattiaux R, de Duve C (1955) Tissue fractionation studies. 5. The association of acid phosphatase with a special class of cytoplasmic granules in rat liver. Biochem J 59:248–254

    Google Scholar 

  3. Bearden JCJr (1978) Quantitation of submicrogram quantities of protein by an improved protein-dye binding assay. Biochim Biophys Acta 533:525–529

    Google Scholar 

  4. Craig LC, Craig D (1956) Laboratory extraction and counter-current distribution. In: A Weissberger, (ed.) Technique of Organic Chemistry, Vol III, Part I, 2nd ed. London, New York: Interscience, pp 149–332

    Google Scholar 

  5. Guha S, Maheshwari SC (1964) In vitro production of embryos from anther cultures of Datura. Nature 204:497

    Google Scholar 

  6. Heberle-Bors E, Reinert J (1980) Isolated pollen cultures and pollen dimorphism. Naturwissenschaften 67:311–312

    Google Scholar 

  7. Horner M, Street HE (1978) Pollen dimorphism: Origin and significance in pollen plant formation by anther culture. Ann Bot 42:763–771

    Google Scholar 

  8. Maheshwari SC, Tyagi AK, Malhotra K (1980) Induction of haploidy from pollen grains in angiosperms: The current status. Theor Appl Genet 58:193–206

    Google Scholar 

  9. Maheshwari SC, Rashid A, Tyagi AK (1982) Haploids from pollen grains: Retrospect and prospect. AM J Bot 69:865–879

    Google Scholar 

  10. Nitsch C, Norreel B (1973) Effet d'un choc thermique sur le pouvoir embryogéne du pollen de Datura innoxia cultivé dans l'anthere. CR Acad Sci Paris 276:303–306

    Google Scholar 

  11. Nitsch JP (1969) Experimental androgenesis in Nicotiana. Phytomorphology 19:389–404

    Google Scholar 

  12. Rashid A (1983) Pollen dimorphism in relation to pollen plant formation. Physiol Plant 58:544–548

    Google Scholar 

  13. Rashid A, Reinert J (1980) Selection of embryogenic pollen from cold-treated buds of Nicotiana tabacum var. Badischer Burley and their development into embryos in cultures. Protoplasma 105: 161–167

    Google Scholar 

  14. Rashid A, Reinert J (1981) High frequency embryogenesis in ab initio pollen cultures of Nicotiana tabacum. Naturwissenschaften 68:378–379

    Google Scholar 

  15. Rashid A, Reinert J (1983) Factors affecting high-frequency embryo formation in ab initio pollen cultures of Nicotiana. Protoplasma 116:155–160

    Google Scholar 

  16. Rashid A, Siddiqui AW, Reinert J (1982) Subcellular aspects of origin and structure of pollen embryos of Nicotiana. Protoplasma 113:202–208

    Google Scholar 

  17. Sangwan-Norreel BS (1977) Androgenic stimulating factors in the anther and isolated pollen grain culture of Datura innoxia Mill. J Exp Bot 28:943–952

    Google Scholar 

  18. Sunderland N, Wicks FM (1971) Embryoid formation in pollen grains of Nicotiana tabacum. J Exp Bot 22:213–226

    Google Scholar 

  19. Wernicke W, Harms CT, Lörz H, Thomas E (1978) Selective enrichment of embryogenic microspore populations. Naturwissenschaften 65:540–541

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Plant Physiology, University of Lund, Box 7007, S-220 07, Lund, Sweden

    Boel K. Mårtensson & Susanne Widell

Authors
  1. Boel K. Mårtensson
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Susanne Widell
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Mårtensson, B.K., Widell, S. Separation of precultured pollen from Nicotiana tabacum by aqueous polymer two-phase partition. Plant Cell Tiss Organ Cult 8, 27–35 (1987). https://doi.org/10.1007/BF00040730

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00040730

Key words

Search

Navigation