Skip to main content
Log in

Engineered fertility control in transgenic Brassica napus L.: Histochemical analysis of anther development

  • Published:
Planta Aims and scope Submit manuscript

Abstract

Cytological and histochemical analyses were performed on developing anthers of wild-type, transgenic male-sterile and fertility-restored Brassica napus plants. Male sterility resulted from the expression of the barnase gene under the control of the tobacco-derived tapetumspecific promoter pTA29. Fertility was restored to male sterile plants by expressing the barstar gene, which encodes a barnase-specific inhibitor protein. In addition, the tissue specificity of the pTA29 promoter in B. napus was studied in transgenic plants expressing pTA29:gus fusions. In B. napus, the pTA29 promoter not only directed the expression of genes to the tapetum, but also, although more weakly, to the vascular tissue region of the anther filament at the late uninucleate and early binucleate stage. The pTA29:barnase gene was expressed in the tapetum at the vacuolated-microspore stage. This resulted in the disappearance of RNA from the tapetum. Degradation of RNA in the tapetum was immediately followed by a complete loss of RNA in the developing microspores. Following lysis of the microspores in the sterile anthers, the pTA29:barnase gene was expressed in the vascular tissue region of the anther filament. This expression resulted in a deposition of wound callose in the phloem, followed by a precocious wilting of the whole anther. Finally, in the fertility-restored plants the cytological and histochemical patterns of anther development were identical to those of wild-type anthers.

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

Access this article

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

Instant access to the full article PDF.

Similar content being viewed by others

Abbreviations

NMS:

nuclear male sterility

WT:

wild type

References

  • Bronner, R., Westphal, E., Dreger, F. (1991) Enhanced peroxidase activity associated with the hypersensitive response of Solanum dulcamara to the gall mite Aceria cladophthirus (Acari:Eriophyoidea). Can. J. Bot. 69, 2192–2196

    Google Scholar 

  • De Block, M., Debrouwer, D. (1992) In-situ enzyme histochemistry on plastic-embedded plant material. The development of an artefact-free β-glucuronidase assay. Plant J. 2, 261–266

    Google Scholar 

  • De Block, M., Debrouwer, D., Tenning, P. (1989) Transformation of Brassica napus and Brassica oleracea using Agrobacterium tumefaciens and the expression of the bar and neo genes in the transgenic plants. Plant. Physiol. 91, 694–701

    Google Scholar 

  • Currier, H.B. (1957) Callose substance in plant cells. Am. J. Bot. 44, 478–488

    Google Scholar 

  • Edreva, A.M. (1991) Stress proteins of plants: PR(b)-proteins. Sov. Plant Physiol. 38, 579–588

    Google Scholar 

  • Eschrich, W., Currier, H.B. (1964) Identification of callose by its diachrome and fluorochrome reactions. Stain Technol. 39, 303–307

    Google Scholar 

  • Evert, R.F., Derr, W.F. (1964) Callose substance in sieve elements. Am. J. Bot. 51, 552–559

    Google Scholar 

  • Frankel, R., Galun, E. (1977) Pollination mechanisms, reproduction and plant breeding (Monographs on theoretical and applied genetics, vol. 2). Springer, New York

    Google Scholar 

  • Grant, I., Beversdorf, W.D, Peterson, R.L. (1986) A comparative light and electron microscopic study of microspore and tapetal development in male fertile and cytoplasmic male sterile oilseed rape (Brassica napus). Can. J. Bot. 64, 1055–1068

    Google Scholar 

  • Hartley, R.W. (1989) Barnase and barstar: two small proteins to fold and fit together. Trends Biochem. Sci. 14, 450–454

    Google Scholar 

  • Jefferson, R.A. (1987) Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol. Biol. Rep. 5, 387–405

    Google Scholar 

  • Jefferson, R.A., Burgess, S.M., Hirsh, D. (1986) β-Glucuronidase from Escherichia coli as a gene-fusion marker. Proc. Natl. Acad. Sci. USA 83, 8447–8451

    Google Scholar 

  • Kaul, M.L.K. (1988) Male sterility in higher plants (Monographs on theoretical and applied genetics, vol. 10). Springer, Berlin Heidelberg New York

    Google Scholar 

  • Koltunow, A.M., Truettner, J., Cox, K.H., Wallroth, M., Goldberg, R.B. (1990) Different temporal and spatial gene expression patterns occur during anther development. Plant Cell 2, 1201–1224

    CAS  PubMed  Google Scholar 

  • Mariani, C., De Beuckeleer, M., Truettner, J., Leemans, J., Goldberg, R.B. (1990) Induction of male sterility in plants by a chimaeric ribonuclease gene. Nature 347, 737–741

    Article  CAS  Google Scholar 

  • Mariani, C., Gossele, V., De Beuckeleer, M., De Block, M., Goldberg, R.B., De Greef, W., Leemans, J. (1992) A chimeric RNase inhibitor gene restores fertility to male sterile plants. Nature 357, 384–387

    Article  CAS  Google Scholar 

  • Nave, E.B., Sawhney, V.K. (1986) Enzymatic changes in postmeiotic anther development in Petunia hybrida. I. Anther ontogeny and isozyme analyses. J. Plant Physiol. 125, 451–465

    Google Scholar 

  • Sawhney, V.K., Nave, E.B. (1986) Enzymatic changes in postmeiotic anther development in Petunia hybrida. II. Histochemical localization of esterase, peroxidase, malate- and alcohol dehydrogenase. J. Plant Physiol. 125, 467–473

    Google Scholar 

  • Smith, L.J., Scarisbrick, D.H. (1990) Reproductive development in oilseed rape (Brassica napus cv. Bienvenu). Ann. Bot. 65, 205–212

    Google Scholar 

  • Theis, R., Röbbelen, G. (1990) Anther and microspore development in different male sterile lines of oilseed rape (Brassica napus). Angew. Botanik 64, 419–434

    Google Scholar 

  • Van Loon, L.C. (1976) Systemic acquired resistance, peroxidase activity and lesion size in tobacco reacting hypersensitively to tobacco mosaic virus. Physiol. Plant Pathol. 8, 231–237

    Google Scholar 

  • Vidhyasekaran, P., ed. (1988) Physiology of disease resistance in plants, vol. 2. CRC Press, Boca Raton, Florida

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

We thank Dr. E. Krebbers (Plant Genetic Systems, Gent, Belgium) for the critical reading of the manuscript.

Rights and permissions

Reprints and permissions

About this article

Cite this article

De Block, M., Debrouwer, D. Engineered fertility control in transgenic Brassica napus L.: Histochemical analysis of anther development. Planta 189, 218–225 (1993). https://doi.org/10.1007/BF00195080

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00195080

Key words

Navigation