Skip to main content
NCBI home page
Plant Physiology logoLink to Plant Physiology
. 1995 Dec;109(4):1151–1158. doi: 10.1104/pp.109.4.1151

Cell Ablation Reveals That Expression from the Phaseolin Promoter Is Confined to Embryogenesis and Microsporogenesis.

AHM Van Der Geest 1, D A Frisch 1, J D Kemp 1, T C Hall 1
PMCID: PMC157645  PMID: 12228659

Abstract

Most previous studies of the [beta]-phaseolin (phas) gene, which encodes the major storage protein in bean (Phaseolus vulgaris L.), have shown its expression to be rigorously confined to the developing seed, both in bean and transgenic tobacco (Nicotiana tabacum L. cv Xanthi) plants. To confirm unequivocally the lack of phas expression in vegetative tissues, we placed the diphtheria toxin A-chain (DT-A) coding region under the control of [beta]-phaseolin promoter sequences. Tobacco plants transgenic for phas/DT-A were phenotypically normal until flowering, when they produced anthers that were externally normal but contained no viable pollen. Microscopic examination of immature anthers revealed a normal tapetum, but the pollen mother cells did not undergo meiosis and subsequently degenerated, resulting in male-sterile plants. This demonstration of phas expression during microsporogenesis was corroborated by the expression of [beta]-glucuronidase in pollen of plants transformed with comparable phas/uidA constructs. Although these findings suggested that similarities in phas expression may exist between seed and pollen maturation, no phas activity could be detected in bean pollen. After fertilization of the DT-A-transformed plants with pollen from wild-type tobacco, 50% of the resulting embryos aborted at the heart stage, defining this as the earliest time for phas expression during embryogenesis.

Full Text

The Full Text of this article is available as a PDF (3.0 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. A simple and general method for transferring genes into plants. Science. 1985 Mar 8;227(4691):1229–1231. doi: 10.1126/science.227.4691.1229. [DOI] [PubMed] [Google Scholar]
  2. Bedinger P. A., Hardeman K. J., Loukides C. A. Travelling in style: the cell biology of pollen. Trends Cell Biol. 1994 Apr;4(4):132–138. doi: 10.1016/0962-8924(94)90068-x. [DOI] [PubMed] [Google Scholar]
  3. Bevan M. Binary Agrobacterium vectors for plant transformation. Nucleic Acids Res. 1984 Nov 26;12(22):8711–8721. doi: 10.1093/nar/12.22.8711. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bustos M. M., Begum D., Kalkan F. A., Battraw M. J., Hall T. C. Positive and negative cis-acting DNA domains are required for spatial and temporal regulation of gene expression by a seed storage protein promoter. EMBO J. 1991 Jun;10(6):1469–1479. doi: 10.1002/j.1460-2075.1991.tb07667.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bustos M. M., Guiltinan M. J., Jordano J., Begum D., Kalkan F. A., Hall T. C. Regulation of beta-glucuronidase expression in transgenic tobacco plants by an A/T-rich, cis-acting sequence found upstream of a French bean beta-phaseolin gene. Plant Cell. 1989 Sep;1(9):839–853. doi: 10.1105/tpc.1.9.839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Czako M., An G. Expression of DNA coding for diphtheria toxin chain a is toxic to plant cells. Plant Physiol. 1991 Mar;95(3):687–692. doi: 10.1104/pp.95.3.687. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Eyal Y., Curie C., McCormick S. Pollen specificity elements reside in 30 bp of the proximal promoters of two pollen-expressed genes. Plant Cell. 1995 Mar;7(3):373–384. doi: 10.1105/tpc.7.3.373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Jefferson R. A., Kavanagh T. A., Bevan M. W. GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J. 1987 Dec 20;6(13):3901–3907. doi: 10.1002/j.1460-2075.1987.tb02730.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kandasamy M. K., Thorsness M. K., Rundle S. J., Goldberg M. L., Nasrallah J. B., Nasrallah M. E. Ablation of Papillar Cell Function in Brassica Flowers Results in the Loss of Stigma Receptivity to Pollination. Plant Cell. 1993 Mar;5(3):263–275. doi: 10.1105/tpc.5.3.263. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kim Y., An G. Pollen-specific expression of the Arabidopsis thaliana alpha 1-tubulin promoter assayed by beta-glucuronidase, chloramphenicol acetyltransferase and diphtheria toxin reporter genes. Transgenic Res. 1992 Jul;1(4):188–194. doi: 10.1007/BF02522538. [DOI] [PubMed] [Google Scholar]
  11. Koltunow A. M., Truettner J., Cox K. H., Wallroth M., Goldberg R. B. Different Temporal and Spatial Gene Expression Patterns Occur during Anther Development. Plant Cell. 1990 Dec;2(12):1201–1224. doi: 10.1105/tpc.2.12.1201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Mascarenhas J. P. The Male Gametophyte of Flowering Plants. Plant Cell. 1989 Jul;1(7):657–664. doi: 10.1105/tpc.1.7.657. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Maxwell I. H., Maxwell F., Glode L. M. Regulated expression of a diphtheria toxin A-chain gene transfected into human cells: possible strategy for inducing cancer cell suicide. Cancer Res. 1986 Sep;46(9):4660–4664. [PubMed] [Google Scholar]
  14. Murai N., Kemp J. D., Sutton D. W., Murray M. G., Slightom J. L., Merlo D. J., Reichert N. A., Sengupta-Gopalan C., Stock C. A., Barker R. F., Hall T. C. Phaseolin gene from bean is expressed after transfer to sunflower via tumor-inducing plasmid vectors. Science. 1983 Nov 4;222(4623):476–482. doi: 10.1126/science.222.4623.476. [DOI] [PubMed] [Google Scholar]
  15. Mutschler M. A., Bliss F. A., Hall T. C. Variation in the Accumulation of Seed Storage Protein Among Genotypes of Phaseolus vulgaris (L.). Plant Physiol. 1980 Apr;65(4):627–630. doi: 10.1104/pp.65.4.627. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Odell J. T., Hoopes J. L., Vermerris W. Seed-specific gene activation mediated by the Cre/lox site-specific recombination system. Plant Physiol. 1994 Oct;106(2):447–458. doi: 10.1104/pp.106.2.447. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Palmiter R. D., Behringer R. R., Quaife C. J., Maxwell F., Maxwell I. H., Brinster R. L. Cell lineage ablation in transgenic mice by cell-specific expression of a toxin gene. Cell. 1987 Jul 31;50(3):435–443. doi: 10.1016/0092-8674(87)90497-1. [DOI] [PubMed] [Google Scholar]
  18. Quattrocchio F., Tolk M. A., Coraggio I., Mol J. N., Viotti A., Koes R. E. The maize zein gene zE19 contains two distinct promoters which are independently activated in endosperm and anthers of transgenic Petunia plants. Plant Mol Biol. 1990 Jul;15(1):81–93. doi: 10.1007/BF00017726. [DOI] [PubMed] [Google Scholar]
  19. Uknes S., Dincher S., Friedrich L., Negrotto D., Williams S., Thompson-Taylor H., Potter S., Ward E., Ryals J. Regulation of pathogenesis-related protein-1a gene expression in tobacco. Plant Cell. 1993 Feb;5(2):159–169. doi: 10.1105/tpc.5.2.159. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Plant Physiology are provided here courtesy of Oxford University Press

RESOURCES