Transgenic Research

, Volume 2, Issue 1, pp 21–28 | Cite as

Apical and lateral shoot apex-specific expression is conferred by promoter of the seed storage protein β-phaseolin gene

  • Partha Sen
  • Caryl A. Chlan
  • Mark D. Burow
  • Woo Sung Lee
  • Norimoto Murai
Article

Abstract

A previous analysis with deletion mutants of the native β-phaseolin gene demonstrated that removal of a negative element 5′ upstream of−107 permitted phaseolin expression in stem cortex and secondary root (Burowet al., 1992). Here we employed the β-glucuronidase (GUS) reporter gene to visualize, by histochemical staining, the cell type-specificity of phaseolin expression in stem and root, and to understand further the spatial control of the β-phaseolin gene. The 782 bp 5′ upstream promoter and its deletion mutants were fused to the GUS gene, and these chimaeric genes were used to transform tobacco. Histochemical staining for GUS activity demonstrated that phaseolin promoters truncated downstream of −227 conferred cell-type specific expression in internal/external phloem and protoxylem of mature stem. Surprisingly, GUS staining was prominent in both apical and lateral shoot apices of plants that contain the full-length −782 promoter and mutant promoters deleted up to −64. GUS expression was extended to all cell types of shoot tips, including epidermis, cortex, vasculature, procambium and pith. Expression in vasculature of petioles was limited to plants with promoters truncated to −106 and −64. The current results are in agreement with our previous findings with the native phaseolin gene: that the major positive element (−295/−228) is sufficient for seed-specific late-temporal expression of the phaseolin gene. We conclude that the 5′ upstream sequence of the β-phaseolin gene directs spatially- and temporally-controlled gene expression in developing seeds during the reproductive phase, but also confers expression in shoot apices during the vegetative phase of plant development.

Keywords

5′-deletion β-glucuronidase histochemical staining phloem xylem 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Anthony, J.L. and Hall, T.C. (1991) Nucleotide sequence of an α-phaseolin gene fromPhaseolus vulgaris.Nucl. Acids Res. 18, 3396.Google Scholar
  2. Bliss, F.A. and Brown, J.W.D. (1982) Genetic control of phaseolin protein expression in seeds of common bean,Phaseolus vulgaris L.Qual. Pl. Foods Hum. Nutr. 31, 269–79.CrossRefGoogle Scholar
  3. Burow, M.D., Chlan, C.A., Sen, P., Lisca, A. and Murai, N. (1990) High-frequency generation of transgenic tobacco plants after modified leak disk cocultivation withAgrobacterium tumefaciens.Pl. Mol. Biol. Rep. 8, 123–38.Google Scholar
  4. Burow, M.D., Sen, P., Chlan, C.A. and Murai, N. (1992) Developmental control of the β-phaseolin gene requires positive, negative, and temporal seed-specific transcriptional regulatory elements, and a negative element for stem and root expression.Pl. J. 2, 537–48.CrossRefGoogle Scholar
  5. Bustos, M.M., Begum, D., Kalkan, F.A., Battrew, M.J. and Hall, T.C. (1991) Positive and negativecis-acting DNA domains are required for spatial and temporal regulation of gene expression by a seed storage promoter.EMBO J. 10, 1469–79.PubMedGoogle Scholar
  6. Bustos, M.M., Guiltinan, M.J., Jordano, J., Begum, D., Kalkan, F.A. and Hall, T.C. (1989) Regulation of β-glucuronidase expression in transgenic tobacco plants by an A/T rich,cis-acting sequence found upstream of a French bean β-phaseolin gene.Pl. Cell 1, 839–53.CrossRefGoogle Scholar
  7. Douglas, C.J., Hauffe, K.D., Ites-Morales, M.-E., Ellard, M., Paszkowski, U., Halbrick, K. and Dangl, J.L. (1991) Exonic sequences are required for elicitor and light activation of a plant defense gene, but promoter sequences are sufficient for tissue specific expression.EMBO J. 10, 1767–75.PubMedGoogle Scholar
  8. Edwards, J.W., Walker, E.L. and Coruzzi, G.M. (1990) Cellspecific expression in transgenic plants reveals non-overlapping roles for chloroplast and cytosolic glutamine synthetase.Proc. Natl Acad. Sci. USA 87, 3459–63.PubMedGoogle Scholar
  9. Greenwood, J.S. and Chrispeels, M.J. (1985) Correct targeting of bean storage protein phaseolin in the seeds of transformed tobacco.Pl. Physiol. 79, 65–71.Google Scholar
  10. Hauffe, K.D., Paszkowski, U., Schulze-Lefert, P., Hahlbrock, K., Dangl, J.L. and Douglas, C.J. (1991) A parsley 4CL-1 promoter fragment specifies complex expression patterns in transgenic tobacco.Pl. Cell 3, 435–43.CrossRefGoogle Scholar
  11. Jefferson, R.A. (1987) Assaying chimeric genes in plants: The GUS gene fusion system.Pl. Mol. Biol. Rep. 5, 387–405.Google Scholar
  12. Keller, B. Templeton, M.D. and Lamb, C.J. (1989) Specific localization of a plant cell wall glycine-rich protein in protoxylem cells of the vascular system.Proc. Natl Acad. Sci. USA 86, 1529–33.Google Scholar
  13. Medford, J.I. Elmer, J.S. and Klee, H. (1991) Molecular cloning and characterization of genes expressed in shoot apical meristem.Pl. Cell 3, 359–70.CrossRefGoogle Scholar
  14. Murai, N., Sutton, D.W., Slightom, J.L., Merlo, D.J., Reichert, N.A., Sengupta-Gopalan, C., Stock, C.A., Barkar, R.J., Kemp, J.D. and Hall, T.C. (1983) Phaseolin gene from bean is expressed after transfer to sunflower via tumor inducing plasmid vectors.Science 222, 476–82.Google Scholar
  15. Murray, M.G. and Kennard, W.C. (1984) Altered chromatin conformation of the higher plant gene phaseolin.Biochemistry 23, 4225–32.CrossRefGoogle Scholar
  16. Mutschler, M.A., Bliss, F.A. and Hall, T.C. (1980) Variation in the accumulation of seed storage protein among genotypes ofPhaseolus vulgaris.Pl. Physiol. 65, 627–30.Google Scholar
  17. Peleman, J., Boerjan, W., Engler, G., Seurinck, J., Botterman, J., Alliotte, T., Van Montagu, M. and Inze, D. (1989) Strong cellular preference in the expression of a housekeeping gene ofArabidopsis thaliana encodingS-adenosylmethionine synthetase.Pl. Cell 1, 81–93.CrossRefGoogle Scholar
  18. Sengupta-Gopalan, C., Reichert, N.A., Barker, R.F., Hall, T.C. and Kemp, J.D. (1985) Developmentally-regulated expression of the bean β-phaseolin gene in tobacco seed.Proc. Natl Acad. Sci. USA 82, 3320–4.Google Scholar
  19. Slightom, J.L., Sun, S.M. and Hall, T.C. (1983) Complete nucleotide sequence of a French bean storage protein gene: Phaseolin.Proc. Natl Acad. Sci. USA 80, 1897–901.Google Scholar
  20. Sun, S.M., Mutschler, M.A., Bliss, F.A. and Hall, T.C. (1978) Protein synthesis and accumulation in bean cotyledons during growth.Pl. Physiol. 61, 918–23.Google Scholar
  21. Walbot, V., Clutter, M. and Sussex, I.M. (1972) Reproductive development and embryogeny inPhaseolus.Develop. Biol. 29, 59–68.CrossRefGoogle Scholar
  22. Young, N.S. and Russell, D. (199) Maize sucrose synthase-1 promoter directs phloem-specific expression ofGus gene in transgenic tobacco plants.Proc. Natl Acad. Sci. USA 87, 4144–8.Google Scholar

Copyright information

© Chapman & Hall 1993

Authors and Affiliations

  • Partha Sen
    • 1
  • Caryl A. Chlan
    • 1
  • Mark D. Burow
    • 1
  • Woo Sung Lee
    • 1
  • Norimoto Murai
    • 1
  1. 1.Department of Plant Pathology and Crop Physiology, College of AgricultureLouisiana State UniversityBaton RougeUSA

Personalised recommendations