Skip to main content
The Plant Cell logoLink to The Plant Cell
. 1999 Feb;11(2):177–189. doi: 10.1105/tpc.11.2.177

Antisense repression of vacuolar and cell wall invertase in transgenic carrot alters early plant development and sucrose partitioning.

G Q Tang 1, M Lüscher 1, A Sturm 1
PMCID: PMC144160  PMID: 9927637

Abstract

To unravel the functions of cell wall and vacuolar invertases in carrot, we used an antisense technique to generate transgenic carrot plants with reduced enzyme activity. Phenotypic alterations appeared at very early stages of development; indeed, the morphology of cotyledon-stage embryos was markedly changed. At the stage at which control plantlets had two to three leaves and one primary root, shoots of transgenic plantlets did not separate into individual leaves but consisted of stunted, interconnected green structures. When transgenic plantlets were grown on media containing a mixture of sucrose, glucose, and fructose rather than sucrose alone, the malformation was alleviated, and plantlets looked normal. Plantlets from hexose-containing media produced mature plants when transferred to soil. Plants expressing antisense mRNA for cell wall invertase had a bushy appearance due to the development of extra leaves, which accumulated elevated levels of sucrose and starch. Simultaneously, tap root development was markedly reduced, and the resulting smaller organs contained lower levels of carbohydrates. Compared with control plants, the dry weight leaf-to-root ratio of cell wall invertase antisense plants was shifted from 1:3 to 17:1. Plants expressing antisense mRNA for vacuolar invertase also had more leaves than did control plants, but tap roots developed normally, although they were smaller, and the leaf-to-root ratio was 1.5:1. Again, the carbohydrate content of leaves was elevated, and that of roots was reduced. Our data suggest that acid invertases play an important role in early plant development, most likely via control of sugar composition and metabolic fluxes. Later in plant development, both isoenzymes seem to have important functions in sucrose partitioning.

Full Text

The Full Text of this article is available as a PDF (782.4 KB).

Selected References

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

  1. Benhamou N., Grenier J., Chrispeels M. J. Accumulation of beta-Fructosidase in the Cell Walls of Tomato Roots following Infection by a Fungal Wilt Pathogen. Plant Physiol. 1991 Oct;97(2):739–750. doi: 10.1104/pp.97.2.739. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Dijkwel P. P., Huijser C., Weisbeek P. J., Chua N. H., Smeekens S. C. Sucrose control of phytochrome A signaling in Arabidopsis. Plant Cell. 1997 Apr;9(4):583–595. doi: 10.1105/tpc.9.4.583. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Herbers K., Meuwly P., Frommer W. B., Metraux J. P., Sonnewald U. Systemic Acquired Resistance Mediated by the Ectopic Expression of Invertase: Possible Hexose Sensing in the Secretory Pathway. Plant Cell. 1996 May;8(5):793–803. doi: 10.1105/tpc.8.5.793. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Höfgen R., Willmitzer L. Storage of competent cells for Agrobacterium transformation. Nucleic Acids Res. 1988 Oct 25;16(20):9877–9877. doi: 10.1093/nar/16.20.9877. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Jang J. C., León P., Zhou L., Sheen J. Hexokinase as a sugar sensor in higher plants. Plant Cell. 1997 Jan;9(1):5–19. doi: 10.1105/tpc.9.1.5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Klann E. M., Chetelat R. T., Bennett A. B. Expression of Acid Invertase Gene Controls Sugar Composition in Tomato (Lycopersicon) Fruit. Plant Physiol. 1993 Nov;103(3):863–870. doi: 10.1104/pp.103.3.863. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Klann E. M., Hall B., Bennett A. B. Antisense acid invertase (TIV1) gene alters soluble sugar composition and size in transgenic tomato fruit. Plant Physiol. 1996 Nov;112(3):1321–1330. doi: 10.1104/pp.112.3.1321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Koch K. E. CARBOHYDRATE-MODULATED GENE EXPRESSION IN PLANTS. Annu Rev Plant Physiol Plant Mol Biol. 1996 Jun;47(NaN):509–540. doi: 10.1146/annurev.arplant.47.1.509. [DOI] [PubMed] [Google Scholar]
  9. Laurière C., Laurière M., Sturm A., Faye L., Chrispeels M. J. Characterization of beta-fructosidase, an extracellular glycoprotein of carrot cells. Biochimie. 1988 Nov;70(11):1483–1491. doi: 10.1016/0300-9084(88)90285-4. [DOI] [PubMed] [Google Scholar]
  10. Lorenz K., Lienhard S., Sturm A. Structural organization and differential expression of carrot beta-fructofuranosidase genes: identification of a gene coding for a flower bud-specific isozyme. Plant Mol Biol. 1995 Apr;28(1):189–194. doi: 10.1007/BF00042049. [DOI] [PubMed] [Google Scholar]
  11. Miller M. E., Chourey P. S. The Maize Invertase-Deficient miniature-1 Seed Mutation Is Associated with Aberrant Pedicel and Endosperm Development. Plant Cell. 1992 Mar;4(3):297–305. doi: 10.1105/tpc.4.3.297. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Murray M. G., Thompson W. F. Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res. 1980 Oct 10;8(19):4321–4325. doi: 10.1093/nar/8.19.4321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ramloch-Lorenz K., Knudsen S., Sturm A. Molecular characterization of the gene for carrot cell wall beta-fructosidase. Plant J. 1993 Sep;4(3):545–554. doi: 10.1046/j.1365-313x.1993.04030545.x. [DOI] [PubMed] [Google Scholar]
  14. SMOGYI M. Notes on sugar determination. J Biol Chem. 1952 Mar;195(1):19–23. [PubMed] [Google Scholar]
  15. Soni R., Carmichael J. P., Shah Z. H., Murray J. A. A family of cyclin D homologs from plants differentially controlled by growth regulators and containing the conserved retinoblastoma protein interaction motif. Plant Cell. 1995 Jan;7(1):85–103. doi: 10.1105/tpc.7.1.85. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Sonnewald U., Brauer M., von Schaewen A., Stitt M., Willmitzer L. Transgenic tobacco plants expressing yeast-derived invertase in either the cytosol, vacuole or apoplast: a powerful tool for studying sucrose metabolism and sink/source interactions. Plant J. 1991 Jul;1(1):95–106. doi: 10.1111/j.1365-313x.1991.00095.x. [DOI] [PubMed] [Google Scholar]
  17. Sturm A., Chrispeels M. J. cDNA cloning of carrot extracellular beta-fructosidase and its expression in response to wounding and bacterial infection. Plant Cell. 1990 Nov;2(11):1107–1119. doi: 10.1105/tpc.2.11.1107. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Unger C., Hardegger M., Lienhard S., Sturm A. cDNA cloning of carrot (Daucus carota) soluble acid beta-fructofuranosidases and comparison with the cell wall isoenzyme. Plant Physiol. 1994 Apr;104(4):1351–1357. doi: 10.1104/pp.104.4.1351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Unger C., Hofsteenge J., Sturm A. Purification and characterization of a soluble beta-fructofuranosidase from Daucus carota. Eur J Biochem. 1992 Mar 1;204(2):915–921. doi: 10.1111/j.1432-1033.1992.tb16712.x. [DOI] [PubMed] [Google Scholar]
  20. Weber H., Borisjuk L., Heim U., Buchner P., Wobus U. Seed coat-associated invertases of fava bean control both unloading and storage functions: cloning of cDNAs and cell type-specific expression. Plant Cell. 1995 Nov;7(11):1835–1846. doi: 10.1105/tpc.7.11.1835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Wyse R. E., Zamski E., Tomos A. D. Turgor regulation of sucrose transport in sugar beet taproot tissue. Plant Physiol. 1986 Jun;81(2):478–481. doi: 10.1104/pp.81.2.478. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Zrenner R., Schüler K., Sonnewald U. Soluble acid invertase determines the hexose-to-sucrose ratio in cold-stored potato tubers. Planta. 1996;198(2):246–252. doi: 10.1007/BF00206250. [DOI] [PubMed] [Google Scholar]

Articles from The Plant Cell are provided here courtesy of Oxford University Press

RESOURCES