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. 1997 Aug 1;16(15):4489–4496. doi: 10.1093/emboj/16.15.4489

Seed-specific immunomodulation of abscisic acid activity induces a developmental switch.

J Phillips 1, O Artsaenko 1, U Fiedler 1, C Horstmann 1, H P Mock 1, K Müntz 1, U Conrad 1
PMCID: PMC1170075  PMID: 9303293

Abstract

A single-chain Fv antibody (scFv) gene, which has previously been used to immunomodulate abscisic acid (ABA) activity in transgenic tobacco to create a 'wilty' phenotype, was put under control of the seed-specific USP promoter from Vicia faba and used to transform tobacco. Transformants were phenotypically similar to wild-type plants apart from their seeds. Anti-ABA scFv embryo development differed markedly from wild-type embryo development. Seeds which accumulated similar levels of a scFv that binds to oxazolone, a hapten absent from plants, developed like wild-type embryos. Anti-ABA scFv embryos developed green cotyledons containing chloroplasts and accumulated photosynthetic pigments but produced less seed storage protein and oil bodies. Anti-ABA scFv seeds germinated precociously if removed from seed capsules during development but were incapable of germination after drying. Total ABA levels were higher than in wild-type seeds but calculated free ABA levels were near-zero until 21 days after pollination. We show for the first time seed-specific immunomodulation and the resulting switch from the seed maturation programme to a germination programme. We conclude that the immunomodulation of hormones can alter the development programme of target organs, allowing the study of the directly blocked endogenous molecules and manipulation of the system concerned.

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Selected References

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  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. Artsaenko O., Peisker M., zur Nieden U., Fiedler U., Weiler E. W., Müntz K., Conrad U. Expression of a single-chain Fv antibody against abscisic acid creates a wilty phenotype in transgenic tobacco. Plant J. 1995 Nov;8(5):745–750. doi: 10.1046/j.1365-313x.1995.08050745.x. [DOI] [PubMed] [Google Scholar]
  3. Beerli R. R., Wels W., Hynes N. E. Intracellular expression of single chain antibodies reverts ErbB-2 transformation. J Biol Chem. 1994 Sep 30;269(39):23931–23936. [PubMed] [Google Scholar]
  4. 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]
  5. Biocca S., Pierandrei-Amaldi P., Cattaneo A. Intracellular expression of anti-p21ras single chain Fv fragments inhibits meiotic maturation of xenopus oocytes. Biochem Biophys Res Commun. 1993 Dec 15;197(2):422–427. doi: 10.1006/bbrc.1993.2496. [DOI] [PubMed] [Google Scholar]
  6. Bäumlein H., Boerjan W., Nagy I., Bassüner R., Van Montagu M., Inzé D., Wobus U. A novel seed protein gene from Vicia faba is developmentally regulated in transgenic tobacco and Arabidopsis plants. Mol Gen Genet. 1991 Mar;225(3):459–467. doi: 10.1007/BF00261688. [DOI] [PubMed] [Google Scholar]
  7. Bäumlein H., Boerjan W., Nagy I., Panitz R., Inzé D., Wobus U. Upstream sequences regulating legumin gene expression in heterologous transgenic plants. Mol Gen Genet. 1991 Jan;225(1):121–128. doi: 10.1007/BF00282650. [DOI] [PubMed] [Google Scholar]
  8. Conrad U., Fiedler U. Expression of engineered antibodies in plant cells. Plant Mol Biol. 1994 Nov;26(4):1023–1030. doi: 10.1007/BF00040685. [DOI] [PubMed] [Google Scholar]
  9. Evan G. I., Lewis G. K., Ramsay G., Bishop J. M. Isolation of monoclonal antibodies specific for human c-myc proto-oncogene product. Mol Cell Biol. 1985 Dec;5(12):3610–3616. doi: 10.1128/mcb.5.12.3610. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fiedler U., Filistein R., Wobus U., Bäumlein H. A complex ensemble of cis-regulatory elements controls the expression of a Vicia faba non-storage seed protein gene. Plant Mol Biol. 1993 Jul;22(4):669–679. doi: 10.1007/BF00047407. [DOI] [PubMed] [Google Scholar]
  11. Giraudat J., Parcy F., Bertauche N., Gosti F., Leung J., Morris P. C., Bouvier-Durand M., Vartanian N. Current advances in abscisic acid action and signalling. Plant Mol Biol. 1994 Dec;26(5):1557–1577. doi: 10.1007/BF00016490. [DOI] [PubMed] [Google Scholar]
  12. Hiatt A., Cafferkey R., Bowdish K. Production of antibodies in transgenic plants. Nature. 1989 Nov 2;342(6245):76–78. doi: 10.1038/342076a0. [DOI] [PubMed] [Google Scholar]
  13. Koornneef M., Hanhart C. J., Hilhorst H. W., Karssen C. M. In Vivo Inhibition of Seed Development and Reserve Protein Accumulation in Recombinants of Abscisic Acid Biosynthesis and Responsiveness Mutants in Arabidopsis thaliana. Plant Physiol. 1989 Jun;90(2):463–469. doi: 10.1104/pp.90.2.463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kruse E., Mock H. P., Grimm B. Reduction of coproporphyrinogen oxidase level by antisense RNA synthesis leads to deregulated gene expression of plastid proteins and affects the oxidative defense system. EMBO J. 1995 Aug 1;14(15):3712–3720. doi: 10.1002/j.1460-2075.1995.tb00041.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  16. Marasco W. A., Haseltine W. A., Chen S. Y. Design, intracellular expression, and activity of a human anti-human immunodeficiency virus type 1 gp120 single-chain antibody. Proc Natl Acad Sci U S A. 1993 Aug 15;90(16):7889–7893. doi: 10.1073/pnas.90.16.7889. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Neri D., Montigiani S., Kirkham P. M. Biophysical methods for the determination of antibody-antigen affinities. Trends Biotechnol. 1996 Dec;14(12):465–470. doi: 10.1016/S0167-7799(96)10067-6. [DOI] [PubMed] [Google Scholar]
  18. Owen M., Gandecha A., Cockburn B., Whitelam G. Synthesis of a functional anti-phytochrome single-chain Fv protein in transgenic tobacco. Biotechnology (N Y) 1992 Jul;10(7):790–794. doi: 10.1038/nbt0792-790. [DOI] [PubMed] [Google Scholar]
  19. Richardson J. H., Sodroski J. G., Waldmann T. A., Marasco W. A. Phenotypic knockout of the high-affinity human interleukin 2 receptor by intracellular single-chain antibodies against the alpha subunit of the receptor. Proc Natl Acad Sci U S A. 1995 Apr 11;92(8):3137–3141. doi: 10.1073/pnas.92.8.3137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Töpfer R., Matzeit V., Gronenborn B., Schell J., Steinbiss H. H. A set of plant expression vectors for transcriptional and translational fusions. Nucleic Acids Res. 1987 Jul 24;15(14):5890–5890. doi: 10.1093/nar/15.14.5890. [DOI] [PMC free article] [PubMed] [Google Scholar]

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