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. 1987 Sep;6(9):2551–2556. doi: 10.1002/j.1460-2075.1987.tb02543.x

Parsley protoplasts retain differential responsiveness to u.v. light and fungal elicitor

Jeffery L Dangl 1, Karl D Hauffe 1, Susanne Lipphardt 2, Klaus Hahlbrock 1, Dierk Scheel 1
PMCID: PMC553673  PMID: 16453792

Abstract

The differential response of cultured parsley cells to u.v. irradiation and elicitor treatment is a paradigm for analysis of specific plant defense responses. We demonstrate that freshly isolated parsley protoplasts, in the absence of detectable cell wall, maintain fully the ability to be activated by these important environmental factors. Stimulated protoplasts synthesize typical qualitative patterns and amounts of potentially protective flavonoid glycosides and coumarin phytoalexins following either u.v. irradiation or treatment with fungal elicitor, respectively. Induced accumulation of mRNAs and enzymes of the phenylpropanoid biosynthetic pathways is nearly identical in protoplasts and cells. Stimulation of protoplasts with elicitor requires only a short period of contact, which is not sufficient for cell wall regeneration. Importantly, there is no activation of these pathways during protoplast preparation. These results establish that parsley protoplasts respond appropriately to two physically distinct stimuli and might serve as an especially suitable system for the analysis of signal transduction and gene activation.

Keywords: coumarins, elicitor, flavonoids, protoplasts, u. v. light

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Selected References

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

  1. Ayers A. R., Ebel J., Valent B., Albersheim P. Host-Pathogen Interactions: X. Fractionation and Biological Activity of an Elicitor Isolated from the Mycelial Walls of Phytophthora megasperma var. sojae. Plant Physiol. 1976 May;57(5):760–765. doi: 10.1104/pp.57.5.760. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  3. Burnette W. N. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 1981 Apr;112(2):195–203. doi: 10.1016/0003-2697(81)90281-5. [DOI] [PubMed] [Google Scholar]
  4. Davis K. R., Lyon G. D., Darvill A. G., Albersheim P. Host-Pathogen Interactions : XXV. Endopolygalacturonic Acid Lyase from Erwinia carotovora Elicits Phytoalexin Accumulation by Releasing Plant Cell Wall Fragments. Plant Physiol. 1984 Jan;74(1):52–60. doi: 10.1104/pp.74.1.52. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Douglas C., Hoffmann H., Schulz W., Hahlbrock K. Structure and elicitor or u.v.-light-stimulated expression of two 4-coumarate:CoA ligase genes in parsley. EMBO J. 1987 May;6(5):1189–1195. doi: 10.1002/j.1460-2075.1987.tb02353.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fromm M. E., Taylor L. P., Walbot V. Stable transformation of maize after gene transfer by electroporation. 1986 Feb 27-Mar 5Nature. 319(6056):791–793. doi: 10.1038/319791a0. [DOI] [PubMed] [Google Scholar]
  7. Fromm M., Taylor L. P., Walbot V. Expression of genes transferred into monocot and dicot plant cells by electroporation. Proc Natl Acad Sci U S A. 1985 Sep;82(17):5824–5828. doi: 10.1073/pnas.82.17.5824. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gamborg O. L., Miller R. A., Ojima K. Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res. 1968 Apr;50(1):151–158. doi: 10.1016/0014-4827(68)90403-5. [DOI] [PubMed] [Google Scholar]
  9. Hahlbrock K., Lamb C. J., Purwin C., Ebel J., Fautz E., Schäfer E. Rapid Response of Suspension-cultured Parsley Cells to the Elicitor from Phytophthora megasperma var. sojae: INDUCTION OF THE ENZYMES OF GENERAL PHENYLPROPANOID METABOLISM. Plant Physiol. 1981 Apr;67(4):768–773. doi: 10.1104/pp.67.4.768. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Jin D. F., West C. A. Characteristics of galacturonic Acid oligomers as elicitors of casbene synthetase activity in castor bean seedlings. Plant Physiol. 1984 Apr;74(4):989–992. doi: 10.1104/pp.74.4.989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kombrink E., Hahlbrock K. Responses of cultured parsley cells to elicitors from phytopathogenic fungi : timing and dose dependency of elicitor-induced reactions. Plant Physiol. 1986 May;81(1):216–221. doi: 10.1104/pp.81.1.216. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kreuzaler F., Ragg H., Fautz E., Kuhn D. N., Hahlbrock K. UV-induction of chalcone synthase mRNA in cell suspension cultures of Petroselinum hortense. Proc Natl Acad Sci U S A. 1983 May;80(9):2591–2593. doi: 10.1073/pnas.80.9.2591. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kuhn D. N., Chappell J., Boudet A., Hahlbrock K. Induction of phenylalanine ammonia-lyase and 4-coumarate:CoA ligase mRNAs in cultured plant cells by UV light or fungal elicitor. Proc Natl Acad Sci U S A. 1984 Feb;81(4):1102–1106. doi: 10.1073/pnas.81.4.1102. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Matern U., Heller W., Himmelspach K. Conformational changes of apigenin 7-O-(6-O-malonylglucoside), a vacuolar pigment from parsley, with solvent composition and proton concentration. Eur J Biochem. 1983 Jun 15;133(2):439–448. doi: 10.1111/j.1432-1033.1983.tb07483.x. [DOI] [PubMed] [Google Scholar]
  15. Muilerman H. G., ter Hart H. G., Van Dijk W. Specific detection of inactive enzyme protein after polyacrylamide gel electrophoresis by a new enzyme-immunoassay method using unspecific antiserum and partially purified active enzyme: application to rat liver phosphodiesterase I. Anal Biochem. 1982 Feb;120(1):46–51. doi: 10.1016/0003-2697(82)90315-3. [DOI] [PubMed] [Google Scholar]
  16. Paszkowski J., Shillito R. D., Saul M., Mandák V., Hohn T., Hohn B., Potrykus I. Direct gene transfer to plants. EMBO J. 1984 Dec 1;3(12):2717–2722. doi: 10.1002/j.1460-2075.1984.tb02201.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Ragg H., Kuhn D. N., Hahlbrock K. Coordinated regulation of 4-coumarate:CoA ligase and phenylalanine ammonia-lyase mRNAs in cultured plant cells. J Biol Chem. 1981 Oct 10;256(19):10061–10065. [PubMed] [Google Scholar]
  18. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  19. Schröder J., Kreuzaler F., Schäfer E., Hahlbrock K. Concomitant induction of phenylalanine ammonia-lyase and flavanone synthase mRNAs in irradiated plant cells. J Biol Chem. 1979 Jan 10;254(1):57–65. [PubMed] [Google Scholar]
  20. Somssich I. E., Schmelzer E., Bollmann J., Hahlbrock K. Rapid activation by fungal elicitor of genes encoding "pathogenesis-related" proteins in cultured parsley cells. Proc Natl Acad Sci U S A. 1986 Apr;83(8):2427–2430. doi: 10.1073/pnas.83.8.2427. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Tietjen K. G., Hunkler D., Matern U. Differential response of cultured parsley cells to elicitors from two non-pathogenic strains of fungi. 1. Identification of induced products as coumarin derivatives. Eur J Biochem. 1983 Mar 15;131(2):401–407. doi: 10.1111/j.1432-1033.1983.tb07277.x. [DOI] [PubMed] [Google Scholar]
  22. Tietjen K. G., Matern U. Differential response of cultured parsley cells to elicitors from two non-pathogenic strains of fungi. 2. Effects on enzyme activities. Eur J Biochem. 1983 Mar 15;131(2):409–413. doi: 10.1111/j.1432-1033.1983.tb07278.x. [DOI] [PubMed] [Google Scholar]
  23. Wendorff H., Matern U. Differential response of cultured parsley cells to elicitors from two non-pathogenic strains of fungi. Microsomal conversion of (+)marmesin into psoralen. Eur J Biochem. 1986 Dec 1;161(2):391–398. doi: 10.1111/j.1432-1033.1986.tb10458.x. [DOI] [PubMed] [Google Scholar]
  24. Yoshikawa M., Keen N. T., Wang M. C. A receptor on soybean membranes for a fungal elicitor of phytoalexin accumulation. Plant Physiol. 1983 Oct;73(2):497–506. doi: 10.1104/pp.73.2.497. [DOI] [PMC free article] [PubMed] [Google Scholar]

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