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High intensity and blue light regulated expression of chimeric chalcone synthase genes in transgenic Arabidopsis thaliana plants

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Summary

To establish a genetic system for dissection of light-mediated signal transduction in plants, we analyzed the light wavelengths and promoter sequences responsible for the light-induced expression of the Arabidopsis thaliana chalcone synthase (CHS) promoter fused to the β-glucuronidase (GUS) marker gene. Transgenic A. thaliana lines carrying 1975, 523, 186, and 17 by of the CHS promoter fused to the GUS gene were generated, and the expression of these chimeric genes was monitored in response to high intensity light in mature plants and to different wavelengths of light in seedlings. Fusion constructs containing 1975 and 523 by of CHS promoter sequence behaved identically to the endogenous CHS gene under all conditions. Expression of these constructs was induced specifically in response to high intensity white light and blue light. The response to blue light was seen in the presence of the Pfr form of phytochrome. Fusion constructs containing 186 by of promoter sequence showed reduced basal levels of expression and only weak stimulation by blue light but were induced significantly by high intensity white light. These analyses showed that the expression of the A. thaliana CHS gene is responsive to a specific blue light receptor and that sequences between — 523 and — 186 by are required for optimal basal and blue light-induced expression of this gene. The experiments lay the foundation for a simple genetic screen for light response mutants.

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References

  • An G (1987) Binary vectors for plant transformation and promoter analysis. Methods Enzymol 153:292–305

    Google Scholar 

  • Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (1989) Current Protocols in Molecular Biology. Green Publishing Associates Wiley Interscience, New York

    Google Scholar 

  • Beggs CJ, Wellmann E (1985) Analysis of light-controlled anthocyanin formation in coleoptiles of Zea mays L.: The role of UV-B, blue, red and far-red light. Photochem Photobiol 41:481–486

    Google Scholar 

  • Bevan M (1984) Binary Agrobacterium vectors for plant transformation. Nucleic Acids Res 12:8711–8721

    Google Scholar 

  • Bruns B, Hahlbrock K, Schäfer E (1986) Fluence dependence of the ultraviolet-light-induced accumulation of chalcone synthase mRNA and effects of blue and far-red light in cultured parsley cells. Planta 169:393–398

    Google Scholar 

  • Chappell J, Hahlbrock K (1984) Transcription of plant defence genes in response to UV light or fungal elicitor. Nature 311:76–78

    Google Scholar 

  • Chaudhury AM, Signer ER (1989) Relative regeneration proficiency of Arabidopsis thaliana ecotypes. Plant Cell Rep 8:368–369

    Google Scholar 

  • Chory J, Peto C, Feinbaum R, Pratt L, Ausubel FM (1989) Arabidopsis thaliana mutant that develops as a light grown plant in the absence of light. Cell 58:991–999

    Google Scholar 

  • Church GM, Gilbert W (1984) Genomic sequencing. Proc Natl Acad Sci USA 81:1991–1995

    Google Scholar 

  • Fenbaum RL, Ausubel FM (1988) Transcriptional regulation of the Arabidopsis thaliana chalcone synthase gene. Mol Cell Biol 8:1985–1992

    CAS  PubMed  Google Scholar 

  • Gamborg OL (1984) Plant cell cultures: nutrition and media. In: Vasil IK (ed) Cell Culture and Somatic Cell Genetics of Plants. Academic Press, Orlando, pp 18–16

    Google Scholar 

  • Horsch RB, Fry JE, Hoffmann NL, Eichholtz D, Rogers SG, Fraley RT (1985) A simple and general method for transferring genes to plants. Science 227:1229–1231

    CAS  Google Scholar 

  • Jefferson RA (1987) Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol Biol Rep 5:387–405

    CAS  Google Scholar 

  • Jefferson RA, Burgess SM, Hirsh D (1986) β-glucuronidase from Escherichia coli as a gene fusion marker. Proc Natl Acad Sci USA 83:8447–8451

    CAS  PubMed  Google Scholar 

  • Karlin-Neumann GA, Sun L, Tobin EM (1988) Expression of lightharvesting chlorophyll a/b-protein genes is phytochrome regulated in etiolated Arabidopsis thaliana seedlings. Plant Physiol 88:1323–1331

    Google Scholar 

  • Kaufman LS, Thompson WF, Briggs WR (1984) Different red light requirements for phytochrome-induced accumulation of cab RNA and rbcS RNA. Science 226:1447–1449

    Google Scholar 

  • Kingston RE (1989) Transcription control and differentiation: the HLH family, c-myc and C/EBP. Curr Opin Cell Biol 1:1081–1087

    Google Scholar 

  • Lipphardt S, Brettschneider R, Kreuzaler R, Schell J, Dangl JL (1988) UV-inducible transient expression in parsley protoplasts identifies regulatory cis-elements of a chimeric Antirrhinum majus chalcone synthase gene. EMBO J 7:4027–4033

    Google Scholar 

  • Ludwig SR, Habera LF, Dellaporta SL, Wessler SR (1989) Lc, a member of the maize R gene family responsible for tissuespecific anthocyanin production, encodes a protein similar to transcriptional activators and contains the myc-homology region. Proc Natl Acad Sci USA 86:7092–7096

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, New York

    Google Scholar 

  • Marrs KA, Kaufman LS (1989) Blue-light regulation of transcription for nuclear genes in pea. Proc Natl Acad Sci USA 86:4492–4495

    Google Scholar 

  • Oelmüller R, Mohr H (1985) Mode of coaction between blue/UV light and light absorbed by phytochrome in light-mediated anthocyanin formation in the milo (Sorghum vulgare Pers.) seedling. Proc Natl Acad Sci USA 82:6124–6128

    Google Scholar 

  • Ohl S, Hahlbrock K, Schafer E (1989) A stable blue-light-derived signal modulates ultraviolet-light-induced activation of the chalcone-synthase gene in cultured parlsey cells. Planta 177:228–236

    Google Scholar 

  • Ooms G, Hooykaas PJJ, van Veen RJM, van Beelen P, Regensburg-Tuink TJG, Schilperoort RA (1982) Octopine Ti-plasmid deletion mutants of Agrobacterium tumefaciens with emphasis on the right side of the T-region. Plasmid 7:15–29

    Google Scholar 

  • Rabino I, Manicinelli AL (1986) Light, temeprature, and anthocyanin production. Plant Physiol 81:992–924

    Google Scholar 

  • Schulze-Lefert P, Becker-Andre M, Schulz W, Hahlbrock K, Dangl JL (1989a) Functional architecture of the light-responsive chalcone synthase promoter from parsley. Plant Cell 1:707–714

    Google Scholar 

  • Schulze-Lefert P, Dangl JL, Becker-Andre M, Hahlbrock K, Schulz W (1989b) Inducible in vivo DNA footprints define sequences necessary for UV light activation of the parsley chalcone synthase gene. EMBO J 8:651–656

    Google Scholar 

  • Sponga F, Deitzer GF, Mancinelli AL (1986) Cryptochrome, phytochrome, and the photoregulation of anthocyanin production under blue light. Plant Physiol 82:952–955

    Google Scholar 

  • Staiger D, Kaulen H, Schell J (1989) A CACGTG motif of the Antirrhinum mains chalcone synthase promoter is recognized by an evolutionarily conserved nuclear protein. Proc Natl Acad Sci USA 86:6930–6934

    Google Scholar 

  • Taylor LP, Briggs WR (1990) Genetic regulation and photocontrol of anthocyanin accumulation in maize seedlings. Plant Cell 2:115–127

    Google Scholar 

  • van Tunen AJ, Koes RE, Spelt CE, van der Krol AR, Stuitje AR, Mol JNM (1988) Cloning of two chalcone flavonone isomerase genes from Petunia hybrida: coordinate, light-regulated, and differential expression. EMBO J 7:1257–1263

    Google Scholar 

  • Valvekens D, Van Montagu M, Van Lijsebettsens M (1988) Agrobacterium tumefaciens mediated transformation of Arabidopsis thaliana root explants by using kanamycin selection. Proc Natl Acad Sci USA 85:5536–5540

    Google Scholar 

  • Wellmann E (1971) Phytochrome-mediated flavone glycoside synthesis in cell suspension cultures of Petroselinum hortense after pre-irradiation with ultaviolet light. Planta 101:283–2886

    Google Scholar 

  • Wellmann E (1975) UV dose-dependent induction of enzymes related to flavonoid biosynthesis in cell suspension cultures of parsley. FEBS Lett 51:105–107

    Google Scholar 

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Author notes

  1. Rhonda L. Feinbaum

    Present address: Department of Cellular and Developmental Biology, Harvard University, 02139, Cambridge, MA, USA

Authors and Affiliations

  1. Department of Genetics, Harvard Medical School, 02114, Boston, Massachusetts, USA

    Rhonda L. Feinbaum, Gisela Storz & Frederick M. Ausubel

  2. Department of Molecular Biology, Massachusetts General Hospital, 02114, Boston, Massachusetts, USA

    Rhonda L. Feinbaum, Gisela Storz & Frederick M. Ausubel

Authors
  1. Rhonda L. Feinbaum
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  2. Gisela Storz
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  3. Frederick M. Ausubel
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Communicated by E. Meyerowitz

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Feinbaum, R.L., Storz, G. & Ausubel, F.M. High intensity and blue light regulated expression of chimeric chalcone synthase genes in transgenic Arabidopsis thaliana plants. Molec. Gen. Genet. 226, 449–456 (1991). https://doi.org/10.1007/BF00260658

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