Abstract
A nuclear factor (SBF-1) has previously been identified in Phaseolus vulgaris L. (bean) suspension cell nuclear extracts that binds in vitro to three DNase I-footprinted elements (SBF-1 boxes I, II, and III, 5′ to 3′) in the 5′ region of the bean CHS15 (chalcone synthase) gene promoter. To define the functional role of the three SBF-1 boxes in development, we examined transgenic tobacco plants carrying a series of nested CHS15 promoter-β-glucuronidase (GUS) fusions for GUS activity by histochemical staining. We show that the CHS15 promoter deleted to position-173 and lacking all three SBF-1 boxes directs the same qualitative pattern of expression in initiating lateral roots and in developing seeds as the full length promoter (-326). Thus, activation of expression in these organs is mediated by sequence elements located downstream of the three SBF-1 boxes. However, specific deletions within the-326 to-173 region modulate expression. Thus, deletion of box II abolishes GUS activity in initiating lateral roots. Further deletion of box III fails to restore expression but subsequent deletion of an additional 43 bp to position-173 re-establishes expression. We show that sequence-specific DNA-binding activities consistent with these results are present in nuclear extracts of bean roots and seeds. These studies reveal cis elements within the CHS15 promoter, and potential trans factors, that permit organ- and tissue-specific developmental patterns of regulation to be combined with a flexible response to environmental cues.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
An G: Binary Ti vectors for plant transformation and promoter analysis. Meth Enzymol 153: 292–305 (1987).
Choudhary AD, Kessman H, Lamb CJ, Dixon RA: Stress responses in alfalfa (Medicago sativa L.). IV. Expression of defense gene constructs in electroporated suspension cell protoplasts. Plant Cell Rep 9: 42–46 (1990).
Doerner PW, Stermer B, Schmid J, Dixon RA, Lamb CJ: Plant defense gene promoter-reporter gene fusions in transgenic plants: tools for identification of novel inducers. Bio technology 8: 845–848 (1990).
Donald RGK, Cashmore AR: Mutation of either G or I box sequences profoundly affects expression from the Arabidopsis rbcS-1A promoter. EMBO J 9: 1717–1726 (1990).
Dron M, Clouse SD, Dixon RA, Lawton MA, Lamb CJ: Glutathione and fungal elicitor regulation of a plant defense gene promoter in electroporated protoplasts. Proc Natl Acad Sci USA 85: 6738–6742 (1988).
Firman JL, Wilson KE, Rossen L, Johnston AWB: Flavonoid activation of nodulation genes in Rhizobium reversed by other compounds present in plants. Nature 324: 90–92 (1986).
Fritze K, Staiger D, Czaja I, Walden R, Schell J, Wing D: Developmental and UV light induction of the snapdragon chalcone synthase promoter. Plant Cell 3: 893–905 (1991).
Green PJ, Yong M-M, Cuozzo M, Kano-Murakami Y, Silverstein P, Chua N-H: Binding site requirements for pea nuclear protein factor GT-1 correlate with sequences required for light-dependent transcriptional activation of the rbcS-3A gene. EMBO J 7: 4035–4044 (1988).
Hahlbrock K, Scheel D: Physiology and molecular biology of phenylpropanoid metabolism. Annu Rev Plant Physiol Mol Biol 40: 347–369 (1989).
Harrison MJ, Choudhary AD, Dubery I, Lamb CJ, Dixon RA: Stress responses in alfalfa (Medicago sativa L.). 8. Cis-elements and trans-acting factors for the quantitative expression of a bean chalcone synthase gene promoter in electroporated alfalfa protoplasts. Plant Mol Biol 16: 877–890 (1991).
Harrison MJ, Lawton MA, Lamb CJ, Dixon RA: Characterization of a nuclear protein that binds to three elements within the silencer region of a bean chalcone synthase promoter. Proc Natl Acad Sci USA 88: 2515–2519 (1991).
Herrera-Estrella L, Simpson J: Foreign gene expression in plants: In: Shaw CH (ed) Plant Molecular Biology: A Practical Approach, pp. 131–160, IRL Press, Oxford (1988).
Jacobs M, Rubery PH: Naturally occurring auxin transport regulators. Science 241: 346–349 (1988).
Jefferson RA: Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol Biol Rep 5: 387–405 (1987).
Jefferson RA, Kavanagh TA, Bevan MW: GUS fusions: β-glucuronidase as a sensitive and versatile gene marker in higher plants. EMBO J 6: 3901–3907 (1987).
Koes RE, vanBlokland R, Quattrocchio F, vanTunen AJ, Mol JNM: Chalcone synthase promoters in petunia are active in pigmented and unpigmented cell types. Plant Cell 2: 379–392 (1990).
Lawton MA, Dean SM, Dron M, Kooter JM, Kragh K, Harrison MJ, Yu L, Tanguay L, Dixon RA, Lamb CJ: Silencer region of a chalcone synthase promoter contains multiple binding sites for a factor, SBF-1, closely related to GT-1. Plant Mol Biol 16: 235–249 (1991).
Leyva A, Liang X, Pintor-Toro JA, Dixon RA, Lamb CJ: Cis-element combinations determine phenylalanine ammonia-lyase gene tissue-specific expression patterns. Plant Cell 4: 263–271 (1992).
Liu C-M, Xu Z-H, Chua N-H: Auxin polar transport is essential for the establishment of bilateral symmetry during early plant embryogenesis. Plant Cell 5: 621–630 (1993).
Loake GJ, Choudhary AD, Harrison MJ, Mavandad M, Lamb CJ, Dixon RA: Phenylpropanoid pathway intermediates regulate transient expression of a chalcone synthase gene promoter. Plant Cell 3: 829–840 (1991).
Loake GJ, Factor O, Lamb CJ, Dixon RA: Combination of H-box [CCTACC(N)7CT] and G-box (CACGTG) cis elements is necessary for feed-forward stimulation of a chalcone synthase promoter by the phenylpropanoid-pathway intermediate p-coumaric acid. Proc Natl Acad Sci USA 89: 9230–9234 (1992).
Peters NK, Frost JW, Long SR: A plant flavone, luteolin, induces expression of Rhizobium meliloti nodulation genes. Science 233: 977–980 (1986).
Redmond JW, Batley M, Djordjevic MA, Ines RW, Krempel PL, Rolfe BG: Flavones induce expression of nodulation genes in Rhizobium. Nature 323: 632–635 (1986).
Rogers SG, Horsch RB, Fraley RT: Gene transfer in plants: Production of transformed plants using Ti plasmid vectors. Meth Enzymol 118: 627–640 (1986).
Ryder TB, Hedrick SA, Bell JN, Liang X, Clouse SD, Lamb CJ: Organization and differential activation of a gene family encoding the plant defense enzyme chalcone synthase in Phaseolus vulgaris. Mol Gen Genet 210: 219–223 (1987).
Sambrook J, Fritsch EF, Maniatis T: Molecular Cloning: A Laboratory Manual. Cold Spring Harbour Laboratory Press, Cold Spring Harbor, NY (1989).
Sanger F, Nicklen S, Coulson AR: DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467 (1977).
Schmid J, Doerner PW, Clouse SD, Dixon RA, Lamb CJ: Developmental and environmental regulation of a bean chalcone synthase promoter in transgenic tobacco. Plant Cell 2: 619–631 (1990).
Staiger D, Becker F, Schell J, Koncz C, Palme K: Purification of tobacco nuclear protein binding to a CACGTG motif of the chalcone synthase promoter by DNA affinity chromatography. Eur J Biochem 199: 519–527 (1991).
Stermer BA, Schmid J, Lamb CJ, Dixon RA: Infection and stress activation of bean chalcone synthase promoters in transgenic tobacco. Mol Plant-Microbe Interact 3: 381–388 (1990).
Yang W-C, Canter Cremers HCJ, Hogendijk P, Katinakis P, Wijffelman CA, Franssen H, vanKammen A, Bisseling T: In-situ localization of chalcone synthase mRNA in pea root nodule tissue. Plant J 2: 143–151 (1992).
Yu LM, Lamb CJ, Dixon RA: Purification and biochemical characterization of proteins which bind to the H-box cis-element implicated in transcriptional activation of plant defense genes. Plant J 3: 805–816 (1993).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Hotter, G.S., Kooter, J., Dubery, I.A. et al. Cis elements and potential trans-acting factors for the developmental regulation of the Phaseolus vulgaris CHS15 promoter. Plant Mol Biol 28, 967–981 (1995). https://doi.org/10.1007/BF00032660
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00032660