Summary
A gene encoding chloroplast fructose-1,6-bisphosphatase (FBPase) was isolated from a genomic library of wheat DNA. Comparison of the gene sequence obtained with that of a wheat cDNA clone revealed the presence of three introns, each less than 100 bases in length. One of these introns lies in a region that may be involved in the light activation of FBPase catalytic activity. Chimeric gene constructs comprising 1673 bp of the upstream FBPase promoter region in a transcriptional fusion to the β-glucuronidase (GUS) reporter gene were used to investigate expression in transgenic tobacco plants. Histochemical localization of GUS activity revealed high levels of expression driven by the FBPase promoter sequences in photosynthetically active tissues and, unexpectedly, also in the meristematic regions of shoots, lateral buds and roots. The biological significance of FBPase expression in meristematic regions is not yet clear but this pattern of expression may be explained by the presence in the FBPase promoter of a short DNA sequence motif which is also found in the CaMV 35S viral promoter.
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References
Benfey PN, Chua N-H (1989) Regulated genes in transgenic plants. Science 244:174–181
Benfey PN, Ren L, Chua N-H (1989) The CaMV 358 enhancer contains at least two domains which can confer different developmental and tissue-specific expression patterns. EMBO J 8:2195–2202
Bevan M (1984) Binary Agrobacterium vectors for plant transformation. Nucleic Acids Res 12:8711–8721
Bouchez D, Tokuhisa JG, Llewellyn DJ, Dennis ES, Ellis JG (1989) The ocs-element is a component of the promoters of several T-DNA and plant viral genes. EMBO J 8:4197–4204
Buchanan BB (1980) Role of light in the regulation of chloroplast enzymes. Annu Rev Plant Physiol 31:341–374
Chao S, Raines CA, Longstaff M, Sharp PJ, Gale MD, Dyer TA (1989) Chromosomal location and copy number in wheat and some of its close relatives of genes for enzymes involved in photosynthesis. Mol Gen Genet 218:423–430
Datta N, Cashmore AR (1989) Binding of a pea nuclear protein to promoters of certain photoregulated genes is modulated by phosphorylation. Plant Cell 1: 1069–1077
Feinberg AP, Vogelstein B (1983) A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132:6–13
Fromm H, Katagiri F, Chua N-H (1989) An octopine synthase enhancer element directs tissue-specific expression and binds ASF-1, a factor from tobacco nuclear extracts. Plant Cell 1:977–984
Gilmartin PM, Sarokin L, Memelink J, Chua N-H (1990) Molecular light switches for plant genes. Plant Cell 2:369–378
Giuliano G, Pichersky E, Malik VS, Timko MP, Scolnik PA, Cashmore AR (1988) An evolutionarily conserved protein binding sequence upstream of a plant light-regulated gene. Proc Natl Acad Sci USA 85:7089–7093
Green PJ, Yong M-H, Cuozzo M, Kano-Murakami Y, Silverstein P. Chua N-H (1988) Binding site requirements for pea nuclear protein factor GT-1 correlate with sequences required for lightdependent transcriptional activation of the rbcS-3A gene. EMBO J 7:4035–4044
Haberlein T, Schimpff-Weiland G, Follman H (1985) Unexpected specificity in the thioredoxin activation of fructose-bis-phosphatases from different plants. Biochem Biophys Res Commun 27:401–446
Henikoff S (1984) Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene 28:351–359
Hershey HP, Barker RF, Idler KB, Murray MG, Quail PH (1987) Nucleotide sequence and characterisation of a gene encoding the phytochrome polypeptide from Arena. Gene 61:339–348
Horsch B, Fry FE, Hoffmann NL, Eichholtz D, Rogers SG, Fraley RT (1985) A simple and general method for transferring genes into plants. Science 227:1229–1231
Jefferson RA (1987) Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol Biol Reporter 5:387–405
Jefferson RA, Kavanagh TA, Bevan MW (1987) GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6:3901–3907
Katagiri F, Lam E, Chua N-H (1989) Two tobacco DNA-binding proteins with homology to the nuclear factor CREB. Nature 340:727–730
Kay R, Chan A, Daly M, McPherson J (1987) Duplication of CaMV 358 promoter sequences creates a strong enhancer for plant genes. Science 236:1299–1302
Kay SA, Keith B, Shinozaki K, Chye M-L, Chua N-H (1989) The rice phytochrome gene: structure, autoregulated expression and binding of GT-1 to a conserved site in the 5′ upstream region. Plant Cell 1:351–360
Ladror US, Latshaw SP, Marcus F (1990) Spinach cytosolic fructose-1,6-bisphosphatase purification, enzyme properties and structural comparisons. Eur J Biochem 189:89–94
Lazurus CM, Baulcombe DC, Martienssen RA (1985) α-Amylase genes of wheat are two multigene families which are differentially expressed. Plant Mol Biol 5:13–24
Loenen WAM, Blattner FR (1983) Lambda Charon vectors (Ch32, 33, 34 and 35) adapted for DNA cloning in recombination deficient hosts. Gene 26:171–179
Longstaff M, Raines CA, McMorrow EM, Bradbeer JW, Dyer TA (1989) Wheat phosphoglycerate kinase: evidence for recombination between the genes for the chloroplastic and cytosolic enzymes. Nucleic Acids Res 17:6569–6580
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY
Marcus F, Moberly L, Latshaw SP (1988) Comparative amino acid sequence of fructose- 1,6-bisphosphatases: identification of a region unique to the light-regulated chloroplast enzyme. Proc Natl Acad Sci USA 85:5379–5383
Mikami K, Tabata T, Kawata T, Nakayama T, Iwabuchi M (1987) Nuclear protein(s) binding to the conserved DNA hexameric sequence postulated to regulate transcription of wheat histone genes. FEBS Lett 223:273–278
Murphy G, Kavanagh T (1988) Speeding-up the sequencing of double-stranded DNA. Nucleic Acids Res 16:5198
Raines CA, Lloyd JC, Longstaff M, Bradley D, Dyer TA (1988) Chloroplast fructose- 1,6-bisphosphatase: the product of a mosaic gene. Nucleic Acids Res 16:7931–7942
Raines CA, Longstaff M, Lloyd JC, Dyer TA (1989) Complete coding sequence of wheat phosphoribulokinase: developmental and light-dependent expression of the mRNA. Mol Gen Genet 220:43–48
Shih M-C, Goodman HM (1988) Differential light regulated expression of nuclear genes encoding chloroplast and cytosolic glyceraldehyde-3-phosphate dehydrogenase in Nicotiana tabacum. EMBO J 7:893–898
Singh K, Tokuhisa JG, Dennis ES, Peacock WJ (1989) Saturation mutagenesis of the octopine synthase enhancer: correlation of mutant phenotypes with binding of a nuclear protein factor. Proc Natl Acad Sci USA 86:3733–3737
Stockhaus J, Schell J, Willmitzer L (1989) Correlation of the expression of the nuclear photosynthetic gene ST-LS1 with the presence of chloroplasts. EMBO J 8:2445–2451
Tabata T, Takase H, Takayama S, Mikami K, Nakatsuka A, Kawata T, Nakayama T, Iwabuchi M (1989) A protein that binds to a cis-acting element of wheat histone genes has a leucine zipper motif. Science 245:965–967
Taylor WC (1989) Regulatory interactions between nuclear and plastid genomes. Annu Rev Plant Physiol Plant Mol Biol 40:211–233
Tokuhisa JG, Singh K, Dennis ES, Peacock WJ (1990) A DNAbinding protein factor recognises two binding domains within the octopine synthase enhancer element. Plant Cell 2:215–224
Zimmerman G, Kelly GJ, Latzko E (1976) Efficient purification and molecular properties of spinach chloroplast fructose-1,6-bisphosphatase. Eur J Biochem 70:361–367
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Communicated by J. Schell
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Lloyd, J.C., Raines, C.A., John, U.P. et al. The chloroplast FBPase gene of wheat: structure and expression of the promoter in photosynthetic and meristematic cells of transgenic tobacco plants. Molec. Gen. Genet. 225, 209–216 (1991). https://doi.org/10.1007/BF00269850
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DOI: https://doi.org/10.1007/BF00269850