Abstract
Deletion analysis of the nitrite-reductase (NiR) promoter from spinach (Spinacia oleracea L.) fused to theβ-glucuronidase (GUS) reporter gene and introduced into tobacco (Nicotiana tabacum L., cv. Coker 176) indicates that basic elements required for light- and nitrate-dependent expression of the reporter are located within the promoter sequence -200/+131 relative to the transcription-initiation site. Detailed analysis indicates that positive regulatory elements exist between -200 and-330 as well as between -1450 and -1730, stimulating the level of GUS gene expression under all experimental conditions. Induction/reversion light-pulse experiments show that the promoter sequence -200/+131 suffices for phytochrome-mediated expression of the reporter gene. The observation that the NiR promoter from spinach exhibits full reversibility in transgenic tobacco confirms the previous conclusion that the NiR promoter from spinach fused to a GUS reporter gene and introduced into tobacco responds to nitrate and phytochrome as would be expected for tobacco (host) and not as would be expected for spinach (donor). When the plastids were damaged by photooxidation in the presence of Norflurazon, GUS activity levels were reduced to the same extent for all NiR-promoter/GUS fusions tested, indicating that the promoter region involved in the action of the ‘plastidic factor’ is between -200 and +131. The GUS gene expression under the control of the CaMV-35S promoter is not affected by light, nitrate or the ‘plastidic factor’.
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Abbreviations
- B:
-
blue light
- cp:
-
cotyledon pair
- GUS:
-
β-glucuronidase (EC 3.2.1.31)
- NiR:
-
nitrite-reductase (EC 1.7.7.1)
- NF:
-
Norflurazon
- R:
-
red light
- RG9:
-
long-wavelength far-red light
References
Back, E., Dunne, W., Schneiderbauer, A., de Framond, A., Rastogi, R., Rothstein, S. (1991) Isolation of the spinach nitritereductase gene promoter which confers nitrate inducibility on GUS gene expression in transgenic tobacco. Plant Mol. Biol.17, 9–18
Baer, G.R., Meyers, S.P., Molin, W.T., Schrader, L. (1982) A simple and sensitive DNA assay for plant extracts. Plant Physiol.70, 999–1003
Bichler, J., Herrmann, R.G. (1990) Analysis of the promoters of the single-copy genes for plastocyanin and subunitδ of the chloroplast ATP synthase from spinach. Eur. J. Biochem.190, 415–426
Börner, T., Mendel, R.R., Schiemann, J. (1986) Nitrate reductase is not accumulated in chloroplast-ribosome-deficient mutants of higher plants. Planta169, 202–207
Chappell, J., Hahlbrock, K. (1984) Transcription of plant defence genes in response to UV light or fungal elicitor. Nature311, 76–78
Elliott, R.C., Dickey, L.F., White, M.J., Thompson, W.F. (1989) Cis-acting elements for light regulation of pea ferredoxin 1 gene expression are located within transcribed sequences. Plant Cell1, 691–698
Feinberg, A.P., Vogelstein, B. (1983) A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity. Anal. Biochem.132, 6–13
Fluhr, R., Chua, N.-H. (1986) Developmental regulation of two genes encoding ribulose-bisphosphate carboxylase small subunit in pea and transgenic petunia plants: phytochrome response and blue light induction. Proc. Natl. Acad. Sci. USA83, 2358–2362
Gardner, R.C., Howarth, A.J., Hahn, P., Brown-Luedi, M., Shepherd, R.J., Messing, J. (1981) The complete nucleotide sequence of an infectious clone of cauliflower mosaic virus by M13mp7 shotgun sequencing. Nucleic Acids Res.9, 2871–2888
Jefferson, R.A. (1987) Assaying chimeric genes in plants: The GUS gene fusion system. Plant Mol. Biol. Rep.5, 387–405
Jefferson, R.A., Kavanagh, T.A., Bevan, M.W. (1987) GUS fusions:β-glucuronidase as a sensitive and versatile gene fusions marker in higher plants. EMBO J.6, 3901–3907
Kuhlemeier, C., Green, P.J., Chua, N.-H. (1987) Regulation of gene expression in higher plants. Annu. Rev. Plant Physiol.38, 221–257
Mohr, H., Drumm-Herrel, H. (1981) Interaction between blue/UV light and light operating through phytochrome in higher plants. In: Plants and the daylight spectrum, pp. 423–441, Shmith, H., ed. Academic Press, London New York Toronto Sydney San Francisco
Mohr, H., Schoser, G. (1959) Eine Interferenzfilter-Monochromatoranlage für photobiologische Zwecke. Planta53, 1–17
Mohr, H., Neininger, A., Seith, B. (1992) Control of nitrate reductase and nitrite reductase gene expression by light, nitrate and a plastidic factor. Bot. Acta105, 81–89
Neininger, A., Kronenberger, J., Mohr, H. (1992) Coaction of light, nitrate and a plastidic factor in controlling nitrite-reductase gene expression in tobacco. Planta187, 381–387
Neininger, A., Bichler, J., Schneiderbauer, A., Mohr, H. (1993) Response of a nitrite-reductase 3.1-kilobase upstream regulatory sequence from spinach to nitrate and light in transgenic tobacco. Planta189, 440–442
Oelmüller, R. (1989) Photooxidative destruction of chloroplasts and its effect on nuclear gene expression and extraplastidic enzyme levels. Photochem. Photobiol.49, 229–239
Oelmüller, R., Levitan, I., Bergfeld, R., Rajasekhar, V.K., Mohr, H. (1986) Expression of nuclear genes as affected by treatments acting on the plastids. Planta168, 482–492
Oelmüller, R., Bolle, C., Tyagi, A.K., Niekrawietz, N., Breit, S., Herrmann, R.G. (1993) Characterization of the promoter from the single-copy gene encoding ferredoxin-NADP+-oxidoreductase from spinach. Mol. Gen. Genet.237, 261–272
Poulsen, C., Chua, N.-H. (1988) Dissection of 5′ upstream sequences for selective expression of theNicotiana plumbaginifolia rbcS-8B gene. Mol. Gen. Genet.214, 16–23
Rajasekhar, V.K., Mohr, H. (1986) Appearance of nitrite reductase in cotyledons of the mustard (Sinapis alba L.) seedling as affected by nitrate, phytochrome and photooxidative damage of plastids. Planta168, 369–376
Rastogi, R., Back, E., Schneiderbauer, A., Bowsher, C.G., Moffatt, B., Rothstein, S. (1993) A 330 bp region of the spinach nitrite reductase gene promoter directs nitrate-inducible tissue-specific expression in transgenic tobacco. Plant J.4, 317–236
Reiß, T., Bergfeld, R., Link, G., Thien, W., Mohr, H. (1983) Photooxidative destruction of chloroplasts and its consequences for cytosolic enzyme levels and plant development. Planta159, 518–528
Rolfe, S.A., Tobin, E.M. (1991) Deletion analysis of a phytochrome regulated monocotrbcS promoter in a transient assay system. Proc. Natl. Acad. Sci. USA88, 2683–2686
Sambrook, J., Fritsch, E.F., Maniatis, T. (1989) Molecular cloning, a laboratory manual, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY
Schuster, C., Mohr, H. (1990) Photooxidative damage to the plastids affects abundance of nitrate reductase mRNA in mustard cotyledons. Planta181, 125–128
Seith, B., Schuster, C., Mohr, H. (1991) Coaction of light, nitrate and a plastidic factor in controlling nitrite-reductase gene expression in spinach. Planta184, 74–80
Sloan, J.S., Schwartz, B.W., Becker, W.M. (1993) Promoter analysis of a light-regulated gene encoding hydroxypyruvate reductase, an enzyme of the photorespiratory glycolate pathway. Plant J.3, 867–874
Vorst, O., van Dam, F., Weisbeek, P., Smeekers, S. (1993) Light-regulated expression of theArabidopsis thaliana ferredoxin A gene involves both transcriptional and post-transcriptional processes. Plant J.3, 793–803
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This paper is dedicated to Professor Hartmut K. Lichtenthaler on the occasion of his 60th birthday
This research was supported by a grant from the Deutsche Forschungsgemeinschaft (SFB 206)
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Neininger, A., Back, E., Bichler, J. et al. Deletion analysis of a nitrite-reductase promoter from spinach in transgenic tobacco. Planta 194, 186–192 (1994). https://doi.org/10.1007/BF01101677
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DOI: https://doi.org/10.1007/BF01101677