Light-induced Expression of the Chimeric Chalcone Synthase-NPTII Gene in Tobacco Cells

Overview

Journal EMBO J

Specialties Biotechnology
Molecular Biology

Date 1986 Jan 1

PMID 16453661

Citations 50

Authors

Hildegard Kaulen

Jeff Schell

Fritz Kreuzaler

Affiliations

Soon will be listed here.

Abstract

A chimeric gene was constructed containing the light-inducible chalcone synthase (chs) promoter from Antirrhinum majus, the neomycin phosphotransferase structural sequence from Tn5 as a reporter gene (NPTII) and the termination region from chs gene 1 from Petroselinum hortense. This gene was introduced into tobacco plants with the help of Ti plasmid-derived plant vectors and NPTII expression was measured. Analysis of the chs promoter sequence indicated the position of several possible regulatory regions. These were deleted to test their influence on the expression of the chs-NPTII gene. The different chimeric genes were all shown to be active after transfer to tobacco with the exception of one, in which the entire 5' upstream sequence from -1200 to -39 was deleted. The transcription of a chimeric gene with a 1.2-kbp 5' upstream promoter sequence was shown to be light inducible in tobacco plants. The analysis of various deletions of this 5' upstream sequence indicates that a number of sequence motifs have a quantitative effect on gene expression. One of these sequence motifs (-564 to -661) contains a direct repeat of 47 bp and the sequence GTGGTTAG which corresponds to the consensus core sequences observed in animal gene enhancer sequences. Deletion of a fragment containing this direct repeat resulted in a reduction of NPTII expression by a factor of 5.

Citing Articles

Pedoclimatic Conditions Influence the Morphological, Phytochemical and Biological Features of L.

Cornara L, Sgro F, Raimondo F, Ingegneri M, Mastracci L, DAngelo V Plants (Basel). 2023; 12(1).

PMID: 36616155 PMC: 9824027. DOI: 10.3390/plants12010024.

Impact of Growth Sites on the Phenolic Contents and Antioxidant Activities of Three Algerian Species ( L., (L.) Huds., and L.).

Brahmi F, Lounis N, Mebarakou S, Guendouze N, Yalaoui-Guellal D, Madani K Front Pharmacol. 2022; 13:886337.

PMID: 35784700 PMC: 9247617. DOI: 10.3389/fphar.2022.886337.

A Proteolytic Regulator Controlling Chalcone Synthase Stability and Flavonoid Biosynthesis in Arabidopsis.

Zhang X, Abrahan C, Colquhoun T, Liu C Plant Cell. 2017; 29(5):1157-1174.

PMID: 28446542 PMC: 5466025. DOI: 10.1105/tpc.16.00855.

UV-B damage and protection at the molecular level in plants.

Strid A, Chow W, Anderson J Photosynth Res. 2013; 39(3):475-89.

PMID: 24311138 DOI: 10.1007/BF00014600.

The chalcone synthase multigene family of Petunia hybrida (V30): differential, light-regulated expression during flower development and UV light induction.

Koes R, Spelt C, Mol J Plant Mol Biol. 2013; 12(2):213-25.

PMID: 24272800 DOI: 10.1007/BF00020506.

References

DHaese P, De Greve H, Decraemer H, Schell J, Van Montagu M . Rapid mapping of transposon insertion and deletion mutations in the large Ti-plasmids of Agrobacterium tumefaciens. Nucleic Acids Res. 1979; 7(7):1837-49. PMC: 342351. DOI: 10.1093/nar/7.7.1837. View

Laemmli U . Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970; 227(5259):680-5. DOI: 10.1038/227680a0. View

Fraley R, Rogers S, Horsch R, Sanders P, Flick J, Adams S . Expression of bacterial genes in plant cells. Proc Natl Acad Sci U S A. 1983; 80(15):4803-7. PMC: 384133. DOI: 10.1073/pnas.80.15.4803. View

Downs R, Siegelman H . Photocontrol of Anthocyanin Synthesis in Milo Seedlings. Plant Physiol. 1963; 38(1):25-30. PMC: 549873. DOI: 10.1104/pp.38.1.25. View

Albersheim P, Valent B . Host-pathogen interactions in plants. Plants, when exposed to oligosaccharides of fungal origin, defend themselves by accumulating antibiotics. J Cell Biol. 1978; 78(3):627-43. PMC: 2110205. DOI: 10.1083/jcb.78.3.627. View

Zambryski P, Joos H, Genetello C, Leemans J, Montagu M, Schell J . Ti plasmid vector for the introduction of DNA into plant cells without alteration of their normal regeneration capacity. EMBO J. 1983; 2(12):2143-50. PMC: 555426. DOI: 10.1002/j.1460-2075.1983.tb01715.x. View

Georgiev O, Birnstiel M . The conserved CAAGAAAGA spacer sequence is an essential element for the formation of 3' termini of the sea urchin H3 histone mRNA by RNA processing. EMBO J. 1985; 4(2):481-9. PMC: 554211. DOI: 10.1002/j.1460-2075.1985.tb03654.x. View

Kreuzaler F, Hahlbrock K . Enzymic synthesis of an aromatic ring from acetate units. Partial purification and some properties of flavanone synthase from cell-suspension cultures of Petroselinum hortense. Eur J Biochem. 1975; 56(1):205-13. DOI: 10.1111/j.1432-1033.1975.tb02223.x. View

Van Haute E, Joos H, Maes M, Warren G, Van Montagu M, Schell J . Intergeneric transfer and exchange recombination of restriction fragments cloned in pBR322: a novel strategy for the reversed genetics of the Ti plasmids of Agrobacterium tumefaciens. EMBO J. 1983; 2(3):411-7. PMC: 555148. DOI: 10.1002/j.1460-2075.1983.tb01438.x. View

10.

Kreuzaler F, Ragg H, Fautz E, Kuhn D, Hahlbrock K . UV-induction of chalcone synthase mRNA in cell suspension cultures of Petroselinum hortense. Proc Natl Acad Sci U S A. 1983; 80(9):2591-3. PMC: 393872. DOI: 10.1073/pnas.80.9.2591. View

11.

Herrera-Estrella L, Van den Broeck G, Maenhaut R, Van Montagu M, Schell J, Timko M . Light-inducible and chloroplast-associated expression of a chimaeric gene introduced into Nicotiana tabacum using a Ti plasmid vector. Nature. 1984; 310(5973):115-20. DOI: 10.1038/310115a0. View

12.

Kreuzaler F, Hahlbrock K . Enzymatic synthesis of aromatic compounds in higher plants. Formation of bis-noryangonin (4-hydroxy-6[4-hydroxystyryl]2-pyrone) from p-coumaroyl-CoA and malonyl-CoA. Arch Biochem Biophys. 1975; 169(1):84-90. DOI: 10.1016/0003-9861(75)90319-7. View

13.

Schroder J, Kreuzaler F, Schafer E, Hahlbrock K . Concomitant induction of phenylalanine ammonia-lyase and flavanone synthase mRNAs in irradiated plant cells. J Biol Chem. 1979; 254(1):57-65. View

14.

Towbin H, Staehelin T, Gordon J . Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979; 76(9):4350-4. PMC: 411572. DOI: 10.1073/pnas.76.9.4350. View

15.

Reimold U, Kroger M, Kreuzaler F, Hahlbrock K . Coding and 3' non-coding nucleotide sequence of chalcone synthase mRNA and assignment of amino acid sequence of the enzyme. EMBO J. 1983; 2(10):1801-5. PMC: 555362. DOI: 10.1002/j.1460-2075.1983.tb01661.x. View

16.

Kreuzaler F, Ragg H, Heller W, Tesch R, Witt I, Hammer D . Flavanone synthase from Petroselinum hortense. Molecular weight, subunit composition, size of messenger RNA, and absence of pantetheinyl residue. Eur J Biochem. 1979; 99(1):89-96. DOI: 10.1111/j.1432-1033.1979.tb13235.x. View

17.

Deblaere R, Bytebier B, De Greve H, Deboeck F, Schell J, Van Montagu M . Efficient octopine Ti plasmid-derived vectors for Agrobacterium-mediated gene transfer to plants. Nucleic Acids Res. 1985; 13(13):4777-88. PMC: 321826. DOI: 10.1093/nar/13.13.4777. View

18.

Siegelman H, Hendricks S . Purification and properties of phytochrome: a chromoprotein regulating plant growth. Fed Proc. 1965; 24(4):863-7. View

19.

Siegelman H, Turner B, Hendricks S . The chromophore of phytochrome. Plant Physiol. 1966; 41(8):1289-92. PMC: 550520. DOI: 10.1104/pp.41.8.1289. View

20.

Willmitzer L, Simons G, Schell J . The TL-DNA in octopine crown-gall tumours codes for seven well-defined polyadenylated transcripts. EMBO J. 1982; 1(1):139-46. PMC: 553008. DOI: 10.1002/j.1460-2075.1982.tb01137.x. View