The Expression Pattern of Alfalfa Flavanone 3-hydroxylase Promoter-gus Fusion in Nicotiana Benthamiana Correlates with the Presence of Flavonoids Detected in Situ

Overview

Journal Plant Mol Biol

Date 1996 Mar 1

PMID 8704126

Citations 4

Authors

B Charrier

C Leroux

A Kondorosi

P Ratet

Affiliations

Soon will be listed here.

Abstract

Flavanone 3-hydroxylase is an enzyme acting in the central part of the flavonoid biosynthesis pathway. It is generally encoded by a single gene and seems to have a key position for the regulation in this pathway. These two features make a single f3h promoter-gus fusion a suitable tool to study both the f3h expression and the regulation of this pathway. We present here the spatial and temporal analysis of the expression of an alfalfa flavanone 3-hydroxylase (f3h) promoter-gus fusion introduced into Nicotiana benthamiana. The Medicago sativa (alfalfa) f3h promoter directed gus expression in flowers, stems, leaves and roots. In flowers, GUS activity was observed in pollen grains, in ovules, in ovary placenta and in the epidermis, medullary parenchyma, trichomes and second cortical cellular layer surrounding the vascular bundles of the peduncle. In stems, GUS activity was detected at the same places as in the peduncle except for the medullary parenchyma. In roots, we found GUS staining in root hairs, epidermis and in the vascular bundles of the elongated zone. Finally, in leaves, the f3h promoter expressed essentially in the stalk cells of the multicellular trichomes. The expression pattern of the f3h-gus fusion was correlated to the presence of flavonoids in situ. These data indicate that this construct can be very useful to study factors controlling the production of flavonoids.

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References

Britsch L, Dedio J, Saedler H, Forkmann G . Molecular characterization of flavanone 3 beta-hydroxylases. Consensus sequence, comparison with related enzymes and the role of conserved histidine residues. Eur J Biochem. 1993; 217(2):745-54. DOI: 10.1111/j.1432-1033.1993.tb18301.x. View

Ditta G, Stanfield S, Corbin D, Helinski D . Broad host range DNA cloning system for gram-negative bacteria: construction of a gene bank of Rhizobium meliloti. Proc Natl Acad Sci U S A. 1980; 77(12):7347-51. PMC: 350500. DOI: 10.1073/pnas.77.12.7347. View

Li J, Ou-Lee T, Raba R, Amundson R, Last R . Arabidopsis Flavonoid Mutants Are Hypersensitive to UV-B Irradiation. Plant Cell. 1993; 5(2):171-179. PMC: 160260. DOI: 10.1105/tpc.5.2.171. View

Meldgaard M . Expression of chalcone synthase, dihydroflavonol reductase, and flavanone-3-hydroxylase in mutants of barley deficient in anthocyanin and proanthocyanidin biosynthesis. Theor Appl Genet. 2013; 83(6-7):695-706. DOI: 10.1007/BF00226687. View

Szabados L, Ratet P, Grunenberg B, de Bruijn F . Functional analysis of the Sesbania rostrata leghemoglobin glb3 gene 5'-upstream region in transgenic Lotus corniculatus and Nicotiana tabacum plants. Plant Cell. 1990; 2(10):973-86. PMC: 159946. DOI: 10.1105/tpc.2.10.973. View

Schmid J, Doerner P, Clouse S, Dixon R, Lamb C . Developmental and environmental regulation of a bean chalcone synthase promoter in transgenic tobacco. Plant Cell. 1990; 2(7):619-31. PMC: 159916. DOI: 10.1105/tpc.2.7.619. View

Peters N, Verma D . Phenolic compounds as regulators of gene expression in plant-microbe relations. Mol Plant Microbe Interact. 1990; 3(1):4-8. DOI: 10.1094/mpmi-3-004. View

Koes R, van Blokland R, Quattrocchio F, van Tunen A, Mol J . Chalcone Synthase Promoters in Petunia Are Active in Pigmented and Unpigmented Cell Types. Plant Cell. 1990; 2(5):379-392. PMC: 159895. DOI: 10.1105/tpc.2.5.379. View

van Tunen A, Mur L, Brouns G, Rienstra J, Koes R, Mol J . Pollen- and anther-specific chi promoters from petunia: tandem promoter regulation of the chiA gene. Plant Cell. 1990; 2(5):393-401. PMC: 159896. DOI: 10.1105/tpc.2.5.393. View

10.

Bevan M . Binary Agrobacterium vectors for plant transformation. Nucleic Acids Res. 1984; 12(22):8711-21. PMC: 320409. DOI: 10.1093/nar/12.22.8711. View

11.

SANGER F, Nicklen S, Coulson A . DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977; 74(12):5463-7. PMC: 431765. DOI: 10.1073/pnas.74.12.5463. View

12.

Murphy L, Herzog C, Rudick J, Fojo A, Bates S . Use of the polymerase chain reaction in the quantitation of mdr-1 gene expression. Biochemistry. 1990; 29(45):10351-6. DOI: 10.1021/bi00497a009. View

13.

Charrier B, Coronado C, Kondorosi A, Ratet P . Molecular characterization and expression of alfalfa (Medicago sativa L.) flavanone-3-hydroxylase and dihydroflavonol-4-reductase encoding genes. Plant Mol Biol. 1995; 29(4):773-86. DOI: 10.1007/BF00041167. View

14.

Sablowski R, Moyano E, Culianez-Macia F, Schuch W, Martin C, Bevan M . A flower-specific Myb protein activates transcription of phenylpropanoid biosynthetic genes. EMBO J. 1994; 13(1):128-37. PMC: 394786. DOI: 10.1002/j.1460-2075.1994.tb06242.x. View

15.

Bevan M, Shufflebottom D, Edwards K, Jefferson R, Schuch W . Tissue- and cell-specific activity of a phenylalanine ammonia-lyase promoter in transgenic plants. EMBO J. 1989; 8(7):1899-906. PMC: 401048. DOI: 10.1002/j.1460-2075.1989.tb03592.x. View

16.

Martin C, Prescott A, Mackay S, Bartlett J, Vrijlandt E . Control of anthocyanin biosynthesis in flowers of Antirrhinum majus. Plant J. 1991; 1(1):37-49. DOI: 10.1111/j.1365-313x.1991.00037.x. View

17.

Britsch L, Forkmann G . Molecular cloning, sequence analysis, and in vitro expression of flavanone 3 beta-hydroxylase from Petunia hybrida. J Biol Chem. 1992; 267(8):5380-7. View

18.

van der Meer I, Stam M, van Tunen A, Mol J, Stuitje A . Antisense inhibition of flavonoid biosynthesis in petunia anthers results in male sterility. Plant Cell. 1992; 4(3):253-62. PMC: 160126. DOI: 10.1105/tpc.4.3.253. View

19.

Deboo G, Albertsen M, Taylor L . Flavanone 3-hydroxylase transcripts and flavonol accumulation are temporally coordinate in maize anthers. Plant J. 1995; 7(5):703-13. DOI: 10.1046/j.1365-313x.1995.07050703.x. View

20.

Sparvoli F, Martin C, Scienza A, Gavazzi G, Tonelli C . Cloning and molecular analysis of structural genes involved in flavonoid and stilbene biosynthesis in grape (Vitis vinifera L.). Plant Mol Biol. 1994; 24(5):743-55. DOI: 10.1007/BF00029856. View