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Plant Physiol. Dec 1996; 112(4): 1617–1624.
PMCID: PMC158095

Overexpression of L-Phenylalanine Ammonia-Lyase in Transgenic Tobacco Plants Reveals Control Points for Flux into Phenylpropanoid Biosynthesis.

Abstract

Transgenic tobacco (Nicotiana tabacum L.) plants overexpressing the enzyme L-phenylalanine ammonia-lyase (PAL; EC 4.3.1.5) were grown from seeds of a primary transformant containing the bean PAL2 gene, which had shown homology-dependent silencing of the endogenous tobacco PAL genes. Analysis of endogenous and transgene-encoded PAL transcripts and protein in the primary transformant (T0) and first-generation (T1) overexpressor plants indicated that the transgene-encoded PAL is the cause of the greater than wild-type levels of PAL activity (up to 5- and 2-fold greater in leaf and stem tissue, respectively) in the T1 plants. Leaves of PAL-overexpressing plants contained increased levels of the hydroxycinnamic acid ester chlorogenic acid but not of the flavonoid rutin, indicating that PAL is the key control point for flux into chlorogenic acid. In addition, levels of the glucoside of 4-coumaric acid increased in the overexpressing plants, suggesting that the 4-coumarate:coenzyme A ligase or coumarate hydroxylase reactions might have become limiting. These results help to define the regulatory architecture of the phenylpropanoid pathway and indicate the possibility of engineering-selective changes in this complex metabolic pathway by overexpression of a single early pathway gene.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Bate NJ, Orr J, Ni W, Meromi A, Nadler-Hassar T, Doerner PW, Dixon RA, Lamb CJ, Elkind Y. Quantitative relationship between phenylalanine ammonia-lyase levels and phenylpropanoid accumulation in transgenic tobacco identifies a rate-determining step in natural product synthesis. Proc Natl Acad Sci U S A. 1994 Aug 2;91(16):7608–7612. [PMC free article] [PubMed]
  • Berry JO, Breiding DE, Klessig DF. Light-mediated control of translational initiation of ribulose-1, 5-bisphosphate carboxylase in amaranth cotyledons. Plant Cell. 1990 Aug;2(8):795–803. [PMC free article] [PubMed]
  • Berry JO, Carr JP, Klessig DF. mRNAs encoding ribulose-1,5-bisphosphate carboxylase remain bound to polysomes but are not translated in amaranth seedlings transferred to darkness. Proc Natl Acad Sci U S A. 1988 Jun;85(12):4190–4194. [PMC free article] [PubMed]
  • Boerjan W, Bauw G, Van Montagu M, Inzé D. Distinct phenotypes generated by overexpression and suppression of S-adenosyl-L-methionine synthetase reveal developmental patterns of gene silencing in tobacco. Plant Cell. 1994 Oct;6(10):1401–1414. [PMC free article] [PubMed]
  • Boutry M, Chua NH. A nuclear gene encoding the beta subunit of the mitochondrial ATP synthase in Nicotiana plumbaginifolia. EMBO J. 1985 Sep;4(9):2159–2165. [PMC free article] [PubMed]
  • Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. [PubMed]
  • Brederode FT, Linthorst HJ, Bol JF. Differential induction of acquired resistance and PR gene expression in tobacco by virus infection, ethephon treatment, UV light and wounding. Plant Mol Biol. 1991 Dec;17(6):1117–1125. [PubMed]
  • Chappell J, Wolf F, Proulx J, Cuellar R, Saunders C. Is the Reaction Catalyzed by 3-Hydroxy-3-Methylglutaryl Coenzyme A Reductase a Rate-Limiting Step for Isoprenoid Biosynthesis in Plants? Plant Physiol. 1995 Dec;109(4):1337–1343. [PMC free article] [PubMed]
  • Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. [PubMed]
  • de Carvalho F, Gheysen G, Kushnir S, Van Montagu M, Inzé D, Castresana C. Suppression of beta-1,3-glucanase transgene expression in homozygous plants. EMBO J. 1992 Jul;11(7):2595–2602. [PMC free article] [PubMed]
  • Dixon RA, Paiva NL. Stress-Induced Phenylpropanoid Metabolism. Plant Cell. 1995 Jul;7(7):1085–1097. [PMC free article] [PubMed]
  • Elkind Y, Edwards R, Mavandad M, Hedrick SA, Ribak O, Dixon RA, Lamb CJ. Abnormal plant development and down-regulation of phenylpropanoid biosynthesis in transgenic tobacco containing a heterologous phenylalanine ammonia-lyase gene. Proc Natl Acad Sci U S A. 1990 Nov;87(22):9057–9061. [PMC free article] [PubMed]
  • Enyedi AJ, Yalpani N, Silverman P, Raskin I. Localization, conjugation, and function of salicylic acid in tobacco during the hypersensitive reaction to tobacco mosaic virus. Proc Natl Acad Sci U S A. 1992 Mar 15;89(6):2480–2484. [PMC free article] [PubMed]
  • Fukasawa-Akada T, Kung SD, Watson JC. Phenylalanine ammonia-lyase gene structure, expression, and evolution in Nicotiana. Plant Mol Biol. 1996 Feb;30(4):711–722. [PubMed]
  • Görlach J, Raesecke HR, Rentsch D, Regenass M, Roy P, Zala M, Keel C, Boller T, Amrhein N, Schmid J. Temporally distinct accumulation of transcripts encoding enzymes of the prechorismate pathway in elicitor-treated, cultured tomato cells. Proc Natl Acad Sci U S A. 1995 Apr 11;92(8):3166–3170. [PMC free article] [PubMed]
  • Harms K, Atzorn R, Brash A, Kuhn H, Wasternack C, Willmitzer L, Pena-Cortes H. Expression of a Flax Allene Oxide Synthase cDNA Leads to Increased Endogenous Jasmonic Acid (JA) Levels in Transgenic Potato Plants but Not to a Corresponding Activation of JA-Responding Genes. Plant Cell. 1995 Oct;7(10):1645–1654. [PMC free article] [PubMed]
  • Hart CM, Fischer B, Neuhaus JM, Meins F., Jr Regulated inactivation of homologous gene expression in transgenic Nicotiana sylvestris plants containing a defense-related tobacco chitinase gene. Mol Gen Genet. 1992 Nov;235(2-3):179–188. [PubMed]
  • Hennig J, Malamy J, Grynkiewicz G, Indulski J, Klessig DF. Interconversion of the salicylic acid signal and its glucoside in tobacco. Plant J. 1993 Oct;4(4):593–600. [PubMed]
  • Klessig DF, Malamy J. The salicylic acid signal in plants. Plant Mol Biol. 1994 Dec;26(5):1439–1458. [PubMed]
  • Logemann E, Parniske M, Hahlbrock K. Modes of expression and common structural features of the complete phenylalanine ammonia-lyase gene family in parsley. Proc Natl Acad Sci U S A. 1995 Jun 20;92(13):5905–5909. [PMC free article] [PubMed]
  • Maher EA, Bate NJ, Ni W, Elkind Y, Dixon RA, Lamb CJ. Increased disease susceptibility of transgenic tobacco plants with suppressed levels of preformed phenylpropanoid products. Proc Natl Acad Sci U S A. 1994 Aug 2;91(16):7802–7806. [PMC free article] [PubMed]
  • Matzke MA, Matzke AJM. How and Why Do Plants Inactivate Homologous (Trans)genes? Plant Physiol. 1995 Mar;107(3):679–685. [PMC free article] [PubMed]
  • Pellegrini L, Rohfritsch O, Fritig B, Legrand M. Phenylalanine ammonia-lyase in tobacco. Molecular cloning and gene expression during the hypersensitive reaction to tobacco mosaic virus and the response to a fungal elicitor. Plant Physiol. 1994 Nov;106(3):877–886. [PMC free article] [PubMed]
  • 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 Sep;76(9):4350–4354. [PMC free article] [PubMed]
  • Tropf S, Kärcher B, Schröder G, Schröder J. Reaction mechanisms of homodimeric plant polyketide synthase (stilbenes and chalcone synthase). A single active site for the condensing reaction is sufficient for synthesis of stilbenes, chalcones, and 6'-deoxychalcones. J Biol Chem. 1995 Apr 7;270(14):7922–7928. [PubMed]
  • Van Soest PJ, Robertson JB, Lewis BA. Methods for dietary fiber, neutral detergent fiber, and nonstarch polysaccharides in relation to animal nutrition. J Dairy Sci. 1991 Oct;74(10):3583–3597. [PubMed]
  • Vayda ME, Shewmaker CK, Morelli JK. Translational arrest in hypoxic potato tubers is correlated with the aberrant association of elongation factor EF-1 alpha with polysomes. Plant Mol Biol. 1995 Jul;28(4):751–757. [PubMed]
  • Wanner LA, Li G, Ware D, Somssich IE, Davis KR. The phenylalanine ammonia-lyase gene family in Arabidopsis thaliana. Plant Mol Biol. 1995 Jan;27(2):327–338. [PubMed]

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