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Expression of bacterial tyrosine ammonia-lyase creates a novel p-coumaric acid pathway in the biosynthesis of phenylpropanoids in Arabidopsis

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Abstract

Some flavonoids are considered as beneficial compounds because they exhibit anticancer or antioxidant activity. In higher plants, flavonoids are secondary metabolites that are derived from phenylpropanoid biosynthetic pathway. A large number of phenylpropanoids are generated from p-coumaric acid, which is a derivative of the primary metabolite, phenylalanine. The first two steps in the phenylpropanoid biosynthetic pathway are catalyzed by phenylalanine ammonia-lyase and cinnamate 4-hydroxylase, and the coupling of these two enzymes forms a rate-limiting step in the pathway. For the generation of p-coumaric acid, the conversion from phenylalanine to p-coumaric acid that is catalyzed by two enzymes can be theoretically performed by a single enzyme, tyrosine ammonia-lyase (TAL) that catalyzes the conversion of tyrosine to p-coumaric acid in certain bacteria. To modify the p-coumaric acid pathway in plants, we isolated a gene encoding TAL from a photosynthetic bacterium, Rhodobacter sphaeroides, and introduced the gene (RsTAL) in Arabidopsis thaliana. Analysis of metabolites revealed that the ectopic over-expression of RsTAL leads to higher accumulation of anthocyanins in transgenic 5-day-old seedlings. On the other hand, 21-day-old seedlings of plants expressing RsTAL showed accumulation of higher amount of quercetin glycosides, sinapoyl and p-coumaroyl derivatives than control. These results indicate that ectopic expression of the RsTAL gene in Arabidopsis enhanced the metabolic flux into the phenylpropanoid pathway and resulted in increased accumulation of flavonoids and phenylpropanoids.

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Abbreviations

C4H:

Cinnamate 4-hydroxylase

4CL:

4-Coumarate:CoA ligase

ORF:

Open reading frame

PAL:

Phenylalanine ammonia-lyase

PCR:

Polymerase chain reaction

RsTAL :

Rhodobacter sphaeroides TAL

RT-PCR:

Reverse transcription polymerase chain reaction

TAL:

Tyrosine ammonia-lyase

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Author notes

  1. Choong-Soo Yun

    Present address: Biotechnology Research Center, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo, 113-8657, Japan

  2. Tadamasa Sasaki

    Present address: Department of Biological Science and Technology, Tokyo University of Science, Yamazaki 2641, Noda, Chiba, 278-8510, Japan

  3. Fumio Matsuda

    Present address: Organization of Advanced Sciences and Technology, Kobe University, 1-1 Rokkodaicho, Nada-ku, Kobe, 657-8501, Japan

Authors and Affiliations

  1. Cell-Free Science and Technology Research Center, and The Venture Business Laboratory, Ehime University, 3 Bunkyo-cho, Matsuyama, Ehime, 890-8577, Japan

    Yasutaka Nishiyama, Choong-Soo Yun, Tadamasa Sasaki & Yuzuru Tozawa

  2. RIKEN Plant Science Center, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama, Kanagawa, 230-0045, Japan

    Fumio Matsuda & Kazuki Saito

  3. Graduate School of Pharmaceutical Sciences, Chiba University, Yayoi-cho 1-33, Inage-ku, Chiba, Chiba, 263-8522, Japan

    Kazuki Saito

Authors
  1. Yasutaka Nishiyama
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  2. Choong-Soo Yun
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  3. Fumio Matsuda
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  4. Tadamasa Sasaki
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  5. Kazuki Saito
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  6. Yuzuru Tozawa
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Corresponding author

Correspondence to Yuzuru Tozawa.

Additional information

Y. Nishiyama and C. S. Yun contributed equally to this work.

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Nishiyama, Y., Yun, CS., Matsuda, F. et al. Expression of bacterial tyrosine ammonia-lyase creates a novel p-coumaric acid pathway in the biosynthesis of phenylpropanoids in Arabidopsis. Planta 232, 209–218 (2010). https://doi.org/10.1007/s00425-010-1166-1

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