Skip to main content
Log in

Phenylalanine ammonia-lyase gene structure, expression, and evolution in Nicotiana

  • Research Article
  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

Abstract

Phenylalanine ammonia-lyase (PAL) catalyzes the first reaction in the general phenylpropanoid pathway leading to the production of phenolic compounds with a significant range of biological functions. A PAL gene we designated gPAL1, cloned from tobacco, consists of two exons separated by an intron of 1932 bp. Exon I, 398 bp, and exon II, 1747 bp, together encode a polypeptide of 715 amino acids. A putative TATA box and polyadenylation signal are found 144 bp upstream of the initiation codon and 193 bp downstream from the stop codon, respectively. Using various parts of gPAL1 as probes, genomic Southern blots indicated the presence of a small family of PAL genes in the tobacco genome that can be divided into two distinct subfamilies, one consisting of pal1 and pal2 and another of pal3 and pal4. Comparative genomic blot analysis of progenitor species (Nicotiana tomentosiformis and N. sylvestris) indicated that each species contains one PAL gene from each of the subfamilies, suggesting that pal1 and pal3 (or pal2 and pal4) diverged prior to the evolution of N. tabacum. Expression of the PAL gene family was examined using RNA gel blots. PAL transcript levels were significantly higher in flowers and roots than in leaves and stems of mature plants. PAL transcripts accumulate differentially during flower and leaf maturation in that mRNA levels decline during flower maturation but increase during leaf maturation. In leaves, PAL transcripts rapidly accumulated after wounding.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Beggs CJ, Schneider-Ziebert U, Wellmann E: Solar ultraviolet radiation and plant life. In: Worrest RC, Caldwell MM (eds) Stratospheric Ozone Reduction, pp. 235–250. Springer-Verlag, Berlin (1986).

    Google Scholar 

  2. Benfey PN, Chua N-H: The cauliflower mosaic virus 35S promoter: combinatorial regulation of transcription in plants. Science 250: 959–966 (1990).

    Google Scholar 

  3. 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 8: 1899–1906 (1989).

    Google Scholar 

  4. Breathnach R, Chambon P: Organization and expression of eucaryotic split genes coding for proteins. Annu Rev Biochem 50: 349–383 (1981).

    Google Scholar 

  5. Cramer CL, Edwards K, Dron M, Liang X, Dildine SL, Bolwell GP, Dixon RA, Lamb CJ, Schuch W: Phenylalanine ammonia-lyase gene organization and structure. Plant Mol Biol 12: 367–383 (1989).

    Google Scholar 

  6. Devereux J, Haeberli P, Smithies O: A comprehensive set of sequence analysis programs for the VAX. Nucl Acids Res 12: 387–395 (1984).

    Google Scholar 

  7. Dixon RA, Harrison MJ: Activation, structure, and organization of genes involved in microbial defense in plants. In: Scandalios JG (eds) Genomic Responses to Environmental Stress, pp. 165–234. Academic Press, San Diego (1990).

    Google Scholar 

  8. Estabrook EM, Sengupta-Gopalan C: Differential expression of phenylalanine ammonia-lyase and chalcone synthase during soybean nodule development. Plant Cell 3: 299–308 (1991).

    Google Scholar 

  9. Frank RL, Vodkin LO: Sequence and structure of a phenylalanine ammonialyase gene from Glycine max. DNA Seq 1: 335–346 (1991).

    Google Scholar 

  10. Fritzemeier K-H, Cretin C, Kombrink E, Rohwer F, Taylor J, Scheel D, Hahlbrock K: Transient induction of phenylalanine ammonia-lyase and 4-coumarate:CoA ligase mRNAs in potato leaves infected with virulent or avirulent races of Phytophthora infestans. Plant Physiol 85: 34–41 (1987).

    Google Scholar 

  11. Gray JC, Kung SD, Wildman SG, Sheen SJ: Origin of Nicotiana tabacum L. detected by polypeptide composition of Fraction I protein. Nature 252: 226–227 (1974).

    Google Scholar 

  12. Hahlbrock K, Grisebach H: Enzymic controls in the biosynthesis of lignin and flavonoids. Annu Rev Plant Physiol 30: 105–30 (1979).

    Google Scholar 

  13. Hauptmann RM, Ashraf M, Vasil V, Hannah LC, Vasil IK, Ferl R: Promoter strength comparisons of maize shrunken 1 and alcohol dehydrogenase 1 and 2 promoters in mono- and dicotyledonous species. Plant Physiol 88: 1063–1066 (1988).

    Google Scholar 

  14. Jacobs M, Rubery PH: Naturally occurring auxin transport regulators. Science 241: 346–349 (1988).

    Google Scholar 

  15. Jones DH: Phenylalanine ammonia-lyase: regulation of its induction, and its role in plant development. Phytochemistry 23: 1349–1359 (1984).

    Google Scholar 

  16. Joos H-J, Hahlbrock K: Phenylalanine ammonia-lyase in potato (Solanum tuberosum L.). Eur J Biochem 204: 621–629 (1992).

    Google Scholar 

  17. Joshi CP: An inspection of the domain between putative TATA box and translation start site in 79 plant genes. Nucl Acids Res 15: 6643–6653 (1987).

    Google Scholar 

  18. Lee S-W, Robb J, Nazar RN: Truncated phenylalanine ammonia-lyase expression in tomato (Lycopersicon esculentum). J Biol Chem 267: 11824–11830 (1992).

    Google Scholar 

  19. Leyva A, Liang X, Pintor-Toro JA, Dixon RA, Lamb CJ: cis-Element combinations determine phenylalanine ammonia-lyase gene tissue-specific expression patterns. Plant Cell 4: 263–271 (1992).

    Google Scholar 

  20. Liang X, Dron M, Schmid J, Dixon RA, Lamb CJ: Developmental and environmental regulation of a phenylalanine ammonia-lyase-β-glucuronidase gene fusion in transgenic tobacco plants. Proc Natl Acad Sci USA 86: 9284–9288 (1989).

    Google Scholar 

  21. Lois R, Dietrich A, Hahlbrock K, Schulz W: A phenylalanine ammonia-lyase gene from parsley: structure, regulation and identification of elicitor and light responsive cis-acting elements. EMBO J 8: 1641–1648 (1989).

    Google Scholar 

  22. Lütcke HA, Chow KC, Mickel FS, Moss KA, Kern HF, Scheele GA: Selection of AUG initiation codons differs in plants and animals. EMBO J 6: 43–48 (1987).

    Google Scholar 

  23. Maniatis T, Fritsch EF, Sambrook J: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1982).

    Google Scholar 

  24. McLauchlan J, Gaffney D, Whitton JL, Clements JB: The consensus sequence YGTGTTYY located downstream from the AATAAA signal is required for efficient formation of mRNA 3′ termini. Nucl Acids Res 13: 1347–1368 (1985).

    Google Scholar 

  25. Minami E, Tanaka Y: Nucleotide sequence of the gene for phenylalanine ammonia-lyase of rice and its deduced amino acid sequence. Biochim Biophys Acta 1171: 321–322 (1993).

    Google Scholar 

  26. Minami E-i, Ozeki Y, Matsuoka M, Koizuka N, Tanaka Y: Structure and some characterization of the gene for phenylalanine ammonia-lyase from rice plants. Eur J Biochem 185: 19–25 (1989).

    Google Scholar 

  27. Nagai N, Kitauchi F, Shimosaka M, Okazaki M: Cloning and sequencing of a full-length cDNA coding for phenylalanine ammonia-lyase from tobacco cell culture. Plant Physiol 104: 1091–1092 (1994).

    Google Scholar 

  28. Ohl S, Hedrick SA, Chory J, Lamb CJ: Functional properties of a phenylalanine ammonia-lyase promoter from Arabidopsis. Plant Cell 2: 837–848 (1990).

    Google Scholar 

  29. Pellegrini L, Rohfritsch O, Fritig B, Legrand M: Phenylalanine ammonia-lyase in tobacco. Plant Physiol 106: 877–886 (1994).

    Google Scholar 

  30. Rhodes MJC: The physiological significance of plant phenolic compounds. In: Sumere CFv, Lea PJ (eds) Annual Proceedings of the Phytochemical Society of Europe: The Biochemistry of Plant Phenolics, vol. 25, pp. 99–118. Oxford University Press, Oxford (1985).

    Google Scholar 

  31. Richards E: Preparation of genomic DNA from plant tissue. In: Ausubel FM, Brent R, Kingston RE, Moore D, Seidman JG, Smith JA, Struhl K (eds) Current Protocols in Molecular Biology, pp. 2.3.1.-2.3.3.. John Wiley, New York (1987).

    Google Scholar 

  32. Sanger F, Nicklen S, Coulson AR: DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467 (1977).

    Google Scholar 

  33. Stafford HA: The metabolism of aromatic compounds. Annu Rev Plant Physiol 25: 459–86 (1974).

    Google Scholar 

  34. Stafford HA: Flavonoid Metabolism. CRC Press, Boca Raton, FL (1990).

    Google Scholar 

  35. Whetten RW, Sederoff RR: Phenylalanine ammonia-lyase from loblolly pine. Plant Physiol 98: 380–386 (1992).

    Google Scholar 

  36. Yamada T, Tanaka Y, Sriprasertsak P, Kato H, Hashimoto T, Kawamata S, Ichinose Y, Kato H, Shiraishi T, Oku H: Phenylalanine ammonia-lyase gene from Pisum sativum: structure, organ-specific expression and regulation by fungal elicitor and suppressor. Plant Cell Physiol 33: 715–725 (1992).

    Google Scholar 

  37. Yamada T, Tanaka Y, Sriprasertsak P, Kato H, Hashimoto T, Kawamata S, Ichinose Y, Kato H, Shiraishi T, Oku H: Phenylalanine ammonia-lyase gene from Pisum sativum: structure, organ-specific expression and regulation by fungal elicitor and suppressor. Plant Mol Biol 33: 715–725 (1992).

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Fukasawa-Akada, T., Kung, Sd. & Watson, J.C. Phenylalanine ammonia-lyase gene structure, expression, and evolution in Nicotiana . Plant Mol Biol 30, 711–722 (1996). https://doi.org/10.1007/BF00019006

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00019006

Key words

Navigation