Skip to main content
SpringerLink
Log in

Characterization of the tissue-specific expression of phenylalanine ammonia-lyase gene promoter from loblolly pine (Pinus taeda) in Nicotiana tabacum

Plant Cell Reports volume 28, pages 1309–1317 (2009)Cite this article

Abstract

We isolated the 5′ flanking region of a gene for phenylalanine ammonia-lyase (PAL; EC 4.3.1.5) from Pinus taeda, PtaPAL. To investigate the tissue-specific expression of the PtaPAL promoter, histochemical assay of GUS activity was performed using the transgenic tobacco expressing the PtaPAL promoter-GUS. The region of −897 to −420 in PtaPAL promoter showed high activities in the secondary xylem and response to bending stress. To characterize the cis-regulatory functions of the promoters for enzymes in phenylpropanoid biosynthesis, we examined the activity of chimeric promoters of PtaPAL and a 4-coumarate CoA ligase, Pta4CLα. The chimeric promoter showed similar activity as the Pta4CLα promoter. Electrophoretic mobility shift assays implicated −897 to –674 of PtaPAL promoter containing cis-elements of the expression in xylem of Pinus taeda. The results suggested that AC elements of PtaPAL have multiple functions in the expression under the various developmental stages and stress conditions in the transgenic tobacco.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

39,95 €

Price includes VAT (Finland)

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

References

  • Armstrong GA, Weisshaar B, Hahlbrock K (1992) Homodimeric and heterodimeric leucine zipper proteins and nuclear factors from parsley recognize diverse promoter elements with ACGT cores. Plant Cell 4:525–537

    Article  PubMed  CAS  Google Scholar 

  • Becker D, Kemper E, Schell J, Masterson R (1992) New plant binary vectors with selectable markers located proximal to the left T-DNA border. Plant Mol Biol 20:1195–1197

    Article  PubMed  CAS  Google Scholar 

  • Bedon F, Grima-Pettenati J, Mackay J (2007) Conifer R2R3-MYB transcription factors: sequence analyses and gene expression in wood-forming tissues of white spruce (Picea glauca). BMC Plant Biol 7:17

    Article  PubMed  Google Scholar 

  • Bell-Lelong DA, Cusumano JC, Meyer K, Chapple C (1997) Cinnamate-4-hydroxylase expression in Arabidopsis. Regulation in response to development and the environment. Plant Physiol 113:729–738

    Article  PubMed  CAS  Google Scholar 

  • Borevitz JO, Xia Y, Blount J, Dixon RA, Lamb C (2000) Activation tagging identifies a conserved MYB regulator of phenylpropanoid biosynthesis. Plant Cell 12:2383–2394

    Article  PubMed  CAS  Google Scholar 

  • Bousquet J, Simon L, Lalonde M (1990) DNA amplification from vegetative and sexual tissues of trees using polymerase chain reaction. Can J For Res 20:254–257

    Article  CAS  Google Scholar 

  • Butland SL, Chow ML, Ellis BE (1998) A diverse family of phenylalanine ammonia-lyase genes expressed in pine trees and cell cultures. Plant Mol Biol 37:15–24

    Article  PubMed  CAS  Google Scholar 

  • Campbell MM, Ellis BE (1992) Fungal elicitor-mediated responses in pine cell cultures: III. Purification and characterization of phenylalanine ammonia-lyase. Plant Physiol 98:62–70

    Article  PubMed  CAS  Google Scholar 

  • Campbell MM, Sederoff RR (1996) Variation in lignin content and composition (mechanisms of control and implications for the genetic improvement of plants). Plant Physiol 110:3–13

    PubMed  CAS  Google Scholar 

  • Chen C, Meyermans H, Burggraeve B, De Rycke RM, Inoue K, De Vleesschauwer V, Steenackers M, Van Montagu MC, Engler GJ, Boerjan WA (2000) Cell-specific and conditional expression of caffeoyl-coenzyme A-3-O-methyltransferase in poplar. Plant Physiol 123:853–867

    Article  PubMed  CAS  Google Scholar 

  • Cochrane FC, Davin LB, Lewis NG (2004) The Arabidopsis phenylalanine ammonia lyase gene family: kinetic characterization of the four PAL isoforms. Phytochemistry 65:1557–1564

    Article  PubMed  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • de Pater S, Katagiri F, Kijne J, Chua NH (1994) bZIP proteins bind to a palindromic sequence without an ACGT core located in a seed-specific element of the pea lectin promoter. Plant J 6:133–140

    Article  PubMed  CAS  Google Scholar 

  • Douglas CJ (1996) Phenylpropanoid metabolism and lignin biosynthesis: from weeds o trees. Trends Plant Sci 1:171–178

    Article  Google Scholar 

  • Grotewold E, Drummond BJ, Bowen B, Peterson T (1994) The myb-homologous P gene controls phlobaphene pigmentation in maize floral organs by directly activating a flavonid biosynthetic gene subset. Cell 76:543–553

    Article  PubMed  CAS  Google Scholar 

  • Hahlbrock K, Scheel D (1989) Physiology and molecular biology of phenylpropanoid metabolism. Annu Rev Plant Mol Biol 40:347–369

    Article  CAS  Google Scholar 

  • Hatton D, Sablowski R, Yung MH, Smith C, Schuch W, Bevan M (1995) Two class of cis sequences contribute to tissue-specific expression of a PAL2 promoter in transgenic tobacco. Plant J 7:859–876

    Article  PubMed  CAS  Google Scholar 

  • Hauffe KD, Lee SP, Subramaniam R, Douglas CJ (1993) Combinatorial interactions between positive and negative cis-acting elements control spatial patterns of 4CL-1 expression in transgenic tobacco. Plant J 4:235–253

    Article  PubMed  CAS  Google Scholar 

  • Jefferson RA (1988) Plant reporter genes: the GUS gene fusion system. Genet Eng 10:247–263

    CAS  Google Scholar 

  • Jin H, Cominelli E, Bailey P, Parr A, Mehrtens F, Jones J, Tonelli C, Weisshaar B, Martin C (2000) Transcriptional repression by AtMYB4 controls production of UV-protecting sunscreens in Arabidopsis. EMBO J 19:6150–6161

    Article  PubMed  CAS  Google Scholar 

  • Joos HJ, Hahlbrock K (1992) Phenylalanine ammonia-lyase in potato (Solanum tuberosum L.). Genomic complexity, structural comparison of two selected genes and modes of expression. Eur J Biochem 204:621–629

    Article  PubMed  CAS  Google Scholar 

  • Kao YY, Harding SA, Tsai CJ (2002) Differential expression of two distinct phenylalanine ammonia-lyase genes in condensed tannin-accumulating and lignifying cells of quaking aspen. Plant Physiol 130:796–807

    Article  PubMed  Google Scholar 

  • Karpinska B, Karlsson M, Srivastava M, Stenberg A, Schrader J, Sterky F, Bhalerao R, Wingsle G (2004) MYB transcription factors are differentially expressed and regulated during secondary vascular tissue development in hybrid aspen. Plant Mol Biol 56:255–270

    Article  PubMed  CAS  Google Scholar 

  • Kumar A, Ellis BE (2001) The phenylalanine ammonia-lyase gene family in raspberry. Structure, expression, and evolution. Plant Physiol 127:230–239

    Article  PubMed  CAS  Google Scholar 

  • Lacombe E, Van Doorsselaere J, Boerjan W, Boudet AM, Grima-Pettenati J (2000) Characterization of cis-elements required for vascular expression of the cinnamoyl CoA reductase gene and for protein-DNA complex formation. Plant J 23:663–676

    Article  PubMed  CAS  Google Scholar 

  • Lauvergeat V, Rech P, Jauneau A, Guez C, Coutos-Thevenot P, Grima-Pettenati J (2002) The vascular expression pattern directed by the Eucalyptus gunnii cinnamyl alcohol dehydrogenase EgCAD2 promoter is conserved among woody and herbaceous plant species. Plant Mol Biol 50:497–509

    Article  PubMed  CAS  Google Scholar 

  • Lewis NG, Yamamoto E (1990) Lignin: occurrence, biogenesis and biodegradation. Annu Rev Plant Physiol Plant Mol Biol 41:455–496

    Article  PubMed  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  • Li L, Osakabe Y, Joshi CP, Chiang VL (1999) Secondary xylem-specific expression of caffeoyl-coenzyme A 3-O-methyltransferase plays an important role in the methylation pathway associated with lignin biosynthesis in loblolly pine. Plant Mol Biol 40:555–565

    Article  PubMed  CAS  Google Scholar 

  • Logemann E, Parniske M, Hahlbrock K (1995) Modes of expression and common structural features of the complete phenylalanine ammonia-lyase gene in parsley. Proc Natl Acd Sci USA 92:5905–5909

    Article  CAS  Google Scholar 

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

    PubMed  CAS  Google Scholar 

  • Martin C, Paz-Ares J (1997) MYB transcription factors in plants. Trends Genet 13:67–73

    Article  PubMed  CAS  Google Scholar 

  • Moyano E, Martinez-Garcia JF, Martin C (1996) Apparent redundancy in myb gene function provides gearing for the control of flavonoid biosynthesis in antirrhinum flowers. Plant Cell 8:1519–1532

    Article  PubMed  CAS  Google Scholar 

  • Nakashima J, Awano T, Takabe K, Fujita M, Saiki H (1997) Immunocytochemical localization of phenylalanine ammonia-lyase and cinnamyl alcohol dehydrogenase in differentiating tracheary elements derived from Zinnia mesophyll cells. Plant Cell Physiol 38:113–123

    CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  • Osakabe Y, Osakabe K, Kawai S, Katayama Y, Morohoshi N (1995) Characterization of the structure and determination of mRNA levels of the phenylalanine ammonia-lyase gene family from Populus kitakamiensis. Plant Mol Biol 28:1133–1141

    Article  PubMed  CAS  Google Scholar 

  • Osakabe Y, Nanto K, Kitamura H, Kawai S, Kondo Y, Fujii T, Takabe K, Katayama Y, Morohoshi N (1996) Immunocytochemical localization of phenylalanine ammonia-lyase in tissues of Populus kitakamiensis. Planta 200:13–19

    Article  PubMed  CAS  Google Scholar 

  • Osakabe Y, Osakabe K, Chiang VL (2009) Isolation of 4-coumarate Co-A ligase gene promoter from loblolly pine (Pinus taeda) and characterization of the tissue-specific activity in transgenic tobacco (in submission)

  • Patzlaff A, McInnis S, Courtenay A, Surman C, Newman LJ, Smith C, Bevan MW, Mansfield S, Whetten RW, Sederoff RR, Campbell MM (2003a) Characterisation of a pine MYB that regulates lignification. Plant J 36:743–754

    Article  PubMed  CAS  Google Scholar 

  • Patzlaff A, Newman LJ, Dubos C, Whetten RW, Smith C, McInnis S, Bevan MW, Sederoff RR, Campbell MM (2003b) Characterisation of PtMYB1, an R2R3-MYB from pine xylem. Plant Mol Biol 53:597–608

    Article  PubMed  CAS  Google Scholar 

  • Pellegrini L, Rohfritsch O, Fritig B, Legrand M (1994) 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 106:877–886

    Article  PubMed  CAS  Google Scholar 

  • Reinold S, Hahlbrock K (1997) In situ localization of phenylpropanoid biosynthetic mRNAs and proteins in parsley (Petroselinum crispum). Bot Acta 110:431–443

    CAS  Google Scholar 

  • Sablowski RW, Moyano E, Culianez-Macia FA, Schuch W, Martin C, Bevan M (1994) A flower-specific Myb protein activates transcription of phenylpropanoid biosynthetic genes. EMBO J 13:128–137

    PubMed  CAS  Google Scholar 

  • Sainz MB, Grotewold E, Chandler VL (1997) Evidence for direct activation of an anthocyanin promoter by the maize C1 protein and comparison of DNA binding by related Myb domain proteins. Plant Cell 9:611–625

    Article  PubMed  CAS  Google Scholar 

  • Seguin A, Laible G, Leyva A, Dixon RA, Lamb CJ (1997) Characterization of a gene encoding a DNA-binding protein that interacts in vitro with vascular specific cis elements of the phenylalanine ammonia-lyase promoter. Plant Mol Biol 35:281–291

    Article  PubMed  CAS  Google Scholar 

  • Stracke R, Werber M, Weisshaar B (2001) The R2R3-MYB gene family in Arabidopsis thaliana. Curr Opin Plant Biol 4:447–456

    Article  PubMed  CAS  Google Scholar 

  • Subramaniam R, Reinold S, Molitor EK, Douglas CJ (1993) Structure, inheritance, and expression of hybrid poplar (Populus trichocarpa × Populus deltoides) phenylalanine ammonia-lyase genes. Plant Physiol 102:71–83

    Article  PubMed  CAS  Google Scholar 

  • Sugimoto K, Takeda S, Hirochika H (2000) MYB-related transcription factor NtMYB2 induced by wounding and elicitors is a regulator of the tobacco retrotransposon Tto1 and defense-related genes. Plant Cell 12:2511–2528

    Article  PubMed  CAS  Google Scholar 

  • Tamagnone L, Merida A, Parr A, Mackay S, Culianez-Macia FA, Roberts K, Martin C (1998) The AmMYB308 and AmMYB330 transcription factors from antirrhinum regulate phenylpropanoid and lignin biosynthesis in transgenic tobacco. Plant Cell 10:135–154

    Article  PubMed  CAS  Google Scholar 

  • Timell TE (1986) Compression wood in gymnosperms. Springer, Heidelberg, pp 360–379

    Google Scholar 

  • Uimari A, Strommer J (1997) Myb26: a MYB-like protein of pea flowers with affinity for promoters of phenylpropanoid genes. Plant J 12:1273–1284

    Article  PubMed  CAS  Google Scholar 

  • Vanholme R, Morreel K, Ralph J, Boerjan W (2008) Lignin engineering. Curr Opin Plant Biol 11:278–285

    Article  PubMed  CAS  Google Scholar 

  • Wagner DB, Furnier GR, Saghai-Maroof MA, Williams SM, Dancik BP, Allard RW (1987) Chloroplast DNA polymorphism in lodgepole and jack pines and their hybrids. Proc Natl Acad Sci USA 84:2097–2100

    Article  PubMed  CAS  Google Scholar 

  • Weisshaar B, Armstrong GA, Block A, e Silva O, Hahlbrock K (1991) Light-inducible and constitutively expressed DNA-binding proteins recognizing a plant promoter elements with functional relevance in light responsiveness. EMBO J 10:1777–1786

    PubMed  CAS  Google Scholar 

  • Whetten RW, Sederoff RR (1992) Phenylalanine ammonia-lyase from loblolly pine: purification of the enzyme and isolation of complementary DNA clones. Plant Physiol 98:380–386

    Article  PubMed  CAS  Google Scholar 

  • Whetten R, Sun Y-H, Zhang Y, Sederoff R (2001) Functional genomics and cell wall biosynthesis in loblolly pine. Plant Mol Biol 47:275–291

    Article  PubMed  CAS  Google Scholar 

  • Yang S, Sweetman JP, Amirsadeghi S, Barghchi M, Huttly AK, Chung WI, Twell D (2001) Novel anther-specific myb genes from tobacco as putative regulators of phenylalanine ammonia-lyase expression. Plant Physiol 126:1738–1753

    Article  PubMed  CAS  Google Scholar 

  • Zhang X-H, Chiang VL (1997) Molecular cloning and 4-coumarate: coenzyme a ligase in loblolly pine and the roles of this enzyme in the biosynthesis of lignin in compression wood. Plant Physiol 113:65–74

    Article  PubMed  CAS  Google Scholar 

  • Zhong R, Ye Z-H (2007) Regulation of cell wall biosynthesis. Curr Opin Plant Biol 10:564–572

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

The authors like to thank Dr. A. Kawaoka, Dr. L. Li, and Dr. F. Qin for the critical comments on the manuscript. The project was supported by a grant from the ArborGen, LLC.

Author information

Author notes

  1. Yuriko Osakabe

    Present address: Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-Ku, Tokyo, 113-8657, Japan

  2. Keishi Osakabe

    Present address: Department of Plant Biotechnology, National Institute of Agrobiological Sciences, 2-1-2 Kannondai, Tsukuba, Ibaraki, 305-8602, Japan

Authors and Affiliations

  1. Plant Biotechnology Research Center, School of Forestry and Wood Products, Michigan Technological University, 1400 Townsend Drive, Houghton, MI, 49931, USA

    Yuriko Osakabe, Keishi Osakabe & Vincent L. Chiang

  2. Forest Biotechnology Group, Department of Forestry and Environmental Resources, North Carolina State University, Raleigh, NC, 27695, USA

    Vincent L. Chiang

Authors
  1. Yuriko Osakabe
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Keishi Osakabe
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Vincent L. Chiang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Yuriko Osakabe.

Additional information

Communicated by H. Ebinuma.

The nucleotide sequence data reported will appear in the EMBL, GenBank, and DDBJ Nucleotide Sequence Databases under the accession number AB449103 (PtaPAL promoter sequence).

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Osakabe, Y., Osakabe, K. & Chiang, V.L. Characterization of the tissue-specific expression of phenylalanine ammonia-lyase gene promoter from loblolly pine (Pinus taeda) in Nicotiana tabacum . Plant Cell Rep 28, 1309–1317 (2009). https://doi.org/10.1007/s00299-009-0707-1

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00299-009-0707-1

Keywords

  • PtaPAL
  • Lignin
  • Pinus taeda
  • AC element

Search

Navigation