Molecular Biology Reports

, Volume 38, Issue 3, pp 1935–1940 | Cite as

Molecular cloning and characterization of a phenylalanine ammonia-lyase gene (LrPAL) from Lycoris radiata

  • Yumei Jiang
  • Nan Xia
  • Xiaodan Li
  • Wenbiao Shen
  • Lijian Liang
  • Chunyan Wang
  • Ren Wang
  • Feng Peng
  • Bing Xia


LrPAL is a novel full-length cDNA isolated from Lycoris radiata by degenerate oligonucleotide primer PCR (DOP-PCR), 3′- and 5′-RACE approaches, harbours an open reading frame (ORF) encoding a 708 amino acid product. Sequence alignment showed that the deduced amino acid sequence of LrPAL shared more than 80% identity with other PAL sequences reported in Arabidopsis thaliana and other plants. RT-PCR revealed that LrPAL transcripts were higher in bud flowers and wilting flowers (5 days after blooming) than in blooming flowers. The transcript levels of LrPAL in leaves were significantly induced by methyl jasmonate (MJ) and nitric oxide (NO), and salicylic acid (SA). Similarly, HPLC analysis showed that galantamine (GAL) content was also higher in bud flowers and wilting flowers than in blooming flowers. The GAL content in leaves was significantly induced by MJ and NO, and inhibited by SA. This study enables us to further elucidate the role of LrPAL in the biosynthesis of GAL in Lycoris radiata at a molecular level.


Phenylalanine ammonia-lyase Galantamine biosynthesis Lycoris radiata 



This study was supported by the National Natural Science Foundation of China (Grant no. 30700057). We wish to thank Dr. Evan Evans from Tasmanian Institute of Agricultural Research at University of Tasmania for critical reading and linguistic help in the preparation of this manuscript.

Supplementary material

11033_2010_314_MOESM1_ESM.doc (422 kb)
(DOC 423 kb)


  1. 1.
    Howes MJR, Houghton PJ (2003) Plants used in Chinese and Indian traditional medicine for improvement of memory and cognitive function. Pharmacol Biochem Behav 75:513–527PubMedCrossRefGoogle Scholar
  2. 2.
    Laurain-Mattar D (2008) Production of alkaloids in plant cell and tissue cultures. In: Ramawat KG, Merillon JM (eds) Bioactive molecules and medicinal plants. Springer, Berlin , pp 165–173CrossRefGoogle Scholar
  3. 3.
    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 106:877–886PubMedCrossRefGoogle Scholar
  4. 4.
    Dixon RA, Paiva NL (1995) Stress-induced phenylpropanoid metabolism. Plant Cell 7:1085–1097PubMedCrossRefGoogle Scholar
  5. 5.
    Liu RR, Xu SH, Li JL, Hu YL, Lin ZP (2006) Expression profile of a PAL gene from Astragalus membranaceus var. Mongholicus and its crucial role in flux into flavonoid biosynthesis. Plant Cell Rep 25:705–710PubMedCrossRefGoogle Scholar
  6. 6.
    Mahesh V, Rakotomalala JJ, Vigne LLH, Kochko A, Noirot SH, Noirot M, Campa C (2006) Isolation and genetic mapping of a Coffea canephora phenylalanine ammonia-lyase gene (CcPAL1) and its involvement in the accumulation of caffeoyl quinic acids. Plant Cell Rep 25:986–992PubMedCrossRefGoogle Scholar
  7. 7.
    Singh K, Kumar S, Gulati ARA, Ahuja PS (2009) Phenylalanine ammonia-lyase (PAL) and cinnamate 4-hydroxylase (C4H) and catechins (flavan-3-ols) accumulation in tea. Funct Integr Genomics 9:125–134PubMedCrossRefGoogle Scholar
  8. 8.
    Chen AH, Chai YR, Li JN, Chen L (2007) Molecular cloning of two genes encoding cinnamate 4-hydroxylase (C4H) from oilseed rape (Brassica napus). J Biochem Mol Biol 40:247–260PubMedGoogle Scholar
  9. 9.
    Barber MS, Mitchell HJ (1997) Regulation of phenylpropanoid metabolism in relation to lignin biosynthesis in plants. Int Rev Cytol 172:243–293CrossRefGoogle Scholar
  10. 10.
    Harakava R (2005) Genes encoding enzymes of the lignin biosynthesis pathway in Eucalyptus. Genet Mol Biol 28:601–607CrossRefGoogle Scholar
  11. 11.
    Song J, Wang Z (2008) Molecular cloning, expression and characterization of a phenylalanine ammonia-lyase gene (SmPAL1) from Salvia miltiorrhiza. Mol Biol Rep 36(5):939–952PubMedCrossRefGoogle Scholar
  12. 12.
    Dixon RA, Lamb CJ, Masoud S, Sewalt VJH, Paiva NL (1996) Metabolic engineering: prospects for crop improvement through the genetic manipulation of phenylpropanoid biosynthesis and defense responses-a review. Gene 179:61–71PubMedCrossRefGoogle Scholar
  13. 13.
    Bate NJ, Orrt J, Nit W, Meromit A, Nadler-hassar T, Doerner PW, Dixon RA, Lamb CJ, Elkind Y (1994) 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 91:7608–7612PubMedCrossRefGoogle Scholar
  14. 14.
    Eichhorn J, Takada T, Kita Y, Zenk MH (1998) Biosynthesis of the Amaryllidaceae alkaloid Galanthamine. Phytochemistry 49:1037–1047CrossRefGoogle Scholar
  15. 15.
    Hahlbrock K, Scheel D (1989) Physiology and molecular biology of phenylpropanoid metabolism. Annu Rev Plant Physiol Plant Mol Biol 40:347–369CrossRefGoogle Scholar
  16. 16.
    Minami E, Ozeki Y, Matsuoka M, Koiruka N, Tanaka Y (1989) Structure and some characterization of the gene for phenylalanine ammonia-lyase from rice plants. Eur J Biochem 185:19–25PubMedCrossRefGoogle Scholar
  17. 17.
    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-acting elements. EMBO J 8:1641–1648PubMedGoogle Scholar
  18. 18.
    Wanner LA, Li G, Ware D, Somssich IE, Davis KR (1995) The phenylalanine ammonialyase gene family in Arabidopsis thaliana. Plant Mol Biol 27:327–338PubMedCrossRefGoogle Scholar
  19. 19.
    Cochrane FC, Davin LB, Lewis NG (2004) The Arabidopsis phenylalanine ammonia lyase gene family: kinetic characterization of the four PAL isoforms. Phytochemistry 65:1557–1564PubMedCrossRefGoogle Scholar
  20. 20.
    Kumar A, Ellis BE (2001) The phenylalanine ammonia-lyase gene family in raspberry: structure, expression, and evolution. Plant Physiol 127:230–239PubMedCrossRefGoogle Scholar
  21. 21.
    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 37:15–24PubMedCrossRefGoogle Scholar
  22. 22.
    Hsieh LS, Hsieh YL, Yeh CS, Cheng CY, Yang CC, Lee PD (2010) Molecular characterization of a phenylalanine ammonia-lyase gene (BoPAL1) from Bambusa oldhamii. Mol Biol Rep. doi:  10.1007/s11033-010-0106-2
  23. 23.
    Colque R, Viladomat F, Bastida J, Codina C (2004) Improved production of galanthamine and related alkaloids by methyl jasmonate in Narcissus confusus shoot-clumps. Planta Med 70:1180–1188PubMedCrossRefGoogle Scholar
  24. 24.
    Li Y, Qi Y, Wu S (2003) RP-HPLC determination of galanthamine hydrobromide and related substances in its oral solution. Chin J Pharm Anal 23:365–367Google Scholar
  25. 25.
    Chaw SM, Zhaekikh A, Sung HM, Lau TC, Li WH (1997) Molecular phylogeny of extant gymnosperms and seed plant evolution: analysis of nuclear 18S rRNA sequences. Mol Biol Evol 14:56–68PubMedGoogle Scholar
  26. 26.
    Okada T, Mikage M, Sekit S (2008) A molecular characterization of the phenylalanine ammonia-lyase from Ephedra sinica. Biol Pharm Bull 31(12):2194–2199PubMedCrossRefGoogle Scholar
  27. 27.
    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 12:367–383CrossRefGoogle Scholar
  28. 28.
    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 130:796–807PubMedCrossRefGoogle Scholar
  29. 29.
    Fukasawa-Akada T, Kung SD, Watson JC (1996) Phenylalanine ammonia-lyase gene structure, expression, and evolution in Nicotiana. Plant Mol Biol 30:711–722PubMedCrossRefGoogle Scholar
  30. 30.
    Nakazawa A, Nozue M, Yasuda H, Takeba G, Kubo H (2001) Expression pattern and gene structure of phenylalanine ammonialyase in Pharbitis nil. J Plant Res 114:323–328CrossRefGoogle Scholar
  31. 31.
    Lawton MA, Lamb CJ (1987) Transcriptional activation of plant defense genes by fungal elicitor, wounding and infection. Mol Cell Biol 7:335–341PubMedGoogle Scholar
  32. 32.
    Leyva A, Jarillo JA, Salinas J, Martinez-Zapater JM (1995) Low temperature induces the accumulation of phenylalanine ammonia-lyase and chalcone synthase mRNAs of Arabidopsis thaliana in a light-dependent manner. Plant Physiol 108:9–46Google Scholar
  33. 33.
    Hu XY, Neill SJ, Cai WM, Tang ZC (2003) Nitric oxide mediates elicitor-induced saponin synthesis in cell cultures of Panax ginseng. Funct Plant Biol 30:901–907CrossRefGoogle Scholar
  34. 34.
    Zheng LP, Wang JW, Guo YT, Tan RX (2008) Nitric oxide potentiates oligosaccharide-induced artemisinin production in Artemisia annua hairy roots. J Integr Plant Biol 50:49–55PubMedCrossRefGoogle Scholar
  35. 35.
    Mu HM, Wang R, Li XD, Jiang YM, Wang CY, Quan JP, Peng F, Xia B (2009) Effect of abiotic and biotic elicitors on growth and alkaloid accumulation of Lycoris chinensis seedlings. Z Naturforsch C 64(7–8):541–550PubMedGoogle Scholar
  36. 36.
    Naoumkina M, Farag MA, Sumner LW, Tang Y, Liu CJ, Dixon RA (2007) Different mechanisms for phytoalexin induction by pathogen and wound signals in Medicago truncatula. Proc Natl Acad Sci USA 104:17909–17915PubMedCrossRefGoogle Scholar
  37. 37.
    Misra P, Pandey A, Tewari S K, Nath P, Trivedi P K (2010) Characterization of isoflavone synthase gene from Psoralea corylifolia: a medicinal plant. Plant Cell Rep. doi: 10.1007/s00299-010-0861-5

Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Yumei Jiang
    • 1
  • Nan Xia
    • 1
    • 2
  • Xiaodan Li
    • 1
  • Wenbiao Shen
    • 3
  • Lijian Liang
    • 1
  • Chunyan Wang
    • 1
  • Ren Wang
    • 1
  • Feng Peng
    • 1
  • Bing Xia
    • 1
  1. 1.Institute of BotanyJiangsu Province and Chinese Academy of SciencesNanjingChina
  2. 2.College of PharmacyNanjing University of Chinese MedicineNanjingChina
  3. 3.College of Life SciencesNanjing Agricultural UniversityNanjingChina

Personalised recommendations