Plant Cell Reports

, Volume 29, Issue 9, pp 955–965 | Cite as

Characterization of five polyamine oxidase isoforms in Arabidopsis thaliana

  • Yoshihiro TakahashiEmail author
  • Runzi Cong
  • G. H. M. Sagor
  • Masaru Niitsu
  • Thomas Berberich
  • Tomonobu KusanoEmail author
Original Paper


The genome of Arabidopsis thaliana contains five genes (AtPAO1 to AtPAO5) encoding polyamine oxidase (PAO) which is an enzyme responsible for polyamine catabolism. To understand the individual roles of the five AtPAOs, here we characterized their tissue-specific and space-temporal expression. AtPAO1 seems to have a specific function in flower organ. AtPAO2 was expressed in shoot meristem and root tip of seedlings, and to a higher extent in the later growth stage within restricted parts of the organs, such as shoot meristem, leaf petiole and also in anther. The expression of AtPAO3 was constitutive, but highest in flower organ. AtPAO3 promoter activity was detected in cotyledon, distal portion of root, boundary region of mature rosette leaf and in filaments of flower. AtPAO4 was expressed at higher level all over young seedlings including roots, and in the mature stage its expression was ubiquitous with rather lower level in stem. AtPAO5 expression was observed in the whole plant body throughout various growth stages. Its highest expression was in flowers, particularly in sepals, but not in petals. Furthermore, we determined the substrate specificity of AtPAO1 to AtPAO4. None of the AtPAO enzymes recognized putrescine (Put). AtPAO2 and AtPAO3 showed almost similar substrate recognition patterns in which the most preferable substrate is spermidine (Spd) followed by less specificity to other tetraamines tested. AtPAO4 seemed to be spermine (Spm)-specific. More interestingly, AtPAO1 preferred thermospermine (T-Spm) and norspermine (NorSpm) to Spm, but did not recognize Spd. Based on the results, the individual function of AtPAOs is discussed.


A. thaliana Gene expression Polyamine oxidase Space-temporal expression Substrate specificity 



Amine oxidase






Polyamine oxidase


Polymerase chain reaction











This work was supported in part by Grant-in-Aids from the Japan Society for the Promotion of Science (JSPS) to TK (21380063) and YT (21780087). TB was supported by a JSPS-BRIDGE fellowship (BR090101).


  1. Alcázar R, Marco F, Cuevas JC, Patron M, Ferrando A, Carrasco P, Tiburcio AF, Altabella T (2006) Involvement of polyamines in plant response to abiotic stress. Biotechnol Lett 28:1867–1876CrossRefPubMedGoogle Scholar
  2. Bagni N, Tassoni A (2001) Biosynthesis, oxidation and conjugation of aliphatic polyamines in higher plants. Amino Acids 20:301–317CrossRefPubMedGoogle Scholar
  3. Binda C, Coda A, Angelini R, Federico R, Ascenzi P, Mattevi A (1999) A 30-Å-long U-shaped catalytic tunnel in the crystal structure of polyamine oxidase. Structure 7:265–276CrossRefPubMedGoogle Scholar
  4. Bolenius FN, Seiler N (1981) Acetyl derivatives as intermediates in polyamine catabolism. Int J Biochem 13:287–292CrossRefGoogle Scholar
  5. Bouchereau A, Aziz A, Larher F, Martin TJ (1999) Polyamines and environmental challenges: recent development. Plant Sci 140:103–125CrossRefGoogle Scholar
  6. Cervelli M, Tavladoraki P, Di Agostino S, Angelini R, Federico R, Mariottini P (2000) Isolation and characterization of three polyamine oxidase genes from Zea mays. Plant Physiol Biochem 38:667–677CrossRefGoogle Scholar
  7. Cervelli M, Cona A, Angelini R, Polticelli F, Federico R, Mariottini P (2001) A barley polyamine oxidase isoform with distinct structural features and subcellular localization. Eur J Biochem 268:3816–3830CrossRefPubMedGoogle Scholar
  8. Cervelli M, Polticelli F, Federico R, Mariottini P (2003) Heterologous expression and characterization of mouse spermine oxidase. J Biol Chem 278:5271–5276CrossRefPubMedGoogle Scholar
  9. Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16:735–743CrossRefPubMedGoogle Scholar
  10. Cohen SS (1998) A guide to the polyamines. Oxford University Press, OxfordGoogle Scholar
  11. Cona A, Moreno S, Cenci F, Federico R, Angelini R (2005) Cellular redistribution of flavin-containing polyamine oxidase in differentiating root and mesocotyl of Zea mays L. seedlings. Planta 221:265–276CrossRefPubMedGoogle Scholar
  12. Cona A, Rea G, Angelini R, Federico R, Tavladoraki P (2006) Functions of amine oxidases in plant development and defence. Trends Plant Sci 11:80–88CrossRefPubMedGoogle Scholar
  13. Del Duca S, Beninati S, Serafini-Fracassini D (1995) Polyamines in chloroplasts: identification of their glutamyl and acetyl derivatives. Biochem J 305:233–237PubMedGoogle Scholar
  14. Federico R, Angelini R (1991) Polyamine catabolism in plants. In: Slocum RD, Flores HE (eds) Biochemistry and physiology of polyamines in plants. CRC Press, Boca Raton, pp 41–56Google Scholar
  15. Fuell C, Elliot KA, Hanfrey CC, Franceschetti M, Michael AJ (2010) Polyamine biosynthetic diversity in plants and algae. Plant Physiol Biochem (in press)Google Scholar
  16. Groppa MD, Benavides MP (2007) Polyamines and abiotic stress: recent advances. Amino Acids 34:35–45CrossRefPubMedGoogle Scholar
  17. Hanzawa Y, Takahashi T, Michael AJ, Burtin D, Long D, Pineiro M, Coupland G, Komeda Y (2000) ACAULIS5, an Arabidopsis gene required for stem elongation, encodes a spermine synthase. EMBO J 19:4248–4256CrossRefPubMedGoogle Scholar
  18. Jefferson RA (1987) Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol Biol Rep 5:387–405CrossRefGoogle Scholar
  19. Kakehi J, Kuwashiro Y, Niitsu M, Takahashi T (2008) Thermospermine is required for stem elongation in Arabidopsis thaliana. Plant Cell Physiol 49:1342–1349CrossRefPubMedGoogle Scholar
  20. Kamada-Nobusada T, Hayashi M, Fukazawa M, Sakakibara H, Nishimura M (2008) A putative peroxisomal polyamine oxidase, AtPAO4, is involved in polyamine catabolism in Arabidopsis thaliana. Plant Cell Physiol 49:1272–1282CrossRefPubMedGoogle Scholar
  21. Knott JM, Romer P, Sumper M (2007) Putative spermine synthases from Thalassiosira pseudonana and Arabidopsis thaliana synthesize thermospermine rather than spermine. FEBS Lett 581:3081–3086CrossRefPubMedGoogle Scholar
  22. Kusano T, Berberich T, Tateda C, Takahashi Y (2008) Polyamines: essential factors for growth and survival. Planta 228:367–381CrossRefPubMedGoogle Scholar
  23. Lim TS, Chitra TR, Han P, Pua EC, Yu H (2006) Cloning and characterization of Arabidopsis and Brassica juncea flavin-containing amine oxidases. J Exp Bot 57:4155–4169CrossRefPubMedGoogle Scholar
  24. Moschou PN, Sanmartin M, Andriopoulou AH, Rojo E, Sanchez-Serrano JJ, Roubelakis-Angelakis KA (2008) Bridging the gap between plant and mammalian polyamine catabolism: a novel peroxisomal polyamine oxidase responsible for a full back-conversion pathway in Arabidopsis. Plant Physiol 147:1845–1857CrossRefPubMedGoogle Scholar
  25. Naka Y, Watanabe K, Sagor G.H.M, Niitsu M, Pillai A, Kusano T, Takahashi Y (2010) Quantitative analysis of plant polyamines including thermospermine during growth and salinity stress. Plant Physiol Biochem (in press)Google Scholar
  26. Page RD (1996) TreeView: an application to display phylogenetic trees on personal computers. Comput Appl Biosci 12:357–358PubMedGoogle Scholar
  27. Seiler N (2004) Catabolism of polyamines. Amino Acids 26:217–233PubMedGoogle Scholar
  28. Tabor CW, Tabor H (1985) Polyamines in microorganisms. Microbiol Rev 49:81–99PubMedGoogle Scholar
  29. Tassoni A, Van Buuren M, Franceschetti M, Fornale IS, Bagni N (2000) Polyamine content and metabolism in Arabidopsis thaliana and effect of spermidine on plant development. Plant Physiol Biochem 38:383–393CrossRefGoogle Scholar
  30. Tavladoraki P, Shinina ME, Cecconi F, Di Agostino S, Manera F, Rea G, Mariottini P, Federico R, Angelini R (1998) Maize polyamine oxidase: primary structure from protein and cDNA sequencing. FEBS Lett 426:62–66CrossRefPubMedGoogle Scholar
  31. Tavladoraki P, Rossi MN, Saccuti G, Perez-Amador MA, Polticelli F, Angelini R, Federico R (2006) Heterologous expression and biochemical characterization of a polyamine oxidase from Arabidopsis involved in polyamine back conversion. Plant Physiol 141:1519–1532CrossRefPubMedGoogle Scholar
  32. Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acid Res 22:4673–4680CrossRefPubMedGoogle Scholar
  33. Vujcic S, Diegelman P, Bacchi CJ, Kramer DL, Porter CW (2002) Identification and characterization of a novel flavin-containing spermine oxidase of mammalian cell origin. Biochem J 367:665–675CrossRefPubMedGoogle Scholar
  34. Vujcic S, Liang P, Diegelman P, Kramer DL, Porter CW (2003) Genomic identification and biochemical characterization of the mammalian polyamine oxidase involved in polyamine back-conversion. Biochem J 370:19–28CrossRefPubMedGoogle Scholar
  35. Wallace HM, Fraser AV, Hughes A (2003) A perspective of polyamine metabolism. Biochem J 376:1–14CrossRefPubMedGoogle Scholar
  36. Walters DR (2003) Polyamines and plant disease. Phytochemistry 64:97–107CrossRefPubMedGoogle Scholar
  37. Wang Y, Devereux W, Woster PM, Stewart TM, Hacker A, Casero RA Jr (2001) Cloning and characterization of a human polyamine oxidase that is inducible by polyamine analogue exposure. Cancer Res 61:5370–5373PubMedGoogle Scholar
  38. Wu T, Yankovskaya V, McIntire WS (2003) Cloning, sequencing, and heterologous expression of the murine peroxisomal flavoprotein, N 1-acetylated polyamine oxidase. J Biol Chem 278:20514–20525CrossRefPubMedGoogle Scholar
  39. Yoda H, Hiroi Y, Sano H (2006) Polyamine oxidase is one of the key elements for oxidative burst to induce programmed cell death in tobacco cultured cells. Plant Physiol 142:193–206CrossRefPubMedGoogle Scholar
  40. Yoda Y, Fujimura K, Takahashi H, Munemura I, Uchimiya H, Sano H (2009) Polyamines as a common source of hydrogen peroxidein host- and nonhost hypersensitive response during pathogen infection. Plant Mol Biol 70:103–112CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Yoshihiro Takahashi
    • 1
    Email author
  • Runzi Cong
    • 1
  • G. H. M. Sagor
    • 1
  • Masaru Niitsu
    • 2
  • Thomas Berberich
    • 1
    • 3
  • Tomonobu Kusano
    • 1
    Email author
  1. 1.Laboratory of Plant Molecular and Cellular Biology, Graduate School of Life SciencesTohoku UniversitySendaiJapan
  2. 2.Faculty of Pharmaceutical SciencesJosai UniversitySakadoJapan
  3. 3.Biodiversity and Climate Research Center FrankfurtFrankfurtGermany

Personalised recommendations