Physiology and Molecular Biology of Plants

, Volume 20, Issue 2, pp 151–159 | Cite as

Arabidopsis mutant plants with diverse defects in polyamine metabolism show unequal sensitivity to exogenous cadaverine probably based on their spermine content

  • Taibo Liu
  • Hayato Dobashi
  • Dong Wook Kim
  • G. H. M. Sagor
  • Masaru Niitsu
  • Thomas Berberich
  • Tomonobu KusanoEmail author
Research Article


Arabidopsis plants do not synthesize the polyamine cadaverine, a five carbon-chain diamine and structural analog of putrescine. Mutants defective in polyamine metabolic genes were exposed to exogenous cadaverine. Spermine-deficient spms mutant grew well while a T-DNA insertion mutant (pao4-1) of polyamine oxidase (PAO) 4 was severely inhibited in root growth compared to wild type (WT) or other pao loss-of-function mutants. To understand the molecular basis of this phenomenon, polyamine contents of WT, spms and pao4-1 plants treated with cadaverine were analyzed. Putrescine contents increased in all the three plants, and spermidine contents decreased in WT and pao4-1 but not in spms. Spermine contents increased in WT and pao4-1. As there were good correlations between putrescine (or spermine) contents and the degree of root growth inhibition, effects of exogenously added putrescine and spermine were examined. Spermine mimicked the original phenomenon, whereas high levels of putrescine evenly inhibited root growth, suggesting that cadaverine-induced spermine accumulation may explain the phenomenon. We also tested growth response of cadaverine-treated WT and pao4-1 plants to NaCl and found that spermine-accumulated pao4-1 plant was not NaCl tolerant. Based on the results, the effect of cadaverine on Arabidopsis growth and the role of PAO during NaCl stress are discussed.


Arabidopsis Cadaverine NaCl response Polyamine oxidase Putrescine Spermidine Spermine 



T-Spm synthase


Arginine decarboxylase




Polyamine oxidase






S-adenosylmethionine decarboxylase




Spd synthase




Spm synthase





We thank to Prof. Taku Takahashi for providing us seeds of spms and acl5 mutants. This work was supported in part by Grant-in-Aids from the Japan Society for the Promotion of Science (JSPS) to TK (21380063), by the research funding programme “LOEWE -Landes-Offensive zur Entwicklung Wissenschaftlich-ökonomischer Exzellenz” of Hesse’s Ministry of Higher Education, Research, and the Arts to TB, and by the grants from The Saito Gratitude Foundation to GHMS (2011) and to DWK (2012) and by Sasagawa Scientific Research Grant to DWK. TL is financially supported by China Scholarship Council. GHMS is a recipient of MEXT fellowship.

Supplementary material

12298_2014_227_Fig9_ESM.jpg (126 kb)
Figure S1

Schematic structure of AtPAO4 gene and the T-DNA insertion site in SALK_133599 line. A. The position of T-DNA insertion of pao4-1 (SALK_133599) is shown as a white rectangle. Box, exon; line, intron; white box, untranslated region; black box, coding region. B. Transcript levels of AtPAO4 in WT (Col-0) and pao4-1 plants. AtPAO4 transcripts were detected by qRT-PCR using total RNAs extracted from 14-day-old seedlings and normalized using CBP20 as an internal control. (JPEG 126 kb)

12298_2014_227_MOESM1_ESM.docx (16 kb)
Table S1 The primers used for qRT-PCR analysis. (DOCX 16 kb)


  1. Alcázar R, Altabella T, Marco F, Bortolotti C, Reymond M, Koncz C, Carrasco P, Tiburcio AF (2010) Polyamines: molecules with regulatory functions in plant abiotic stress tolerance. Planta 231:1237–1249PubMedCrossRefGoogle Scholar
  2. Bagni N, Tassoni A (2001) Biosynthesis, oxidation and conjugation of aliphatic polyamines in higher plants. Amino Acids 20:301–317PubMedCrossRefGoogle Scholar
  3. Binda C, Cona A, Angelini R, Federico R, Ascenzi P, Mattevi A (1999) A 30-angstrom-long U-shaped catalytic tunnel in the crystal structure of polyamine oxidase. Structure 7:265–276PubMedCrossRefGoogle Scholar
  4. Bunsupa S, Katayama K, Ikeura E, Oikawa A, Toyooka K, Saito K, Yamazaki M (2012) Lysine decarboxylase catalyzes the first step of quinolizidine alkaloid biosynthesis and coevolved with alkaloid production in luguminosae. Plant Cell 24:1202–1216PubMedCentralPubMedCrossRefGoogle Scholar
  5. Cervelli M, Polticelli F, Federico R, Mariottini P (2003) Heterologous expression and characterization of mouse spermine oxidase. J Biol Chem 278:271–5276CrossRefGoogle Scholar
  6. Cervelli M, Bianchi M, Cona A, Crosatti C, Stanca M, Angelini R, Federico R, Mariottini P (2006) Barley polyamine oxidase isoforms 1 and 2, a peculiar case of gene duplication. FEBS J 273:3990–4002PubMedCrossRefGoogle Scholar
  7. Cohen SS (1998) A guide to the polyamines. Oxford University Press, New YorkGoogle Scholar
  8. Cona A, Moreno S, Cenci F, Federico R, Angelini R (2005) Cellular re-distribution of flavin-containing polyamine oxidase in differentiating root and mescotyl of Zea mays L. seedlings. Planta 221:265–276PubMedCrossRefGoogle Scholar
  9. Cona A, Rea G, Angelini R, Federico R, Tavladoraki P (2006) Functions of amine oxidases in plant development and defence. Trends Plant Sci 11:80–88PubMedCrossRefGoogle Scholar
  10. Federico R, Cona A, Angelini R, Schininà ME, Giartosio A (1990) Characterization of maize polyamine oxidase. Phytochemistry 29:2411–2414PubMedCrossRefGoogle Scholar
  11. Fincato P, Moschou PN, Spedaletti V, Tavazza R, Angelini R, Federico R, Roubelakis-Angelakis KA, Tavladoraki P (2011) Functional diversity inside the Arabidopsis polyamine oxidase gene family. J Exp Bot 62:1155–1168PubMedCrossRefGoogle Scholar
  12. Fryer MJ, Ball L, Oxborough K, Karpinski S, Mullineaux PM, Baker NR (2003) Control of Ascorbate Peroxidase2 expression by hydrogen peroxide and leaf water status during excess light stress reveals a functional organisation of Arabidopsis leaves. Plant J 33:691–705PubMedCrossRefGoogle Scholar
  13. Fuell C, Elliot KA, Hanfrey CC, Franceschetti M, Michael AJ (2010) Polyamine biosynthetic diversity in plants and algae. Plant Physiol Biochem 48:513–520PubMedCrossRefGoogle Scholar
  14. Gamarnik A, Frydman RB (1991) Cadaverine, an essential diamine for the normal root development of germinating soybean (Glycine max) seeds. Plant Physiol 97:778–785PubMedCentralPubMedCrossRefGoogle Scholar
  15. Groppa MD, Benavides MP (2007) Polyamines and abiotic stress: recent advances. Amino Acids 34:35–45PubMedCrossRefGoogle Scholar
  16. Kakehi JI, Kuwashiro Y, Niitsu M, Takahashi Y (2008) Thermospermine is required for stem elongation in Arabidopsis thaliana. Plant Cell Physiol 49:1342–1349PubMedCrossRefGoogle Scholar
  17. 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 49:1272–1282PubMedCrossRefGoogle Scholar
  18. Knott JM, Romer P, Sumper M (2007) Putative spermine synthases from Thalassiosira pseudonana and Arabidopsis thaliana synthesize thermospermine rather than spermine. FEBS Lett 58:3081–3086CrossRefGoogle Scholar
  19. Kusano T, Berberich T, Tateda C, Takahashi Y (2008) Polyamines: essential factors for growth and survival. Planta 228:367–381PubMedCrossRefGoogle Scholar
  20. Kuznetsov V, Shorina M, Aronova E, Stetsenko L, Rakitin V, Shevyakova N (2007) NaCl- and ethylene-dependent cadaverine accumulation and its possible protective role in the adaptation of the common ice plant to salt stress. Plant Sci 172:363–370CrossRefGoogle Scholar
  21. Kuznetsov VV, Stetsenko LA, Shevyakova NI (2009) Exogenous cadaverine induces oxidative burst and reduces cadaverine conjugate content in the common ice plant. J Plant Physiol 166:40–51PubMedCrossRefGoogle Scholar
  22. 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 147:1845–1857PubMedCentralPubMedCrossRefGoogle Scholar
  23. Naka Y, Watanabe K, Sagor GHM, Niitsu M, Pillai A, Kusano T, Takahashi Y (2010) Quantitative analysis of plant polyamines including thermospermine during growth and salinity stress. Plant Physiol Biochem 48:527–533PubMedCrossRefGoogle Scholar
  24. Niitsu M, Samejima K (1986) Syntheses of a series of linear pentaamines with three and four methylene chain intervals. Chem Pharm Bull 34:1032–1038CrossRefGoogle Scholar
  25. Ono Y, Kim DW, Watanabe K, Sasaki A, Niitsu M, Berberich T, Kusano T, Takahashi Y (2012) Constitutively and highly expressed Oryza sativa polyamine oxidases localize in peroxisomes and catalyze polyamine back conversion. Amino Acids 42:867–876PubMedCrossRefGoogle Scholar
  26. Samejima K, Takeda Y, Kawase M, Okada M, Kyogoku Y (1984) Syntheses of 15N-enriched polyamines. Chem Pharm Bull 32:3428–3435PubMedCrossRefGoogle Scholar
  27. Seiler N (2004) Catabolism of polyamines. Amino Acids 26:217–233PubMedGoogle Scholar
  28. Shevyakova NI, Rakitin VY, Dam DB, Kuznetsov VV (2000) Cadaverine as a signal of heat shock in plants. Dokl Biol Sci 375:657–659PubMedCrossRefGoogle Scholar
  29. Tabor CW, Tabor H (1984) Polyamines. Annu Rev Biochem 53:749–790PubMedCrossRefGoogle Scholar
  30. Takahashi Y, Cong R, Sagor GHM, Niitsu M, Berberich T, Kusano T (2010) Characterization of five polyamine oxidase isoforms in Arabidopsis thaliana. Plant Cell Rep 29:955–965PubMedCrossRefGoogle Scholar
  31. Takano A, Kakehi J, Takahashi T (2012) Thermospermine is not a minor polyamine in the plant kingdom. Plant Cell Physiol 53:606–616PubMedCrossRefGoogle Scholar
  32. Tavladoraki P, Schininà 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 426:62–66PubMedCrossRefGoogle Scholar
  33. Tavladoraki P, Rossi MN, Saccut G, Perez-Amador MA, Polticelli F, Angelini R, Ferderico R (2006) Heterologous expression and biochemical characterization of a polyamine oxidase from Arabidopsis involved in polyamine back conversion. Plant Physiol 141:1519–1532PubMedCentralPubMedCrossRefGoogle Scholar
  34. Thordal-Christensen H, Zhang Z, Wei Y, Collinge DB (1997) Subcellular localization of H2O2 in plants: H2O2 accumulation in papillae and hypersensitive response during the barley powdery mildew interaction. Plant J 11:1187–1194CrossRefGoogle Scholar
  35. Vujcic S, Diegelmann P, Bacchi CJ, Kramer DL, Porter CW (2002) Identification and characterization of a novel flavin-containing spermine oxidase of mammalian cell origin. Biochem J 367:665–675PubMedCentralPubMedCrossRefGoogle Scholar
  36. Vujcic S, Liang P, Diegelmann P, Kramer DL, Porter CW (2003) Genomic identification and biochemical characterization of the mammalian polyamine oxidase involved in polyamine back-conversion. Biochem J 370:19–28PubMedCentralPubMedCrossRefGoogle 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, Mclntire WS (2003) Cloning, sequencing and heterologous expression of the murine poroxisomal flavoprotein, N1-acetylated polyamine oxidase. J Biol Chem 278:20514–20525PubMedCrossRefGoogle Scholar
  39. Yamaguchi K, Takahashi Y, Berberich T, Imai A, Miyazaki A, Takahashi T, Michael A, Kusano T (2006) The polyamine spermine protects against high salt stress in Arabidopsis thaliana. FEBS Lett 580:6783–6788PubMedCrossRefGoogle Scholar

Copyright information

© Prof. H.S. Srivastava Foundation for Science and Society 2014

Authors and Affiliations

  • Taibo Liu
    • 1
  • Hayato Dobashi
    • 1
  • Dong Wook Kim
    • 1
  • G. H. M. Sagor
    • 1
  • Masaru Niitsu
    • 2
  • Thomas Berberich
    • 3
  • Tomonobu Kusano
    • 1
    Email author
  1. 1.Graduate School of Life SciencesTohoku UniversitySendaiJapan
  2. 2.Faculty of Pharmaceutical SciencesJosai UniversitySakadoJapan
  3. 3.Biodiversity and Climate Research Center (BiK-F)Frankfurt am MainGermany

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