Plant Cell Reports

, Volume 32, Issue 9, pp 1477–1488 | Cite as

Longer uncommon polyamines have a stronger defense gene-induction activity and a higher suppressing activity of Cucumber mosaic virus multiplication compared to that of spermine in Arabidopsis thaliana

  • G. H. M. Sagor
  • Taibo Liu
  • Hideki Takahashi
  • Masaru Niitsu
  • Thomas Berberich
  • Tomonobu KusanoEmail author
Original Paper


Key message

Our work suggests that long chain polyamines and their derivatives are potential chemicals to control viral pathogens for crop production.


Previously we showed that two tetraamines, spermine (Spm) and thermospermine (T-Spm), induce the expression of a subset of defense-related genes and repress proliferation of Cucumber mosaic virus (CMV) in Arabidopsis. Here we tested whether the longer uncommon polyamines (LUPAs) such as caldopentamine, caldohexamine, homocaldopentamine and homocaldohexamine have such the activity. LUPAs had higher gene induction activity than Spm and T-Spm. Interestingly the genes induced by LUPAs could be classified into two groups: the one group was most responsive to caldohexamine while the other one was most responsive to homocaldopentamine. In both the cases, the inducing activity was dose-dependent. LUPAs caused local cell death and repressed CMV multiplication more efficiently as compared to Spm. LUPAs inhibited the viral multiplication of not only avirulent CMV but also of virulent CMV in a dose-dependent manner. Furthermore, LUPAs can activate the systemic acquired resistance against CMV more efficiently as compared to Spm. When Arabidopsis leaves were incubated with LUPAs, the putative polyamine oxidase (PAO)-mediated catabolites were detected even though the conversion rate was very low. In addition, we found that LUPAs induced the expression of three NADPH oxidase genes (rbohC, rbohE and rbohH) among ten isoforms. Taken together, we propose that LUPAs activate two alternative reactive oxygen species evoked pathways, a PAO-mediated one and an NADPH-oxidase-mediated one, which lead to induce defense-related genes and restrict CMV multiplication.


Arabidopsis thaliana Cucumber mosaic virus  Longer uncommon polyamines  Signaling activity  Spermine 



This work was supported in part by Grant-in-Aids from the Japan Society for the Promotion of Science (JSPS) to TK (21380063), and by the grant from The Saito Gratitude Foundation (to GHMS) and 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). GHMS is a recipient of MEXT scholarship.

Supplementary material

299_2013_1459_MOESM1_ESM.docx (17 kb)
Supplementary material 1 (DOC × 17 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. Breusegem FV, Dat JF (2006) Reactive oxygen species in plant cell death. Plant Physiol 141:384–390PubMedCrossRefGoogle Scholar
  4. Cona A, Rea G, Angelini R, Federico R, Tavladoraki P (2006) Functions of amine oxidases in plant development and defence. Trends Plant Sci 11:1360–1685CrossRefGoogle Scholar
  5. Durrant WE, Dong X (2004) Systemic acquired resistance. Annu Rev Phytopathol 42:185–209PubMedCrossRefGoogle Scholar
  6. 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
  7. Flores HE, Protacio CM, Signs MW (1989) Primary and secondary metabolism of polyamines in plants. Recent Adv Phytochem 23:329–393Google Scholar
  8. Gao F, Su Q, Fan Y, Wang L (2010) Expression pattern and core region analysis of AtMPK3 promoter in response to environmental stresses. Sci China Life Sci 53:1315–1321PubMedCrossRefGoogle Scholar
  9. Iwata Y, Koizumi N (2005) An Arabidopsis transcription factor, AtbZIP60, regulates the endoplasmic reticulum stress response in a manner unique to plants. Proc Natl Acad Sci USA 102:5280–5285PubMedCrossRefGoogle Scholar
  10. Kakehi J, Kuwashiro Y, Niitsu M, Takahashi T (2008) Thermospermine is required for stem elongation in Arabidopsis thaliana. Plant Cell Physiol 49:1342–1349PubMedCrossRefGoogle Scholar
  11. Kuehn GD, Bagga S, Rodriguez-Garay B, Phillips GC (1990a) Biosynthesis of uncommon polyamines in higher plants and their relationship to abiotic stress responses. In: Flores HE, Arteca RN (eds) Polyamines and ethylene: biosynthesis, physiology and interactions. American Society of Plant Physiologists, RockvilleGoogle Scholar
  12. Kuehn GD, Rodriguez-Garay B, Bagga S, Phillips GC (1990b) Novel occurrence of uncommon polyamines in higher plants. Plant Physiol 94:855–857PubMedCrossRefGoogle Scholar
  13. Kusano T, Berberich T, Tateda C, Takahashi Y (2008) Polyamines: essential factors for growth and survival. Planta 228:367–381PubMedCrossRefGoogle Scholar
  14. Langebartels C, Schraudner M, Heller W, Ernst D, Sandermann H (2002) Oxidative stress and defense reactions in plants exposed to air pollutants and UV-B radiation. In: Inzé D, Van Montagu M (eds) Oxidative stress in plants. Taylor and Francis, London, pp 105–135Google Scholar
  15. Lindsay GS, Wallace HM (1999) Changes in polyamine catabolism in HL-60 human promyelogenous leukaemic cells in response to etoposide-induced apoptosis. Biochem J 337:83–87PubMedCrossRefGoogle Scholar
  16. Mackey D, Holt BF, Wiig A, Dangl JL (2002) RIN4 interacts with Pseudomonas syringae type III effector molecules and is required for RPM1-mediated resistance in Arabidopsis. Cell 108:743–754PubMedCrossRefGoogle Scholar
  17. Marina M, Maiale SJ, Rossi FR, Romero MF, Rivas EI, Gárriz A, Ruiz OA, Pieckenstain FL (2008) Apoplastic polyamine oxidation plays different roles in local responses of tobacco to infection by the necrotrophic fungus Sclerotinia sclerotiorum and the biotrophic bacterium Pseudomonas viridiflava. Plant Physiol 147:2164–2178PubMedCrossRefGoogle Scholar
  18. Marina M, Sirera FV, Rambla JL, Gonzalez ME, Blázquez MA, Carbonell J, Pieckenstain FL, Ruiz OA (2013) Thermospermine catabolism increases Arabidopsis thaliana resistance to Pseudomonas viridiflava. J Exp Bot. doi: 10.1093/jxb/ert012
  19. Mitsuya Y, Takahashi Y, Berberich T, Miyazaki A, Matsumura H, Takahashi H, Terauchi R, Kusano T (2009) Spermine signaling plays a significant role in the defense response of Arabidopsis thaliana to cucumber mosaic virus. J Plant Physiol 166:626–643PubMedCrossRefGoogle Scholar
  20. Mittler R (2002) Oxidative stress, antioxidants and stress tolerance. Trends Plant Sci 7:405–410PubMedCrossRefGoogle Scholar
  21. 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
  22. Niitsu M, Samejima K (1986) Synthesis of a series of linear pentaamines with three and four methylene chain intervals. Chem Pharm Bull (Tokyo) 34:1032–1103CrossRefGoogle Scholar
  23. Ohno-Iwashita Y, Oshima T, Imahori K (1975) In vitro protein synthesis at elevated temperature by an extract of an extreme thermophile. Arch Biochem Biophys 171:490–499CrossRefGoogle Scholar
  24. 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
  25. Oshima T (1978) Novel polyamines of extremely thermophilic bacteria. In: Friedman SM (ed) Biochemistry of thermophily. Academic Press, New York, pp 211–220CrossRefGoogle Scholar
  26. Oshima T (1979) Molecular basis for unusual thermostabilities of cell constituents from an extreme thermophile, Thermus thermophilus. In: Shilo M (ed) Strategies of microbial life in extreme environments. Dahlem Konferenzen, Berlin, pp 455–469Google Scholar
  27. Oshima T (2007) Unique polyamines produced by an extreme thermophile, Thermus thermophilus. Amino Acids 33:367–372PubMedCrossRefGoogle Scholar
  28. Oshima T (2010) Enigmas of biosyntheses of unusual polyamines in an extreme thermophile, Thermus thermophilus. Plant Physiol Biochem 48(7):521–526PubMedCrossRefGoogle Scholar
  29. Oshima T, Kawahata S (1983) Homocaldopentamine: a new naturally occurring pentaamine. J Biochem 93:1455–1456PubMedGoogle Scholar
  30. Parchment RE (1993) The implications of a unified theory of programmed cell death, polyamines, oxyradicals and histogenesis in the embryo. Int J Dev Biol 37:75–83PubMedGoogle Scholar
  31. Rodriguez-Garay B, Phillips GC, Kuehn GD (1989) Detection of norspermidine and norspermine in Medicago sativa L. (alfalfa). Plant Physiol 89:525–529PubMedCrossRefGoogle Scholar
  32. Sagor GHM, Yamaguchi K, Watanabe K, Berberich T, Kusano T, Takahashi Y (2011) Spatio-temporal expression analysis of Arabidopsis thaliana spermine synthase gene promoter. Plant Biotechnol 28:407–411CrossRefGoogle Scholar
  33. Sagor GHM, Berberich T, Takahashi Y, Niitsu M, Kusano T (2012a) The polyamine spermine protects Arabidopsis from heat stress-induced damage by increasing expression of heat shock-related genes. Transg Res. doi: 10.1007/s11248-012-9666-3 Google Scholar
  34. Sagor GHM, Takahashi H, Niitsu M, Takahashi Y, Berberich T, Kusano T (2012b) Exogenous thermospermine has an activity to induce a subset of the defense genes and restrict cucumber mosaic virus multiplication in Arabidopsis thaliana. Plant Cell Rep 31:1227–1232PubMedCrossRefGoogle Scholar
  35. Sakamoto H, Maruyama K, Sakuma Y, Meshi T, Iwabuchi M, Shinozaki K, Yamaguchi-Shinozaki K (2004) Arabidopsis Cys2/His2-type zinc-finger proteins function as transcription repressors under drought, cold, and high-salinity stress conditions. Plant Physiol 136:2734–2746PubMedCrossRefGoogle Scholar
  36. Samejima K, Yamauchi H, Asghar A, Yasui T (1984) Role of myosin heavy chains from rabbit skeletal muscle in the heat- induced gelation mechanisms. Agric Biol Chem 48:2225–2228CrossRefGoogle Scholar
  37. Seo S, Okamoto M, Seto H, Ishizuka K, Sano H, Ohashi Y (1995) Tobacco MAP kinase: a possible mediator in wound signal transduction pathways. Science 270:1988–1992PubMedCrossRefGoogle Scholar
  38. Takahashi H, Goto N, Ehara Y (1994) Hypersensitive response in cucumber mosaic virus-inoculated Arabidopsis thaliana. Plant J 6:369–377CrossRefGoogle Scholar
  39. Takahashi H, Miller J, Nozaki Y, Sukamto, Takeda M, Shah J, Hase S, Ikegami M, Ehara Y, Dinesh-Kumar SP (2002) RCY1, an Arabidopsis thaliana RPP8/HRT family resistance gene, conferring resistance to cucumber mosaic virus requires salicylic acid, ethylene and a novel signal transduction mechanism. Plant J 32:655–667Google Scholar
  40. Takahashi Y, Berberich T, Miyazaki A, Seo S, Ohashi Y, Kusano T (2003) Spermine signalling in tobacco: activation of mitogen-activated protein kinases by spermine is mediated through mitochondrial dysfunction. Plant J 36:820–829PubMedCrossRefGoogle Scholar
  41. Takahashi Y, Uehara Y, Berberich T, Ito A, Saitoh H, Miyazaki A, Terauchi R, Kusano T (2004) A subset of the hypersensitive response marker genes including HSR203J is downstream target of a spermine-signal transduction pathway in tobacco. Plant J 40:586–595PubMedCrossRefGoogle Scholar
  42. Takano A, Kakehi JI, Takahashi T (2012) Thermospermine is not a minor polyamine in the plant kingdom. Plant Cell Physiol 53:606–616PubMedCrossRefGoogle Scholar
  43. Takatsuji H (1999) Zinc-finger proteins: the classical zinc finger emerges in contemporary plant science. Plant Mol Biol 39:1073–1078PubMedCrossRefGoogle Scholar
  44. 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 141:1519–1532PubMedCrossRefGoogle Scholar
  45. Thurau T, Kifle S, Jung C, Cai D (2003) The promoter of the nematode resistance gene Hs1pro-1 activates a nematode responsive and feeding site specific gene expression in sugar beet (Beta vulgaris L.) and Arabidopsis thaliana. Plant Mol Biol 52:643–660PubMedCrossRefGoogle Scholar
  46. Torres MA, Joned JDG, Dangle JL (2006) Reactive oxygen species signaling in response to pathogens. Plant Physiol 141:373–378PubMedCrossRefGoogle Scholar
  47. Wang X, Xu Y, Han Y, Bao S, Du J, Yuan M, Xu Z, Chong K (2006) Overexpression of RAN1 in rice and Arabidopsis alter primordial meristem, mitotic progress and sensitivity to auxin. Plant Physiol 140:91–101PubMedCrossRefGoogle Scholar
  48. 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
  49. Yamaguchi K, Takahashi Y, Berberich T, Imai A, Miyazaki A, Takahashi T, Michael A, Kusano T (2007) A protective role for the polyamine spermine against drought stress in Arabidopsis. Biochem Biophys Res Commun 352:486–490PubMedCrossRefGoogle Scholar
  50. Yamakawa H, Kamada H, Satoh M, Ohashi Y (1998) Spermine is a salicylate-independent endogenous inducer for both tobacco acidic pathogenesis-related proteins and resistance against Tobacco mosaic virus infection. Plant Physiol 118:1213–1222PubMedCrossRefGoogle Scholar
  51. 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 142:193–206PubMedCrossRefGoogle Scholar
  52. Yoda H, Fujimura K, Takahashi H, Munemura I, Uchimiya H, Sano H (2009) Polyamines as a common source of hydrogen peroxide in host- and non-host hypersensitive response during pathogen infection. Plant Mol Biol 70:103–112PubMedCrossRefGoogle Scholar
  53. Zellner G, Kneifel H (1993) Caldopentamine and caldohexamine in cells of Thermotoga species, a possible adaptation to the growth at high temperatures. Arch Microbiol 159:472–476CrossRefGoogle Scholar
  54. Zhang S, Klessig DF (1997) Salicylic acid activates a 48-kDa MAP kinase in tobacco. Plant Cell 9:809–824PubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • G. H. M. Sagor
    • 1
  • Taibo Liu
    • 1
  • Hideki Takahashi
    • 2
  • Masaru Niitsu
    • 3
  • Thomas Berberich
    • 4
  • Tomonobu Kusano
    • 1
    Email author
  1. 1.Graduate School of Life SciencesTohoku UniversitySendaiJapan
  2. 2.Graduate School of Agricultural SciencesTohoku UniversitySendaiJapan
  3. 3.Faculty of Pharmaceutical SciencesJosai UniversitySakadoJapan
  4. 4.Biodiversity and Climate Research Center (BiK-F)Frankfurt am MainGermany

Personalised recommendations