The Physiological Relationship Between Abscisic Acid and Gibberellin During Seed Germination of Trichocline catharinensis (Asteraceae) Is Associated with Polyamine and Antioxidant Enzymes

  • Ana P. Lando
  • W. G. Viana
  • R. A. da Silva
  • C. D. D. Costa
  • Hugo P. F. Fraga
  • Marisa Santos
  • Paulo T. Mioto
  • Miguel P. Guerra
  • N. SteinerEmail author


An improved understanding of seed quality and germination control can contribute effectively to the use and conservation of neglected native species with ecological and economic value, such as Trichocline catharinensis, an endemic Asteraceae species from southern Brazil. We investigated the effects of applying gibberellin (GA3), abscisic acid (ABA) and their biosynthesis inhibitors, paclobutrazol (PAC) and fluridone (FLU), respectively, on T. catharinensis seed germination, and on polyamine (PA) content and antioxidant enzyme activities in germinating seeds. FLU and GA3 increased seed germination speed compared to treatment with H2O only. ABA inhibited both germination speed index and percentage, while PAC severely inhibited seed germination. The stimulatory effect of GA3 and FLU was associated with increased contents of putrescine (PUT) and spermidine (SPD) relative to spermine (SPM). As a result, high ratio (PUT + SPD/SPM) as well as superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX) enzyme activities were observed when seed germination occurs. In contrast, in low or no seed germination treatment (ABA and PAC), low ratio (PUT + SPD/SPM) was observed, while the antioxidant enzymes, mainly SOD activity, tended to increase. Application of PAs at 200 μM stimulated germination through improving the speed and uniformity, and this effect was associated with antioxidant enzyme activity. Our results suggest a relationship between PA and the antioxidant system with the physiological mechanism of seed germination. These results improve the physiological knowledge of seed germination control in Asteraceae and contribute to the biological groundwork for future studies on the use and conservation of native species.


Antioxidant enzymes Biodiversity Fluridone Paclobutrazol Polyamines Seed germination 



Abscisic acid


Arginine decarboxylase


Ascorbate peroxidase




Days after imbibition


Diamine oxidases






Glutathione reductase


Hydrochloric acid


High-performance liquid chromatography




Indole-3-acetic acid


Germination speed index


Ornithine decarboxylase


Nitro tetrazolium Blue chloride






Polyamine oxidases


Plant growth regulators






Reactive oxygen species


Superoxide dismutase







The authors thank the Plant Developmental Physiology and Genetics Laboratory of the Federal University of Santa Catarina, Brazil. The authors also thank the Laboratory of Morphogenesis and Plant Biochemistry of the Federal University of Santa Catarina, Brazil. This study was financed in part by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior—Brazil (CAPES)-Finance Code 001.


This work was supported by the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq, Brazil) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES, Brazil). Grant Number of Neusa Steiner (311156/2017-7 457940/2014-0).

Compliance with Ethical Standards

Conflict of interest

The authors declare that no conflict exists among the authors.

Supplementary material

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  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 231:1237–1249. Google Scholar
  2. Apel K, Hirt H (2004) Reactive oxygen species: metabolism, oxidative stress, and signal transduction. Annu Rev Plant Biol 55:373–399Google Scholar
  3. Bailly C, Kranner I (2011) Analyses of reactive oxygen species and antioxidants in relation to seed longevity and germination. Methods Mol Biol 773:343–367. Google Scholar
  4. Bailly C, Leymarie J, Lehner A, Rousseau S, Côme D, Corbineau F (2004) Catalase activity and expression in developing sunflower seeds as related to drying. J Exp Bot 55:475–483. Google Scholar
  5. Bailly C, El-Maarouf-Bouteau H, Corbineau F (2008) From intracellular signaling networks to cell death: the dual role of reactive oxygen species in seed physiology. C. R. Biologies 331:806–814. Google Scholar
  6. Barba-Espin G, Diaz-Vivancos P, Clemente-Moreno M, Albacete A, Faize L, Faize M, Pérez-Alfocea F, Hernández J (2010) Interaction between hydrogen peroxide and plant hormones during germination and the early growth of pea seedlings. Plant Cell Environ 33:981–994. Google Scholar
  7. Bewley DJ, Bradford K, Hilhorst H (2013) Seeds: physiology of development, germination and dormancy. Springer, LondonGoogle Scholar
  8. Bombo AB, Oliveira TSD, Appezzato-Da-Glória B, Novembre ADDLC (2015) Seed germination of Brazilian Aldama species (Asteraceae). J. Seed Sci. 37:185–191. Google Scholar
  9. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254. Google Scholar
  10. Brown RF, Mayer DG (1988) Representing cumulative germination. 1. A critical analysis of single-value germination indices. Ann Bot 61:117–125. Google Scholar
  11. Bueno M, Matilla A (1992) Abscisic acid increases the content of free polyamines and delays mitotic activity induced by spermine in isolated embryonic axes of chick-pea seeds. Physiol Plant 85:531–536Google Scholar
  12. Bull JW, Maron M (2016) How humans drive speciation as well as extinction. Proc R Soc Lond B 283:1–10. Google Scholar
  13. Cabrera AL, Klein RM (1973) Compostas tribo Mutisieae. In: Cabrera AL, Klein RM (eds) Flora ilustrada catarinense. Herbário Barbosa Rodrigues, Itajaí, p 94Google Scholar
  14. Cao D, Hu J, Zhu S, Hu W, Knapp A (2010) Relationship between changes in endogenous polyamines and seed quality during development of sh2 sweet corn (Zea mays L.) seed. Sci Hortic 123:301–307. Google Scholar
  15. Chen QL, Guo Y, Jiang Y, Tu P (2016) Mechanism of fluridone-induced seed germination of Cistanche tubulosa. Pak J Bot 48:971–976Google Scholar
  16. Coradin L, Siminski A, Reis A (2011) Espécies Nativas da Flora Brasileira de Valor Econômico Atual ou Potencial. Ministério do Meio Ambiente, BrasíliaGoogle Scholar
  17. Cury G, Novembre ADDLC, Glória BAD (2010) Seed germination of Chresta sphaerocephala DC. and Lessingianthus bardanoides (Less.) H. Rob. (asteraceae) from Cerrado. Braz Arch Biol Technol 53:1299–1308. Google Scholar
  18. Davide AC, Silva CSJ, Silva EAAD, Pinto LVA, Faria JMR (2008) Morpho-anatomical, biochemical and physiological studies in seeds of Eremanthus erythropappus (DC.) MacLeish during germination. Rev. Bras. Sementes 30:171–176. Google Scholar
  19. Diaz-Vivancos P, Barba-Espín G, Hernández JA (2013) Elucidating hormonal/ROS networks during seed germination: insights and perspectives. Plant Cell Rep 32:1491–1502. Google Scholar
  20. El-Maarouf-Bouteau H, Bailly C (2008) Oxidative signaling in seed germination and dormancy. Plant Signal Behav 3:175–182. Google Scholar
  21. El-Maarouf-Bouteau H, Sajjad Y, Bazin J, Langlade N, Cristescu SM, Balzergue S, Baudouin E, Bailly C (2015) Reactive oxygen species, abscisic acid and ethylene interact to regulate sunflower seed germination. Plant Cell Environ 38:364–374Google Scholar
  22. Ferreira AG, Cassol B, Rosa SGTD, Silveira TSD, Stival AL, Silva AA (2001) Germination of seeds of Asteraceae natives of Rio Grande do Sul. Brazil Acta Bot Bras 15:231–242. Google Scholar
  23. Finch-Savage WE, Leubner-Metzger G (2006) Seed dormancy and the control of germination. New Phytol 171:501–523. Google Scholar
  24. Foyer CH, Noctor G (2005) Redox homeostasis and antioxidant signaling: a metabolic interface between stress perception and physiological responses. Plant Cell 17:1866–1875. Google Scholar
  25. Funk VA, Susanna A, Stuessy TF, Robinson H (2009) Classification of compositae. In: Funk VA, Susanna A, Stuessy T, Bayer R (eds) Systematics, evolution and biogeography of the Compositae. International Association for Plant Taxonomy, Michigan, pp 171–189Google Scholar
  26. Galston AW, Sawhney RK (1990) Polyamines in plant physiology. Plant Physiol 94:406–410Google Scholar
  27. Giannopolitis CN, Ries SK (1977) Superoxide dismutases: I Occurrence in higher plants. Plant Physiol 59:309–314Google Scholar
  28. Goggin DE, Steadman KJ, Emery RJN, Farrow SC, Benech-Arnold RL, Powles SB (2009) ABA inhibits germination but not dormancy release in mature imbibed seeds of Lolium rigidum Gaud. J Exp Bot 60:3387–3396. Google Scholar
  29. Gomes V, Fernandes GW (2002) Germination of Baccharis dracunculifolia DC (Asteraceae) achene. Acta Bot Bras 16:421–427. Google Scholar
  30. Gomes MP, Garcia QS (2013) Reactive oxygen species and seed germination. Biologia 68:351–357. Google Scholar
  31. Gordin CRB, Marques RF, Masetto TE, Scalon SDPQ (2012) Germination, seed biometrics and seedling morphology of Guizotia abyssinica Cass. Rev Bras Sementes 34:619–627. Google Scholar
  32. Grappin P, Bouinot D, Sotta B, Miginiac E, Jullien M (2000) Control of seed dormancy in Nicotiana plumbaginifolia: post-imbibition abscisic acid synthesis imposes dormancy maintenance. Planta 210:279–285Google Scholar
  33. Gupta K, Sengupta A, Chakraborty M, Gupta B (2016) Hydrogen peroxide and polyamines act as double edged swords in plant abiotic stress responses. Front Plant Sci 7:1343. Google Scholar
  34. Hedden P, Graebe JE (1985) Inhibition of gibberellin biosynthesis by paclobutrazol in cell-free homogenates of Cucurbita maxima endosperm and Malus pumila embryos. J Plant Growth Regul 4:111Google Scholar
  35. Holdsworth MJ, Bentsink L, Soppe WJJ (2008) Molecular networks regulating Arabidopsis seed maturation, after-ripening, dormancy and germination. New Phytol 179:33–54Google Scholar
  36. Hu X, Zhang A, Zhang J, Jiang M (2006) Abscisic acid is a key inducer of hydrogen peroxide production in leaves of maize plants exposed to water stress. Plant Cell Physiol 47:1484–1495Google Scholar
  37. Hu XW, Huang XH, Wang YR (2012) Hormonal and temperature regulation of seed dormancy and germination in Leymus chinensis. Plant Growth Regul 67:199–207Google Scholar
  38. Huang Y, Lin C, He F, Li Z, Guan Y, Hu Q, Hu J (2017) Exogenous spermidine improves seed germination of sweet corn via involvement in phytohormone interactions, H2O2 and relevant gene expression. BMC Plant Biol 17:1–16Google Scholar
  39. Huarte HR, Benech-Arnold RL (2010) Hormonal nature of seed responses to fluctuating temperatures in Cynara cardunculus (L.). Seed Sci Res 20:39–45Google Scholar
  40. Igarashi K, Kashiwagi K (2000) Polyamines: mysterious modulators of cellular functions. Biochem Biophys Res Commun 271:559–564Google Scholar
  41. Jaleel CA, Manivannan P, Wahid A, Farooq M, Al-Juburi HJ, Somasundaram R, Panneerselvam R (2009) Drought stress in plants: a review on morphological characteristics and pigments composition. Int J Agric Biol 11:100–105Google Scholar
  42. Jiang M, Zhang J (2003) Cross-talk between calcium and reactive oxygen species originated from NADPH oxidase in abscisic acid-induced antioxidant defence in leaves of maize seedlings. Plant Cell Environ 26:929–939Google Scholar
  43. Jiménez-Bremont JF, Marina M, Guerrero-González Mde L, Rossi FR, Sánchez-Rangel D, Rodríguez-Kessler M, Ruiz OA, Gárriz A (2014) Physiological and molecular implications of plant polyamine metabolism during biotic interactions. Front Plant Sci 5:95. Google Scholar
  44. Job C, Rajjou L, Lovigny Y, Belghazi M, Job D (2005) Patterns of protein oxidation in Arabidopsis seeds and during germination. Plant Physiol 138:790–802Google Scholar
  45. Justo CF, Alvarenga AAD, Nery FC, Delu Filho N (2007) Chemical composition, imbibition curve and temperature effect on seed germination of Eugenia pyriformis Camb. (Myrtaceae). Revista Brasileira Biociências 5:510–512Google Scholar
  46. Kaur-Sawhney R, Tiburcio AF, Altabella T, Galston AW (2003) Polyamines in plants: an overview. J Cell Mol Biol 2:1–12Google Scholar
  47. Kim ST, Kang SY, Wang Y, Kim SG, Hwang DH, Kang KY (2008) Analysis of embryonic proteome modulation by GA and ABA from germinating rice seeds. Proteomics 8:3577–3587Google Scholar
  48. Koshiba T (1993) Cytosolic ascorbate peroxidase in seedlings and leaves of maize (Zea mays). Plant Cell Physiol 34:713–721. Google Scholar
  49. Krasuska U, Gniazdowska A (2012) Nitric oxide and hydrogen cyanide as regulating factors of enzymatic antioxidant system in germinating apple embryos. Acta Physiol Plant 34:683–692. Google Scholar
  50. Krasuska U, Ciacka K, Bogatek R, Gniazdowska A (2014) Polyamines and nitric oxide link in regulation of dormancy removal and germination of apple (Malus domestica Borkh.) Embryos. J Plant Growth Regul 33:590–601. Google Scholar
  51. Krasuska U, Ciacka K, Gniazdowska A (2017) Nitric oxide-polyamines cross-talk during dormancy release and germination of apple embryos. Nitric Oxide 68:38–50Google Scholar
  52. Kucera B, Cohn MA, Leubner-Metzger G (2005) Plant hormone interactions during seed dormancy release and germination. Seed Sci Res 15:281–307Google Scholar
  53. Kusano T, Suzuki H (2015) Polyamines: a universal molecular nexus for growth, survival, and specialized metabolism. Springer, New YorkGoogle Scholar
  54. Kusumoto D, Chae SH, Mukaida K, Yoneyama K, Yoneyama K, Joel DM, Takeuchi Y (2006) Effects of fluridone and norflurazon on conditioning and germination of Striga asiatica seeds. Plant Growth Regul 48:73–78Google Scholar
  55. Kuznetsov VV, Radyukina N, Shevyakova N (2006) Polyamines and stress: biological role, metabolism, and regulation. Russ J Plant Physiol 53:583Google Scholar
  56. Leubner-Metzger G, Knight C, Linkies A, Graeber K (2010) The evolution of seeds. New Phytol 186:817–831Google Scholar
  57. Leymarie J et al (2011) Role of reactive oxygen species in the regulation of Arabidopsis seed dormancy. Plant Cell Physiol 53:96–106Google Scholar
  58. Li Z et al (2015) Polyamine regulates tolerance to water stress in leaves of white clover associated with antioxidant defense and dehydrin genes via involvement in calcium messenger system and hydrogen peroxide signaling. Front Physiol 6:280. Google Scholar
  59. Li S, Jin H, Zhang Q (2016) The Effect of Exogenous Spermidine Concentration on Polyamine Metabolism and Salt Tolerance in Zoysiagrass (Zoysia japonica Steud) Subjected to Short-Term Salinity Stress. Front Plant Sci. Google Scholar
  60. Liu X, Hou X (2018) Antagonistic regulation of ABA and GA in metabolism and signaling pathways. Front Plant Sci 9:251. Google Scholar
  61. Maguire JD (1962) Speed of germination—aid in selection and evaluation for seedling emergence and vigor. Crop Sci 2:176–177Google Scholar
  62. Mattoo AK, Handa AK (2008) Higher polyamines restore and enhance metabolic memory in ripening fruit. Plant Sci 174:386–393Google Scholar
  63. Miransari M, Smith D (2014) Plant hormones and seed germination. Environ Exp Bot 99:110–121Google Scholar
  64. Mirza JI, Rehman A (1998) A spermine-resistant mutant of Arabidopsis thaliana displays precocious germination. Acta Physiol Plant 20:235–240Google Scholar
  65. Moschou PN, Paschalidis KA, Roubelakis-Angelakis KA (2008) Plant polyamine catabolism: the state of the art. Plant Signal Behav 3:1061–1066Google Scholar
  66. Niedzwiedz-Siegien I, Bogatek-Leszczynska R, Côme D, Corbineau F (2004) Effects of drying rate on dehydration sensitivity of excised wheat seedling shoots as related to sucrose metabolism and antioxidant enzyme activities. Plant Sci 167:879–888. Google Scholar
  67. Nieuwland J, Stamm P, Wen B, Randall RS, Murray JA, Bassel GW (2016) Re-induction of the cell cycle in the Arabidopsis post-embryonic root meristem is ABA-insensitive, GA-dependent and repressed by KRP6. Sci Rep 6:23586Google Scholar
  68. Palavan N, Galston AW (1982) Polyamine biosynthesis and titer during various developmental stages of Phaseolus vulgaris. Physiol Plant 55:438–444Google Scholar
  69. Pasini E, Ritter MR (2012) O gênero Trichocline Cass. (Asteraceae, Mutisieae) no Rio Grande do Sul, Brasil. Revista Brasileira Biociências 10:490–506Google Scholar
  70. Peixoto PHP, Cambraia J, Sant’Anna R, Mosquim PR, Moreira MA (1999) Aluminum effects on lipid peroxidation and on the activities of enzymes of oxidative metabolism in sorghum. Rev Bras Fisiol Veg 11:137–143Google Scholar
  71. Peng J, Harberd NP (2002) The role of GA-mediated signalling in the control of seed germination. Curr Opin Plant Biol 5:376–381Google Scholar
  72. Piskurewicz U, Lopez-Molina L (2009) The GA-signaling repressor RGL3 represses testa rupture in response to changes in GA and ABA levels. Plant Signal Behav. 4:63–65Google Scholar
  73. Piskurewicz U, Jikumaru Y, Kinoshita N, Nambara E, Kamiya Y, Lopez-Molina L (2008) The gibberellic acid signaling repressor RGL2 inhibits Arabidopsis seed germination by stimulating abscisic acid synthesis and ABI5 activity. Plant Cell 20:2729–2745Google Scholar
  74. Rademacher W (2000) Growth Retardants: effects on Gibberellin. Annu Rev Plant Biol 51:501–531Google Scholar
  75. Radhakrishnan R, Lee I-J (2013) Spermine promotes acclimation to osmotic stress by modifying antioxidant, abscisic acid, and jasmonic acid signals in soybean. J Plant Growth Regul 32:22–30Google Scholar
  76. Ranal MA, Santana DGd (2006) How and why to measure the germination process? Braz J Bot 29:1–11Google Scholar
  77. Saha J, Brauer EK, Sengupta A, Popescu SC, Gupta K, Gupta B (2015) Polyamines as redox homeostasis regulators during salt stress in plants. Front Environ Sci 3:21. Google Scholar
  78. Sánchez-Rangel D, Chávez-Martínez AI, Rodríguez-Hernández AA, Maruri-López I, Urano K, Shinozaki K, Jiménez-Bremont JF (2016) Simultaneous silencing of two arginine decarboxylase genes alters development in Arabidopsis. Front Plant Sci. Google Scholar
  79. Shiozaki S, Ogata T, Horiuchi S, Zhuo X (1998) Involvement of polyamines in gibberellin-induced development of seedless grape berries. Plant Growth Regul 25:187–193. Google Scholar
  80. Silveira V, Balbuena TS, Santa-Catarina C, Floh EIS, Guerra MP, Handro W (2004) Biochemical changes during seed development in Pinus taeda L. Plant Growth Regul 44:147–156. Google Scholar
  81. Skubacz A, Daszkowska-Golec A (2017) Seed dormancy: the complex process regulated by abscisic acid, gibberellins, and other phytohormones that makes seed germination work. IntechOpen, LondonGoogle Scholar
  82. Sokal RR, Rohlf FJ (1995) Biometry: the principles and practice of statistics in Biological Research, 3rd edn. W. H. Freeman and Company, New YorkGoogle Scholar
  83. Steiner N, Santa-Catarina C, Silveira V, Floh EI, Guerra MP (2007) Polyamine effects on growth and endogenous hormones levels in Araucaria angustifolia embryogenic cultures. Plant Cell Tissue Organ Cult 89:55–62. Google Scholar
  84. Tanou G, Job C, Rajjou L, Arc E, Belghazi M, Diamantidis G, Molassiotis A, Job D (2009) Proteomics reveals the overlapping roles of hydrogen peroxide and nitric oxide in the acclimation of citrus plants to salinity. Plant J 60:795–804Google Scholar
  85. Tassoni A, van Buuren M, Franceschetti M, Fornalè S, Bagni N (2000) Polyamine content and metabolism in Arabidopsis thaliana and effect of spermidine on plant development. Plant Physiol Biochem 38:383–393Google Scholar
  86. Team RC (2014) R: A language and environment for statistical computing. R Foundation for Statistical Computing.
  87. Tiburcio A, Besford R, Capell T, Borrell A, Testillano P, Risueno M (1994) Mechanisms of polyamine action during senescence responses induced by osmotic stress. J Exp Bot 45:1789–1800Google Scholar
  88. Urano K, Hobo T, Shinozaki K (2005) Arabidopsis ADC genes involved in polyamine biosynthesis are essential for seed development. FEBS Lett 579:1557–1564. Google Scholar
  89. Vieira B, Bicalho E, Munné-Bosch S, Garcia Q (2017) Abscisic acid regulates seed germination of Vellozia species in response to temperature. Plant Biol (Stuttg) 19:211–216. Google Scholar
  90. Wang L, Hua D, He J, Duan Y, Chen Z, Hong X, Gong Z (2011) Auxin Response Factor2 (ARF2) and its regulated homeodomain gene HB33 mediate abscisic acid response in Arabidopsis. PLoS Genet 7:1–14Google Scholar
  91. Wojtyla Ł, Lechowska K, Kubala S, Garnczarska M (2016) Different modes of hydrogen peroxide action during seed germination. Front Plant Sci. Google Scholar
  92. Yang L, Hong X, Wen XX, Liao YC (2016) Effect of polyamine on seed germination of wheat under drought stress is related to changes in hormones and carbohydrates. J Integr Agric 15:1–17. Google Scholar

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Authors and Affiliations

  1. 1.Plant Physiology Laboratory, Department of BotanyFederal University of Santa CatarinaFlorianópolisBrazil
  2. 2.Department of BotanyFederal University of ParanáCuritibaBrazil
  3. 3.Plant Developmental Physiology and Genetics Laboratory, Department of Plant ScienceFederal University of Santa CatarinaFlorianópolisBrazil

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