Regulation of Ethylene Biosynthesis by Phytohormones in Etiolated Rice (Oryza sativa L.) Seedlings

  • Lee, Han Yong (Department of Botany and Plant Pathology, Purdue University) ;
  • Yoon, Gyeong Mee (Department of Botany and Plant Pathology, Purdue University)
  • Received : 2017.09.26
  • Accepted : 2018.01.04
  • Published : 2018.04.30


The gaseous hormone ethylene influences many aspects of plant growth, development, and responses to a variety of stresses. The biosynthesis of ethylene is tightly regulated by various internal and external stimuli, and the primary target of the regulation is the enzyme 1-aminocyclopropane-1-carboxylic acid (ACC) synthase (ACS), which catalyzes the rate-limiting step of ethylene biosynthesis. We have previously demonstrated that the regulation of ethylene biosynthesis is a common feature of most of the phytohormones in etiolated Arabidopsis seedlings via the modulation of the protein stability of ACS. Here, we show that various phytohormones also regulate ethylene biosynthesis from etiolated rice seedlings in a similar manner to those in Arabidopsis. Cytokinin, brassinosteroids, and gibberellic acid increase ethylene biosynthesis without changing the transcript levels of neither OsACS nor ACC oxidases (OsACO), a family of enzymes catalyzing the final step of the ethylene biosynthetic pathway. Likewise, salicylic acid and abscisic acid do not alter the gene expression of OsACS, but both hormones downregulate the transcript levels of a subset of ACO genes, resulting in a decrease in ethylene biosynthesis. In addition, we show that the treatment of the phytohormones results in distinct etiolated seedling phenotypes, some of which resemble ethylene-responsive phenotypes, while others display ethylene-independent morphologies, indicating a complicated hormone crosstalk in rice. Together, our study brings a new insight into crosstalk between ethylene biosynthesis and other phytohormones, and provides evidence that rice ethylene biosynthesis could be regulated by the post-transcriptional regulation of ACS proteins.


ethylene biosynthesis;hormone;OsACO;OsACS;post-transcriptional regulation;rice


  1. Abeles, F.B. (1973). Ethylene in plant biology. Academic, New York 302.
  2. Abeles, F.B., Morgan, P.W., and Saltveit, M.E.J. (1992). Ethylene in plant biology. San Diego, CA: Academic Press.
  3. Argueso, C.T., Hansen, M., and Kieber, J.J. (2007). Regulation of ethylene biosynthesis. J. Plant Growth Regul. 262, 92-105.
  4. Barry, C.S., Blume, B., Bouzayen, M., Cooper, W., Hamilton, A.J., and Grierson, D. (1996). Differential expression of the 1-aminocyclopropane-1-carboxylate oxidase gene family of tomato. Plant J. 9, 525-535.
  5. Bidonde, S., Ferrer, M.A., Zegzouti, H., Ramassamy, S., Latche, A., Pech, J.C., Hamilton, A.J., Grierson, D., and Bouzayen, M. (1998). Expression and characterization of three tomato 1-aminocyclopropane-1-carboxylate oxidase cDNAs in yeast. Eur. J. Biochem. 253, 20-26.
  6. Binnie, J.E., and McManus, M.T. (2009). Characterization of the 1-aminocyclopropane-1-carboxylic acid (ACC) oxidase multigene family of Malus domestica Borkh. Phytochemistry 70, 348-360.
  7. Chae, H.S., Cho, Y.G., Park, M.Y., Lee, M.C., Eun, M.Y., Kang, B.G., and Kim, W.T. (2000). Hormonal cross-talk between auxin and ethylene differentially regulates the expression of two members of the 1-aminocyclopropane-1-carboxylate oxidase gene family in rice (Oryza sativa L.). Plant Cell Physiol. 41, 354-362.
  8. Chae, H.S., and Kieber, J.J. (2005). Eto Brute? Role of ACS turnover in regulating ethylene biosynthesis. Trend. Plant Sci. 10, 291-296.
  9. Chae, H.S., Faure, F., and Kieber, J.J. (2003). The eto1, eto2, and eto3 mutations and cytokinin treatment increase ethylene biosynthesis in Arabidopsis by increasing the stability of ACS protein. Plant Cell 15, 545-559.
  10. Depuydt, S., and Hardtke, C.S. (2011). Hormone signalling crosstalk in plant growth regulation. Curr. Biol. 21, R365-373.
  11. Du, H., Wu, N., Cui, F., You, L., Li, X., and Xiong, L. (2014). A homolog of ETHYLENE OVERPRODUCER, OsETOL1, differentially modulates drought and submergence tolerance in rice. Plant J. 78, 834-849.
  12. Fukao, T., and Bailey-Serres, J. (2008). Submergence tolerance conferred by Sub1A is mediated by SLR1 and SLRL1 restriction of gibberellin responses in rice. Proc. Natl. Acad. Sci. USA 105, 16814-16819.
  13. Gomez-Jimenez, M.C., Matilla, A.J., and Garrido, D. (1998). Isolation and characterization of a cDNA encoding an ACC oxidase from Cicer arietinum and its expression during embryogenesis and seed germination. Australian J. Plant Physiol. 25, 765-773
  14. Guzman, P., and Ecker, J.R. (1990). Exploiting the triple response of Arabidopsis to identify ethylene-related mutants. Plant Cell 2, 513-523.
  15. Hansen, M., Chae, H.S., and Kieber, J.J. (2009). Regulation of ACS protein stability by cytokinin and brassinosteroid. Plant J. 57, 606-614.
  16. Hazman, M., Hause, B., Eiche, E., Riemann, M., and Nick, P. (2016). Different forms of osmotic stress evoke qualitatively different responses in rice. J. Plant Physiol. 202, 45-56.
  17. Huang, Y.F., Chen, C.T., and Kao, C.H. (1993). Salicylic acid inhibits the biosynthesis of ethylene in detached rice leaves. Plant Growth Regul. 12, 79-82.
  18. Iwai, T., Miyasaka, A., Seo, S., and Ohashi, Y. (2006). Contribution of ethylene biosynthesis for resistance to blast fungus infection in young rice plants. Plant Physiol. 142, 1202-1215.
  19. Iwamoto, M., Baba-Kasai, A., Kiyota, S., Hara, N., and Takano, M. (2010). ACO1, a gene for aminocyclopropane-1-carboxylate oxidase: effects on internode elongation at the heading stage in rice. Plant Cell Environ. 33, 805-815.
  20. Jackson, M.B. (2008). Ethylene-promoted elongation: an adaptation to submergence stress. Ann. Bot. 101, 229-248.
  21. Jaspert, N., Throm, C., and Oecking, C. (2011). Arabidopsis 14-3-3 proteins: fascinating and less fascinating aspects. Front. Plant Sci. 2, 96.
  22. Kim, J., Wilson, R.L., Case, J.B., and Binder, B.M. (2012). A comparative study of ethylene growth response kinetics in eudicots and monocots reveals a role for gibberellin in growth inhibition and recovery. Plant Physiol. 160, 1567-1580.
  23. Larsen, P.B., and Cancel, J.D. (2004). A recessive mutation in the RUB1-conjugating enzyme, RCE1, reveals a requirement for RUB modification for control of ethylene biosynthesis and proper induction of basic chitinase and PDF1.2 in Arabidopsis. Plant J. 38, 626-638.
  24. Lee, H.Y., Chen, Y.C., Kieber, J.J., and Yoon, G.M. (2017). Regulation of the turnover of ACC synthases by phytohormones and heterodimerization in Arabidopsis. Plant J. 91, 491-504.
  25. Lin, Z., Zhong, S., and Grierson, D. (2009). Recent advances in ethylene research. J. Exp. Bot. 60, 3311-3336.
  26. Linkies, A., and Leubner-Metzger, G. (2012). Beyond gibberellins and abscisic acid: how ethylene and jasmonates control seed germination. Plant Cell Rep. 31, 253-270.
  27. Liu, Y., and Zhang, S. (2004). Phosphorylation of 1-aminocyclopropane-1-carboxylic acid synthase by MPK6, a stressresponsive mitogen-activated protein kinase, induces ethylene biosynthesis in Arabidopsis. Plant Cell 16, 3386-3399.
  28. Lu, J., Li, J., Ju, H., Liu, X., Erb, M., Wang, X., and Lou, Y. (2014). Contrasting effects of ethylene biosynthesis on induced plant resistance against a chewing and a piercing-sucking herbivore in rice. Mol. Plant 7, 1670-1682.
  29. Ma, B., Chen, S.Y., and Zhang, J.S. (2010). Ethylene signaling in rice. Chinese Sci. Bull. 55, 2204-2210.
  30. Matillaa, A.J., and Matilla-Vazquezb, M.A. (2008). Involvement of ethylene in seed physiology. Plant Sci. 176, 87-97.
  31. Mekhedov, S.I., and Kende, H. (1996). Submergence enhances expression of a gene encoding 1-aminocyclopropane-1-carboxylate oxidase in deepwater rice. Plant Cell Physiol. 37, 531-537.
  32. Miro, B., and Ismail, A.M. (2013). Tolerance of anaerobic conditions caused by flooding during germination and early growth in rice (Oryza sativa L.). Front. Plant Sci. 4, 269.
  33. Morgan, P.W., and Drew, C.D. (1997). Ethylene and plant responses to stress. Physiologia Plantarum 100, 620-630.
  34. Nadeau, J.A., Zhang, X.S., Nair, H., and O'Neill, S.D. (1993). Temporal and spatial regulation of 1-aminocyclopropane-1-carboxylate oxidase in the pollination-induced senescence of orchid flowers. Plant Physiol. 103, 31-39.
  35. Nie, X., Singh, R.P., and Tai, G.C. (2002). Molecular characterization and expression analysis of 1-aminocyclopropane-1-carboxylate oxidase homologs from potato under abiotic and biotic stresses. Genome 45, 905-913.
  36. Petruzzelli, L., Coraggio, I., and Leubner-Metzger, G. (2000). Ethylene promotes ethylene biosynthesis during pea seed germination by positive feedback regulation of 1-aminocyclopropane-1-carboxylic acid oxidase. Planta 211, 144-149.
  37. Sahi, C., Singh, A., Kumar, K., Blumwald, E., and Grover, A. (2006). Salt stress response in rice: genetics, molecular biology, and comparative genomics. Funct. Integr. Genomics 6, 263-284.
  38. Shimamoto, K. (1999). Molecular biology of rice. Springer-Verlag, Tokyo.
  39. Tang, X., Wang, H., Brandt, A.S., and Woodson, W.R. (1993). Organization and structure of the 1-aminocyclopropane-1-carboxylate oxidase gene family from Petunia hybrida. Plant Mol. Biol. 23, 1151-1164.
  40. Tsuchisaka, A., and Theologis, A. (2004). Unique and overlapping expression patterns among the Arabidopsis 1-amino-cyclopropane-1-carboxylate synthase gene family members. Plant Physiol. 136, 2982-3000.
  41. Vriezen, W.H., Zhou, Z., and Van Der Straeten, D. (2003). Regulation of submergence-induced enhanced shoot elongation in Oryza sativa L. Ann. Bot. 91 Spec No, 263-270.
  42. Wang, K.L., Yoshida, H., Lurin, C., and Ecker, J.R. (2004). Regulation of ethylene gas biosynthesis by the Arabidopsis ETO1 protein. Nature 428, 945-950.
  43. Wang, N.N., Shih, M.C., and Li, N. (2005). The GUS reporter-aided analysis of the promoter activities of Arabidopsis ACC synthase genes AtACS4, AtACS5, and AtACS7 induced by hormones and stresses. J. Exp. Bot. 56, 909-920.
  44. Watanabe, H., Hase, S., and Saigusa, M. (2007). The effect of ethylene and other regulators on coleoptile growth of rice under anoxia Plant Prod. Sci 10, 468-472.
  45. Yamagami, T., Tsuchisaka, A., Yamada, K., Haddon, W.F., Harden, L.A., and Theologis, A. (2003). Biochemical diversity among the 1-amino-cyclopropane-1-carboxylate synthase isozymes encoded by the Arabidopsis gene family. J. Biol. Chem. 278, 49102-49112.
  46. Yang, S.F., and Hoffman, N.E. (1984). Ethylene biosynthesis and its regulation in higher plants. Ann. Rev. Plant Physiol. 35, 155-189.
  47. Yao, Y., Du, Y., Jiang, L., and Liu, J.Y. (2007). Interaction between ACC synthase 1 and 14-3-3 proteins in rice: a new insight. Biochemistry 72, 1003-1007.
  48. Yoon, G.M. (2015). New insights into the protein turnover regulation in ethylene biosynthesis. Mol. Cells 38, 597-603.
  49. Yoon, G.M., and Kieber, J.J. (2013). 14-3-3 regulates 1-aminocyclopropane-1-carboxylate synthase protein turnover in Arabidopsis. Plant Cell 25, 1016-1028.
  50. Yoshida, H., Nagata, M., Saito, K., Wang, K.L., and Ecker, J.R. (2005). Arabidopsis ETO1 specifically interacts with and negatively regulates type 2 1-aminocyclopropane-1-carboxylate synthases. BMC Plant Biol. 5, 14.
  51. Yoshida, H., Wang, K.L., Chang, C.M., Mori, K., Uchida, E., and Ecker, J.R. (2006). The ACC synthase TOE sequence is required for interaction with ETO1 family proteins and destabilization of target proteins. Plant Mol. Biol. 62, 427-437.
  52. Zarembinski, T.I., and Theologis, A. (1994). Ethylene biosynthesis and action: a case of conservation. Plant Mol. Biol. 26, 1579-1597.
  53. Zarembinski, T.I., and Theologis, A. (1997). Expression characteristics of OS-ACS1 and OS-ACS2, two members of the 1-aminocyclopropane-1-carboxylate synthase gene family in rice (Oryza sativa L. cv. Habiganj Aman II) during partial submergence. Plant Mol. Biol. 33, 71-77.