Plant Molecular Biology Reporter

, Volume 36, Issue 5–6, pp 710–724 | Cite as

Ectopic Expression of Two FOREVER YOUNG FLOWER Orthologues from Cattleya Orchid Suppresses Ethylene Signaling and DELLA Results in Delayed Flower Senescence/Abscission and Reduced Flower Organ Elongation in Arabidopsis

  • Wei-Han Chen
  • Yung-I Lee
  • Chang-Hsien YangEmail author
Original Paper


Two orthologues of Arabidopsis FOREVER YOUNG FLOWER (FYF), CaFYF1 and CaFYF2, were identified from Cattleya intermedia. To investigate the function of these two genes, we performed ectopic expression of CaFYF1/2 in Arabidopsis. Delayed flower senescence and abscission were observed in 35S::CaFYF1/2 transgenic Arabidopsis plants. Furthermore, once CaFYF1/2 was fused with the strong repressor domain SRDX, severe delayed flower senescence and abscission were observed in 35S::CaFYF1/2+SRDX transgenic Arabidopsis plants. In contrast, when 35S::CaFYF1/2 was converted to a potent activator by fusion with the VP16-AD motif, flower senescence and abscission were promoted in these 35S::CaFYF1/2+VP16 transgenic dominant-negative mutant Arabidopsis plants. These results indicated that similar to Arabidopsis FYF, CaFYF1/2 also act as repressors in controlling floral organ senescence and abscission in transgenic Arabidopsis plants. The delayed senescence and abscission of the flower organs in 35S::CaFYF1/2 and 35S::CaFYF1/2+SRDX transgenic Arabidopsis plants were unaffected by ethylene treatment. Genes of the ethylene signaling and abscission-associated pathways, such as EDF1/2/3/4, BOP1/2, and IDA, were repressed in 35S::CaFYF1/2 and 35S::CaFYF1/2+SRDX transgenic Arabidopsis plants. Furthermore, 35S::CaFYF1/2 and 35S::CaFYF1/2+SRDX transgenic Arabidopsis plants showed additional morphological defects, such as short sepals and petals, which were correlated with the upregulation of the DELLA genes RGA, GAI, RGL1, and RGL2. These results suggested a possible role for Cattleya orchid CaFYF1/2 in controlling floral senescence/abscission by suppressing ethylene signaling and abscission-associated genes as well as controlling flower organ elongation through negative regulation of GA response by activating the expression of the DELLA genes during flower development.


Senescence Abscission Cattleya intermedia Ethylene responses FOREVER YOUNG FLOWER MADS box gene Repressor 



This work was supported by grants to C-H Y from the Ministry of Science and Technology, Taiwan, ROC, grant numbers MOST 106-2321-B-005-003 and MOST 106-2321-B-005-010. This work was also financially supported (in part) by the Advanced Plant Biotechnology Center from The Featured Areas Research Center Program within the framework of the Higher Education Sprout Project by the Ministry of Education (MOE) in Taiwan.

Supplementary material

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Table S1 (PDF 14 kb)
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ESM 3 (PDF 29.7 kb)
11105_2018_1114_MOESM4_ESM.pdf (141 kb)
ESM 4 (PDF 140 kb)


  1. Aalen RB, Wildhagen M, Sto IM, Butenko MA (2013) IDA: a peptide ligand regulating cell separation processes in Arabidopsis. J Exp Bot 64:5253–5261CrossRefGoogle Scholar
  2. Adamczyk BJ, Lehti-Shiu MD, Fernandez DE (2007) The MADS domain factors AGL15 and AGL18 act redundantly as repressors of the floral transition in Arabidopsis. Plant J 50:1007–1019CrossRefGoogle Scholar
  3. Alonso JM, Stepanova AN, Leisse TJ, Kim CJ, Chen H, Shinn P, Stevenson DK, Zimmerman J, Barajas P, Cheuk R, Gadrinab C, Heller C, Jeske A, Koesema E, Meyers CC, Parker H, Prednis L, Ansari Y, Choy N, Deen H, Geralt M, Hazari N, Hom E, Karnes M, Mulholland C, Ndubaku R, Schmidt I, Guzman P, Aguilar-Henonin L, Schmid M, Weigel D, Carter DE, Marchand T, Risseeuw E, Brogden D, Zeko A, Crosby WL, Berry CC, Ecker JR (2003) Genome-wide insertional mutagenesis of Arabidopsis thaliana. Science 301:653–657CrossRefGoogle Scholar
  4. Alvarez-Buylla ER, Liljegren SJ, Pelaz S, Gold SE, Burgeff C, Ditta GS, Vergara-Silva F, Yanofsky MF (2000) MADS-box gene evolution beyond flowers: expression in pollen, endosperm, guard cells, roots and trichomes. Plant J 24:457–466CrossRefGoogle Scholar
  5. Arora A (2005) Ethylene receptors and molecular mechanism of ethylene sensitivity in plants. Curr Sci 89:1348–1361Google Scholar
  6. Avila-Ospina L, Moison M, Yoshimoto K, Masclaux-Daubresse C (2014) Autophagy, plant senescence, and nutrient recycling. J Exp Bot 65:3799–3811CrossRefGoogle Scholar
  7. Brumos J, Alonso JM, Stepanova AN (2014) Genetic aspects of auxin biosynthesis and its regulation. Physiol Plant 151:3–12CrossRefGoogle Scholar
  8. Butenko MA, Patterson SE, Grini PE, Stenvik GE, Amundsen SS, Mandal A, Aalen RB (2003) Inflorescence deficient in abscission controls floral organ abscission in Arabidopsis and identifies a novel family of putative ligands in plants. Plant Cell 15:2296–2307CrossRefGoogle Scholar
  9. Castillejo C, Pelaz S (2008) The balance between CONSTANS and TEMPRANILLO activities determines FT expression to trigger flowering. Curr Biol 18:1338–1343CrossRefGoogle Scholar
  10. Chen QG, Bleecker AB (1995) Analysis of ethylene signal-transduction kinetics associated with seedling-growth response and chitinase induction in wild-type and mutant Arabidopsis. Plant Physiol 108:597–607CrossRefGoogle Scholar
  11. Chen YF, Etheridge N, Schaller GE (2005) Ethylene signal transduction. Ann Bot 95:901–915CrossRefGoogle Scholar
  12. Chen MK, Hsu WH, Lee PF, Thiruvengadam M, Chen HI, Yang CH (2011a) The MADS box gene, FOREVER YOUNG FLOWER, acts as a repressor controlling floral organ senescence and abscission in Arabidopsis. Plant J 68:168–185CrossRefGoogle Scholar
  13. Chen MK, Lee PF, Yang CH (2011b) Delay of flower senescence and abscission in Arabidopsis transformed with an FOREVER YOUNG FLOWER homolog from Oncidium orchid. Plant Signal Behav 6:1841–1843CrossRefGoogle Scholar
  14. Chen WH, Li PF, Chen MK, Lee YI, Yang CH (2015) FOREVER YOUNG FLOWER negatively regulates ethylene response DNA-binding factors by activating an ethylene-responsive factor to control Arabidopsis floral organ senescence and abscission. Plant Physiol 168:1666–1683CrossRefGoogle Scholar
  15. Cheng H, Qin L, Lee S, Fu X, Richards DE, Cao D, Luo D, Harberd NP, Peng J (2004) Gibberellin regulates Arabidopsis floral development via suppression of DELLA protein function. Development 131:1055–1064CrossRefGoogle Scholar
  16. Cho SK, Larue CT, Chevalier D, Wang H, Jinn TL, Zhang S, Walker JC (2008) Regulation of floral organ abscission in Arabidopsis thaliana. Proc Natl Acad Sci U S A 105:15629–15634CrossRefGoogle Scholar
  17. Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16:735–743CrossRefGoogle Scholar
  18. Dill A, Sun T (2001) Synergistic derepression of gibberellin signaling by removing RGA and GAI function in Arabidopsis thaliana. Genetics 159:777–785PubMedPubMedCentralGoogle Scholar
  19. Ellis CM, Nagpal P, Young JC, Hagen G, Guilfoyle TJ, Reed JW (2005) AUXIN RESPONSE FACTOR1 and AUXIN RESPONSE FACTOR2 regulate senescence and floral organ abscission in Arabidopsis thaliana. Development 132:4563–4574CrossRefGoogle Scholar
  20. Fernandez DE, Heck GR, Perry SE, Patterson SE, Bleecker AB, Fang SC (2000) The embryo MADS domain factor AGL15 acts postembryonically: inhibition of perianth senescence and abscission via constitutive expression. Plant Cell 12:183–198CrossRefGoogle Scholar
  21. Fernandez DE, Wang CT, Zheng Y, Adamczyk BJ, Singhal R, Hall PK, Perry SE (2014) The MADS-domain factors AGAMOUS-LIKE15 and AGAMOUS-LIKE18, along with SHORT VEGETATIVE PHASE and AGAMOUS-LIKE24, are necessary to block floral gene expression during the vegetative phase. Plant Physiol 165:1591–1603CrossRefGoogle Scholar
  22. Hepworth SR, Zhang Y, McKim S, Li X, Haughn GW (2005) BLADE-ON-PETIOLE-dependent signaling controls leaf and floral patterning in Arabidopsis. Plant Cell 17:1434–1448CrossRefGoogle Scholar
  23. Huang H, Mizukami Y, Hu Y, Ma H (1993) Isolation and characterization of the binding sequences for the product of the Arabidopsis floral homeotic gene AGAMOUS. Nucleic Acids Res 21:4769–4776CrossRefGoogle Scholar
  24. Hwang I, Sheen J, Muller B (2012) Cytokinin signaling networks. Annu Rev Plant Biol 63:353–380CrossRefGoogle Scholar
  25. Jinn TL, Stone JM, Walker JC (2000) HAESA, an Arabidopsis leucine-rich repeat receptor kinase, controls floral organ abscission. Genes Dev 14:108–117PubMedPubMedCentralGoogle Scholar
  26. Kater MM, Dreni L, Colombo L (2006) Functional conservation of MADS-box factors controlling floral organ identity in rice and Arabidopsis. J Exp Bot 57:3433–3444CrossRefGoogle Scholar
  27. Kim JI, Murphy AS, Baek D, Lee SW, Yun DJ, Bressan RA, Narasimhan ML (2011) YUCCA6 over-expression demonstrates auxin function in delaying leaf senescence in Arabidopsis thaliana. J Exp Bot 62:3981–3992CrossRefGoogle Scholar
  28. Koornneef M, van der Veen JH (1980) Induction and analysis of gibberellin-insensitive mutants in Arabidopsis thaliana (L.) Heynh. Theor Appl Genet 58:257–263CrossRefGoogle Scholar
  29. Koyama T (2014) The roles of ethylene and transcription factors in the regulation of onset of leaf senescence. Front Plant Sci 5:650CrossRefGoogle Scholar
  30. Lee JH, Chung KS, Kim SK, Ahn JH (2014) Post-translational regulation of short vegetative phase as a major mechanism for thermoregulation of flowering. Plant Signal Behav 9:e28193CrossRefGoogle Scholar
  31. Lewis MW, Leslie ME, Liljegren SJ (2006) Plant separation: 50 ways to leave your mother. Curr Opin Plant Biol 9:59–65CrossRefGoogle Scholar
  32. McKim SM, Stenvik GE, Butenko MA, Kristiansen W, Cho SK, Hepworth SR, Aalen RB, Haughn GW (2008) The BLADE-ON-PETIOLE genes are essential for abscission zone formation in Arabidopsis. Development 135:1537–1546CrossRefGoogle Scholar
  33. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–479CrossRefGoogle Scholar
  34. Noh YS, Amasino RM (1999) Identification of a promoter region responsible for the senescence-specific expression of SAG12. Plant Mol Biol 41:181–194CrossRefGoogle Scholar
  35. Norberg M, Holmlund M, Nilsson O (2005) The BLADE ON PETIOLE genes act redundantly to control the growth and development of lateral organs. Development 132:2203–2213CrossRefGoogle Scholar
  36. Patterson SE, Bleecker AB (2004) Ethylene-dependent and -independent processes associated with floral organ abscission in Arabidopsis. Plant Physiol 134:194–203CrossRefGoogle Scholar
  37. Peng J, Carol P, Richards DE, King KE, Cowling RJ, Murphy GP, Harberd NP (1997) The Arabidopsis GAI gene defines a signaling pathway that negatively regulates gibberellin responses. Genes Dev 11:3194–3205CrossRefGoogle Scholar
  38. Riefler M, Novak O, Strnad M, Schmülling T (2006) Arabidopsis cytokinin receptor mutants reveal functions in shoot growth, leaf senescence, seed size, germination, root development, and cytokinin metabolism. Plant Cell 18:40–54CrossRefGoogle Scholar
  39. Rogers HJ (2013) From models to ornamentals: how is flower senescence regulated? Plant Mol Biol 82:563–574CrossRefGoogle Scholar
  40. Schenk PM, Kazan K, Rusu AG, Manners JM, Maclean DJ (2005) The SEN1 gene of Arabidopsis is regulated by signals that link plant defence responses and senescence. Plant Physiol Biochem 43:997–1105CrossRefGoogle Scholar
  41. Sekhon RS, Childs KL, Santoro N, Foster CE, Buell CR, de Leon N, Kaeppler SM (2012) Transcriptional and metabolic analysis of senescence induced by preventing pollination in maize. Plant Physiol 159:1730–1744CrossRefGoogle Scholar
  42. Shi H, Reiter RJ, Tan DX, Chan Z (2015) INDOLE-3-ACETIC ACID INDUCIBLE 17 positively modulates natural leaf senescence through melatonin-mediated pathway in Arabidopsis. J Pineal Res 58:26–33CrossRefGoogle Scholar
  43. Silverstone AL, Ciampaglio CN, Sun T (1998) The Arabidopsis RGA gene encodes a transcriptional regulator repressing the gibberellin signal transduction pathway. Plant Cell 10:155–169CrossRefGoogle Scholar
  44. Stenvik GE, Tandstad NM, Guo Y, Shi CL, Kristiansen W, Holmgren A, Clark SE, Aalen RB, Butenko MA (2008) The EPIP peptide of INFLORESCENCE DEFICIENT IN ABSCISSION is sufficient to induce abscission in Arabidopsis through the receptor-like kinases HAESA and HAESA-LIKE2. Plant Cell 20:1805–1817CrossRefGoogle Scholar
  45. Stepanova AN, Alonso JM (2005) Arabidopsis ethylene signaling pathway. Sci STKE 2005:cm4PubMedGoogle Scholar
  46. Tao Z, Shen L, Liu C, Liu L, Yan Y, Yu H (2012) Genome-wide identification of SOC1 and SVP targets during the floral transition in Arabidopsis. Plant J 70:549–561CrossRefGoogle Scholar
  47. Theissen G (2001) Development of floral organ identity: stories from the MADS house. Curr Opin Plant Biol 4:75–85CrossRefGoogle Scholar
  48. Theissen G, Kim JT, Saedler H (1996) Classification and phylogeny of the MADS-box multigene family suggest defined roles of MADS-box gene subfamilies in the morphological evolution of eukaryotes. J Mol Evol 43:484–516CrossRefGoogle Scholar
  49. Tyler L, Thomas SG, Hu J, Dill A, Alonso JM, Ecker JR, Sun TP (2004) DELLA proteins and gibberellin-regulated seed germination and floral development in Arabidopsis. Plant Physiol 135:1008–1019CrossRefGoogle Scholar
  50. van Doorn WG, Kamdee C (2014) Flower opening and closure: an update. J Exp Bot 65:5749–5757CrossRefGoogle Scholar
  51. Wen CK, Chang C (2002) Arabidopsis RGL1 encodes a negative regulator of gibberellin responses. Plant Cell 14:87–100CrossRefGoogle Scholar
  52. Woltering EJ, Somhorst D, Van Der Veer P (1995) The role of ethylene in interorgan signaling during flower senescence. Plant Physiol 109:1219–1225CrossRefGoogle Scholar
  53. Yu H, Ito T, Zhao Y, Peng J, Kumar P, Meyerowitz EM (2004) Floral homeotic genes are targets of gibberellin signaling in flower development. Proc Natl Acad Sci U S A 101:7827–7832CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Institute of BiotechnologyNational Chung Hsing UniversityTaichungRepublic of China
  2. 2.Botany DepartmentNational Museum of Natural ScienceTaichungRepublic of China
  3. 3.Advanced Plant Biotechnology CenterNational Chung Hsing UniversityTaichungRepublic of China

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