Plant Molecular Biology

, Volume 68, Issue 1–2, pp 17–30 | Cite as

Functional analysis reveals pleiotropic effects of rice RING-H2 finger protein gene OsBIRF1 on regulation of growth and defense responses against abiotic and biotic stresses

  • Huizhi Liu
  • Huijuan Zhang
  • Yayun Yang
  • Guojun Li
  • Yuxia Yang
  • Xiao’e Wang
  • B. M. Vindhya S. Basnayake
  • Dayong Li
  • Fengming SongEmail author


RING finger proteins comprise a large family and play key roles in regulating growth/developmental processes, hormone signaling and responses to biotic and abiotic stresses in plants. A rice gene, OsBIRF1, encoding a putative RING-H2 finger protein, was cloned and identified. OsBIRF1 encodes a 396 amino acid protein belonging to the ATL family characterized by a conserved RING-H2 finger domain (C-X2-C-X15-C-X1-H-X2-H-X2-C-X10-C-X2-C), a transmembrane domain at the N-terminal, a basic amino acid rich region and a characteristic GLD region. Expression of OsBIRF1 was up-regulated in rice seedlings after treatment with benzothaidiazole, salicylic acid, l-aminocyclopropane-1-carboxylic acid and jasmonic acid, and was induced differentially in incompatible but not compatible interactions between rice and Magnaporthe grisea, the causal agent of blast disease. Transgenic tobacco plants that constitutively express OsBIRF1 exhibit enhanced disease resistance against tobacco mosaic virus and Pseudomonas syringae pv. tabaci and elevated expression levels of defense-related genes, e.g. PR-1, PR-2, PR-3 and PR-5. The OsBIRF1-overexpressing transgenic tobacco plants show increased oxidative stress tolerance to exogenous treatment with methyl viologen and H2O2, and up-regulate expression of oxidative stress-related genes. Reduced ABA sensitivity in root elongation and increased drought tolerance in seed germination were also observed in OsBIRF1 transgenic tobacco plants. Furthermore, the transgenic tobacco plants show longer roots and higher plant heights as compared with the wild-type plants, suggesting that overexpression of OsBIRF1 promote plant growth. These results demonstrate that OsBIRF1 has pleiotropic effects on growth and defense response against multiple abiotic and biotic stresses.


Rice OsBIRF1 RING finger proteins Disease resistance Abiotic stress 



Abscisic acid


1-Amino cyclopropane-1-carboxylic acid




Cauliflower mosaic virus




Hydrogen peroxide


Jasmonic acid


Methyl viologen


Open reading frame


Polymerase chain reaction


Polyethylene glycol




Reverse transcription-PCR


Salicylic acid


Tobacco mosaic virus



We are grateful to Dr Zuhua He, Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Science, for the rice near-isogenic lines H8R and H8S, and Mr Rongyao Chai, Zhejiang Academy of Agricultural Science, for the Magnaporthe grisea isolate 85-14B1. This study was supported by National Natural Science Foundation of China (grants no. 30571209 and 30771399), the National High-Tech (“863”) Project (2006AA10Z430), the National Key Basic Research and Development Program (2006CB101903) and the Fund for the New Century Talent Program from MOE of China.


  1. Cheung MY, Zeng NY, Tong SW, Li FWY, Zhao KJ, Zhang Q, Sun SM, Lam HM (2007) Expression of a RING-HC protein from rice improves resistance to Pseudomonas syringae pv. tomato DC3000 in transgenic Arabidopsis thaliana. J Exp Bot 58:4147–4159PubMedCrossRefGoogle Scholar
  2. Disch S, Anastasiou E, Sharma VK, Laux T, Fletcher JC, Lenhard M (2006) The E3 ubiquitin ligase BIG BROTHER controls Arabidopsis organ size in a dosage-dependent manner. Curr Biol 16:272–279PubMedCrossRefGoogle Scholar
  3. Dong CH, Agarwal M, Zhang Y, Xie Q, Zhu JK (2006) The negative regulator of plant cold responses, HOS1, is a RING E3 ligase that mediates the ubiquitination and degradation of ICE1. Proc Natl Acad Sci USA 103:8281–8286PubMedCrossRefGoogle Scholar
  4. Duek PD, Elmer MV, van Oosten VR, Fankhauser C (2004) The degradation of HFR1, a putative bHLH class transcription factor involved in light signaling, is regulated by phosphorylation and requires COP1. Curr Biol 14:2296–2301PubMedCrossRefGoogle Scholar
  5. Durrant WE, Rowland O, Piedras P, Hammond-Kosack KE, Jones JDG (2000) cDNA-AFLP reveals a striking overlap in race specific resistance and wound response gene expression profiles. Plant Cell 12:963–977PubMedCrossRefGoogle Scholar
  6. Fang S, Weissman AM (2004) A field guide to ubiquitylation. Cell Mol Life Sci 61:1546–1561PubMedCrossRefGoogle Scholar
  7. Finkelstein RR, Gampala SS, Rock CD (2002) Abscisic acid signaling in seeds and seedlings. Plant Cell 14(suppl):S15–S45PubMedGoogle Scholar
  8. Freemont PS (2000) RING for destruction? Curr Biol 10:R84–R87PubMedCrossRefGoogle Scholar
  9. Glazebrook J (2005) Contrasting mechanisms of defense against biotrophic and necrotrophic pathogens. Annu Rev Phytopathol 43:205–227PubMedCrossRefGoogle Scholar
  10. Glickman MH, Ciechanover A (2002) The ubiquitin-proteasome proteolytic pathway: destruction for the sake of construction. Physiol Rev 82:373–428PubMedGoogle Scholar
  11. Hardtke CS, Gohda K, Osterlund MT, Oyama T, Okada K, Deng XW (2000) HY5 stability and activity in Arabidopsis is regulated by phosphorylation in its COP1 binding domain. EMBO J 19:4997–5006PubMedCrossRefGoogle Scholar
  12. Hardtke CS, Okamoto H, Stoop-Myer C, Deng XW (2002) Biochemical evidence for ubiquitin ligase activity of the Arabidopsis COP1 interacting protein 8 (CIP8). Plant J 30:385–394PubMedCrossRefGoogle Scholar
  13. Hershko A, Ciechanover A (1998) The ubiquitin system. Ann Rev Biochem 67:425–479PubMedCrossRefGoogle Scholar
  14. Holm M, Ma LG, Qu LJ, Deng XW (2002) Two interacting bZIP proteins are direct targets of COP1-mediated control of light-dependent gene expression in Arabidopsis. Genes Dev 16:1247–1259PubMedCrossRefGoogle Scholar
  15. Hondo D, Hase S, Kanayama Y, Yoshikawa N, Takenaka S, Takahashi H (2007) The LeATL6-associated ubiquitin/proteasome system may contribute to fungal elicitor-activated defense response via the jasmonic acid-dependent signaling pathway in tomato. Mol Plant Microbe Interact 20:72–81PubMedCrossRefGoogle Scholar
  16. Hong JK, Choi HW, Hwang IS, Hwang BK (2007) Role of a novel pathogen-induced pepper C3-H-C4 type RING-finger protein gene, CaRFP1, in disease susceptibility and osmotic stress tolerance. Plant Mol Biol 63:571–588PubMedCrossRefGoogle Scholar
  17. Jang IC, Yang JY, Seo HS, Chua NH (2005) HFR1 is targeted by COP1 E3 ligase for post-translational proteolysis during phytochrome A signaling. Genes Dev 19:593–602PubMedCrossRefGoogle Scholar
  18. Joazeiro CA, Weissman AM (2000) RING finger proteins: mediators of ubiquitin ligase activity. Cell 102:549–552PubMedCrossRefGoogle Scholar
  19. Kam J, Gresshoff P, Shorter R, Xue G-P (2007) Expression analysis of RING zinc finger genes from Triticum aestivum and identification of TaRZF70 that contains four RING-H2 domains and differentially responds to water deficit between leaf and root. Plant Sci 173:650–659CrossRefGoogle Scholar
  20. Karlowski WM, Hirsch AM (2003) The over-expression of an alfalfa RING-H2 gene induces pleiotropic effects on plant growth and development. Plant Mol Biol 52:121–133PubMedCrossRefGoogle Scholar
  21. Katoh S, Hong C, Tsunoda Y, Murata K, Takai R, Minami E, Yamazaki T, Katoh E (2003) High precision NMR structure and function of the RING-H2 finger domain of EL5, a rice protein whose expression is increased upon exposure to pathogen-derived oligosaccharides. J Biol Chem 278:15341–15348PubMedCrossRefGoogle Scholar
  22. Katoh S, Tsunoda Y, Murata K, Minami E, Katoh E (2005) Active site residues and amino acid specificity of the ubiquitin carrier protein-binding RING-H2 finger domain. J Biol Chem 280:41015–41024PubMedCrossRefGoogle Scholar
  23. Kawasaki T, Nam J, Boyes DC, Holt 3rd BF, Hubert DA, Wiig A, Dangl JL (2005) A duplicated pair of Arabidopsis RING-finger E3 ligases contribute to the RPM1- and RPS2-mediated hypersensitive response. Plant J 44:258–270PubMedCrossRefGoogle Scholar
  24. Ko JH, Yang SH, Han KH (2006) Upregulation of an Arabidopsis RING-H2 gene, XERICO, confers drought tolerance through increased abscisic acid biosynthesis. Plant J 47:343–355PubMedCrossRefGoogle Scholar
  25. Koiwai H, Tagiri A, Katoh S, Katoh E, Ichikawa H, Minami E, Nishizawa Y (2007) RING-H2 type ubiquitin ligase EL5 is involved in root development through the maintenance of cell viability in rice. Plant J 51:92–104PubMedCrossRefGoogle Scholar
  26. Lechner E, Goloubinoff P, Genschik P, Shen WH (2002) A gene trap dissociation insertion line, associated with a RING-H2 finger gene, shows tissue specific and developmental regulated expression of the gene in Arabidopsis. Gene 290:63–71PubMedCrossRefGoogle Scholar
  27. Lee H, Xiong L, Gong Z, Ishitani M, Stevenson B, Zhu JK (2001) The Arabidopsis HOS1 gene negatively regulates cold signal transduction and encodes a RING finger protein that displays cold-regulated nucleo-cytoplasmic partitioning. Genes Dev 15:912–924PubMedCrossRefGoogle Scholar
  28. Liu K, Wang L, Xu Y, Chen N, Ma Q, Li F, Chong K (2007) Overexpression of OsCOIN, a putative cold inducible zinc finger protein, increased tolerance to chilling, salt and drought, and enhanced proline level in rice. Planta 226:1007–1116PubMedCrossRefGoogle Scholar
  29. Luo H, Song F, Goodman RM, Zheng Z (2005a) Up-regulation of OsBIHD1, a rice gene encoding BELL homeodomain transcriptional factor, in disease resistance responses. Plant Biol 7:459–468PubMedCrossRefGoogle Scholar
  30. Luo H, Song F, Zheng Z (2005b) Overexpression in transgenic tobacco reveals different roles for the rice homeodomain gene OsBIHD1 in biotic and abiotic stress responses. J Exp Bot 56:2673–2682PubMedCrossRefGoogle Scholar
  31. Ma L, Gao Y, Qu L, Chen Z, Li J, Zhao H, Deng XW (2002) Genomic evidence for COP1 as a repressor of light-regulated gene expression and development in Arabidopsis. Plant Cell 14:2383–2398PubMedCrossRefGoogle Scholar
  32. Meng XB, Zhao WS, Lin RM, Wang M, Peng YL (2006) Molecular cloning and characterization of a rice blast-inducible RING-H2 type zinc finger gene. DNA Seq 17:41–48PubMedGoogle Scholar
  33. Mittler R, Vanderauwera S, Gollery M, Van Breusegem F (2004) The reactive oxygen gene network of plants. Trends Plant Sci 9:490–498PubMedCrossRefGoogle Scholar
  34. Mittler R, Kim Y, Song L, Coutu J, Coutu A, Ciftci-Yilmaz S, Lee H, Stevenson B, Zhu JK (2006) Gain- and loss-of-function mutations in Zat10 enhance the tolerance of plants to abiotic stress. FEBS Lett 580:6537–6542PubMedCrossRefGoogle Scholar
  35. Molnar G, Bancoş S, Nagy F, Szekeres M (2002) Characterisation of BRH1, a brassinosteroid-responsive RING-H2 gene from Arabidopsis thaliana. Planta 215:127–133PubMedCrossRefGoogle Scholar
  36. Moon J, Parry G, Estelle M (2004) The ubiquitin-proteasome pathway and plant development. Plant Cell 16:3181–3195PubMedCrossRefGoogle Scholar
  37. Nishimura R, Ohmori M, Fujita H, Kawaguchi M (2002) A Lotus basic leucine zipper protein with a RING-finger motif negatively regulates the developmental program of nodulation. Proc Natl Acad Sci USA 99:15206–15210PubMedCrossRefGoogle Scholar
  38. Nodzon LA, Xu WH, Wang YS, Pi LY, Chakrabarty PK, Song WY (2004) The ubiquitin ligase XBAT32 regulates lateral root development in Arabidopsis. Plant J 40:996–1006PubMedCrossRefGoogle Scholar
  39. Osterlund MT, Hardtke CS, Wei N, Deng XW (2000) Targeted destabilization of HY5 during light-regulated development of Arabidopsis. Nature 405:462–466PubMedCrossRefGoogle Scholar
  40. Ramonell K, Berrocal-Lobo M, Koh S, Wan J, Edwards H, Stacey G, Somerville S (2005) Loss-of-function mutations in chitin responsive genes show increased susceptibility to the powdery mildew pathogen Erysiphe cichoracearum. Plant Physiol 138:1027–1036PubMedCrossRefGoogle Scholar
  41. Sahin-Cevik M, Moore GA (2007) Isolation and characterization of a novel RING-H2 finger gene induced in response to cold and drought in the interfertile Citrus relative Poncirus trifoliate. Physiol Plant 126:153–161CrossRefGoogle Scholar
  42. Saijo Y, Sullivan JA, Wang H, Yang J, Shen Y, Rubio V, Ma L, Hoecker U, Deng XW (2003) The COP1-SPA1 interaction defines a critical step in phytochrome A-mediated regulation of HY5 activity. Genes Dev 17:2642–2647PubMedCrossRefGoogle Scholar
  43. Salinas-Mondragon RE, Garciduenas-Pina C, Guzman P (1999) Early elicitor induction in members of a novel multigene family coding for highly related RING-H2 proteins in Arabidopsis thaliana. Plant Mol Biol 40:579–590PubMedCrossRefGoogle Scholar
  44. Saurin AJ, Borden KL, Boddy MN, Freemont PS (1996) Does this have a familiar RING? Trends Biochem Sci 21:208–214PubMedGoogle Scholar
  45. Schumann U, Prestele J, O’Geen H, Brueggeman R, Wanner G, Gietl C (2007) Requirement of the C3HC4 zinc RING finger of the Arabidopsis PEX10 for photorespiration and leaf peroxisome contact with chloroplasts. Proc Natl Acad Sci USA 104:1069–1074PubMedCrossRefGoogle Scholar
  46. Seo HS, Yang JY, Ishikawa M, Bolle C, Ballesteros ML, Chua NH (2003) LAF1 ubiquitination by COP1 controls photomorphogenesis and is stimulated by SPA1. Nature 423:995–999PubMedCrossRefGoogle Scholar
  47. Seo HS, Watanabe E, Tokutomi S, Nagatani A, Chua NH (2004) Photoreceptor ubiquitination by COP1 E3 ligase desensitizes phytochrome A signaling. Genes Dev 18:617–622PubMedCrossRefGoogle Scholar
  48. Serrano M, Guzman P (2004) Isolation and gene expression analysis of Arabidopsis thaliana mutants with constitutive expression of ATL2, an early elicitor-response RING-H2 zinc-finger gene. Genetics 167:919–929PubMedCrossRefGoogle Scholar
  49. Serrano M, Parra S, Alcaraz LD, Guzman P (2006) The ATL gene family from Arabidopsis thaliana and Oryza sativa comprises a large number of putative ubiquitin ligases of the RING-H2 type. J Mol Evol 62:434–445PubMedCrossRefGoogle Scholar
  50. Shimomura K, Nomura M, Tajima S, Kouchi H (2006) LjnsRING, a novel RING finger protein, is required for symbiotic interactions between Mesorhizobium loti and Lotus japonicus. Plant Cell Physiol 47:1572–1581PubMedCrossRefGoogle Scholar
  51. Smalle J, Vierstra RD (2004) The ubiquitin 26s proteasome proteolytic pathway. Ann Rev Plant Biol 55:555–590CrossRefGoogle Scholar
  52. Song F, Goodman RM (2002) OsBIMK1, a rice MAP kinase gene involved in disease resistance responses. Planta 215:997–1005PubMedCrossRefGoogle Scholar
  53. Sonoda Y, Yao SG, Sako K, Sato T, Kato W, Ohto MA, Ichikawa T, Matsui M, Yamaguchi J, Ikeda A (2007) SHA1, a novel RING finger protein, functions in shoot apical meristem maintenance in Arabidopsis. Plant J 50:586–596PubMedCrossRefGoogle Scholar
  54. Stone SL, Hauksdottir H, Troy A, Herschleb J, Kraft E, Callis J (2005) Functional analysis of the RING-type ubiquitin ligase family of Arabidopsis. Plant Physiol 137:13–30PubMedCrossRefGoogle Scholar
  55. Takai R, Hasegawa K, Kaku H, Shibuya N, Minami E (2001) Isolation and analysis of expression mechanisms of a rice gene, EL5, which shows structural similarity to ATL family from Arabidopsis, in response to N-acetylchitooligosaccharide elicitor. Plant Sci 160:577–583PubMedCrossRefGoogle Scholar
  56. Takai R, Matsuda N, Nakano A, Hasegawa K, Akimoto C, Shibuya N, Minami E (2002) EL5, a rice N-acetylchitooligosaccharide elicitor-responsive RING-H2 finger protein, is a ubiquitin ligase which functions in vitro in co-operation with an elicitor-responsive ubiquitin-conjugating enzyme, OsUBC5b. Plant J 30:447–455PubMedCrossRefGoogle Scholar
  57. Veronese P, Narasimhan ML, Stevenson RA, Zhu JK, Weller SC, Subbarao KV, Bressan RA (2003) Identification of a locus controlling Verticillium disease symptom response in Arabidopsis thaliana. Plant J 35:574–587PubMedCrossRefGoogle Scholar
  58. Xie Q, Guo HS, Dallman G, Fang SY, Weissman AM, Chua NH (2002) SINAT5 promotes ubiquitin-related degradation of NAC1 to attenuate auxin signals. Nature 419:167–170PubMedCrossRefGoogle Scholar
  59. Xu R, Li QQ (2003) A RING-H2 zinc-finger protein gene RIE1 is essential for seed development in Arabidopsis. Plant Mol Biol 53:37–50PubMedCrossRefGoogle Scholar
  60. Yang J, Lin R, Sullivan J, Hoecker U, Liu B, Xu L, Deng XW, Wang H (2005) Light regulates COP1-mediated degradation of HFR1, a transcription factor essential for light signaling in Arabidopsis. Plant Cell 17:804–821PubMedCrossRefGoogle Scholar
  61. Zhang X, Garreton V, Chua NH (2005) The AIP2 E3 ligase acts as a novel negative regulator of ABA signaling by promoting ABI3 degradation. Genes Dev 19:1532–1543PubMedCrossRefGoogle Scholar
  62. Zhang Y, Yang C, Li Y, Zheng N, Chen H, Zhao Q, Gao T, Guo H, Xie Q (2007) SDIR1 is a RING finger E3 ligase that positively regulates stress-responsive abscisic acid signaling in Arabidopsis. Plant Cell 19:1912–1929PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2008

Authors and Affiliations

  • Huizhi Liu
    • 1
  • Huijuan Zhang
    • 1
  • Yayun Yang
    • 1
  • Guojun Li
    • 1
  • Yuxia Yang
    • 1
  • Xiao’e Wang
    • 1
  • B. M. Vindhya S. Basnayake
    • 1
  • Dayong Li
    • 1
  • Fengming Song
    • 1
    • 2
    Email author
  1. 1.State Key Laboratory for Rice Biology, Institute of BiotechnologyZhejiang UniversityHangzhouPeople’s Republic of China
  2. 2.Department of Plant ProtectionZhejiang UniversityHangzhouPeople’s Republic of China

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