, Volume 224, Issue 5, pp 1209–1225 | Cite as

Functional roles of the pepper pathogen-induced bZIP transcription factor, CAbZIP1, in enhanced resistance to pathogen infection and environmental stresses

  • Sung Chul Lee
  • Hyong Woo Choi
  • In Sun Hwang
  • Du Seok Choi
  • Byung Kook HwangEmail author
Original Article


Transcription factors often belong to multigene families and their individual contribution in a particular regulatory network remains difficult to assess. We identify and functionally characterize the pepper bZIP transcription factor CAbZIP1 gene isolated from pepper leaves infected with Xanthomonas campestris pv. vesicatoria. Transient expression analysis of the CAbZIP1–GFP fusion protein in Arabidopsis protoplasts revealed that the CAbZIP1 protein is localized in the nucleus. The N-terminal region of CAbZIP1 fused to the GAL4 DNA-binding domain is required to activate transcription of reporter genes in yeast. The CAbZIP1 transcripts are constitutively expressed in the pepper root and flower, but not in the leaf, stem and fruit. The CAbZIP1 gene is locally or systemically induced in pepper plants infected by either X. campestris pv. vesicatoria or Pseudomonas fluorescens. The CAbZIP1 gene is also induced by abiotic elicitors and environmental stresses. The CAbZIP1 transgenic Arabidopsis exhibits a dwarf phenotype, indicating that CAbZIP1 may be involved in plant development. The CAbZIP1 overexpression in the transgenic Arabidopsis plants confers enhanced resistance to Pseudomonas syringae pv. tomato DC3000, accompanied by expression of the AtPR-4 and AtRD29A. The transgenic plants also exhibit increased drought and salt tolerance during all growth stages. Moreover, the transgenic plants are tolerant to methyl viologen-oxidative stress. Together, these data suggest that the CAbZIP1 transcription factor function as a possible regulator in enhanced disease resistance and environmental stress tolerance.


bZIP transcription factor Disease resistance Environmental stress Transactivation Transgenic plant 



Abscisic acid


Hypersensitive response


Jasmonic acid


Nitric oxide


Methyl viologen


Pathogenesis related


Salicylic acid


Systemic acquired resistance



This research was financially supported by a grant (CG1432) from the Crop Functional Genomics Center of the 21st Century, Frontier Research Program, funded by the Ministry of Science and Technology of the Republic of Korea, as well as a grant from the Center for Plant Molecular Genetics and Breeding Research, Korea Science and Engineering Foundation.


  1. Abe H, Yamaguchi-Shinozaki K, Urao T, Iwasaki T, Hosokawa D, Shinozaki K (1997) Role of Arabidopsis MYC and MYB homologs in drought- and abscisic acid-regulated gene expression. Plant Cell 9:1859–1868CrossRefPubMedGoogle Scholar
  2. Asada K (1999) The water–water cycle in chloroplasts: scanvenging of active oxygen and dissipation of excess photons. Annu Rev Plant Physiol Plant Mol Biol 50:601–639CrossRefPubMedGoogle Scholar
  3. Boch J, Verbsky ML, Robertson T, Larkin JC, Kunkel BN (1998) Analysis of resistance gene-mediated defense response in Arabidopsis thaliana plants carrying a mutation in CPR5. Mol Plant Microbe Interact 12:1196–1206CrossRefGoogle Scholar
  4. Busk PK, Pages M (1998) Regulation of abscisic acid induced transcription. Plant Mol Biol 37:425–435CrossRefPubMedGoogle Scholar
  5. Chakravarthy S, Tuori RP, D’Ascenzo MD, Fobert PR, Despres C, Martin GB (2003) The tomato transcription factor Pti4 regulates defense-related gene expression via GCC box and non-GCC box cis elements. Plant Cell 2:3033–3050CrossRefGoogle Scholar
  6. Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16:735–743CrossRefPubMedGoogle Scholar
  7. Després C, DeLong C, Glaze S, Liu E, Fobert PR (2000) The Arabidopsis NPR1/NIM1 protein enhances the DNA binding activity of a subgroup of the TGA family of bZIP transcription factors. Plant Cell 12:279–290CrossRefPubMedGoogle Scholar
  8. Després C, Chubak C, Rochon A, Clark R, Bethune T, Desveaux D, Fobert PR (2003) The Arabidopsis NPR1 disease resistance protein is a novel cofactor that confers redox regulation of DNA binding activity to the basic domain/leucine zipper transcription factor TGA1. Plant Cell 15:2181–2191CrossRefPubMedGoogle Scholar
  9. Epple P, Klaus A, Bohlmann H (1997) Overexpression of an endogenous thionin enhances resistance of Arabidopsis against Fusarium oxysporum. Plant Cell 9:509–520CrossRefPubMedGoogle Scholar
  10. Eulgem T, Rushton PJ, Robatzek S, Somssich IE (2000) The WRKY superfamily of plant transcription factors. Trends Plant Sci 5:199–206CrossRefPubMedGoogle Scholar
  11. Fan W, Dong X (2002) In vivo interaction between NPR1 and transcription factor TGA2 leads to salicylic acid-mediated gene activation in Arabidopsis. Plant Cell 14:1377–1389CrossRefPubMedGoogle Scholar
  12. Finkelstein R, Lynch T (2000) The Arabidopsis abscisic acid response gene ABI5 encodes a basic leucine zipper transcription factor. Plant Cell 12:599–609CrossRefPubMedGoogle Scholar
  13. Glazebrook J (1999) Genes controlling expression of defense responses in Arabidopsis. Curr Opin Plant Biol 2:280–286CrossRefPubMedGoogle Scholar
  14. Gu Y, Wildermuth MC, Chakravarthy S, Loh Y, Yang C, He X, Han Y, Martin GB (2002) Tomato transcription factors Pti4, Pti5, and Pti6 activate defense responses when expressed in Arabidopsis. Plant Cell 14:817–831CrossRefPubMedGoogle Scholar
  15. Hagen G, Guilfoyle T (2002) Auxin-responsive gene expression: genes, promoters and regulatory factors. Plant Mol Biol 49:373–385CrossRefPubMedGoogle Scholar
  16. Hahn S (1993) Structure (?) and function of acidic transcription activators. Cell 72:481–483CrossRefPubMedGoogle Scholar
  17. Heinekamp T, Kuhlmann M, Lenk A, Strathmann A, Dröge-Laser W (2002) The tobacco bZIP transcription factor BZI-1 binds to G-box elements in the promoters of phenylpropanoid pathway genes in vitro, but it is not involved in their regulation in vivo. Mol Genet Genomics 267:16–26CrossRefPubMedGoogle Scholar
  18. Heinekamp T, Lenk A, Strathmann A, Kuhlmann M, Froissard M, Müller A, Perrot-Rechenmann C, Dröge-Laser W (2004) The tobacco bZIP transcription factor BZI-1 binds the GH3 promoter in vivo and modulates auxin-induced transcription. Plant J 38:298–309CrossRefPubMedGoogle Scholar
  19. Hobo T, Kowyama Y, Hattori T (1999) A bZIP factor, TRAB1, interacts with VP1 and mediates abscisic acid-induced transcription. Proc Natl Acad Sci USA 96:15348–15353CrossRefPubMedGoogle Scholar
  20. Hong JK, Hwang BK (2005) Functional characterization of PR-1 protein, β-1,3-glucanase and chitinase genes during defense response to biotic and abiotic stresses in Capsicum annuum. Plant Pathol J 21:195–206Google Scholar
  21. Hunter T, Karin M (1992) The regulation of transcription by phosphorylation. Cell 70:375–387CrossRefPubMedGoogle Scholar
  22. Hurst HC (1995) Transcription factors. 1. bZIP proteins. Protein Profile 2:105–168Google Scholar
  23. Ishitani M, Xiong L, Lee H, Stevenson B, Zhu JK (1998) HOS1, a genetic locus involved in cold-responsive gene expression in Arabidopsis. Plant Cell 10:1151–1161PubMedCrossRefGoogle Scholar
  24. 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 102:5280–5285CrossRefPubMedGoogle Scholar
  25. Jakoby M, Weisshaar B, Dröge-Laser W, Tiedemann J, Kroij T, Parcy F (2002) The family of bZIP transcription factors in Arabidopsis thaliana. Trends Plant Sci 7:106–111CrossRefPubMedGoogle Scholar
  26. Johnson RR, Wagner RL, Verhey SD, Walker-Simmons MK (2002) The abscisic acid-responsive kinase PKABA1 interacts with a seed-specific abscisic acid response element-binding factor, TaABF, and phosphorylates TaABF peptide sequences. Plant Physiol 130:837–846CrossRefPubMedGoogle Scholar
  27. Jung HW, Hwang BK (2000) Isolation, partial sequencing, and expression of pathogenesis-related cDNA genes from pepper leaves infected by Xanthomonas campestis pv. vesicatoria. Mol Plant Microbe Interact 13:136–142PubMedCrossRefGoogle Scholar
  28. Kang SG, Jin JB, Piao HL, Pih KT, Jang HJ, Lim JH, Hwang I (1998) Molecular cloning of an Arabidopsis cDNA encoding a dynamin-like protein that is localized to plastids. Plant Mol Biol 38:437–447CrossRefPubMedGoogle Scholar
  29. Kang J, Choi H, Im M, Kim SY (2002) Arabidopsis basic leucine zipper proteins mediate stress-responsive abscisic acid signaling. Plant Cell 14:343–357CrossRefPubMedGoogle Scholar
  30. 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 52:121–133CrossRefPubMedGoogle Scholar
  31. Karpinski S, Wingsle G, Karpinska B, Hällogren JE (2001) Redox sensing of photooxidative stress and acclamatory mechanisms in plants. In: Aro EM, Andersson B (eds) Regulation of photosynthesis. Kluwer, Dordrecht, pp 469–486Google Scholar
  32. Kim YJ, Martin GB (2004) Molecular mechanisms involved in bacterial speck disease resistance of tomato. Plant Pathol J 20:7–12Google Scholar
  33. Kim SY, Chung H-J, Thomas TL (1997) Isolation of a novel class of bZIP transcription factors that interact with ABA-responsive and embryo-specification elements in the Dc3 promoter using a modified yeast one-hybrid system. Plant J 11:1237–1251CrossRefPubMedGoogle Scholar
  34. Kim SH, Hong JK, Lee SC, Sohn KH, Jung HW, Hwang BK (2004) CAZFP1, Cys2/His2-type zinc-finger transcription factor gene functions as a novel pathogen-induced early-defense gene in Capsicum annuum. Plant Mol Biol 55:883–904PubMedGoogle Scholar
  35. Kranner I, Beckett RP, Wornik S, Zorn M, Pfeifhofer HW (2002) Revival of a resurrection plant correlates with its antioxidant status. Plant J 31:13–24PubMedCrossRefGoogle Scholar
  36. Kuhlmann M, Horvay K, Stathmann A, Heinekamp T, Fischer U, Böttner S, Dröge-Laser W (2003) The α-helical D1 domain of the bZIP transcription factor BZI-1 interacts with the ankyrin-repeat protein ANK1, and is essential for BZI-1 function, both in auxin signaling and pathogen response. J Biol Chem 278:8786–8794CrossRefPubMedGoogle Scholar
  37. Lamb CJ, Lawton MA, Dixon RA (1989) Signal and transduction mechanisms for activation of plant defenses against microbial attack. Cell 56:215–224CrossRefPubMedGoogle Scholar
  38. Landschulz WH, Johnson PF, McKnight SL (1988) The leucine zipper: a hypothetical structure common to a new class of DNA binding proteins. Science 240:1759–1764PubMedCrossRefGoogle Scholar
  39. Lee SC, Hwang BK (2003) Identification of the pepper SAR8.2 gene as a molecular marker for pathogen infection, abiotic elicitors and environmental stresses in Capsicum annuum. Planta 216:387–396PubMedGoogle Scholar
  40. Lee YK, Hwang BK (1996) Differential induction and accumulation of β-1,3-glucanase and chitinase isoforms in the intercellular space and leaf tissues of pepper by Xanthomonas campestris pv. vesicatoria infection. J Phytopathol 144:79–87CrossRefGoogle Scholar
  41. Lee SC, Hong JK, Kim YJ, Hwang BK (2000) Pepper gene encoding thionin is differentially induced by pathogens, ethylene and methyl jasmonate. Physiol Mol Plant Pathol 56:207–216CrossRefGoogle Scholar
  42. Lee SC, Kim YJ, Hwang BK (2001) A pathogen-induced chitin-binding protein gene from pepper: its isolation and differential expression in pepper tissues treated with pathogens, ethephon, methyl jasmonate or wounding. Plant Cell Physiol 42:1321–1330CrossRefPubMedGoogle Scholar
  43. Lee J-H, Kim S-H, Jung Y-H, Kim J-A, Lee M-O, Choi P-G, Choi W, Kim K-N, Jwa N-S (2005) Molecular cloning and functional analysis of rice (Oryza sativa L.) OsNDR1 on defense signaling pathway. Plant Pathol J 21:149–157Google Scholar
  44. Leung J, Giraudat J (1998) Abscisic acid signal transduction. Annu Rev Plant Physiol Plant Mol Biol 49:199–222CrossRefPubMedGoogle Scholar
  45. Li J, Brader G, Palva ET (2004) The WRKY70 transcription factor: a node of convergence for jasmonate-mediated and salicylate-mediated signals in plant defense. Plant Cell 16:319–331CrossRefPubMedGoogle Scholar
  46. Liu L, White MJ, MacRae TH (1999) Transcription factors and their genes in higher plants. Eur J Biochem 262:247–257PubMedCrossRefGoogle Scholar
  47. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar
  48. Murgia I, Tarantino D, Vannini C, Bracale M, Carravieri S, Soave C (2004) Arabidopsis thaliana plants overexpressing thylakoidal ascorbate peroxidase show increased resistance to paraquat-induced photooxidative stress and to nitric oxide-induced cell death. Plant J 38:940–953CrossRefPubMedGoogle Scholar
  49. Nakayama T, Okanami M, Meshi T, Iwabuchi M (1997) Dissection of the wheat transcription factor HBP-1a (17) reveals a modular structure for the activation domain. Mol Gen Genet 253:553–561CrossRefPubMedGoogle Scholar
  50. Noack E, Feelisch M (1991) Molecular mechanisms of nitrovasodilator bioactivation. Basic Res Cardiol 86(Suppl 2):37–50PubMedGoogle Scholar
  51. Oh S-K, Lee S-W, Kwon S, Choi D (2005) Development of a screening system for plant defense-inducing agent using transgenic tobacco plant with PR-1a promoter and GUS gene. Plant Pathol J 21:288–292Google Scholar
  52. Park JM (2005) The hypersensitive response. A cell death during disease resistance. Plant Pathol J 21:99–101Google Scholar
  53. Penninckx IA, Thomma BP, Buchala A, Metraux JP, Broekaert WF (1998) Concomitant activation of jasmonate and ethylene response pathways is required for induction of a plant defensin gene in Arabidopsis. Plant Cell 10:2103–2113CrossRefPubMedGoogle Scholar
  54. Piao HL, Lim JH, Kim SJ, Cheong GW, Hwang I (2001) Constitutive over-expression of AtGSK1 induces NaCl stress responses in the absence of NaCl stress and results in enhanced NaCl tolerance in Arabidopsis. Plant J 27:305–314PubMedCrossRefGoogle Scholar
  55. Reintanz B, Lehnen M, Reichelt M, Gershenzon J, Kowalczyk M, Sandberg G, Godde M, Uhl R, Palme K (2001) Bus, a bushy Arabidopsis CYP79F1 knockout mutant with abolished synthesis of short-chain aliphatic glucosinolates. Plant Cell 13:351–367CrossRefPubMedGoogle Scholar
  56. Rickauer M, Brodschelm W, Bottin A, Veronesi C, Grimal H, Esquerre-Tugaye MT (1997) The jasmonate pathway is involved differentially in the regulation of different defense responses in tobacco cells. Planta 202:155–162CrossRefGoogle Scholar
  57. Sainz MB, Goff SA, Chandler VL (1997) Extensive mutagenesis of a transcriptional activation domain identifies single hydrophobic and acidic amino acids important for activation in vivo. Mol Cell Biol 17:115–122PubMedGoogle Scholar
  58. Sano T, Nagata T (2002) The possible involvement of a phosphate-induced transcription factor encoded by phi-2 gene from tobacco in ABA-signaling pathways. Plant Cell Physiol 43:12–20 CrossRefPubMedGoogle Scholar
  59. Schwechheimer C, Bevan M (1998) The regulation of transcription factor activity in plants. Trends Plant Sci 3:378–383CrossRefGoogle Scholar
  60. Singh KB, Foley RC, Onate-Sanchez L (2002) Transcription factors in plant defense and stress responses. Curr Opin Plant Biol 5:430–436CrossRefPubMedGoogle Scholar
  61. Sohn KH, Lee SC, Jung HW, Hong JK, Hwang BK (2006) Overexpression of the pepper CARAV1 pathogen-induced gene encoding a RAV transcription factor induces pathogenesis-related genes and enhances resistance to bacterial pathogen in Arabidopsis. Plant Mol Biol (in press)Google Scholar
  62. Sprenger-Haussels M, Weisshaar B (2000) Transactivation properties of parsley proline-rich bZIP transcription factors. Plant J 22:1–8CrossRefPubMedGoogle Scholar
  63. Suntres ZE (2002) Role of antioxidants in paraquat toxicity. Toxicology 180:65–77CrossRefPubMedGoogle Scholar
  64. Triezenberg SJ (1995) Structure and function of transcriptional activation domains. Curr Opin Gen Dev 5:190–196CrossRefGoogle Scholar
  65. Tsugane K, Kobayashi K, Niwa Y, Ohba Y, Wada K, Kobayashi H (1999) A recessive Arabidopsis mutant that grows photoautotrophically under salt stress shows enhanced active oxygen detoxification. Plant Cell 11:1195–1206CrossRefPubMedGoogle Scholar
  66. Ukness S, Mauch-Mani B, Moyer M, Potter S, Williams S, Dincher S, Chandler D, Slusarenko A, Ward E, Ryals J (1992) Acquired resistance in Arabidopsis. Plant Cell 4:645–656CrossRefGoogle Scholar
  67. Uno Y, Furihata T, Abe H, Yoshida R, Shinozaki K, Yamaguchi-Shinozaki Y (2000) Arabidopsis basic leucine zipper transcription factors involved in an absicsic acid-dependent signal transduction pathway under drought and high-salinity conditions. Proc Natl Acad Sci USA 97:11632–11637CrossRefPubMedGoogle Scholar
  68. Varet A, Hause B, Hause G, Scheel D, Lee J (2003) The Arabidopsis NHL3 gene encodes a plasma membrane protein and its overexpression correlates with increased resistance to Pseudomonas syringae pv. tomato DC3000. Plant Physiol 132:2023–2033CrossRefPubMedGoogle Scholar
  69. Ward ER, Uknes SJ, Williams SC, Dincher SS, Wiederhold DL, Alexander DC, Ahl-Goy P, Metraux J, Ryals JA (1991) Coordinate gene activity in response to agents that induce systemic acquired resistance. Plant Cell 3:1085–1094CrossRefPubMedGoogle Scholar
  70. Whalen MC, Innes RW, Bent AF, Staskawicz BJ (1991) Identification of Pseudomonas syringae pathogens of Arabidopsis and a bacterial locus determining avirulence on both Arabidopsis and soybean. Plant Cell 3:49–59CrossRefPubMedGoogle Scholar
  71. Yamaguchi-Shinozaki K, Shinozaki K (1994) A novel cis-acting element in an Arabidopsis gene is involved in responsiveness to drought, low-temperature, or high-salt stress. Plant Cell 6:251–264CrossRefPubMedGoogle Scholar
  72. Yang Y, Shah J, Klessig DF (1997) Signal perception and transduction in plant defense responses. Genes Dev 11:1621–1639PubMedCrossRefGoogle Scholar
  73. Zeier J, Delledonne M, Mishina T, Severi E, Sonoda M, Lamb C (2004) Genetic elucidation of nitric oxide signaling in incompatible plant–pathogen interactions. Plant Physiol 136:2875–2886CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Sung Chul Lee
    • 1
  • Hyong Woo Choi
    • 1
  • In Sun Hwang
    • 1
  • Du Seok Choi
    • 1
  • Byung Kook Hwang
    • 1
    Email author
  1. 1.Laboratory of Molecular Plant Pathology, College of Life Sciences and BiotechnologyKorea UniversitySeoulSouth Korea

Personalised recommendations