, Volume 235, Issue 6, pp 1157–1169 | Cite as

bZIP transcription factor OsbZIP52/RISBZ5: a potential negative regulator of cold and drought stress response in rice

  • Citao Liu
  • Yanbin Wu
  • Xiping Wang
Original Article


OsbZIP52/RISBZ5 is a member of the basic leucine zipper (bZIP) transcription factor (TF) family in rice (Oryza sativa) isolated from rice (Zhonghua11) panicles. Expression of the OsbZIP52 gene was strongly induced by low temperature (4°C) but not by drought, PEG, salt, or ABA. The subcellular localization of OsbZIP52-GFP in onion (Allium cepa) epidermis cells revealed that OsbZIP52 is a nuclear localized protein. A transactivation assay in yeast demonstrated that OsbZIP52 functions as a transcriptional activator and can specifically bind to the G-box promoter motif. In a yeast two-hybrid (Y-2-H) experiment, OsbZIP52 was able to form homodimeric complexes. Rice plants overexpressing OsbZIP52 showed significantly increased sensitivity to cold and drought stress. Real-time PCR analysis revealed that some abiotic stress-related genes, such as OsLEA3, OsTPP1, Rab25, gp1 precursor, β-gal, LOC_Os05g11910 and LOC_Os05g39250, were down-regulated in OsbZIP52 overexpression lines. These results suggest that OsbZIP52/RISBZ5 could function as a negative regulator in cold and drought stress environments.


Abiotic stress bZIP Transcription factor G-box Transgenic rice (Oryza sativa



Basic leucine zipper


Rice late embryogenesis abundant protein


Trehalose-6-phosphate phosphatase


Glycoprotein 1


Responsive to abscisic acid




Rice transcription activator-1


Rice seed b-Zipper


G-box/H-box binding factors


Basic leucine zipper O2 homolog


Low-temperature-induced protein 19



This research was supported by grants from the Agricultural Ministry of China (2011ZX08009-003-002), and the National Natural Science Foundation of China (2010 No. 31071378).

Supplementary material

425_2011_1564_MOESM1_ESM.doc (2 mb)
Suppl. Fig. S1 Seedling development of OsbZIP52 overexpression transgenic rice plants under ABA treatment. ab The seed germination rate and seedling growth of OsbZIP52-overexpression rice lines with and without ABA. c Shoot and root lengths and root number at different ABA concentrations in the WT control (ZH11) and the transgenic rice lines. d Seedlings grown at different concentrations of ABA for 12 days. All the experiments were repeated at least three times (150 seeds for each replicate). ZH11, wild-type Zhonghua11 rice variety. 197-1, -3, -16, -24, -27 are independent transgenic lines of OsbZIP52 (DOC 2055 kb)
425_2011_1564_MOESM2_ESM.doc (334 kb)
Suppl. Fig. S2 OsbZIP52 expression in three rice varieties. a OsbZIP52 expression patterns in different tissues of rice variety Nipponbare. bd Expression of OsbZIP52 in three rice varieties grown at low temperature (4°C) measured by real-time PCR. DAF, days after flowering (DOC 333 kb)
425_2011_1564_MOESM3_ESM.doc (44 kb)
Suppl. Table S1. Primers used in semi-quantitative RT-PCR and real-time PCR analysis and constructs for vectors (DOC 44 kb)
425_2011_1564_MOESM4_ESM.doc (32 kb)
Suppl. Table S2 Cis-acting elements analysis in promoters of stress-regulated genes in OsbZIP52 overexpression lines. (DOC 31 kb)


  1. Aguan K, Sugawara K, Suzuki N, Kusano T (1993) Low-temperature-dependent expression of a rice gene encoding a protein with a leucine-zipper motif. Mol Gen Genet 240:1–8PubMedCrossRefGoogle Scholar
  2. Bartels D, Sunkar R (2005) Drought and salt tolerance in plants. Crit Rev Plant Sci 24:23–58CrossRefGoogle Scholar
  3. Choi H, Hong J, Ha J, Kang J, Kim SY (2000) ABFs, a family of ABA-responsive element binding factors. J Biol Chem 275:1723–1730PubMedCrossRefGoogle Scholar
  4. Correa LG, Riano-Pachon DM, Schrago CG, dos Santos RV, Mueller-Roeber B, Vincentz M (2008) The role of bZIP transcription factors in green plant evolution: adaptive features emerging from four founder genes. PLoS One 3:e2944PubMedCrossRefGoogle Scholar
  5. Degenkolbe T, Do PT, Zuther E, Repsilber D, Walther D, Hincha DK, Köhl KI (2009) Expression profiling of rice cultivars differing in their tolerance to long-term drought stress. Plant Mol Biol 69:133–153PubMedCrossRefGoogle Scholar
  6. Droge-Laser W, Kaiser A, Lindsay WP, Halkier BA, Loake GJ, Doerner P, Dixon RA, Lamb C (1997) Rapid stimulation of a soybean protein-serine kinase that phosphorylates a novel bZIP DNA-binding protein, G/HBF-1, during the induction of early transcription-dependent defenses. EMBO J 16:726–738PubMedCrossRefGoogle Scholar
  7. Ehlert A, Weltmeier F, Wang X, Mayer CS, Smeekens S, Vicente-Carbajosa J, Droge-Laser W (2006) Two-hybrid protein–protein interaction analysis in Arabidopsis protoplasts: establishment of a heterodimerization map of group C and group S bZIP transcription factors. Plant J 46:890–900PubMedCrossRefGoogle Scholar
  8. Finkelstein RR, Lynch TJ (2000) The Arabidopsis abscisic acid response gene ABI5 encodes a basic leucine zipper transcription factor. Plant Cell 12:599–609PubMedCrossRefGoogle Scholar
  9. Ge LF, Chao DY, Shi M, Zhu MZ, Gao JP, Lin HX (2008) Overexpression of the trehalose-6-phosphate phosphatase gene OsTPP1 confers stress tolerance in rice and results in the activation of stress responsive genes. Planta 228:191–201PubMedCrossRefGoogle Scholar
  10. Goyal K, Walton LJ, Tunnacliffe A (2005) LEA proteins prevent protein aggregation due to water stress. Biochem J 388:151–157PubMedCrossRefGoogle Scholar
  11. Grelet J, Benamar A, Teyssier E, Avelange-Macherel MH, Grunwald D, Macherel D (2005) Identification in pea seed mitochondria of a late-embryogenesis abundant protein able to protect enzymes from drying. Plant Physiol 137:157–167PubMedCrossRefGoogle Scholar
  12. 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 96:15348–15353PubMedCrossRefGoogle Scholar
  13. Honjoh K, Matsumoto H, Shimizu H, Ooyama K, Tanaka K, Oda Y, Takata R, Joh T, Suga K, Miyamoto T (2000) Cryoprotective activities of group 3 late embryogenesis abundant proteins from Chlorella vulgaris C-27. Biosci Biotechnol Biochem 64:1656–1663PubMedCrossRefGoogle Scholar
  14. Hossain MA, Lee Y, Cho JI, Ahn CH, Lee SK, Jeon JS, Kang H, Lee CH, An G, Park PB (2010) The bZIP transcription factor OsABF1 is an ABA responsive element binding factor that enhances abiotic stress signaling in rice. Plant Mol Biol 72:557–566CrossRefGoogle Scholar
  15. Hu TZ (2008) OsLEA3, a late embryogenesis abundant protein gene from rice, confers tolerance to water deficit and salt stress to transgenic rice. Russ J Plant Physiol 55:530–537CrossRefGoogle Scholar
  16. Hu H, You J, Fang Y, Zhu X, Qi Z, Xiong L (2008) Characterization of transcription factor gene SNAC2 conferring cold and salt tolerance in rice. Plant Mol Biol 67:169–181PubMedCrossRefGoogle Scholar
  17. Huang XY, Chao DY, Gao JP, Zhu MZ, Shi M, Lin HX (2009) A previously unknown zinc finger protein, DST, regulates drought and salt tolerance in rice via stomatal aperture control. Genes Dev 23:1805–1817PubMedCrossRefGoogle Scholar
  18. Iwaguro H, Yamaguchi J, Kalka C, Murasawa S, Masuda H, Hayashi S, Silver M, Li T, Isner JM, Asahara T (2002) Endothelial progenitor cell vascular endothelial growth factor gene transfer for vascular regeneration. Circulation 105:732–738PubMedCrossRefGoogle Scholar
  19. Izawa T, Foster R, Nakajima M, Shimamoto K, Chua NH (1994) The rice bZIP transcriptional activator RITA-1 is highly expressed during seed development. Plant Cell 6:1277–1287PubMedCrossRefGoogle Scholar
  20. Jakoby M, Weisshaar B, Droge-Laser W, Vicente-Carbajosa J, Tiedemann J, Kroj T, Parcy F (2002) bZIP transcription factors in Arabidopsis. Trends Plant Sci 7:106–111PubMedCrossRefGoogle Scholar
  21. Kaminaka H, Nake C, Epple P, Dittgen J, Schutze K, Chaban C, Holt BF 3rd, Merkle T, Schafer E, Harter K, Dangl JL (2006) bZIP10-LSD1 antagonism modulates basal defense and cell death in Arabidopsis following infection. EMBO J 25:4400–4411PubMedCrossRefGoogle Scholar
  22. Kang SG, Price J, Lin PC, Hong JC, Jang JC (2010) The Arabidopsis bZIP1 transcription factor is involved in sugar signaling, protein networking, and DNA binding. Mol Plant 3:361–373PubMedCrossRefGoogle Scholar
  23. Karimi M, Inzé D, Depicker A (2002) GATEWAYTM vectors for Agrobacterium-mediated plant transformation. Trends Plant Sci 7:193–195PubMedCrossRefGoogle Scholar
  24. Kasuga M, Liu Q, Miura S, Yamaguchi-Shinozaki K, Shinozaki K (1999) Improving plant drought, salt, and freezing tolerance by gene transfer of a single stress-inducible transcription factor. Nat Biotechnol 17:287–291PubMedCrossRefGoogle Scholar
  25. Kawakatsu T, Takaiwa F (2010) Differences in transcriptional regulatory mechanisms functioning for free lysine content and seed storage protein accumulation in rice grain. Plant Cell Physiol 51:1964–1974PubMedCrossRefGoogle Scholar
  26. Kim SOOY, Thomas TL (1998) A family of novel basic leucine zipper proteins binds to seed-specification elements in the carrot Dc3 gene promoter. J Plant Physiol 152:607–613CrossRefGoogle Scholar
  27. Kusano T, Aguan K, Abe M, Sugawara K (1992) Nucleotide sequence of a rice rab16 homologue gene. Plant Mol Biol 18:127–129PubMedCrossRefGoogle Scholar
  28. Lee SC, Lee MY, Kim SJ, Jun SH, An G, Kim SR (2005) Characterization of an abiotic stress-inducible dehydrin gene, OsDhn1 in rice (Oryza sativa L.). Mol Cells 19:212–218PubMedGoogle Scholar
  29. Liu X, Bai X, Qian Q, Wang X, Chen M, Chu C (2005) OsWRKY03, a rice transcriptional activator that functions in defense signaling pathway upstream of OsNPR1. Cell Res 15:593–603PubMedCrossRefGoogle Scholar
  30. Liu X, Bai X, Wang X, Chu C (2007) OsWRKY71, a rice transcription factor, is involved in rice defense response. J Plant Physiol 164:969–979PubMedCrossRefGoogle Scholar
  31. Locatelli S, Piatti P, Motto M, Rossi V (2009) Chromatin and DNA modifications in the Opaque2-mediated regulation of gene transcription during maize endosperm development. Plant Cell 21:1410–1427PubMedCrossRefGoogle Scholar
  32. Martínez-García JF, Moyano E, Alcocer MJC, Martin C (1998) Two bZIP proteins from Antirrhinum flowers preferentially bind a hybrid C-box/G-box motif and help to define a new sub-family of bZIP transcription factors. Plant J 13:489–505PubMedCrossRefGoogle Scholar
  33. Menkens AE, Schindler U, Cashmore AR (1995) The G-box: a ubiquitous regulatory DNA element in plants bound by the GBF family of bZIP proteins. Trends Biochem Sci 20:506–510PubMedCrossRefGoogle Scholar
  34. Nakase M, Aoki N, Matsuda T, Adachi T (1997) Characterization of a novel rice bZIP protein which binds to the alpha-globulin promoter. Plant Mol Biol 33:513–522PubMedCrossRefGoogle Scholar
  35. Nijhawan A, Jain M, Tyagi AK, Khurana JP (2008) Genomic survey and gene expression analysis of the basic leucine zipper transcription factor family in rice. Plant Physiol 146:333–350PubMedCrossRefGoogle Scholar
  36. Niu X, Renshaw-Gegg L, Miller L, Guiltinan MJ (1999) Bipartite determinants of DNA-binding specificity of plant basic leucine zipper proteins. Plant Mol Biol 41:1–13PubMedCrossRefGoogle Scholar
  37. Onate L, Vicente-Carbajosa J, Lara P, Diaz I, Carbonero P (1999) Barley BLZ2, a seed-specific bZIP protein that interacts with BLZ1 in vivo and activates transcription from the GCN4-like motif of B-hordein promoters in barley endosperm. J Biol Chem 274:9175–9182PubMedCrossRefGoogle Scholar
  38. Onodera Y, Suzuki A, Wu CY, Washida H, Takaiwa F (2001) A rice functional transcriptional activator, RISBZ1, responsible for endosperm-specific expression of storage protein genes through GCN4 motif. J Biol Chem 276:14139–14152PubMedGoogle Scholar
  39. Oono Y, Wakasa Y, Hirose S, Yang L, Sakuta C, Takaiwa F (2010) Analysis of ER stress in developing rice endosperm accumulating β-amyloid peptide. Plant Biotechnol J 8:691–718PubMedCrossRefGoogle Scholar
  40. Pfaffl MW (2001) A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 29:e45PubMedCrossRefGoogle Scholar
  41. Pramanik MH, Imai R (2005) Functional identification of a trehalose 6-phosphate phosphatase gene that is involved in transient induction of trehalose biosynthesis during chilling stress in rice. Plant Mol Biol 58:751–762PubMedCrossRefGoogle Scholar
  42. Pysh LD, Schmidt RJ (1996) Characterization of the maize OHP1 gene: evidence of gene copy variability among inbreds. Gene 177:203–208PubMedCrossRefGoogle Scholar
  43. Rabbani MA, Maruyama K, Abe H, Khan MA, Katsura K, Ito Y, Yoshiwara K, Seki M, Shinozaki K, Yamaguchi-Shinozaki K (2003) Monitoring expression profiles of rice genes under cold, drought, and high-salinity stresses and abscisic acid application using cDNA microarray and RNA gel-blot analyses. Plant Physiol 133:1755–1767PubMedCrossRefGoogle Scholar
  44. Ren W, Beard RW (2005) Consensus seeking in multiagent systems under dynamically changing interaction topologies. IEEE Trans Automatic Control 50:655–661CrossRefGoogle Scholar
  45. Rombauts S, Déhais P, Van Montagu M, Rouzé P (1999) PlantCARE, a plant cis-acting regulatory element database. Nucleic Acids Res 27:295–296PubMedCrossRefGoogle Scholar
  46. Schmidt RJ, Ketudat M, Aukerman MJ, Hoschek G (1992) Opaque-2 is a transcriptional activator that recognizes a specific target site in 22-kD zein genes. Plant Cell 4:689–700PubMedCrossRefGoogle Scholar
  47. Shimizu H, Sato K, Berberich T, Miyazaki A, Ozaki R, Imai R, Kusano T (2005) LIP19, a basic region leucine zipper protein, is a Fos-like molecular switch in the cold signaling of rice plants. Plant Cell Physiol 46:1623–1634PubMedCrossRefGoogle Scholar
  48. Shinozaki K, Yamaguchi-Shinozaki K (1996) Molecular responses to drought and cold stress. Curr Opin Biotechnol 7:161–167PubMedCrossRefGoogle Scholar
  49. Shinozaki K, Yamaguchi-Shinozaki K (1997) Gene expression and signal transduction in water-stress response. Plant Physiol 115:327–334PubMedCrossRefGoogle Scholar
  50. Siberil Y, Doireau P, Gantet P (2001) Plant bZIP G-box binding factors: modular structure and activation mechanisms. Eur J Biochem 268:5655–5666PubMedCrossRefGoogle Scholar
  51. Sohlenkamp C, Wood CC, Roeb GW, Udvardi MK (2002) Characterization of Arabidopsis AtAMT2, a high-affinity ammonium transporter of the plasma membrane. Plant Physiol 130:1788–1796PubMedCrossRefGoogle Scholar
  52. Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG (1997) The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 25:4876–4882PubMedCrossRefGoogle Scholar
  53. Vincentz M, Bandeira-Kobarg C, Gauer L, Schlögl P, Leite A (2003) Evolutionary pattern of angiosperm bZIP factors homologous to the maize Opaque2 regulatory protein. J Mol Evol 56:105–116PubMedCrossRefGoogle Scholar
  54. Weltmeier F, Ehlert A, Mayer CS, Dietrich K, Wang X, Schütze K, Alonso R, Harter K, Vicente-Carbajosa J, Dröge-Laser W (2006) Combinatorial control of Arabidopsis proline dehydrogenase transcription by specific heterodimerisation of bZIP transcription factors. EMBO J 25:3133–3143PubMedCrossRefGoogle Scholar
  55. Wise EA (2004) POPP the question: what do LEA proteins do? Trends Plant Sci 9:13–17PubMedCrossRefGoogle Scholar
  56. Xiao B, Huang Y, Tang N, Xiong L (2007) Over-expression of a LEA gene in rice improves drought resistance under the field conditions. Theor Appl Genet 115:35–46PubMedCrossRefGoogle Scholar
  57. Xiong Y, Tang N, Du H, Ye H, Xiong L (2008) Characterization of OsbZIP23 as a key player of the basic leucine zipper transcription factor family for conferring abscisic acid sensitivity and salinity and drought tolerance in rice. Plant Physiol 148:1938–1952PubMedCrossRefGoogle Scholar
  58. Yamamoto MP, Onodera Y, Touno SM, Takaiwa F (2006) Synergism between RPBF Dof and RISBZ1 bZIP activators in the regulation of rice seed expression genes. Plant Physiol 141:1694–1707Google Scholar
  59. Zhou B, Zhao X, Kawabata S, Li Y (2009) Transient expression of a foreign gene by direct incorporation of DNA into intact plant tissue through vacuum infiltration. Biotechnol Lett 31:1811–1815PubMedCrossRefGoogle Scholar
  60. Zou M, Guan Y, Ren H, Zhang F, Chen F (2008) A bZIP transcription factor, OsABI5, is involved in rice fertility and stress tolerance. Plant Mol Biol 66:675–683PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  1. 1.Key Laboratory of Gene Engineering Drug and Biotechnology, Key Laboratory of Cell Proliferation and Regulation of Ministry of EducationCollege of Life Sciences, Beijing Normal UniversityBeijingPeople’s Republic of China
  2. 2.National Center for Molecular Crop DesignWeiming Kaituo Agriculture Biotech Co., LtdBeijingPeople’s Republic of China

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