Acta Physiologiae Plantarum

, 40:179 | Cite as

Responsiveness comparison of three stress inducible promoters in transgenic rice

  • Haiyan Teng
  • Boran Shen
  • Ee Liu
  • Jianjun Zhang
  • Xinxiang PengEmail author
Original Article


Although dozens of stress inducible promoters have been identified in rice, detailed and comparative investigations under uniform condition are still limited. In this study, we selected eight previously reported drought-inducible genes [Oshox24, hypothetical protein (hp), OsLEA3-1, OsNAC5, OsNCED3, Rab21, RRJ1 and Wsi18] and analyzed their transcriptional responses to drought and osmotic stresses in rice. Based on their transcription patterns and the required standards for inducible promoters, Oshox24, Rab21 and OsLEA3-1 were chosen for cloning of their promoters for detailed analyses at both the RNA and protein/activity levels, with rice oxalate oxidase gene (OsOxO3) as a reporter gene. By generating transgenic plants and determining oxalate oxidase activity that is activated under different stresses, we defined more quantitatively that the promoters of Oshox24, OsLEA3-1 and Rab21 are ideally applicable to transgene expression for inducibly controlling drought resistance genes or other functional genes in rice. The results provided a quantitative assessment of the strength, sensitivity, stress specificity and time course of the three promoters. Such detailed information is essential for the selection of promoters for use in improving stress resistance.


Abscisic acid (ABA) Drought Oxalate oxidase gene Rice Stress inducible promoter 



Abscisic acid


ABA-responsive element


Stage when all the leaves had just completely rolled


Late embryogenesis abundant protein


Oxalate oxidase


Stage when leaves started rolling



This work was supported by the National Natural Science Foundation of China (31470343) and Science and Technology Project of Guangzhou City (201607020006).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

11738_2018_2753_MOESM1_ESM.rar (1.3 mb)
Supplementary material 1 (RAR 1346 KB)


  1. Agalou A, Purwantomo S et al (2008) A genome-wide survey of HD-Zip genes in rice and analysis of drought-responsive family members. Plant Mol Biol 66:87–103. CrossRefPubMedGoogle Scholar
  2. Bhatnagar-Mathur P, Vadez V, Sharma K (2008) Transgenic approaches for abiotic stress tolerance in plants: retrospect and prospects. Plant Cell Rep 27:411–424. CrossRefPubMedGoogle Scholar
  3. Butaye K, Cammue D, Delauré S, De Bolle M (2005) Approaches to minimize variation of transgene expression in plants. Mol Breed 16:79–91. CrossRefGoogle Scholar
  4. Cornejo M, Luth D, Blankenship K, Anderson O, Blechl A (1993) Activity of a maize ubiquitin promoter in transgenic rice. Plant Mol Biol 23:567–581. CrossRefPubMedGoogle Scholar
  5. Fang Y, Xiong L (2015) General mechanisms of drought response and their application in drought resistance improvement in plants. Cell Mol Life Sci 72:673–689. CrossRefPubMedGoogle Scholar
  6. Feng Y, Cao C, Vikram M, Park S, Kim H, Hong J, Cisneros-Zevallos L, Koiwa H (2011) A three-component gene expression system and its application for inducible flavonoid overproduction in transgenic Arabidopsis thaliana. Plos One 6(3):e17603. CrossRefPubMedPubMedCentralGoogle Scholar
  7. Gupta P, Raghuvanshi S, K Tyagi A (2001) Assessment of the efficiency of various gene promoters via biolistics in leaf and regenerating seed callus of Millets, Eleusine coracana and Echinochloa crusgalli. Plant Biotechnol 18:275–282. CrossRefGoogle Scholar
  8. Hasthanasombut S, Supaibulwatana K, Mii M, Nakamura I (2011) Genetic manipulation of Japonica rice using the OsBADH1 gene from Indica rice to improve salinity tolerance. Plant Cell Tissue Organ Cult (PCTOC) 104:79–89. CrossRefGoogle Scholar
  9. Hu C, Chee P, Chesney R, Zhou J, Miller P, O’Brien W (1990) Intrinsic GUS-like activities in seed plants. Plant Cell Rep 9:1–5. CrossRefPubMedGoogle Scholar
  10. Ito Y, Katsura K, Kyonoshin M (2006) Functional analysis of rice DREB1/CBF-type transcription factors involved in cold-responsive gene expression in transgenic rice. Plant Cell Physiol 47:141–153. CrossRefPubMedGoogle Scholar
  11. 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. CrossRefPubMedGoogle Scholar
  12. Lane BG (1994) Oxalate, germin, and the extracellular matrix of higher plants. FASEB J 8:8294–8301Google Scholar
  13. Lawlar DW (2012) Genetic engineering to improve plant performance under drought: physiological evaluation of achievements, limitations, and possibilities. J Exp Bot 64:83–108. CrossRefGoogle Scholar
  14. Li J, Gong X, Lin H, Song Q, Chen J, Wang X (2005) DGP1, a drought-induced guard cell-specific promoter and its function analysis in tobacco plants. Sci China Ser c-Life Sci 48:181–186CrossRefGoogle Scholar
  15. Li X, Liao Y, Leung D, Wang H, Chen B, Peng X, Liu E (2015) Divergent biochemical and enzymatic properties of oxalate oxidase isoforms encoded by four similar genes in rice. Phytochemistry 118:216–223. CrossRefPubMedGoogle Scholar
  16. Matzke AJ, Matzke MA (1998) Position effects and epigenetic silencing of plant transgenes. Curr Opin Plant Biol 1:142–148CrossRefGoogle Scholar
  17. Msanne J, Lin J, Stone JM, Awada T (2011) Characterization of abiotic stress-responsive Arabidopsis thaliana RD29A and RD29B genes and evaluation of transgenes. Planta 234:97–107. CrossRefPubMedGoogle Scholar
  18. Mundy J, Chua NH (1988) Abscisic acid and water-stress induce the expression of a novel rice gene. EMBO J 7:2279–2286CrossRefGoogle Scholar
  19. Nakashima K, Tran LP et al (2007) Functional analysis of a NAC-type transcription factor OsNAC6 involved in abiotic and biotic stress-responsive gene expression in rice. Plant J 51:617–630. CrossRefPubMedGoogle Scholar
  20. Nakashima K, Jan A, Todaka D, Maruyama K, Goto S, Shinozaki K, Yamaguchi-Shinozaki K (2014) Comparative functional analysis of six drought-responsive promoters in transgenic rice. Planta 239:47–60. CrossRefPubMedGoogle Scholar
  21. Ozawa K (2009) Establishment of a high efficiency Agrobacterium-mediated transformation system of rice (Oryza sativa L.). Plant Sci 176:522–527. CrossRefPubMedGoogle Scholar
  22. Peng X, Ma X et al (2011) Improved drought and salt tolerance of Arabidopsis thaliana by transgenic expression of a novel DREB gene from Leymus chinensis. Plant Cell Rep 30:1493–1502. CrossRefGoogle Scholar
  23. Rai M, He C, Wu R (2009) Comparative functional analysis of three abiotic stress-inducible promoters in transgenic rice. Transgenic Res 18:787–799. CrossRefPubMedGoogle Scholar
  24. Schmittgen TD, Livak KJ (2008) Analyzing real-time PCR data by the comparative CT method. Nat Protoc 3:1101–1108. CrossRefPubMedGoogle Scholar
  25. Shen Q, Zhang P, Ho TH (1996) Modular nature of abscisic acid (ABA) response complexes: composite promoter units that are necessary and sufficient for ABA induction of gene expression in barley. Plant Cell 8:1107–1109. CrossRefPubMedPubMedCentralGoogle Scholar
  26. Simmonds J, Cass L, Routly E, Hubbard K, Donaldson P, Bancroft B, Davidson A, Hubbard S, Simmonds D (2004) Oxalate oxidase: a novel reporter gene for monocot and dicot transformations. Mol Breed 13:79–91. CrossRefGoogle Scholar
  27. Straub PF, Shen Q, Ho TD (1994) Structure and promoter analysis of an ABA- and stress-regulated barley gene, HVA1. Plant Mol Biol 26:617–630. CrossRefPubMedGoogle Scholar
  28. Su J, Sheng Q, Ho TD, Wu R (1998) Dehydration-stress-regulated transgene expression in stably transformed rice plants. Plant Physiol 117:913–922CrossRefGoogle Scholar
  29. Svedružić D, Jónsson S, Toyota CG, Reinhardt LA, Ricagno S, Lindqvist Y, Richards NGJ (2005) The enzymes of oxalate metabolism: unexpected structures and mechanisms. Arch Biochem Biophys 433:176–192. CrossRefPubMedGoogle Scholar
  30. Tester M, Langridge P (2010) Breeding technologies to increase crop production in a changing world. Science 327:818–822. CrossRefPubMedGoogle Scholar
  31. Todaka D, Shinozaki K, Yamaguchi-Shinozaki K (2015) Recent advances in the dissection of drought-stress regulatory networks and strategies for development of drought-tolerant transgenic rice plants. Front Plant Sci 6:84. CrossRefPubMedPubMedCentralGoogle Scholar
  32. Toki S (1997) Rapid and efficient Agrobacterium-mediated transformation in rice. Plant Mol Biol Rep 15:16–21. CrossRefGoogle Scholar
  33. Tör M, Mantell S, Ainsworth C (1992) Endophytic bacteria expressing β-glucuronidase cause false positives in transformation of Dioscorea species. Plant Cell Rep 11:452–456. CrossRefPubMedGoogle Scholar
  34. Wang X, Zhu H, Jin G, Liu H, Wu W, Zhu J (2007) Genome-scale identification and analysis of LEA genes in rice (Oryza sativa L.). Plant Sci 172:414–420. CrossRefGoogle Scholar
  35. Xiao F, Xue G (2001) Analysis of the promoter activity of late embryogenesis abundant protein genes in barley seedlings under conditions of water deficit. Plant Cell Rep 20:667–673. CrossRefGoogle Scholar
  36. 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 115:35–46. CrossRefPubMedGoogle Scholar
  37. 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 6:251–264. CrossRefPubMedPubMedCentralGoogle Scholar
  38. Yi N, Kim YS, Jeong M, Oh S, Jeong JS, Park S, Jung H, Choi YD, Kim J (2010) Functional analysis of six drought-inducible promoters in transgenic rice plants throughout all stages of plant growth. Planta 232:743–754. CrossRefPubMedGoogle Scholar
  39. Yoshida T, Mogami J, Yamaguchi-Shinozaki K (2015) Omics approaches toward defining the comprehensive abscisic acid signaling network in plants. Plant Cell Physiol 56:1043–1052. CrossRefPubMedGoogle Scholar
  40. Zhang Y, Su J et al (2011) A highly efficient rice green tissue protoplast system for transient gene expression and studying light/chloroplast-related processes. Plant Methods 7:30. CrossRefPubMedPubMedCentralGoogle Scholar
  41. Zhang X, Nie Z, Wang W, Leung D, Xu D, Chen B, Chen Z, Zeng L, Liu E (2013) Relationship between disease resistance and rice oxalate oxidases in transgenic rice. Plos One 8:e78348. CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Franciszek Górski Institute of Plant Physiology, Polish Academy of Sciences, Kraków 2018

Authors and Affiliations

  • Haiyan Teng
    • 1
    • 2
  • Boran Shen
    • 1
  • Ee Liu
    • 1
  • Jianjun Zhang
    • 1
  • Xinxiang Peng
    • 1
    Email author
  1. 1.State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, College of Life SciencesSouth China Agricultural UniversityGuangzhouChina
  2. 2.College of Chemistry and BioengineeringYichun UniversityYichunChina

Personalised recommendations