Plant Cell Reports

, Volume 29, Issue 5, pp 449–460

Functional identification and regulation of the PtDrl02 gene promoter from triploid white poplar

  • Huiquan Zheng
  • Shanzhi Lin
  • Qian Zhang
  • Yang Lei
  • Lu Hou
  • Zhiyi Zhang
Original Paper


The PtDrl02 gene belongs to the TIR-NBS gene family in triploid white poplar (Populus tomentosa × P. bolleana) × P. tomentosa. Its expression pattern displays tissue-specificity, and the transcript level can be induced by wounding, methyl jasmonate (MeJA), and salicylic acid (SA). To understand the regulatory mechanism controlling PtDrl02 gene expression, we functionally characterized the PtDrl02 promoter region. Using the β-glucuronidase as a reporter, we found that the PtDrl02 promoter directed gene expression mainly in the aerial parts of the plants and was confined to the cortex tissues of leaf veins, petioles, stems, and stem piths, showing a typical tissue-specific expression pattern. Deletion analysis revealed two positive regulatory regions (−985 to −669 and −669 to −467) responsible for the basal activity of the PtDrl02 promoter. Impressively, the sequence from –669 to –467 was shown to contain cis-element (s) responding to wounding and MeJA, while the promoter region between –244 and 0 could individually display wounding-responsiveness, and the fragment from –467 to –244 was required for SA- and NaCl-inducible expression of the PtDrl02 promoter. Additionally, it was found that the –985 to –669 sequence was the ABA-responding promoter fragment. These results suggested that the PtDrl02 promoter was modulated by multiple cis-regulatory elements in distinct and complex patterns to regulate PtDrl02 gene expression. Our study also suggested that the PtDrl02 gene 5′ untranslated region, as well as a Populus WRKY transcription factor, PtWRKY1, was involved in the regulation of PtDrl02 promoter activities.


Populus TIR-NBS cis-element Untranslated region Transcription factor 

Supplementary material

299_2010_834_MOESM1_ESM.tif (2 mb)
Supplemental Fig. 1 Sequence alignment of the W-box-containing fragments within PtDrl02 promoter. The W-box core sequence (TGAC) is marked with double underlines (TIFF 2070 kb)
299_2010_834_MOESM2_ESM.tif (86 kb)
Supplemental Fig. 2 The identified WRKY domains in PtWRKY1 protein. The NCBI () BLASTP program was employed to identify the conserved WRKY domains (TIF 85 kb)


  1. An G (1987) Binary Ti vectors for plant transformation and promoter analysis. Methods Enzymol 153:292–305CrossRefGoogle Scholar
  2. Anderson PA, Lawrence GJ, Morrish BC, Ayliffe MA, Finnegan EJ, Ellis JG (1997) Inactivation of the flax rust resistance gene M associated with loss of a repeated unit within the leucine-rich repeat coding region. Plant Cell 9(4):641–651PubMedCrossRefGoogle Scholar
  3. Bradford MM (1976) A rapid sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254PubMedCrossRefGoogle Scholar
  4. Burch-Smith TM, Dinesh-Kumar SP (2007) The functions of plant TIR domains. Sci STKE 401:pe46Google Scholar
  5. Burch-Smith TM, Schiff M, Caplan JL, Tsao J, Czymmek K, Dinesh-Kumar SP (2007) A novel role for the TIR domain in association with pathogen-derived elicitors. PLoS Biol 5(3):e68PubMedCrossRefGoogle Scholar
  6. Ciolkowski I, Wanke D, Birkenbihl RP, Somssich IE (2008) Studies on DNA-binding selectivity of WRKY transcription factors lend structural clues into WRKY-domain function. Plant Mol Biol 68(1–2):81–92PubMedCrossRefGoogle Scholar
  7. Eitas TK, Nimchuk ZL, Dangl JL (2008) Arabidopsis TAO1 is a TIR-NB-LRR protein that contributes to disease resistance induced by the Pseudomonas syringae effector AvrB. Proc Natl Acad Sci USA 105(17):6475–6480PubMedCrossRefGoogle Scholar
  8. Eulgem T, Rushton PJ, Schmelzer E, Hahlbrock K, Somssich IE (1999) Early nuclear events in plant defence signalling: rapid gene activation by WRKY transcription factors. EMBO J 18(17):4689–4699PubMedCrossRefGoogle Scholar
  9. Gassmann W, Hinsch ME, Staskawicz BJ (1999) The Arabidopsis RPS4 bacterial-resistance gene is a member of the TIR-NBS-LRR family of disease-resistance genes. Plant J 20(3):265–277PubMedCrossRefGoogle Scholar
  10. Higashi K, Ishiga Y, Inagaki Y, Toyoda K, Shiraishi T, Ichinose Y (2008) Modulation of defense signal transduction by flagellin-induced WRKY41 transcription factor in Arabidopsis thaliana. Mol Genet Genomics 279(3):303–312PubMedCrossRefGoogle Scholar
  11. Higo K, Ugawa Y, Iwamoto M, Korenaga T (1999) Plant cis-acting regulatory DNA elements (PLACE) database. Nucleic Acids Res 27(1):297–300PubMedCrossRefGoogle Scholar
  12. Hiroyuki K, Terauchi R (2008) Regulation of expression of rice thaumatin-like protein: inducibility by elicitor requires promoter W-box elements. Plant Cell Rep 27(9):1521–1528PubMedCrossRefGoogle Scholar
  13. Horsch RB, Fry JE, Hoffmann NL, Eichholtz D, Rogers SG, Fraley RT (1985) A simple and general method for transferring genes into plants. Science 227:1229–1231CrossRefGoogle Scholar
  14. Hwang SH, Lee IA, Yie SW, Hwang DJ (2008) Identification of an OsPR10a promoter region responsive to salicylic acid. Planta 227(5):1141–1150PubMedCrossRefGoogle Scholar
  15. Jebanathirajah JA, Peri S, Pandey A (2002) Toll and interleukin-1 receptor (TIR) domain-containing proteins in plants: a genomic perspective. Trends Plant Sci 7(9):388–391PubMedCrossRefGoogle Scholar
  16. Jefferson RA, Kavanagh TA, Bevan MW (1987) GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6(13):3901–3907PubMedGoogle Scholar
  17. Kohler A, Rinaldi C, Duplessis S, Baucher M, Geelen D, Duchaussoy F, Meyers BC, Boerjan W, Martin F (2008) Genome-wide identification of NBS resistance genes in Populus trichocarpa. Plant Mol Biol 66(6):619–636PubMedCrossRefGoogle Scholar
  18. Laloi C, Mestres-Ortega D, Marco Y, Meyer Y, Reichheld JP (2004) The Arabidopsis cytosolic thioredoxin h5 gene induction by oxidative stress and its W-box-mediated response to pathogen elicitor. Plant Physiol 134(3):1006–1016PubMedCrossRefGoogle Scholar
  19. Lawrence GJ, Finnegan EJ, Ayliffe MA, Ellis JG (1995) The L6 gene for flax rust resistance is related to the Arabidopsis bacterial resistance gene RPS2 and the tobacco viral resistance gene N. Plant Cell 7(8):1195–1206PubMedCrossRefGoogle Scholar
  20. Lescot M, Déhais P, Thijs G, Marchal K, Moreau Y, Van de Peer Y, Rouzé P, Rombauts S (2002) PlantCARE, a database of plant cis-acting regulatory elements and a portal to tools for in silico analysis of promoter sequences. Nucleic Acids Res 30(1):325–327PubMedCrossRefGoogle Scholar
  21. Li Y, Yang S, Yang H, Hua J (2007) The TIR-NB-LRR gene SNC1 is regulated at the transcript level by multiple factors. Mol Plant Microbe Interact 20(11):1449–1456PubMedCrossRefGoogle Scholar
  22. Marathe R, Guan Z, Anandalakshmi R, Zhao HY, Dinesh-Kumar SP (2004) Study of Arabidopsis thaliana resistome in response to cucumber mosaic virus infection using whole genome microarray. Plant Mol Biol 55:501–520PubMedCrossRefGoogle Scholar
  23. Meyers BC, Morgante M, Michelmore RW (2002) TIR-X and TIR-NBS proteins: two new families related to disease resistance TIR-NBS-LRR proteins encoded in Arabidopsis and other plant genomes. Plant J 32(1):77–92PubMedCrossRefGoogle Scholar
  24. Meyers BC, Kozik A, Griego A, Kuang H, Michelmore RW (2003) Genome-wide analysis of NBS-LRR-encoding genes in Arabidopsis. Plant Cell 15(4):809–834PubMedCrossRefGoogle Scholar
  25. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar
  26. Nishiuchi T, Shinshi H, Suzuki K (2004) Rapid and transient activation of transcription of the ERF3 gene by wounding in tobacco leaves: possible involvement of NtWRKYs and autorepression. J Biol Chem 279(53):55355–55361PubMedCrossRefGoogle Scholar
  27. Park HC, Kim ML, Kang YH, Jeon JM, Yoo JH, Kim MC, Park CY, Jeong JC, Moon BC, Lee JH, Yoon HW, Lee SH, Chung WS, Lim CO, Lee SY, Hong JC, Cho MJ (2004) Pathogen- and NaCl-induced expression of the SCaM-4 promoter is mediated in part by a GT-1 box that interacts with a GT-1-like transcription factor. Plant Physiol 135(4):2150–2161PubMedCrossRefGoogle Scholar
  28. Parker JE, Coleman MJ, Szabò V, Frost LN, Schmidt R, van der Biezen EA, Moores T, Dean C, Daniels MJ, Jones JD (1997) The Arabidopsis downy mildew resistance gene RPP5 shares similarity to the toll and interleukin-1 receptors with N and L6. Plant Cell 9(6):879–894PubMedCrossRefGoogle Scholar
  29. Robatzek S, Somssich IE (2002) Targets of AtWRKY6 regulation during plant senescence and pathogen defense. Genes Dev 16(9):1139–1149PubMedCrossRefGoogle Scholar
  30. Rocher A, Dumas C, Cock JM (2005) A W-box is required for full expression of the SA-responsive gene SFR2. Gene 344:181–192PubMedCrossRefGoogle Scholar
  31. Rouster J, Leah R, Mundy J, Cameron-Mills V (1997) Identification of a methyl jasmonate-responsive region in the promoter of a lipoxygenase 1 gene expressed in barley grain. Plant J 11(3):513–523PubMedCrossRefGoogle Scholar
  32. Rushton PJ, Somssich IE (1998) Transcriptional control of plant genes responsive to pathogens. Curr Opin Plant Biol 1(4):311–315PubMedCrossRefGoogle Scholar
  33. Rushton PJ, Torres JT, Parniske M, Wernert P, Hahlbrock K, Somssich IE (1996) Interaction of elicitor-induced DNA-binding proteins with elicitor response elements in the promoters of parsley PR1 genes. EMBO J 15(20):5690–5700PubMedGoogle Scholar
  34. Rushton PJ, Reinstädler A, Lipka V, Lippok B, Somssich IE (2002) Synthetic plant promoters containing defined regulatory elements provide novel insights into pathogen- and wound-induced signaling. Plant Cell 14(4):749–762PubMedCrossRefGoogle Scholar
  35. Schmidt S, Lombardi M, Gardiner DM, Ayliffe M, Anderson PA (2007) The M flax rust resistance pre-mRNA is alternatively spliced and contains a complex upstream untranslated region. Theor Appl Genet 115(3):373–382PubMedCrossRefGoogle Scholar
  36. Sobajima H, Tani T, Chujo T, Okada K, Suzuki K, Mori S, Minami E, Nishiyama M, Nojiri H, Yamane H (2007) Identification of a jasmonic acid-responsive region in the promoter of the rice 12-oxophytodienoic acid reductase 1 gene OsOPR1. Biosci Biotechnol Biochem 71(12):3110–3115PubMedCrossRefGoogle Scholar
  37. Takken FL, Albrecht M, Tameling WI (2006) Resistance proteins: molecular switches of plant defence. Curr Opin Plant Biol 9(4):383–390PubMedCrossRefGoogle Scholar
  38. Tan X, Meyers BC, Kozik A, West MA, Morgante M, St Clair DA, Bent AF, Michelmore RW (2007) Global expression analysis of nucleotide binding site-leucine rich repeat-encoding and related genes in Arabidopsis. BMC Plant Biol 7:56PubMedCrossRefGoogle Scholar
  39. Whitham S, Dinesh-Kumar SP, Choi D, Hehl R, Corr C, Baker B (1994) The product of the tobacco mosaic virus resistance gene N: similarity to toll and the interleukin-1 receptor. Cell 78(6):1101–1115PubMedCrossRefGoogle Scholar
  40. Yang S, Zhang X, Yue JX, Tian D, Chen JQ (2008) Recent duplications dominate NBS-encoding gene expansion in two woody species. Mol Genet Genomics 280(3):187–198PubMedCrossRefGoogle Scholar
  41. Yi H, Richards EJ (2007) A cluster of disease resistance genes in Arabidopsis is coordinately regulated by transcriptional activation and RNA silencing. Plant Cell 19(9):2929–2939PubMedCrossRefGoogle Scholar
  42. Zhang ZY, Li FL, Zhu ZT (1992) Chromosome doubling and triploid breeding of Populus tomentosa Carr.and its hybrid. J Beijing For Univ 14(Suppl):52–58Google Scholar
  43. Zhang ZY, Li FL, Zhu ZT (1997) Doubling technology of pollen chromosome of Populus tomentosa and its hybrids. J Beijing For Univ (English edn) 6:9–20Google Scholar
  44. Zhang Q, Zhang ZY, Lin SZ, Zheng HQ, Lin YZ, An XM, Li Y, Li HX (2008) Characterization of resistance gene analogs with a nucleotide binding site isolated from a triploid white poplar. Plant Biol 10:310–322PubMedCrossRefGoogle Scholar
  45. Zheng HQ, Lin SZ, Zhang Q, Zhang ZZ, Zhang ZY, Lei Y, Hou L (2007) Isolation and analysis of a TIR-specific promoter from poplar. For Stud China 9(2):95–106Google Scholar
  46. Zheng HQ, Lin SZ, Zhang Q, Lei Y, Zhang ZY (2009) Functional analysis of 5′ untranslated region of a TIR-NBS-encoding gene from triploid white poplar. Mol Genet Genomics 282(4):381–394PubMedCrossRefGoogle Scholar
  47. Zipfel C, Robatzek S, Navarro L, Oakeley EJ, Jones JD, Felix G, Boller T (2004) Bacterial disease resistance in Arabidopsis through flagellin perception. Nature 428(6984):764–767PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Huiquan Zheng
    • 1
  • Shanzhi Lin
    • 1
  • Qian Zhang
    • 1
    • 2
  • Yang Lei
    • 1
  • Lu Hou
    • 1
  • Zhiyi Zhang
    • 1
  1. 1.National Engineering Laboratory for Tree Breeding, Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants, Ministry of Education, The Tree and Ornamental Plant Breeding and Biotechnology Laboratory of State Forestry AdministrationBeijing Forestry UniversityBeijingPeople’s Republic of China
  2. 2.Guangdong Academy of ForestryGuangzhouPeople’s Republic of China

Personalised recommendations