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Identification of pathogen-responsive regions in the promoter of a pepper lipid transfer protein gene (CALTPI) and the enhanced resistance of the CALTPI transgenic Arabidopsis against pathogen and environmental stresses

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Abstract

The 5′ flanking region of the CALTPI gene, which encodes a basic lipid transfer protein, was isolated and characterized from the genomic DNA of Capsicum annuum. Four different regions of the promoter sequence of the CALTPI gene were fused to the β-glucuronidase (GUS) coding region. In an Agrobacterium-mediated transient expression assay, the transcriptional activations of the promoter deletions were examined in tobacco leaves after infection with Pseudomonas syringae pv. tabaci, and treatment with ethylene and salicylic acid. The −808 bp region of the CALTPI gene promoter sequence exhibited full promoter activity. The W-box and ERE-box elements, which are essential for induction by all signals, were localized in the region between −555 bp and −391 bp upstream of the translation initiation site. A CALTPI transgene was then introduced under the control of the 35S promoter into the Arabidopsis ecotype Col-0. Transgenic Arabidopsis lines expressing the CALTPI gene developed rapidly compared to the wild-type plants, indicating that CALTPI may be involved in plant development. Overexpression of the CALTPI gene enhanced the resistance against infection by P. syringae pv. tomato and Botrytis cinerea. The transgenic plants expressing the CALTPI gene also showed high levels of tolerance to NaCl and drought stresses at various vegetative growth stages. No transcription of the PR-1, PR-2, PR-5, thionin, and RD29A genes was observed in untreated leaf tissues of the transgenic plants. The enhanced resistance to pathogen and environmental stresses in transgenic Arabidopsis correlated with the enhanced expression of the CALTPI gene.

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References

  • Berrocal-Lobo M, Molina A, Solano R (2002) Constitutive expression of ethylene-response-factor 1 in Arabidopsis confers resistance to several necrophic fungi. Plant J 29:23–32

    Article  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  CAS  PubMed  Google Scholar 

  • Canevascini S, Caderas D, Mandel T, Fleming AJ, Dupuis I, Kuhlemeier C (1996) Tissue-specific expression and promoter analysis of the tobacco ltp1 gene. Plant Physiol 112:513–524

    Article  Google Scholar 

  • Clough SH, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16:735–743

    Article  CAS  PubMed  Google Scholar 

  • Colmenero-Flores JM, Campos F, Garciarrubio A, Covarrubias AA (1997) Characterization of Phaseolus vulgaris cDNA clones responsive to water deficit: identification of a novel late embryogenesis abundant-like protein. Plant Mol Biol 35:393–405

    Article  Google Scholar 

  • Despres C, Subramaniam R, Matton DP, Brisson N (1995) The activation of the potato PR-10a gene requires the phosphorylation of the nuclear factor PBF-1. Plant Cell 7:589–598

    Article  Google Scholar 

  • Dunn MA, White AJ, Vural S, Hughes MA (1998) Identification of promoter elements in a low-temperature-responsive gene (blt4.9) from barley (Hordeum vulgare L.). Plant Mol Biol 38:551–564

    Article  Google Scholar 

  • Fleming AJ, Mandel T, Hofmann S, Sterk P, de Vries SC, Kuhlemeier C (1992) Expression pattern of a tobacco lipid transfer protein gene within the shoot apex. Plant J 2:855–86

    Article  Google Scholar 

  • Ge X, Chen J, Sun C, Cao K (2003) Preliminary study on the structural basis of the antifungal activity of a rice lipid transfer protein. Protein Eng 16:387–390

    Article  CAS  PubMed  Google Scholar 

  • Gomès E, Sagot E, Gaillard C, Laquitaine L, Poinssot B, Sanejouand Y, Delrop S, Coutos-Thévenot P (2003) Nonspecific lipid transfer protein genes expression in grape (Vitis sp.) cells in response to fungal elicitor treatments. Mol Plant Microbe Interact 16:456–464

    Google Scholar 

  • Guiderdoni E, Cordero MJ, Vignols F, Garcia-Garrido JM, Lescot M, Tharreau D, Meynard D, Ferrière N, Notteghem J, Deldeny M (2002) Inducibility by pathogen attack and developmental regulation of the rice Ltp1 gene. Plant Mol Biol 49:683–699

    Article  Google Scholar 

  • Herbers K, Meuwly P, Frommer WB, Métraux JP, Sonnewald U (1996) Systemic acquired resistance mediated by the ectopic expression of invertase: possible hexose sensing in the secretory pathway. Plant Cell 8:793–803

    Article  Google Scholar 

  • Higo K, Ugawa Y, Iwamoto M, Korenaga T (1999) Plant cis-acting regulatory DNA elements (PLACE) database. Nucleic Acids Res 27:297–300

    Google Scholar 

  • Horvath BM, Bachem CWB, Trindade LM, Oortwijn MEP, Visser RGF (2002) Expression analysis of a family of ns LTP genes tissue specifically expressed throughout the plant and during potato tuber life cycle. Plant Physiol 129:1494–1506

    Article  Google Scholar 

  • Hughes MA, Dunn MA, Pearce RS, White AJ, Zhang L (1992) An abscisic acid-responsive, low temperature barley gene has homology with a maize phospholipid transfer protein. Plant Cell Environ 15:861–865

    Google Scholar 

  • Iraki NM, Singh N, Bressen RA, Carpita NC (1989) Cell walls of tobacco cells and changes in composition associated with reduced growth upon adaptation to water and saline stress. Plant Physiol 91:48–53

    CAS  Google Scholar 

  • Jefferson RA, Kavanagh TA, Bevan MW (1987) GUS fusions beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6:3901–3907

    CAS  PubMed  Google Scholar 

  • Joshi CP (1987) An inspection of the domain between putative TATA box and translation start site in 79 plant genes. Nucleic Acids Res 15:6643–6653

    Google Scholar 

  • Jung HW, Hwang BK (2000) Pepper gene encoding a basic β-1,3-glucanase is differentially expressed in pepper tissues upon pathogen infection and ethephon or methyl jasmonate treatment. Plant Sci 159:97–106

    Article  Google Scholar 

  • Jung HW, Kim W, Hwang BK (2003) Three pathogen-inducible genes encoding lipid transfer protein from pepper are differentially activated by pathogens, abiotic, and environmental stresses. Plant Cell Environ 26:915–928

    Article  Google Scholar 

  • Kader JC (1996) Lipid transfer proteins in plants. Annu Rev Plant Physiol Plant Mol Biol 47:627–654

    Article  Google Scholar 

  • Katagiri F, Thilmony R, He SY (2002) The Arabidopsis thaliana-Pseudomonas syringae interaction. In: Somerville CR, Meyerowitz EM (ed) The Arabidopsis book. American Society of Plant Biologists, Rockville

  • Kim SY, Chung HJ, 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 11:1237–1251

    Article  CAS  PubMed  Google Scholar 

  • Knight H, Brandt S, Knight MR (1998) A history of stress alters drought calcium signaling pathways in Arabidopsis. Plant J 16:681–687

    Article  Google Scholar 

  • Maldonodo AM, Doerner P, Dixon RA, Lamb CJ, Cameron RK (2002) A putative lipid transfer protein involved in systemic resistance signaling in Arabidopsis. Nature 419:399–403

    Article  CAS  PubMed  Google Scholar 

  • Meshi T, Iwabuchi M (1995) Plant transcription factors. Plant Cell Physiol 36:1405–1420

    Google Scholar 

  • Mittler R, Shulaev V, Lam E (1995) Coordinated activation of programmed cell death and defense mechanisms in transgenic tobacco plants expressing a bacterial proton pump. Plant Cell 7:29–42

    Article  Google Scholar 

  • Molina A, Garcia-Olmedo F (1993) Developmental and pathogen-induced expression of three barley genes encoding lipid transfer protein. Plant J 4:983–991

    Article  Google Scholar 

  • Molina A, Garcia-Olmedo F (1997) Enhanced tolerance to bacterial pathogens caused by the transgenic expression of barley lipid transfer protein LTP2. Plant J 12:669–675

    Article  Google Scholar 

  • Molina A, Segura A, Garcia-Olmedo F (1993) Lipid transfer protein (nsLTPs) from barley and maize leaves are potent inhibitors of bacterial and fungal plant pathogens. FEBS Lett 316:119–122

    Article  CAS  PubMed  Google Scholar 

  • Molina A, Diaz I, Vasil IK, Carbonero P, Garcia-Olmedo F (1996) Two cold-inducible genes encoding lipid transfer protein LTP4 from barley show differential responses to bacterial pathogens. Mol Gen Genet 252:162–168

    Article  Google Scholar 

  • Montgomery J, Goldman S, Deikman J, Margossian L, Fischer RL (1993) Identification of an ethylene-responsive region in the promoter of a fruit ripening gene. Proc Natl Acad Sci USA 90:5939–5943

    CAS  PubMed  Google Scholar 

  • Nielson KK, Nielson JE, Madrid SM, Mikkelsen JD (1996) New antifungal proteins from sugar beet (Beta vulgaris L.) showing homology to non-specific lipid transfer proteins. Plant Mol Biol 31:539–552

    Article  Google Scholar 

  • Oh SA, Park J, Lee GI, Paek KH, Park SK, Nam HG (1997) Identification of three genetic loci controlling leaf senescence in Arabidopsis thaliana. Plant J 12:527–535

    Article  CAS  PubMed  Google Scholar 

  • Penninckx IAMA, Eggermont K, Terras FRG, Thomma BPHJ, De Samblanx GW, Buchala A, Metrux JP, Manners JM, Broekaert WF (1996) Pathogen-induced systemic activation of a plant defensin gene in Arabidopsis follows a salicylic acid-independent pathway. Plant Cell 8:2309–2323

    CAS  PubMed  Google Scholar 

  • Rathjen J, Chang JH, Staskawicz B, Michelmore RW (1999) Constitutively active Pto induces a Prf-dependent hypersensitive response in the absence of avrPto. EMBO J 18:3232–3240

    Article  Google Scholar 

  • Rogers EE, Ausubel FM (1997) Arabidopsiss enhanced disease susceptibility mutant exhibit enhanced susceptibility to several bacterial pathogens and alterations in PR-1 gene expression. Plant Cell 9:305–316

    Article  CAS  PubMed  Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis R (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor

    Google Scholar 

  • SAS Institute (1990) SAS user’s guide, version 6, 4th edn, vol 1. SAS institute, Cary

  • Segura A, Moreno M, Garcia-Olmedo F (1993) Purification and antipathogenic activity of lipid transfer proteins (LTPs) from the leaves of Arabidopsis and spinach. FEBS Lett 332:243–246

    Article  Google Scholar 

  • Sohal AK, Pallas JA, Jenkins GI (1999) The promoter of a Brassica napus lipid transfer protein gene is active in a range of tissues and stimulated by light and viral infection in transgenic Arabidopsis. Plant Mol Biol 41:75–87

    Article  Google Scholar 

  • Staehelin LA, Newcomb EH (2000) Membrane structure and membranous organelles. In: Buchanan B, Gruissem W, Jones RL (eds) Biochemistry and molecular biology of plants. ASPP Press, Rockville, pp 2–50

    Google Scholar 

  • Stalberg K, Ellerstom M, Ezcurra I, Ablov S, Rask L (1996) Disruption of an overlapping E-box/ABRE motif abolished high transcription of the napA storage-protein promoter in transgenic Brassica napus seeds. Planta 199:515–519

    PubMed  Google Scholar 

  • Sterk P, Booij H, Schellekens GA, Van Kammen A, De Vries SC (1991) Cell-specific expression of the carrot EP2 lipid transfer protein gene. Plant Cell 3:907–921

    Article  CAS  PubMed  Google Scholar 

  • Thoma S, Hecht U, Kippers A, Botella J, De Varies S, Somerville C (1994). Tissue-specific expression of a gene encoding a cell wall-localized lipid transfer protein from Arabidopsis. Plant Physiol 105:35–45

    Article  Google Scholar 

  • Torres-Schumann S, Godoy JA, Pintor-Toro JA (1992) A probable lipid transfer protein gene is induced by NaCl in stems of tomato plants. Plant Mol Biol 18:749–757

    Article  Google Scholar 

  • Trevino MB, O’Connell MA (1998) Three drought-responsive members of the nonspecific lipid-transfer protein gene family in Lyco persicon pennellii show different developmental patterns of expression. Plant Physiol 116:1461–1468

    Article  Google Scholar 

  • 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

    Article  CAS  PubMed  Google Scholar 

  • Yang Y, Li R, Qi M (2000) In vivo analysis of plant promoters and transcription factors by agroinfiltration of tobacco leaves. Plant J 22:543–551

    Article  CAS  PubMed  Google Scholar 

  • Yu D, Chen C, Chen Z (2001) Evidence for an important role of WRKY DNA binding proteins in the regulation of NPR1 gene expression. Plant Cell 13:1527–1540

    Article  Google Scholar 

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Acknowledgements

This work was supported by the Korea Science and Engineering Foundation (grant no. R01-2000-000-00086-0), Crop Functional Genomics Center of the 21st Century Frontier Research Program (grant no. CG1432), and the Center for Plant Molecular Genetics and Breeding Research, Seoul National University, Korea. We thank Stuart Timmis for critically reading the manuscript.

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  1. Division of Bioscience and Technology, College of Life and Environmental Sciences, Korea University, Anam-dong, Sungbuk-ku, Seoul, 136-713, Korea

    Ho Won Jung, Ki Deok Kim & Byung Kook Hwang

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Correspondence to Byung Kook Hwang.

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Jung, H.W., Kim, K.D. & Hwang, B.K. Identification of pathogen-responsive regions in the promoter of a pepper lipid transfer protein gene (CALTPI) and the enhanced resistance of the CALTPI transgenic Arabidopsis against pathogen and environmental stresses. Planta 221, 361–373 (2005). https://doi.org/10.1007/s00425-004-1461-9

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