Cloning and Characterization of Pathogenesis-related Gene 10a (OgPR10a) Derived from Wild Rice (Oryza grandiglumis)

  • Shin, Sang Hyun (National Crop Experiment Station, Rural Development Administration) ;
  • Pak, Jung-Hun (College of Life Science and Natural Resources, Dong-A University) ;
  • Kim, Mi Jin (College of Life Science and Natural Resources, Dong-A University) ;
  • Kim, Hye Jeong (College of Life Science and Natural Resources, Dong-A University) ;
  • Lee, Jai Heon (College of Life Science and Natural Resources, Dong-A University) ;
  • Kim, Doh Hoon (College of Life Science and Natural Resources, Dong-A University) ;
  • Choi, Hong Kyu (College of Life Science and Natural Resources, Dong-A University) ;
  • Kang, Kyung Ho (National Crop Experiment Station, Rural Development Administration) ;
  • Jeong, Ji Ung (National Crop Experiment Station, Rural Development Administration) ;
  • Kang, Chon-Sik (National Crop Experiment Station, Rural Development Administration) ;
  • Jung, Ho Won (College of Life Science and Natural Resources, Dong-A University) ;
  • Chung, Young Soo (College of Life Science and Natural Resources, Dong-A University)
  • Published : 2012.03.31

Abstract

Wild rice might have previously unidentified genes important for disease resistance and stress tolerance in response to biotic and abiotic stresses. A set of subtractive library was constructed both from leaves of wild rice plants, Oryza grandiglumis (CCDD, 2n=48), treated with fungal elicitor and from wounded leaves. A partial fragment that was homologous to PR10 genes from other plant species was identified via suppression subtractive hybridization and cDNA macroarray. The obtained full-length cDNA sequence (OgPR10) contains an open reading frame of 480 bp nucleotide, encoding 160 amino acids with a predicted molecular mass of 16.944 kDa and an isoelectric point (pI) of 4.91. The multiple alignment analyses showed the higher sequence homology of OgPR10 with PR10 genes identified in rice plants at amino acid level. The OgPR10 mRNA was not expressed by treatment with wounding, jasmonic acid, and salicylic acid, but markedly expressed in leaves treated with protein phosphatase inhibitors cantharidin and endothall, and yeast extract. In addition, the expression of OgPR10 mRNA was induced within 72 h after treatment with probenazole, one of well-known chemical elicitors, and reached the highest level at 144 h. Heterologous expression of OgPR10 caused growth inhibition and seedling lethality in E. coli and Arabidopsis, respectively. Chemically induced OgPR10 expression with glucocorticoid-mediated transcriptional induction system further reconfirmed its lethality on Arabidopsis seedling. In addition, OgPR10-expressing rice plants, Oryzae sativar were resistant against the infection of rice blast fungus, Magnaporthe grisea. These results indicate that OgPR10 is involved in probenazole- and microbe associated molecular patterns-mediated disease resistance responses in plants and is a potential gene for developing disease resistance crop plants.

Keywords

References

  1. Agrawal GK, Rakwal R, Yonekura M, Kubo A, Saji H. 2002. Proteome analysis of differentially displayed proteins as a tool for investigating ozone stress in rice (Oryza sativa L.) seedlings. Proteomics. 2:947-957.
  2. Akimoto M. 1998. Population genetic structure of the wild rice Oryza glumaepatula Steud. distributed in the amazon flood area influenced by-history traits. Mol. Ecol. 7:1371-1381
  3. Alexander D, Goodman RM, Gut-Rella M, Glascock C, Weymann K, Friedrich L, Maddox D, Ahl-Goy P, Luntz T, Ward E, Ryals J. 1993. Increased tolerance to two oomycete pathogens in transgenic tobacco expressing pathogenesis-related 1a. Pro. Natl. Acad. Sci. U.S.A. 90: 7327-7331.
  4. Aoyama T, Chua NH. 1997. A glucocorticoid-mediated transcriptional induction system in transgenic plants. Plant J. 11:605-612.
  5. Bantignies B, Seguin J, Muzac I, Dedaldechamp F, Gulick P, Ibrahim R. 2000. Direct evidence for ribonucleolytic activity of a PR-10-like protein from white lupin roots. Plant Mol. Biol. 42:871-881.
  6. Barratt DHP, Clark JA. 1991. Proteins arising during the late stages of embryogenesis in Pisum sativum L. Planta. 184:14-23.
  7. Bol JF, Linthorst HJM, Cornelissen BJC. 1990. Plant pathogenesis-related proteins induced by virus infection. Annu. Rev. Phytopathol. 28:113-138.
  8. Breiteneder H, Pettenburger K, Bito A, Valenta R, Dietrich K, Rumpold H, Scheiner O, Breitenbach M. 1989. The gene coding for the major birch pollen allergen Betv1, is highly homologous to a pea disease response gene. EMBO J. 8:1935-1938.
  9. Clough SJ, Bent AF. 1998. Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16:735-743.
  10. Colditz F, Niehaus K, Krajinski F. 2007. Silencing of PR-10-like proteins in Medicago truncatula results in an antagonistic induction of other PR proteins and in an increased tolerance upon infection with the oomycete Aphanomyces euteiches. Planta 226:57-71.
  11. Crowell DN, John ME, Russell D, Amasino RM. 1992. Characterization of a stress-induced, developmentally regulated gene family from soybean. Plant Mol. Biol. 18:459-466.
  12. Datta K, Velazhahan R, Oliva N, Ona I, Mew T, Khush GS, Muthukrishnan S, Datta SK. 1999. Over-expression of the cloned rice thaumatin-like protein (PR-5) gene in transgenic rice plants enhances environmental friendly resistance to Rhizoctonia solani causing sheath blight disease. Theor. Appl. Genet. 98:1138-1145.
  13. Fujimoto Y, Nagata R, Fukasawa H, Yano K, Azuma M, Iida A, Sugimoto S, Shudo K, Hashimoto Y. 1998. Purification and cDNA cloning of cytokinin-specific binding protein from mung bean (Vigna radiata). Eur. J. Biochem. 258:794-802.
  14. Gajhede M, Osmark P, Poulse FM, Ipsen H, Larsen JN, van Neerven RJJ, Schou C, Lowenstein H, Spangfort MD. 1996. X-ray and NMR structure of Bet v1, the origin of birch pollen allergy. Nat. Struct. Biol. 3:1040-1045.
  15. Horton P, Park KJ, Obayashi T, Fujita N, Harada H, Adams-Collier CJ, Nakai K. 2007. WoLF PSORT: protein localization predictor. Nucleic Acids Res. 35(Web Server issue):W585-7.
  16. Jeon EH, Chung ES, Pak JH, Nam JS, Cho SK, Shin SH, Kim DH, Kim GT, Lee JH, Kang KH, Chung YS. 2008. Overexpression of OgPAE1 from wild rice confers fungal resistance against Botrytis cinerea. J. Plant Res. 121:435-440.
  17. Jwa NS, Agrawal GK, Rakwal R, Park CH, Agrawal VP. 2001. Molecular cloning and characterization of a novel jasmonate inducible pathogenesis-related class 10 protein gene, JIOsPR10, from rice (Oryza sativa L.) seedling leaves. Bioche. Biophys. Res. Commun. 286:973-983.
  18. Kim KM,Cho SK, Shin SH, Kim GT, Lee JH, Oh, BJ, Kang KH, Hong JC, Choi JY, Shin JS, Chung YS. 2005. Analysis of differentially expressed transcripts of fungal elicitor-and wound-treated wild rice (Oryza grandiglumis). J. Plant Res. 118:347-354.
  19. Lee SC, Hwang BK. 2006. CASAR82A, a Pathogeninduced Pepper SAR8.2, Exhibits an antifungal activity and its overexpression enhances disease resistance and stress tolerance. Plant Mol. Biol. 61:95-109.
  20. Linthorst HJM. 1991. Pathogenesis-related proteins of plants. Crit. Rev. Plant Sci. 10:123-150.
  21. Lo SCC, Hipskind JD. Nicholson RL. 1999. cDNA cloning of a sorghum pathogenesis-related protein (PR-10) and differential expression of defense-related genes following inoculation with Cochliobolus heterostrophus of Colletotrichum sublineolum. Mol. Plant-Microbe Interact. 12:479-489.
  22. MacKintosh C, MacKintosh RW. 1994. Inhibitors of protein kinases and phosphatases. Trends Biochem Sci. 19:444-448.
  23. Matton DP, Brission N. 1989. Cloning, expression, and sequence conservation of pathogenesis-related gene transcripts of potato. Mol. Plant-Microbe Interact. 2:325-331.
  24. McGee JD, Hamer JE, Hodges TK. 2001. Characterization of a PR-10 Pathogenesis-related gene family induced in Rice during infection with Magnaporthe grisea. Mol. Plant-Microbe Interact. 14:877-886.
  25. Midoh N, Iwata M. 1996. Cloning and characterization of a probenazole-inducible gene for an intracellular pathogenesisrelated protein in rice. Plant Cell Physiol. 37:9-18.
  26. Niderman T, Genetet I, Bruyere T, Gees T, Stintzi A, Legrand M, Fritig B, Mosinger E. 1995. Pathogenesisrelated PR-1 proteins are antifungal. Plant Physiol. 108: 17-27.
  27. Park CJ, Kim KJ, Shin R, Park JM, Shin YC, Paek KH. 2004. Pathogenesis-related protein 10 isolated from hot pepper functions as a ribonuclease in an antiviral pathway. Plant J. 37:186-198.
  28. Rakwal R, Agrawal GK, Yonekura M. 1999. Separation of proteins from stressed rice (Oryza sativa L.) leaf tissues by two-dimensional polyacrylamide gel electrophoresis: induction of pathogenesis-related and cellular protectant proteins by jasmonic acid, UV irradiation and copper chloride. Electrophoresis. 20:3472-3478.
  29. Rakwal R, Agrawal GK, Yonekura M. 2001. Lightdependent induction of OsPR10 in rice (Oryza sativa L.) seedlings by the global stress signaling molecule jasmonic acid and protein phosphatase 2A inhibitors. Plant Sci. 161:469-479.
  30. Reymond P, Farmer EE. 1998. Jasmonate and salicylate as global signals for defense gene expression. Curr. Opin. Plant. Biol. 1:404-411.
  31. Somssich IE, Schmelzer E, Kawalleck P, Hahlbrock K. 1988. Gene structure and in situ transcript localization of pathogenesis-related protein 1 in parsley. Mol. Gen. Genet. 312:93-98.
  32. Srivastava S, Eristensky B, Kav NNV. 2004. Constitutive Expression of a PR10 Protein Enhances the Germination of Brassica napus under Saline Conditions. Plant Cell Physiol. 45:1320-1324.
  33. Tanaka N, Che FS, Watanabe N, Fujiwara S, Takayama S, Isogai A. 2003. Flagellin from an incompatible strain of Acidovorax avenae mediates $H_2O_2$ generation accompanying hypersensitive cell death and expression of PAL, Cht-1, and PBZ1, but not of LOX in rice. Mol. Plant-Microbe Interact. 16:422-428.
  34. Van Loon LC, Van Kammen A. 1970. Polyacrylamide disc electrophoresis of the soluble leaf proteins from Nicotiana tabacum var. 'Samsun' and 'Samsun NN'.II. Changes in protein constitution after infection with tobacco mosaic virus. Virology 40:199-211.
  35. Van Loon LC. 1985. Pathogenesis-related proteins. Plant Mol. Biol. 4:111-116.
  36. Van Loon LC, Van Strien EA. 1999. The families of pathogenesis-related proteins, their activities, and comparative analysis of PR-1 type proteins. Physiol. Mol. Plant Pathol. 55:85-97.
  37. Vaughan DA. 1994. The wild relatives of rice-a genetic resources handbook. IRRI, Manila, pp 3-24.
  38. Wang Y, Nowak G, Culley D, Hadwiger LA, Fristensky B. 1999. Constitutive expression of pea defense gene DRR206 confers resistance to blackleg (Leptosphaeria maculans) disease in transgenic canola (Brassica napus). Mol. Plant-Microbe Interact. 12:410-418.
  39. Warner SAJ, Scott R, Draper J. 1993. Isolation of an asparagus intracellular PR gene (AoPR1) wound-responsive promoter by the inverse polymerase chain reaction and its characterization in transgenic tobacco. Plant J. 3:191-201.