Functional & Integrative Genomics

, Volume 12, Issue 2, pp 265–275 | Cite as

LOX genes in blast fungus (Magnaporthe grisea) resistance in rice

  • Soma S. Marla
  • V. K. Singh
Original Paper


Plant Lipoxygenases (LOX) are known to play major role in plant immunity by providing front-line defense against pathogen-induced injury. To verify this, we isolated a full-length OsLOX3 gene and also 12 OsLOX cDNA clones from Oryza sativa indica (cultivar Pusa Basmati 1). We have examined the role played by LOXs in plant development and during attack by blast pathogen Magnaporthe grisea. Gene expression, promoter region analysis, and biochemical and protein structure analysis of isolated OsLOX3 revealed significant homology with LOX super family. Protein sequence comparison of OsLOXs revealed high levels of homology when compared with japonica rice (up to100%) and Arabidopsis (up to 64%). Isolated LOX3 gene and 12 OsLOX cDNAs contained the catalytic LOX domains much required for oxygen binding and synthesis of oxylipins. Amino acid composition, protein secondary structure, and promoter region analysis (with abundance of motifs CGTCA and TGACG) support the role of OsLOX3 gene in providing resistance to diseases in rice plants. OsLOX3 gene expression analysis of root, shoot, flag leaf, and developing and mature seed revealed organ specific patterns during rice plant development and gave evidence to association between tissue location and physiological roles played by individual OsLOXs. Increased defense activity of oxylipins was observed as demonstrated by PCR amplification of OsLOX3 gene and upon inoculation with virulent strains of M. grisea and ectopic application of methyl jasmonate in the injured leaf tissue in adult rice plants.


Lipoxygenase Rice Blast disease Defense response Methyl jasmonate Expression 



The authors are grateful to Bioinformatics Information Network under Department of Biotechnology, Government of India for grant provided to support experimental costs and fellowship to VKS. Also, the authors are thankful to B. Murugan and A. Dev for help in disease inoculation experiments in the green house and to Department of Molecular Biology and Genetic engineering, G.B. Pant University of Agriculture and Technology, Pantngar, India for providing all laboratory and green house facilities.

Supplementary material

10142_2012_268_MOESM1_ESM.doc (538 kb)
ESM 1 (DOC 537 kb)


  1. Altschul SF, Warren GW, Miller W, Myers EW, Lipmanl DJ (1990) Basic local alignment search tool. J Mol Biol 215(3):403–410PubMedGoogle Scholar
  2. Altschul SF, Madden TL, Alejandro A, Schäffer AA, Zhang J, Zheng Z, Miller W, Lipman D (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucl Acids Res 25(17):3389–3402PubMedCrossRefGoogle Scholar
  3. Bailey TM, Elkan C (1994) Fitting a mixture model by expectation maximization to discover motifs in biopolymers. In: Proceedings of the Second International Conference on Intelligent Systems for Molecular Biology. AAAI Press, Menlo Park, California, pp. 28–36Google Scholar
  4. Bell E, Mullet JE (1993) Characterization of an Arabidopsis lipoxygenase gene responsive to methyl jasmonate and wounding. Plant Physiol 103(4):1133–1137PubMedCrossRefGoogle Scholar
  5. Brash AR (1999) Lipoxygenases: occurrence, functions, catalysis, and acquisition of substrate. J Biol Chem 274(34):23679–23682PubMedCrossRefGoogle Scholar
  6. Bu Q, Jiang H, Li CB, Zhai Q, Zhang J, Wu X, Sun J, Xie Q, Li C (2008) Role of the Arabidopsis thaliana NAC transcription factors ANAC019 and ANAC055 in regulating jasmonic acid-signaled defense responses. Cell Res 18:756–767PubMedCrossRefGoogle Scholar
  7. Burow GB, Gardner HW, Keller NP (2000) A peanut seed lipoxygenase responsive to Aspergillus colonization. Plant Mol Biol 42(5):689–701PubMedCrossRefGoogle Scholar
  8. Carver T, Berriman M, Tivey A, Patel C, Böhme U, Barrell BG, Parkhill J, Rajandream MA (2008) Artemis and ACT: viewing, annotating and comparing sequences stored in a relational database. Bioinformatics 24(23):2672–2676PubMedCrossRefGoogle Scholar
  9. Cheong JJ, Choi YDJ (2003) Methyl jasmonate as a vital substance in plants. Trends Genet 19(7):409–413PubMedCrossRefGoogle Scholar
  10. Creelman RA, Tierneyt ML, Mullet JE (1992) Jasmonic acid/methyl jasmonate accumulate in wounded soybean hypocotyls and modulate wound gene expression. Proc Natl Acad Sci USA 89:4938–4941PubMedCrossRefGoogle Scholar
  11. Després C, Chubak C, Rochon A, Clark R, Bethune T, Desveaux D, Fobert PR (2003) The Arabidopsis NPR1 disease resistance protein is a novel cofactor that confers redox regulation of DNA binding activity to the basic domain/leucine zipper transcription factor TGA1. Plant Cell 15:2181–2191PubMedCrossRefGoogle Scholar
  12. Eckardt NA (2008) Oxylipin signaling in plant stress responses. Plant Cell 20(3):495–497PubMedCrossRefGoogle Scholar
  13. Farmer EE, Ryan CA (1990) Interplant communication: airborne methyl jasmonate induces synthesis of proteinase inhibitors in plant leaves. Proc Natl Acad Sci USA 87:7713–7716PubMedCrossRefGoogle Scholar
  14. Feussner I, Wasternack C (2002) The lipoxygenase pathway. Annu Rev Plant Biol 53:275–297PubMedCrossRefGoogle Scholar
  15. Gao G, Zhang S, Wang C, Yang X, Wang Y (2011) Arabidopsis CPR5 independently regulates seed germination and postgermination arrest of development through LOX pathway and ABA signaling. PLoS One 6(4):19406CrossRefGoogle Scholar
  16. Griffiths A, Barry C, Alpuche-Solis AG, Grierson D (1999) Ethylene and developmental signals regulate expression of lipoxygenase genes during tomato fruit ripening. J Exp Bot 50:793–798Google Scholar
  17. He X, Chen J, Zhang Z, Zhang J, Chen S (2002) Identification of salt-stress responsive genes in rice (Oryza sativa L.) by cDNA array. Sci China C Life Sci 45(5):477–484PubMedCrossRefGoogle Scholar
  18. 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
  19. Kachroo A, Kachroo P (2009) Fatty acid derived signals in plant defense. Annu Rev Phytopathol 47:153–157PubMedCrossRefGoogle Scholar
  20. Kenta S, Takeuchi Y, Ebitani T, Suzuki Y (2008) Identification of gene for rice (Oryza sativa) seed lipoxigenase-3 involved in the generation of stale flavor development of SNP markers for lipoxigenase-3 deficiency. Breed Sci 58(2):169–175CrossRefGoogle Scholar
  21. Kesarwani M, Yoo J, Dong X (2007) Genetic interactions of TGA transcription factors in the regulation of pathogenesis-related genes and disease resistance in Arabidopsis. Plant Physiol 144:336–346PubMedCrossRefGoogle Scholar
  22. Kolomites MV, Hao CH, Richard J, Gladon RJ, Braun EJ, Hannapel DJ (2000) A leaf lipoxygenase of potato induced specifically by pathogen infection. Plant Physiol 124(3):1121–1130CrossRefGoogle Scholar
  23. 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
  24. Liavonchanka A, Feussner I (2006) Lipoxygenases: occurrence, functions and catalysis. J Plant Physiol 163(3):348–357PubMedCrossRefGoogle Scholar
  25. Lütteke T, Krieg P, Fürstenberger G, von der Lieth CW (2003) OSLOX-DB-database on lipoxygenases. Bioinformatics 19(18):2482–2483PubMedCrossRefGoogle Scholar
  26. Melan MA, Dong X, Endara ME, Davis KR, Ausubel FM, Peterman TK (1993) An Arabidopsis thaliana lipoxygenase gene can be induced by pathogens, abscisic acid, and methyl jasmonate. Plant Physiol 101(2):441–450PubMedCrossRefGoogle Scholar
  27. Minor W, Steczko J, Stec B, Otwinowski Z, Bolin JT, Walter R, Axelrod B (1996) Crystal structure of soybean lipoxygenase L-1 at 1.4 A resolution. Biochemistry 35:10687–10701PubMedCrossRefGoogle Scholar
  28. Murray MG, Thompson WF (1980) Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res 8(19):4321–4325PubMedCrossRefGoogle Scholar
  29. Ohta H, Shirano Y, Tanaka K, Morita Y, Shibata D (1992) cDNA cloning of rice lipoxygenase L-2 and characterization using an active enzyme expressed from the cDNA in Escherichia coli. Eur J Biochem 206:331–336PubMedCrossRefGoogle Scholar
  30. Peng YL, Shirano Y, Ohta H, Hibino T, Tanaka K, Shibata D (1994) A novel lipoxygenase from rice. Primary structure and specific expression upon incompatible infection with rice blast fungus. J Biol Chem 269:3755–3761PubMedGoogle Scholar
  31. Porta H, Rocha-Sosa M (2002) Plant lipoxygenases, physiological and molecular features. Plant Physiol 130(1):15–21PubMedCrossRefGoogle Scholar
  32. Porta H, Rueda-Benítez P, Campos F, Colmenero-Flores JM, Colorado JM, Carmona MEJ, Covarrubias AA, Rocha-Sosa M (1999) Analysis of lipoxygenase mRNA accumulation in the common bean (Phaseolus vulgaris L.) during development and under stress conditions. Plant Cell Physiol 40(8):850–858PubMedCrossRefGoogle Scholar
  33. Quevillon E, Silventoinen V, Pillai S, Harte N, Mulder N, Apweiler R, Lopez R (2005) Interproscan: protein domains identifier. Nucleic Acids Res 33(Web Server issue):W116–W120PubMedCrossRefGoogle Scholar
  34. Saravitz DM, Siedeow JN (1995) The lipoxygenase isozymes in soybean [Glycine max (L.) Merr.] leaves (changes during leaf development, after wounding, and following reproductive sink removal). Plant Physiol 107(2):535–543PubMedGoogle Scholar
  35. Shen H, Hu H, Wang F, Wu Q, Huang Q, Wang H (2008) Coevolving residues of (beta/alpha)(8)-barrel proteins play roles in stabilizing active site architecture and coordinating protein dynamics. J Struct Biol 164(3):281–292PubMedCrossRefGoogle Scholar
  36. Siedeow JN (1991) The lipoxygenase-structure and function. Annu Rev Plant Physiol Plant Mol Biol 42:145–188CrossRefGoogle Scholar
  37. Solovyev V, Kosarev P, Seledsov I, Vorobyev D (2006) Automatic annotation of eukaryotic genes, pseudogenes and promoters. Genome Biol 7 Suppl 1(S10):1–12Google Scholar
  38. Stormo GD (2000) Gene-finding approaches for eukaryotes. Genome Res 10(4):394–397PubMedCrossRefGoogle Scholar
  39. Suzuki Y, Ise K, Li C, Honda I, Iwai Y, Matsukura U (1999) Volatile components in stored rice [Oryza sativa (L.)] of varieties with and without lipoxygenase-3 in seeds. J Agric Food Chem 47(3):1119–1124PubMedCrossRefGoogle Scholar
  40. Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22(22):4673–4680PubMedCrossRefGoogle Scholar
  41. Vellosillo T, Martínez M, López MA, Jorge Vicente J, Cascón T, Dolan L, Hamberg M, Castresana C (2007) Oxylipins produced by the 9-lipoxygenase pathway in Arabidopsis regulate lateral root development and defense responses through a specific signaling cascade. Plant Cell 19:831–846PubMedCrossRefGoogle Scholar
  42. Wang R, Shen W, Liu L, Liu L, Jiang L, Liu Y, Su N, Wan J (2008) A novel lipoxygenase gene from developing rice seeds confers dual position specificity and responds to wounding and insect attack. Plant Mol Biol 66:401–414PubMedCrossRefGoogle Scholar
  43. Zhang YY, Radmark O, Samuelsson B (1992) Mutagenesis of some conserved residues in human 5-lipoxygenase: effects on enzyme activity. Proc Natl Acad Sci USA 89:485–489PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  1. 1.Molecular Biology and Genetic EngineeringG.B. Pant University of Agriculture and TechnologyPantnagarIndia
  2. 2.National Genomic Resources and Bioinformatics, National Bureau of Plant Genetic ResourcesICARNew DelhiIndia
  3. 3.Bioinformatics CentreBanaras Hindu UniversityVaranasiIndia

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