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Planta

, Volume 227, Issue 1, pp 13–24 | Cite as

Analysis of gene expression profiles in response to Sclerotinia sclerotiorum in Brassica napus

  • Jianwei ZhaoEmail author
  • Jianlin Wang
  • Lingling An
  • R. W. Doerge
  • Z. Jeffrey Chen
  • Craig R. Grau
  • Jinling Meng
  • Thomas C. Osborn
Original Article

Abstract

Sclerotinia sclerotiorum is a necrotrophic plant pathogen which causes serious disease in agronomically important crop species. The molecular basis of plant defense to this pathogen is poorly understood. We investigated gene expression changes associated with S. sclerotiorum infection in a partially resistant and a susceptible genotype of oilseed Brassica napus using a whole genome microarray from Arabidopsis. A total of 686 and 1,547 genes were found to be differentially expressed after infection in the resistant and susceptible genotypes, respectively. The number of differentially expressed genes increased over infection time with the majority being up-regulated in both genotypes. The putative functions of the differentially expressed genes included pathogenesis-related (PR) proteins, proteins involved in the oxidative burst, protein kinase, molecule transporters, cell maintenance and development, abiotic stress, as well as proteins with unknown functions. The gene regulation patterns indicated that a large part of the defense response exhibited as a temporal and quantitative difference between the two genotypes. Genes associated with jasmonic acid (JA) and ethylene signal transduction pathways were induced, but no salicylic acid (SA) responsive genes were identified. Candidate defense genes were identified by integration of the early response genes in the partially resistant line with previously mapped quantitative trait loci (QTL). Expression levels of these genes were verified by Northern blot analyses. These results indicate that genes encoding various proteins involved in diverse roles, particularly WRKY transcription factors and plant cell wall related proteins may play an important role in the defense response to S. sclerotiorum disease.

Keywords

Brassica Gene expression profile Microarray Sclerotinia Arabidopsis 

Abbreviations

ET

Ethylene

hpi

Hours post-inoculation

JA

Jasmonic acid

RT-PCR

Reverse-transcriptase polymerase chain reaction

SA

Salicylic acid

Notes

Acknowledgments

We thank Drs. D. H. Hegedus of Saskatoon Research Center and Chris Pires of University of Missouri-Columbia for critical review of the manuscript. This work was funded by the United States Department of Agriculture Sclertotinia Initiative to T. C. Osborn, and a NSF Plant Genome Research Program (grant DBI0077774) to R. W. Doerge, Z. J. Chen, and T. C. Osborn. This work was also partially supported by National Natural Science Foundation of China to Zhao Jianwei (30170496).

Supplementary material

References

  1. Ali GS, Reddy VS, Lindgren PB, Jakobek JL, Reddy ASN (2003) Differential expression of genes encoding calmodulin-binding proteins in response to bacterial pathogens and inducers of defense responses. Plant Mol Biol 51:803–815PubMedCrossRefGoogle Scholar
  2. Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215:403–410PubMedGoogle Scholar
  3. Benjamini Y, Hochberg Y (1995) Controlling the false discovery rate: a practical and powerful approach to multiple testing. J R Stat Soc Ser B 57:289–300Google Scholar
  4. Campbell EJ, Schenk PM, Kazan K, Penninckx IAMA, Anderson JP, Maclean DJ, Cammue BPA, Ebert PR, Manners JM (2003) Pathogen-responsive expression of a putative ATP-binding cassette transporter gene conferring resistance to the diterpenoid sclareol is regulated by multiple defense signaling pathways in Arabidopsis. Plant Physiol 133:1272–1284PubMedCrossRefGoogle Scholar
  5. Cavell AC, Lydiate DJ, Parkin IA, Dean C, Trick M (1998) Collinearity between a 30-centimorgan segment of Arabidopsis thaliana chromosome 4 and duplicated regions within the Brassica napus genome. Genome 41:62–69PubMedCrossRefGoogle Scholar
  6. Chen W, Provart NJ, Glazebrook J, Katagiri F, Chang HS, Eulgem T, Mauch F, Luan S, Zou G, Whitham SA, Budworth PR, Tao Y, Xie Z, Chen X, Lam S, Kreps JA, Harper JF, Si-Ammour A, Mauch-Mani B, Heinlein M, Kobayashi K, Hohn T, Dangl JL, Wang X, Zhu T (2002) Expression profile matrix of Arabidopsis transcription factor genes suggests their putative functions in response to environmental stresses. Plant Cell 14:559–574PubMedCrossRefGoogle Scholar
  7. Cheong YH, Chang HS, Gupta R, Wang X, Zhu T, Luan S (2002) Transcriptional profiling reveals novel interactions between wounding, pathogen, abiotic stress, and hormonal responses in Arabidopsis. Plant Physiol 129:661–677PubMedCrossRefGoogle Scholar
  8. Cleveland WS (1979) Robust locally weighted regression and smoothing scatterplots. J Am Stat Assoc 74:829–836CrossRefGoogle Scholar
  9. Craig BA, Black MA, Doerge RW (2003) Gene expression data: the technology and statistical analysis. J Agri Biol Environ Stat (JABES) 8:1–28CrossRefGoogle Scholar
  10. Dong X (1998) SA, JA, ethylene, and disease resistance in plants. Curr Opin Plant Biol 1:316–323PubMedCrossRefGoogle Scholar
  11. Dowd C, Wilson IW, McFadden H (2004) Gene expression profile changes in cotton root and hypocotyl tissues in response to infection with Fusarium oxysporum f. sp. vasinfectum. Mol Plant Microbe Interact 17:654–667PubMedCrossRefGoogle Scholar
  12. Ferreira ME, Williams PH, Osborn TC (1994) RFLP mapping of Brassica napus using doubled haploid lines. Theor Appl Genet 89:615–621CrossRefGoogle Scholar
  13. Feys BJ, Parker JE (2000) Interplay of signaling pathways in plant disease resistance. Trends Genet 16:449–455PubMedCrossRefGoogle Scholar
  14. Fotopoulos V, Gilbert MJ, Pittman JK, Marvier AC, Buchanan AJ, Sauer N, Hall JL, Williams LE (2003) The monosaccharide transporter gene, AtSTP4, and the cell-wall invertase, Atβfruct1, are induced in Arabidopsis during infection with the fungal biotroph Erysiphe cichoracearum. Plant Physiol 132:821–829PubMedCrossRefGoogle Scholar
  15. Girke T, Todd J, Ruuska S, White J, Benning C, Ohlrogge J (2000) Microarray analysis of developing Arabidopsis seeds. Plant Physiol 124:1570–1581PubMedCrossRefGoogle Scholar
  16. Glazebrook J (2005) Contrasting mechanisms of defense against biotrophic and necrotrophic pathogens. Annu Rev Phytopathol 43:205–227PubMedCrossRefGoogle Scholar
  17. Horvath DP, Schaffer R, West M, Wisman E (2003) Arabidopsis microarrays identify conserved and differentially expressed genes involved in shoot growth and development from distantly related plant species. Plant J 34:125–134PubMedCrossRefGoogle Scholar
  18. Jasinski M, Ducos E, Martinoia E, Boutry M (2003) The ATP-binding cassette transporters: structure, function, and gene family comparison between rice and Arabidopsis. Plant Physiol 131:1169–1177PubMedCrossRefGoogle Scholar
  19. Jiang H (2004) A two-step procedure for multiple pairwise comparisons in microarray experiments. Ph.D. Dissertation, Department of Statistics, Purdue University, West Lafayette, INGoogle Scholar
  20. Kachroo A, Kachroo P (2007) Salicylic acid-, jasmonic acid- and ethylene-mediated regulation of plant defense signaling. Genetic engineering: principles and methods. Plenum, New York, pp 28:55–75Google Scholar
  21. Lee H, Wang J, Tian L, Jiang H, Black M, Madlung A, Watson B, Lukens, Pires JC, Wang J, Comai L, Osborn TC, Doerge RW, Chen J (2004) Sensitivity of 70-mer oligonucleotides and cDNAs for microarray analysis of gene expression in Arabidopsis and its related species. Plant Biotechnol J 2:45–57PubMedCrossRefGoogle Scholar
  22. Maleck K, Levine A, Eulgem T, Morgan A, Schmid J, Lawton KA, Dangl JL, Dietrich RA (2000) The transcriptome of Arabidopsis thaliana during systemic acquired resistance. Nat Genet 26:403–410PubMedCrossRefGoogle Scholar
  23. Marathe R, Guan Z, Anandalakshmi R, Zhao H, 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
  24. Martinez C, Blanc F, Le Claire E, Besnard O, Nicole M, Baccou J-C (2001) Salicylic acid and ethylene pathways are differentially activated in melon cotyledons by active or heat-denatured cellulase from Trichoderma longibrachiatum. Plant Physiol 127:334–344PubMedCrossRefGoogle Scholar
  25. McDowell JM, Dangl JL (2000) Signal transduction in the plant immune response. Trends Biochem Sci 25:79–82PubMedCrossRefGoogle Scholar
  26. Parkin IAP, Gulden SM, Sharpe AG, Lukens L, Trick M, Osborn TC, Lydiate DJ (2005) Segmental structure of the Brassica napus genome based on comparative analysis with Arabidopsis thaliana. Genetics 171:765–781PubMedCrossRefGoogle Scholar
  27. Pieterse CMJ, van Wees SCM, van Pelt JA, Knoester M, Laan R, Gerrits H, Weisbeek PJ, van Loon LC (1998) A novel signaling pathway controlling induced systemic resistance in Arabidopsis. Plant Cell 10:1571–1580PubMedCrossRefGoogle Scholar
  28. Rathjen JP, Moffett P (2003) Early signal transduction events in specific plant disease resistance. Curr Opin Plant Biol 6:300–306PubMedCrossRefGoogle Scholar
  29. Reymond P, Bodenhausen N, Van Poecke RMP, Krishnamurthy V, Dicke M, Farmer EE (2004) A conserved transcript pattern in response to a specialist and a generalist herbivore. Plant Cell 16:3132–3147PubMedCrossRefGoogle Scholar
  30. Riechmann JL, Heard J, Martin G, Reuber L, Jiang ZC, Keddie J, Adam L, Pineda O, Ratcliffe OJ, Samaha RR, Creelman R, Pilgrim M, Broun P, Zhang JZ, Ghandehari D, Sherman BK, Yu LG (2000) Arabidopsis transcription factors: genome-wide comparative analysis among eukaryotes. Science 290:2105–2110Google Scholar
  31. Schenk PM, Kazan K, Wilson I, Anderson JP, Richmond T, Somerville SC, Manners JM (2000) Coordinated plant defense responses in Arabidopsis revealed by microarray analysis. Proc Natl Acad Sci USA 97:11655–11660PubMedCrossRefGoogle Scholar
  32. Tao Y, Xie Z, Chen W, Glazebrook J, Chang H-S, Han B, Zhu T, Zou G, Katagiri F (2003) Quantitative nature of Arabidopsis responses during compatible and incompatible interactions with the bacterial pathogen Pseudomonas syringae. Plant Cell 15:317–330PubMedCrossRefGoogle Scholar
  33. Truernit E, Schmid J, Epple P, Illig J, Sauer N (1996) The sink-specific and stress-regulated Arabidopsis STP4 gene: enhanced expression of a gene encoding a monosaccharide transporter by wounding, elicitors, and pathogen challenge. Plant Cell 8:2169–2182PubMedCrossRefGoogle Scholar
  34. Udall JA, Quijada PA, Osborn TC (2005) Detection of chromosomal rearrangements derived from homeologous recombination in four mapping populations of Brassica napus L. Genetics 169:967–979PubMedCrossRefGoogle Scholar
  35. Wang J, Lee JJ, Tian L, Lee H-S, Chen M, Rao S, Wei EN, Doerge RW, Comai L, Chen ZJ (2005) Methods for genome-wide analysis of gene expression changes in polyploids. Methods Enzymol 395:570–596PubMedCrossRefGoogle Scholar
  36. Xia Y, Suzuki H, Borevitz J, Blount J, Guo Z, Patel K, Dixon RA, Lamb C (2004) An extracellular aspartic protease functions in Arabidopsis disease resistance signaling. EMBO J 23:980–988PubMedCrossRefGoogle Scholar
  37. Zhao J, Peltier AJ, Meng J, Osborn T, Grau CR (2004) Evaluation of Sclerotinia stem rot resistance in oilseed Brassica napus using a petiole inoculation technique under greenhouse conditions. Plant Dis 88:1033–1039CrossRefGoogle Scholar
  38. Zhao J, Udall JA, Quijada P, Grau C, Meng J, Osborn T (2006) Quantitative trait loci for resistance to Sclerotinia sclerotiorum and its association with a homeologous non-reciprocal transposition in Brassica napus L. Theor Appl Genet 112:509–516PubMedCrossRefGoogle Scholar
  39. Zheng Z, Qamar S, Chen Z, Mengiste T (2006) Arabidopsis WRKY33 transcription factor is required for resistance to necrotrophic fungal pathogens. Plant J 48:592–605PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Jianwei Zhao
    • 1
    • 5
    • 6
    Email author
  • Jianlin Wang
    • 3
  • Lingling An
    • 4
  • R. W. Doerge
    • 4
  • Z. Jeffrey Chen
    • 3
    • 7
  • Craig R. Grau
    • 2
  • Jinling Meng
    • 5
  • Thomas C. Osborn
    • 1
    • 8
  1. 1.Department of AgronomyUniversity of WisconsinMadisonUSA
  2. 2.Department of Plant PathologyUniversity of WisconsinMadisonUSA
  3. 3.Plant Genetics and Genomics Laboratory, Department of Soil and Crop SciencesTexas A&M UniversityCollege StationUSA
  4. 4.Department of StatisticsPurdue UniversityWest LafayetteUSA
  5. 5.National Key Laboratory of Crop Genetic ImprovementHuazhong Agricultural UniversityWuhanChina
  6. 6.Agriculture and Agri-Food CanadaSaskatoon Research CentreSaskatoonCanada
  7. 7.Institute for Cellular and Molecular BiologyUniversity of TexasAustinUSA
  8. 8.Seminis Vegetable SeedsWoodlandUSA

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