Analysis of gene expression profiles in response to Sclerotinia sclerotiorum in Brassica napus
- 1.3k Downloads
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.
KeywordsBrassica Gene expression profile Microarray Sclerotinia Arabidopsis
Reverse-transcriptase polymerase chain reaction
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).
- 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
- 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
- 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
- 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
- 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
- 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
- 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
- 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