Transcriptional profiling by cDNA-AFLP reveals novel insights during methyl jasmonate, wounding and insect attack in Brassica napus.

Plants exploit a broad range of defense mechanisms to effectively combat invasion by pathogens or herbivores. Each environmental stress activates multiple signal transduction pathways to ensure an effective spatial and temporal defense response. A detailed transcriptome analysis using the cDNA-AFLP technique was performed to identify genes that are differentially expressed in oilseed rape (Brassica napus cv. Westar) leaves upon treatment with methyl jasmonate, mechanical wounding, or feeding by diamondback moth larvae (Plutella xylostella). In total, 16 different primer combinations were used, generating cDNA fragments ranging from 50 bp to 500 bp in size. This technique generated an average of 60 amplification products per reaction and therefore a total number of 5,600 fragments per treatment. Out of 16,800 bands, 124 showed qualitative differences among the treated and their respective control samples, including 95 up-regulated and 29 down-regulated bands. Expression of a selected subset of differentially expressed genes was confirmed by Northern blot analysis. Sequencing of fragments grouped many of the expressed genes in the categories of signaling and wound or pathogen response with examples like Jacalin, Strictosidine synthase and MD-2-LPS homologs. Genes with altered expression in distal tissue included those involved in cellular housekeeping functions, suggesting modified resource allocation needed to respond to different stress conditions. Differences in local and systemic response as well as among the three different challenges were observed. Several new transcripts were identified that may play a role in insect attack and other signal transduction pathways.