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

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Abstract

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.

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Abbreviations

AFLP:

Amplified fragment length polymorphism

DBM:

Diamond back moth

JA:

Jasmonic acid

LPS:

Lipopolysaccharide

MBP:

Myrosinase binding protein

MeJa:

Methyl jasmonate

SA:

Salicylic acid

TDF:

Transcript derived fragment

TLR:

Toll-like receptor

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Acknowledgements

We thank Barbara Ekbom and Carol Högfeldt (Dept. of Entomology, Swedish University of Agricultural Sciences) for rearing of the insects, Mattias Persson for help with electrophoresis setup, and Jens Staal and Jens Sundström for comments on the manuscript. This work was supported by grants from Carl Tryggers Stiftelse för Vetenskaplig Forskning, the Swedish Research Council for Environment, Agricultural Sciences and Spatial Planning, the Helge Ax:son Johnsons Stiftelse, the Persons Fund and the Nilsson-Ehle Fund.

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  1. Department of Plant Biology and Forest Genetics, Uppsala BioCenter, Swedish University of Agricultural Sciences, P.O. Box 7080, Uppsala, 750 07, Sweden

    Bejai R. Sarosh & Johan Meijer

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  1. Bejai R. Sarosh
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  2. Johan Meijer
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Correspondence to Bejai R. Sarosh.

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Sarosh, B.R., Meijer, J. Transcriptional profiling by cDNA-AFLP reveals novel insights during methyl jasmonate, wounding and insect attack in Brassica napus . Plant Mol Biol 64, 425–438 (2007). https://doi.org/10.1007/s11103-007-9164-9

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