ARS | Publication request: TRANSCRIPTIONAL PROFILING OF HESSIAN FLY-RESPONSIVE GENES IN WHEAT

Submitted to: Plant and Animal Genome Conference
Publication Type: Abstract Only
Publication Acceptance Date: November 1, 2004
Publication Date: November 1, 2004
Citation: Sardesai, N., Subramanyam, S., Williams, C.E. 2004. Transcriptional profiling of Hessian fly-responsive genes in wheat. Plant and Animal Genome Conference. http://www.intl-pag.org/13/abstracts/PAG13_P687.html.

Technical Abstract: At the level of plant phenotypes, gene-for-gene interactions between wheat (Triticum aestivum L. em Thell.) and Hessian fly [Mayetiola destructor (Say)] are well-defined. However, at the molecular level, only limited information is available about defense responses in wheat. In order to identify genes that are differentially regulated during incompatible and compatible interactions of the Hessian fly with wheat, we performed suppression subtractive hybridization for the construction of three cDNA libraries. Quantitative real-time PCR was employed to select a time point during the process of infestation of wheat by Hessian fly for expression profiling. We monitored the expression of WCI-2 (lipoxygenase), WCI-5 (PR17-like protein), WCK-1 (MAP kinase) and WIR1 (integral membrane protein) and selected tissue 6 hours after egg hatch, soon after early plant resistance responses were initiated. The forward subtraction used the incompatible interaction as the tester and the compatible interaction as the driver, whereas the reverse subtraction was the opposite. The third library was generated from the subtraction between incompatible interaction and uninfested control plants. From the three libraries approximately 9000 clones were picked and a colony macroarray printed. Differential gene expression is being confirmed through dot blot analyses and positive clones are being confirmed by quantitative real-time PCR. Candidate genes thus identified are to be sequenced and used for the generation of a cDNA macroarray.