Virus Induced Gene Silencing for Functional Characterization of Genes Expressing in Various Wheat Tissues

Authors: BENNYPAUL, HARVINDER - WASHINGTON STATE UNIV.; GILL, UPINDER - WASHINGTON STATE UNIV.; Okubara, Patricia; GILL, KULVINDER - WASHINGTON STATE UNIV.

Submitted to: Plant and Animal Genome VX Conference Abstracts
Publication Type: Abstract Only
Publication Acceptance Date: 1/13/2007
Publication Date: 1/13/2007
Citation: Bennypaul, H.S., Gill, U.S., Okubara, P.A., Gill, K.S. 2007. Virus Induced Gene Silencing for Functional Characterization of Genes Expressing in Various Wheat Tissues. Plant and Animal Genome VX Conference Abstracts.

Interpretive Summary:

Technical Abstract: Virus induced gene silencing (VIGS) has a great potential as a functional genomics tool. Methodological simplicity, robustness and speedy results make VIGS an ideal technique for high-throughput functional analysis of genes. In monocots, like wheat and barley, this technique has been shown to work in leaf tissue. Applicability of this technique has not been shown in other important tissues of monocots. We have successfully applied this technique in silencing genes expressing in root, spike, and shoot tissues of wheat. Virus movement in various plant tissues was tested using GFP before targeting the endogenous genes. VIGS constructs were designed for Granule bound starch synthase (GBSSI) and TaCOI1 genes expressing in seeds and roots, respectively. Plants infected at boot stage using hairpin and anti-sense GBSSI-VIGS constructs showed 35 to 50% reduction in amylase content compared to the control. Transmission efficiency of VIGS to the next generation was tested and found to be 8-12% in the progenies. The VIGS approach was found to be effective for gene expression studies in roots by analyzing GFP expression. The experiments to knockdown TaCOI1 expression in wheat roots are underway. The most current data from these experiments will be presented.