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2006 Awardee

Genetic Dissection of the LignocellulosicPathway of Wheat to Improve Biomass Quality of Grasses as aFeedstock for Biofuels

INVESTIGATOR(S): Gill, B.; Li, W.

INSTITUTION: Kansas State University

NON-TECHNICAL SUMMARY: As fossil fuelreserves dwindle, we are about to transition fromnonrenewable energy to renewable bioenergy. Demand forethanol is increasing steadily as an alternative fuel aswell as an octane-boosting and pollution-reducing additiveto gasoline. Keeping in step with the demand will requirehigher quantity and quality of biomass. In the GreatPlains, cereal crops (wheat, corn and sorghum), as well asnative grasses (switchgrass, bluestem) predominate and areimportant but as yet relatively untapped resources forbioenergy.

OBJECTIVES: 1) Investigate the expressionof ~80 candidate genes for lignin biosynthesis, theirenzymatic activities, and lignin content and composition indifferent organs at different stages of diploid wheatplant; 2) Silence these 80 genes individually by VIGS; 3)Identify knockout mutants of these genes using TILLING and4) Characterize the silenced tissues and knockout mutantsby metabolite profiling.

APPROACH: obj. 1. Plants will be grown atseedling stage (Feekes stages F1-2), leaf sheath/stemelongation stages F4-5 and heading stages F8-9. The leaf,sheath, stem and spike tissues will be assayed for theexpression of the 80 genes by QRT-PCR to decide thedevelopmental phase and tissue on which all experimentswill be done. Obj. 2. Conserved sequences will be used tosilence all 80 genes individually by VIGS and the silencedtissues will be verified by QRT-PCR and positively silencedsamples will be subjected to metabolite profiling,enzymatic assays and histochemical staining to determinethe consequences of the genetic block in thelignocellulosic pathway. Obj. 3. We will do TILLING toscreen for mutations for genes with major effect on thelignin content based on VIGS results. The homozygous plantscontaining genetic lesions for lignocellulosic pathwaygenes will be phenotyped with regard to growth anddevelopment, lignin content and composition, andlignocellulose degradability. Obj. 4. We will determine thelignin content of wheat silenced tissues, knockout mutantsand controls by the acetyl bromide method, lignincomposition by thioacidolysis methods, soluble andwall-bound phenolic compounds by HPLC, and polysaccharidecontent and composition by the phenol-sulfuric acid method.We will also histochemically characterize these plantmaterials by Mauls and Wiesner staining.

PROJECT CONTACT:

Name: Gill, B.
Phone: 785-532-2364
Fax: 785-532-5692
Email: bsgill@ksu.edu

 

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