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HalgrenAnneSupplemental3.xls

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  • Ethylene regulates multiple developmental processes during a plant life cycle, but the effect of ethylene on the upregulation of senescence-, stress-, and post-harvest-related genes in forage grasses is poorly understood. In this work, we used quantitative PCR to determine whether ethylene application affected the expression of selected cell-wall degradation related genes that are typically upregulated post-harvest. The expression levels of beta-D-glucan exohydrolase isoenzyme, alpha glucosidase, and arabinoxylan arabinofuranohydrolase isoenzyme, all putative cell wall degrading enzymes, were quantified at six points in the life cycle of the model grass species Darnel ryegrass (Lolium temulentum L.). We also quantified the expression of ACC oxidase and ACC synthase in response to ethylene application to determine if endogenous upregulation of ethylene biosynthesis occurred. Grass developmental stage had a significant impact on gene expression response to ethylene-treatment, indicating that discrete life cycle stages present different ethylene-responsive windows for treatment. Under our experimental conditions, ACC oxidase and ACC synthase expression were downregulated in response to ethylene-treatment, suggesting that exogenous ethylene served an auto-inhibitory role. Transcripts corresponding to the three cell wall degradation related genes increased significantly in response to ethylene treatment, suggesting that ethylene may have future utility in the pretreatment of lignocellulosic biomass. To our knowledge, this is the first report of a life cycle analysis of ethylene-induced genes in forage grasses.
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