Modulation of flavonoid metabolism in Arabidopsis using a phage-derived antibody |
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Authors: | Santos Michael O Crosby William L Winkel Brenda SJ |
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Institution: | (1) Department of Biology and Fralin Biotechnology Center, Virginia Tech, Blacksburg, VA, 24061;(2) Gene Expression Group, Plant Biotechnology Institute, National Research Council of Canada, Saskatoon, Saskatchewan, Canada, S7N 0W9;(3) Present address: Hong Kong University – Pasteur Research Centre, Pokfulam, Hong Kong SAR |
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Abstract: | The utility of recombinant antibodies for immunomodulation of plant metabolism was tested using the Arabidopsis flavonoid biosynthetic pathway as a target. Two genes encoding antibodies against chalcone isomerase (CHI) were isolated from a human synthetic single chain variable fragment (scFv) library and expressed in the cytoplasm of transgenic plants. In one line, low-level expression of the scFv resulted in reduced visible pigmentation in seedlings as well as lower accumulation of phenolics in plants throughout development. Surprisingly, other transgenic lines with much higher expression of the same antibody showed no phenotype. Protein mobility shift assays indicated that the scFv is bound to the target enzyme, but only in the affected plants. This work shows that recombinant scFv antibody technology offers a viable approach to disrupting protein activity in plants, but that further refinement is required before it will be of general utility for metabolic engineering and other antibody-based applications in plants. |
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Keywords: | Chalcone synthase Chalcone isomerase scFv antibody Arabidopsis Flavonoid Transgenic plants |
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