Designing a Molecular Switch to Optimize Phenylpropanoid Neutraceuticals in Vegetables
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Phenylpropanoids and many small phenolic molecules such as flavonoids, lignins, coumarins have various functions in structural support, pigmentation, defense mechanism and signaling. Antioxidant properties of dietary phenylpropanoids provide protection against cancer, aging, injury, inflammation and other chronic diseases. Arabidopsis PAP1 is a regulator of the multiple enzymes in the phenylpropanoid biosynthesis pathway. Previously, overproduction of phenylpropanoids in plants resulted in retardation of vegetative growth. To optimize the level of phenylpropanoid accumulation in vegetables without compromising their growth, we designed a molecular switch that activates PAP1 via osmotic stress, thus the production of phytochemicals can be induced as pre-and/or post-harvest processes. Arabidopsis cDNA encoding PAP1 and CBF3 were placed downstream of low temperature and osmotic stress-inducible RD29a promoter. Application of osmotic stress will induce expression of RD29a-CBF3 and RD29a-PAP1. Since CBF3 is an activator of RD29a promoter, induced CBF3 further amplifies the transcription level of RD29a-PAP1 transgene. In order to test this system, RD29a-CBF3 and RD29a-PAP1 were co-transformed to Arabidopsis plants (WT and fry2-1 background) through Agrobacterium mediated transformation. Transgenic plants were selected using Hygromycin (on MS media) and phosphinothricine (on soil) resistant genes as genetic markers. Molecular and phytochemical analyses are in progress using T3 double homozygous transgenic lines.
