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January 19, 2007

Title: Real-time RT-PCR Analyses of Carotenoid Genes in Daucus carota L.

Candie Bautista

Transcriptional and translational regulation of carotenoid genes, which code for the synthesis of biomolecules possessing provitamin and antioxidant properties in Daucus carota L., has become a novel research interest with future health and agricultural implications. Real-time Revererse Transcription PCR (RT-PCR) is a highly innovative and sensitive technique that allows multiple gene analyses by detection of micro-scale target gene transcript levels.  Target genes LCYCB and PSY2 of the carotenoid pathway were selected for transcriptional regulation analyses in the white, yellow and pale orange color varieties and ubiquitin [ubi3] and actin were selected as housekeeping genes. The expression of PSY2 in the white color variety was observed to be 6.5-fold less than the pale orange and yellow color varieties. The expression of LCYCB in the yellow color variety was perhaps significantly less than that of the pale orange color variety, but further data analyses of significant statistical variation using ANOVA is needed.