Transcriptome analysis using Arabidopsis leaves after exogenous FAD treatment

Riboflavin (RF) and its cofactors (FMN and FAD) are essential molecules for vital metabolic processes in all organisms. Thus, the flavin metabolism must be tightly regulated in the cells. We previously demonstrated that the intracellular levels of flavins were tightly maintained by the enzymes involved in their synthesis and degradation, inducing a plastidic FAD hydrolase (AtNUDX23), in Arabidopsis. However, information of regulatory factors involved in the flavin homeostasis and transporters in each organelle are mostly limited. Terefore, we tried to identify novel factors involved in the flavin metabolism, such as transcription factors and transporters by a transcriptome analysis after exogenous FAD treatment. Wild-type Arabidopsis plants were grown on soil in a growth chamber kept at 23°C during 16 h of light (100 µmol/m2/s) and 8 h of darkness for 3 weeks. At 4 h after the illumination, detached rosette leaves were floated on water (mock) or 0.1 mM FAD solution for 1h. After 1h, collected leaves were washed and used for microarray analysis. Two biological replicates were analyzed.