A Simple and Rapid Oxidative Stress Screening Method of Small Molecules for Functional Studies of Transcription Factor

摘要

Structure-assisted reverse chemical discovery approaches are becoming attractive due to target based screening.However,Small molecules(SMs)identified by reverse chemical genetic approach require robust phenotyping assays to study the plant responses.We optimized a screening method in rice seedlings using methyl viologen(MV)induced oxidative stress under high light conditions.This method was suitable for studying the efficacy of SM targeting oxidative stress related genes.Structure-assisted drug designing approach was used to identify SM targeting DREB2A transcription factor(TF).Rice seedlings treated with pronetalol showed susceptible phenotype when exposed to oxidative stress.Pronetalol inhibited the DREB2A activity and suppressed the expression of its target LEA7 and HSFA3 genes under oxidative stress.The assay is quite simple and robust and can be expanded to high throughput screening method for functional validation of TF.