AUTHOR=Zhu Ling , Xin Ruijiao , Huq Enamul 

TITLE=A Protein-Based Genetic Screening Uncovers Mutants Involved in Phytochrome Signaling in Arabidopsis

JOURNAL=Frontiers in Plant Science

VOLUME=7

YEAR=2016

URL=https://www.frontiersin.org/journals/plant-science/articles/10.3389/fpls.2016.01086

DOI=10.3389/fpls.2016.01086

ISSN=1664-462X

ABSTRACT=<p>Plants perceive red and far-red region of the light spectrum to regulate photomorphogenesis through a family of photoreceptors called phytochromes. Phytochromes transduce the light signals to trigger a cascade of downstream gene regulation in part via a subfamily of bHLH transcription factors called Phytochrome Interacting Factors (PIFs). As the repressors of light signaling pathways, most PIFs are phosphorylated and degraded through the ubiquitin/26S proteasome pathway in response to light. The mechanisms involved in the phosphorylation and degradation of PIFs have not been fully understood yet. Here we used an EMS mutagenesis and luminescent imaging system to identify mutants defective in the degradation of one of the PIFs, called PIF1. We identified five mutants named <underline>s</underline>table <underline>P</underline>I<underline>F</underline> (<italic>spf</italic>) that showed reduced degradation of PIF1 under light treatment in both luminescent imaging and immunoblot assays. The amounts of PIF1 in <italic>spf3</italic>, <italic>spf4</italic>, and <italic>spf5</italic> were similar to a PIF1 missense mutant (PIF1–3M) that lacks interactions between PIF1 and phyA/phyB under light. The hypocotyl lengths of <italic>spf1</italic> and <italic>spf2</italic> were slightly longer under red light compared to the LUC-PIF1 control, while only <italic>spf1</italic> displayed weak phenotype under far-red light conditions. Interestingly, the <italic>spf3</italic>, <italic>spf4</italic>, and <italic>spf5</italic> displayed high abundance of PIF1, yet the hypocotyl lengths were similar to the wild type under these conditions. Cloning and characterization of these mutants will help identify key players in the light signaling pathways including, the light-regulated kinase(s) and the E3 ligase(s) necessary for the light-induced degradation of PIFs.</p>