Microarray Identifies Transcription Factors Potentially Involved in Gravitropic Signal Transduction in Arabidopsis

Abstract

ABSTRACT Gravity is a fundamental stimulus that affects plant growth and development. The gravity persistent signal (GPS) treatment uses a cold treatment to isolate the events of signal transduction. Plants are reoriented horizontally in the dark at 4°C for 1 hour and then returned to vertical at room temperature. A gene expression microarray was designed to identify genes that are regulated during the GPS treatment. Arabidopsis thaliana var. Columbia was grown to maturity with inflorescence stems of 8-10 cm. Total mRNA was collected from inflorescence stems at 2, 4, 10, and 30 min after reorientation in the cold. cDNA was synthesized from the mRNA and then probed against an Arabidopsis gene expression array with 4 replicates per time point. Analyses presented here focus on transcription factors because of their regulatory functions in response pathways. Five transcription factors (AtAIB, WRKY18, WRKY26, WRKY33, and BT2) were selected for further study based on their expression at 4 min. Quantitative real-time polymerase chain reaction ((PCR) RT-qPCR) was performed to confirm expression seen in the microarray data. Seeds of Arabidopsis lines containing T-DNA insertions in the genes were obtained, plants bred to homozygosity, and the mutants analyzed for GPS phenotype. Mutant analysis shows significant differences in curvature of inflorescence stems between mutants and wild type.