Project description:Desiccation tolerance (DT) allowed seed plants to conquer ecosystems with long periods of limited water availability. This adaptive features allows seeds to remain dried for very long times without losing their ability to germinate. There is little information about all the signaling components required to achieve DT and on how transcription factors (TFs) modulate global DT processes. We performed RNA-seq experiment and carbohydrates profiles of lec1, lec2, fus3 and abi3, as well as their corresponding wild types, at three stages of seed development 15, 17 and 21 DAF (day after open flower) belonging to the seed desiccation period. A complex experimental design approach and regulatory networks prediction were used to identify differentially expressed genes specifically involved in DT process. In order to identify mechanisms involved in the acquisition of DT during seed development, we designed a comparative transcriptomic analysis between the seed desiccation intolerant (DI) mutants lec1-1, abi3-5 and fus3-3, the desiccation tolerant mutant lec2-1 and the desiccation tolerant weak allele of abi3 (abi3-1) with their respective wild type controls. This analysis should allow to identify genes that are differentially expressed in the desiccation intolerant mutants respect to tolerant mutants and WT controls.
Project description:LEAFY COTYLEDON1 (LEC1), a subunit of the NF-Y CCAAT binding transcription factor, is a key regulator that controls many aspects of seed development including embryo growth and seed storage accumulation. To identified genomic region bound by LEC1 and target genes of LEC1, we expressed a FLAG-tagged form of LEC1 under the control of its native promoter and terminator in lec1-4 mutant seeds. Using an anti-FLAG antibody (Sigma), we performed chromatin immunoprecipitation followed by sequencing (ChIP-Seq) using chromatin isolated from mature stage seeds to identify the genes bound by LEC1. These data reveal that the transcription regulation network of LEC1.