Project description:Several pathways conferring environmental flowering responses in Arabidopsis converge on developmental processes that mediate floral transition in the shoot apical meristem. Many characterized mutations disrupt these environmental responses, but downstream developmental processes have been more refractory to mutagenesis. We constructed a quintuple mutant impaired in several environmental pathways and showed that it possesses severely reduced flowering responses to changes in photoperiod and ambient temperature. RNA-seq analysis of the quintuple mutant showed that the expression of genes encoding gibberellin biosynthesis enzymes and transcription factors involved in the age pathway correlates with flowering. Mutagenesis of the quintuple mutant generated two late-flowering mutants, quintuple ems 1 (qem1) and qem2. The mutated genes were identified by isogenic mapping and transgenic complementation. The qem1 mutant was an allele of ga20ox2, confirming the importance of gibberellin for flowering in the absence of environmental responses. By contrast, the qem2 mutation is in CHROMATIN REMODELING 4 (CHR4), which has not been genetically implicated in floral induction. Using co-immunoprecipitation, RNA-seq and ChIP-seq, we show that CHR4 interacts with transcription factors involved in floral meristem identity and affects expression of key floral regulators. We conclude that CHR4 mediates the response to endogenous flowering pathways in the inflorescence meristem to promote floral identity.
Project description:How plants control the transition to flowering in response to ambient temperature is only beginning to be understood. In Arabidopsis thaliana, the MADS-box transcription factor genes FLOWERING LOCUS M (FLM) and SHORT VEGETATIVE PHASE (SVP) have key roles in this process. FLM is subject to temperature-dependent alternative splicing, producing two splice variants, FLM-M-NM-2 and FLM-M-NM-4, which compete for interaction with the floral repressor SVP. The SVP/FLM-M-NM-2 complex is predominately formed at low temperatures and prevents precocious flowering. In contrast, the competing SVP FLM-M-NM-4 complex is impaired in DNA binding and acts as a dominant negative activator of flowering at higher temperatures. Our results demonstrate the importance of temperature-dependent alternative splicing in modulating the timing of the floral transition in response to environmental change. ChIP-seq A. thaliana FLM (3 replicates for gFLM and 2 replicates for FLM splice variants)
Project description:Flower development is a dynamics process in which floral organs are produced from pools of stem cells residing in meristems (Smyth et al., 1990). In order to obtain a high resolution map of the changes of miRNA gene expression during this process thus to provide insights into specific expression patterns and their underlying gene regulatory networks, an inducible system which allows us to obtain synchronized flowers (Wellmer et al., 2006) was used to collect seedlings as well as stage-specific floral tissues at four stages (stages 0, 2, 4 and 8) for transcriptome profiling by miRNA-seq .
Project description:Flower development is a dynamics process in which floral organs are produced from pools of stem cells residing in meristems (Smyth et al., 1990). In order to obtain a high resolution map of the changes of gene expression during this process thus to provide insights into specific expression patterns and their underlying gene regulatory networks, an inducible system which allows us to obtain synchronized flowers (Wellmer et al., 2006) was used to collect stage-specific floral tissues at four stages (stages 0, 2, 4 and 8) for transcriptome profiling by RNA-seq . These stages represent the status of inflorescence meristem, floral meristem specification, floral organ specification and floral organ differentiation, respectively during Arabidopsis flower development.
Project description:ChIP-seq of ASY1 was carried out on meiotic-stage floral buds of Arabidopsis using an a-ASY1 antibody. The experiment aims to determine the genome-wide profile of ASY1. ASY1 is a component of the chromosome axis and is expressed exclusively during meiosis. Two negative controls were used to test the specificity of the ChIP experiment. First, ChIP-seq using the pre-immune on floral buds was carried out. Second, ChIP-seq using an a-ASY1 antibody was performed on leaf tissue where ASY1 is not expressed.
Project description:In a recent study, we showed that a T-DNA insertional mutation in a mitochondrial PPR protein, POCO1, led to the earlier floral transition (Emami and Kempken 2019). We used RNA-seq analysis to provide an overview of the global transcriptome changes in poco1 mutant during different developmental stages.