Project description:Here we use bisulfite conversion of rRNA depleted RNA combined with high-throughput Illumina sequencing (RBS-seq) to identify single-nucleotide resolution of m5C sites transcriptome-wide in Arabidopsis thaliana roots. m5C sites were analyzed in wild type (WT) and an Arabidopsis T-DNA KO mutant for the RNA methyltransferase TRM4B.
Project description:The goal of the experiments is to identify differential genes between the roots of Arabidopsis thaliana and roots treated for 3 h with 1-aminocyclopropane-1-carboxylic acid (ACC). ACC blocks the maximal cell elongation of the root cells. - Arabidopsis plants are grown for 5 days on MS medium and then we transfer them to a medium supplemented with ACC (5uM) or without ACC. After 3 h of growth, we isolate the RNA of the roots. Keywords: treated vs untreated comparison
Project description:The scarcity of accessible sites that are dynamic or cell type-specific in plants may be due in part to tissue heterogeneity in bulk studies. To assess the effects of tissue heterogeneity, we apply single-cell ATAC-seq to Arabidopsis thaliana roots and identify thousands of differentially accessible sites, sufficient to resolve all major cell types of the root. We find that the entirety of a cell's regulatory landscape and its transcriptome independently capture cell type identity. We leverage this shared information on cell identity to integrate accessibility and transcriptome data to characterize developmental progression, endoreduplication and cell division. We further use the combined data to characterize cell type-specific motif enrichments of transcription factor families and link the expression of family members to changing accessibility at specific loci, resolving direct and indirect effects that shape expression. Our approach provides an analytical framework to infer the gene regulatory networks that execute plant development.
Project description:This study evaluates the transcriptome of Arabidopsis thaliana roots exposed to the MAMP flg22 in the presence of a 35-member bacterial SynCom
Project description:Increasing soil salinization has led to severe losses of plant yield and quality. Researching the formation of plant salt tolerance and the molecular mechanism of the salt stress response is therefore urgent. We here take advantage of recent progress in single-cell transcriptomics technology to systematically analyze plant roots response to salt stress, and 57,185 high-quality cells were totally obtained from 5-day-old lateral root tips of Gossypium arboreum under natural growth and different salt-treatment conditions. Eleven cell types with an array of novel marker genes were identified and confirmed with RNA in situ hybridization. Pseudotime analysis on epidermal and root hair cells indicated the differentiation trajectory, and differential root-type cells responding to salt stress resulted in more epidermal and less endodermal cells. Abundant differentially expressed genes (DEGs) were found to be engaged in glycometabolism, reactive oxygen species hemeostasis, and phosphatidylinositol and MAKP signal pathways through functional enrichment analyses. Some candidate DEGs related with transcription factors and plant hormones responding to salt stress were also screened, which requires further functional verification to reveal the regulatory model of the plant roots response to salt stress. Combined with bulk RNA-seq data, a common DEG (Ga08G2497) annotated as Aldo-keto reductase-1 (AKR1) was selected to perform virus-induced gene silencing verification, and the silencing of GaAKR1 gene in G. arboreum resulted in severe stress-susceptibility phenotype, shorter root length, and less number of lateral roots. Physiological and biochemical detection also demonstrated that GaAKR1-silenced plants suffered more serious oxidative damages and, indicating that GaAKR1 might participates in salt tolerance of cotton by oxidation-reduction process. For the first time, we characterized a transcriptional atlas of plant roots under salt stress at a single-cell resolution, which explored the cellular heterogeneity, differential root-type cells, and differentiation trajectory, while providing valuable insights into the molecular mechanism underlying stress tolerance in plants.