Project description:To determine the extent to which the major small RNA pathways functions across the Arabidopsis thaliana genome, small RNA populations from several tissues of wild-type (wt) and mutant plants were amplified by RT-PCR and sequenced using high-throughput 454 sequencing technology. Keywords: small RNAs, high-throughput sequencing
Project description:Cell proliferation and division are fundamental processes of plant development and homeostasis. With the feature of high homogeneity, tobacco Bright Yellow-2 (BY-2) cells, a so-called “HeLa” cell line in plants, provide a useful system for study the cell cycle of plants, particularly when in combination with the latest high-throughput single-cell RNA sequencing (scRNA-seq) technology. Here, we generated both scRNA-seq and bulk RNA-seq data from the BY-2 cells and found that the enzymatic hydrolysis during protoplasting was the major factor contributing to cell clustering noise. We introduced the ppScoring concept to filter out confounding cells due to protoplasting and clustered the remaining cells into clusters corresponding to G0-G1, S, and G2-M phases as well as clusters with phase transition arrested or cell cycle exit cells. The clustering results were supported by cell cycle phase specific marker genes and reconstruction of the continuous full cell cycle phases, i.e., from G0-G1 to S to G2-M, based on pseudotime trajectory analysis. The clustering results also identified a set of 896 cell cycle marker genes. This work demonstrated cell cycle-dependent transcriptional heterogeneity of the BY-2 cell population, provided marker genes for study of the cell cycle of plants and new insights into the progression of cell division.