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Time-lapse RNA-seq analysis to study the gene networks in cell fate transition in de novo root regeneration from detached Arabidospis leaves

ABSTRACT: Detached Arabidopsis leaves can regenerate adventitious roots, providing a platform to study de novo root regeneration (DNRR). We performed time-lapse RNA-seq within 5 d revealed activation of gene networks in cell fate transition.

ORGANISM(S): Arabidopsis thaliana

PROVIDER: GSE147296 | GEO | 2022/02/10

REPOSITORIES: GEO

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Time-lapse RNA-seq analysis to study transcriptional regulation by early signals in De Novo Root Regeneration in Arabidopsis

Project description:Detached Arabidopsis leaves can regenerate adventitious roots, providing a platform to study de novo root regeneration (DNRR). We performed time-lapse RNA-seq within 12 h to reveal transcriptional changes in response to early signals in DNRR.

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Single-Cell RNA-seq analysis of cell lineages in De Novo Root Regeneration

Project description:Detached Arabidopsis leaves can regenerate adventitious roots, providing a platform to study de novo root regeneration (DNRR). We performed single-cell RNA-seq analysis and revealed that regeneration primarily originated from vascular stem-cell organizer within procambium, followed by step-by-step changes of transcriptome in cell fate transition, including gradually erasing the vascular stem-cell organizer identity and recruiting the root development program.

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Transcriptional landscapes of de novo root regeneration from detached Arabidopsis leaves revealed by time-lapse and single-cell RNA sequencing analyses.

Project description:Detached Arabidopsis thaliana leaves can regenerate adventitious roots, providing a platform for studying de novo root regeneration (DNRR). However, the comprehensive transcriptional framework of DNRR remains elusive. Here, we provide a high-resolution landscape of transcriptome reprogramming from wound response to root organogenesis in DNRR and show key factors involved in DNRR. Time-lapse RNA sequencing (RNA-seq) of the entire leaf within 12 h of leaf detachment revealed rapid activation of jasmonate, ethylene, and reactive oxygen species (ROS) pathways in response to wounding. Genetic analyses confirmed that ethylene and ROS may serve as wound signals to promote DNRR. Next, time-lapse RNA-seq within 5 d of leaf detachment revealed the activation of genes involved in organogenesis, wound-induced regeneration, and resource allocation in the wounded region of detached leaves during adventitious rooting. Genetic studies showed that BLADE-ON-PETIOLE1/2, which control aboveground organs, PLETHORA3/5/7, which control root organogenesis, and ETHYLENE RESPONSE FACTOR115, which controls wound-induced regeneration, are involved in DNRR. Furthermore, single-cell RNA-seq data revealed gene expression patterns in the wounded region of detached leaves during adventitious rooting. Overall, our study not only provides transcriptome tools but also reveals key factors involved in DNRR from detached Arabidopsis leaves.

| S-EPMC9284295 | biostudies-literature

Time-lapse RNA-seq analysis to study the gene networks in cell fate transition in de novo root regeneration from detached Arabidospis leaves

Project description:Time-lapse RNA-seq analysis to study the gene networks in cell fate transition in de novo root regeneration from detached Arabidospis leaves

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effect of sucrose feeding to detached leaves

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Methanol is an endogenous elicitor molecule upon senescence of detached rice leaves

Project description:During senescence of detached rice leaves, tryptophan (Trp) and Trp-derived secondary metabolites such as serotonin and 4-coumaroylserotonin accumulated in concert with methanol (MeOH) production. This senescence-induced MeOH induction was closely associated with levels of pectin methylesterase (PME)1 mRNA and PME enzyme activity. Exogenous challenge of detached rice leaves with 1% MeOH accelerated Trp and serotonin biosynthesis with induction of the corresponding genes. No other solvents including ethanol resulted in a Trp-inducing effect. This MeOH-induced Trp synthesis was positively regulated by abscisic acid but negatively regulated by cytokinin, suggesting hormonal involvement on the action of MeOH. Endogenous overproduction or suppression of MeOH either by PME1 overexpression or RNAi gene silencing revealed that PME1 overexpressing lines produced twofold higher Trp levels with elevated Trp biosynthetic gene expression, whereas RNAi lines showed twofold reduction in Trp level in healthy control rice leaves, suggesting that MeOH acts as an endogenous elicitor to enhance Trp biosynthesis. Among many transcription factors induced following MeOH treatment, the WRKY family showed significant induction patterns of which WRKY14 appeared to play a key regulatory role in MeOH-induced Trp and Trp-derived secondary metabolite biosynthesis. Total RNAs were extracted from the detached rice leaves with 1% MeOH or distilled water for 1 d, and gene expression was compared between the two groups with two replicates. DW, detached leaves in distilled water for 1 day; MeOH (2-replications), methanol treated detached leaves at the same time point as control. 2 sets of separately normalized data; DW-MeOH(1) and MeOH(2).

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Transcription profiling of Arabidopsis response to sucrose feeding to detached leaves

Project description:The effect of sucrose feeding on gene expression in Arabidopsis thaliana leaves was investigated using affymetrix ATH1 microarrays. For this, petioles of detached leaves were put in a solution containing either sucrose or sorbitol (control). Sugars were taken up into the leaf via the respiration stream for 13 hours. After that, leaves were frozen in liquid nitrogen and RNA was extracted for analysis. Experiment Overall Design: The effect of sucrose feeding on gene expression was investigated using affymetrix ATH1 microarrays. For this, petioles of detached leaves were put in a solution containing either sucrose or sorbitol (control). Sugars were taken up into the leaf via the respiration stream for 13 hours. After that, leaves were frozen in liquid nitrogen and RNA was extracted for analysis.

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Methanol is an endogenous elicitor molecule upon senescence of detached rice leaves

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Time-lapse RNA-seq analysis to study transcriptional regulation by early signals in De Novo Root Regeneration in Arabidopsis

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Histone H3K27me3 demethylase act as a regulator of stemness in hypoxic cancer cells that are detached from extracellular matrix

Project description:Metastasis of cancer cells requires detachment from Extra Cellular Matrix (ECM) to seed cancer cells in a distant organ. Hypoxia is prevalent in this matrix detached cancer cells. Studies have established hypoxia as a chromatin modifier, as it transcriptionally controls expression of various histone demethylases (KDMs); therefore, we hypothesized that the presence of hypoxia could modulate the expression of KDMs in matrix detached cancer cells. Our study showed that in matrix detached cancer cells, both hypoxia and one of the hypoxia regulated H3K27me3 histone demethylase, namely KDM6B, was increased. Simultaneously, we found increased expression of stemness-associated genes, namely SOX-2, SOX-9, and CD44, in hypoxic matrix detached cancer cells. We discovered that KDM6B occupies the promoter region of both SOX-2 and CD44 to regulate their expression epigenetically. Targeting KDM6B reduces its occupancy, thereby expressing the stemness-related genes in matrix detached cancer cells. Further, we noticed increased occupancy of HIF1α promoter by KDM6B, suggesting its regulatory role in maintaining hypoxia in matrix detached cancer cells. This observation was further strengthened as we found a significant positive association between KDM6B and HIF1α in various cancer types. Overall, we found that matrix detachment modulates epigenome by inducing KDM6B activity to regulate the expression of stemness-related genes and hypoxia primarily through HIF1α in matrix detached conditions. KDM6B can be developed as a therapeutic target to eliminate matrix detached cancer cells bound for metastatic events

2021-06-11 | GSE176558 | GEO

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