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Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Cell type specific gene expression map of Arabidopsis thaliana SAM.

ABSTRACT: Shoot apical meristem (SAM) of higher plant composed of a few distinct cell types. All the cells in a mature plant’s SAM derived from 30~35 stem cells reservoir which are located at the tip of the apex. Plants ability to give rise diverse cell types from a pool of pluripotent stem cells requires orchestrated gene network that controls the cell fate commitment during the meristem development. To understand, how gene regulatory networks control cell identities switches during cell differentiation requires resolution in recording their gene expression pattern at single cell resolution. An earlier expression map involving three-cell population of stem cell niche revealed complex expression pattern among the cell types1. We developed this approach further and report here a gene expression map using cell-sorting methods for fluorescent protein marked cells in Arabidopsis shoot. The map covered 10 cell populations. This gene expression map represents data from 10 different cell types from Arabidopsis SAM. It will be first step in defining the function of many unknown genes in model plant Arabidopsis. Based on the in situ hybridization we identified 7 new cell types specific gene expression patterns. The promoters of these genes were used to generate fluorescent reporters. After treating the SAM with protoplasting cocktail, we sorted the fluorescent protein tagged cells using fluorescent activated cell sorter (FACS). The purified cell population was used to isolate RNA. Two round of RNA amplification was performed before microarray hybridization.

ORGANISM(S): Arabidopsis thaliana

SUBMITTER: Ram Yadav

PROVIDER: E-GEOD-28109 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Cell type specific gene expression map of Arabidopsis thaliana SAM.

Project description:Shoot apical meristem (SAM) of higher plant composed of a few distinct cell types. All the cells in a mature plant’s SAM derived from 30~35 stem cells reservoir which are located at the tip of the apex. Plants ability to give rise diverse cell types from a pool of pluripotent stem cells requires orchestrated gene network that controls the cell fate commitment during the meristem development. To understand, how gene regulatory networks control cell identities switches during cell differentiation requires resolution in recording their gene expression pattern at single cell resolution. An earlier expression map involving three-cell population of stem cell niche revealed complex expression pattern among the cell types1. We developed this approach further and report here a gene expression map using cell-sorting methods for fluorescent protein marked cells in Arabidopsis shoot. The map covered 10 cell populations. This gene expression map represents data from 10 different cell types from Arabidopsis SAM. It will be first step in defining the function of many unknown genes in model plant Arabidopsis.

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DEAD-box RNA helicase 27-Mediated MicroRNA Biogenesis Is Essential for Zygote Division and Stem Cell Homeostasis in Arabidopsis

Project description:Zygote division and stem cell homeostasis are crucial events for plant development. Here, we found that mutations in DEAD-box RNA helicase 27 (RH27) affected zygote division and stem cell homeostasis in Arabidopsis (Arabidopsis thaliana). The strong mutant allele rh27-1 showed a zygote-lethal phenotype, while the weak mutant allele rh27-2 showed defective embryogenesis and compromised stem cell homeostasis in both the shoot apical meristem (SAM) and root apical meristem (RAM). RH27 was expressed in embryos from the zygote stage, and in the SAM and RAM. Cytological analyses revealed that stem cell homeostasis in the SAM and RAM was disrupted in rh27-2. Then, the expression levels of genes related to stem cell homeostasis were elevated in rh27-2, which coincides with down-regulations of their corresponding negative regulatory microRNAs. Furthermore, small RNA-sequencing indicated that the reduction of miRNAs was a global effect in both shoot apices and root tips of rh27-2. Additionally, the accumulation of pri-miRNAs was detected to be decreased. Biochemical studies showed that the nucleus-localized RH27 interacted directly with SERRATE, a component of the microprocessor complex, and DAWDLE, which facilitates the recruitment of substrates to the complex, suggesting that RH27 is a subunit of the microprocessor complex. Overall, this work provides evidences that RH27 regulates zygote division and stem cell homeostasis in Arabidopsis through miRNA biogenesis.

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