Differential expression of microRNAs in wildtype versus DCL1 mutants in Arabidopsis thaliana
Ontology highlight
ABSTRACT:
ORGANISM(S): Arabidopsis thaliana
PROVIDER: GSE90771 | GEO | 2017/04/17
SECONDARY ACCESSION(S): PRJNA355875
REPOSITORIES: GEO
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Project description:DICER-LIKE1 (DCL1) is required for miRNA biogenesis and embryonic pattern formation. Presumably the patterning defects observed in dcl1-5 null embryos are due to the loss of miRNAs and the consequent up-regulation of their respective targets. To test which miRNA targets were up-regulated in dcl1 embryos relative to wild-type embryos, we performed genome-wide transcript profiling of wild-type and dcl1-5 early globular embryos using directional mRNA-Seq. For RNA isolation, pools of 30 Col-0 (wild-type) and dcl1-5 early globular embryos were hand-dissected in water, immediately transferred to 30 ul of RNAlater (Ambion), incubated at 60M-BM-0 C with 500 ul TRIzol Reagent (Invitrogen) for 30 minutes, and then purified according to the TRIzol Reagent protocol for RNA isolation from small quantities of tissue (Invitrogen). Two rounds of linear amplification of poly(A) RNA were performed using the Arcturus RiboAmp HS Plus kit (Molecular Devices). Strand-specific mRNA-Seq libraries were generated as previously described (Guo et al., Nature, 2010). After removing adaptor sequences, sequences were mapped to the Arabidopsis thaliana genome (TAIR9 assembly) using the Bowtie short read aligner allowing up to two mismatches within a 25 nt M-bM-^@M-^XseedM-bM-^@M-^Y sequence and retaining reads mapping uniquely to the genome. Since the Arcuturus RiboAmp HS Plus kit generates amplified RNAs in the antisense orientation, reads were counted as matching TAIR9 annotated genes if they overlapped the antisense strand.
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Project description:DICER-LIKE1 (DCL1) is required for miRNA biogenesis and embryonic pattern formation. Presumably the patterning defects observed in dcl1-5 null embryos are due to the loss of miRNAs and the consequent up-regulation of their respective targets. To test which miRNA targets were up-regulated in dcl1 embryos relative to wild-type embryos, we performed genome-wide transcript profiling of wild-type and dcl1-5 early globular embryos using directional mRNA-Seq.
2010-12-01 | GSE25404 | GEO
Project description:Belonging to the Carmovirus family, Turnip crinkle virus (TCV) is a positive-strand RNA virus that can infect Arabidopsis. Most Arabidopsis ecotypes are highly susceptible to TCV, except for the TCV resistant line Di-17 derived from ecotype Dijon. Previous studies showed that many of the stress related genes have changed significantly after TCV infection. Besides the virus-triggered genes, small RNAs also play critical roles in plant defense by triggering either transcriptional and/or post-transcriptional gene silencing. In this study, TCV-infected wildtype Arabidopsis thaliana and dcl1-9 mutant plants were subjected to transcriptome and small RNA analysis to investigate the role of DCL1 in virus defense network.
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Project description:Belonging to the Carmovirus family, Turnip crinkle virus (TCV) is a positive-strand RNA virus that can infect Arabidopsis. Most Arabidopsis ecotypes are highly susceptible to TCV, except for the TCV resistant line Di-17 derived from ecotype Dijon. Previous studies showed that many of the stress related genes have changed significantly after TCV infection. Besides the virus-triggered genes, small RNAs also play critical roles in plant defense by triggering either transcriptional and/or post-transcriptional gene silencing. In this study, TCV-infected wildtype Arabidopsis thaliana and dcl1-9 mutant plants were subjected to transcriptome and small RNA analysis to investigate the role of DCL1 in virus defense network.
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Project description:The SnRK1 protein kinase, the plant ortholog of mammalian AMPK and yeast Snf1, is activated by the energy depletion caused by adverse environmental conditions. Upon activation, SnRK1 triggers extensive transcriptional changes to restore homeostasis and promote stress tolerance and survival partly through the inhibition of anabolism and the activation of catabolism. Despite the identification of a few bZIP transcription factors as downstream effectors, the mechanisms underlying gene regulation, and in particular gene repression by SnRK1, remain mostly unknown. microRNAs (miRNAs) are 20-24nt RNAs that regulate gene expression post-transcriptionally by driving the cleavage and/or translation attenuation of complementary mRNA targets. In addition to the well-established role of miRNAs as regulators of plant development, mounting evidence implicates miRNAs in the response to environmental stress. Given the involvement of miRNAs in stress responses and the fact that some of the SnRK1-regulated genes are miRNA targets, we postulated that miRNAs drive part of the transcriptional reprogramming triggered by SnRK1 activation. To test this we have performed comparative analyses of the transcriptional response to energy deprivation between WT and dcl1-9, a mutant deficient in miRNA biogenesis. To assess the impact of miRNA deficiency on the starvation response we performed transcriptomics analyses of WT and dcl1-9 plants by subjecting leaves to 6h of light (control) or darkness (starvation)
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Project description:m6A-seq and RNA-seq in Arabidopsis dcl1 mutant
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