Gene Expression Nebulas
A data portal of transcriptomic profiles analyzed by a unified pipeline across multiple species

Gene Expression Nebulas

A data portal of transcriptome profiles across multiple species

PRJNA552367: DNA methylation of a 3' cis-element promotes gene activation

Source: NCBI / GSE133741
Submission Date: Jul 02, 2019
Release Date: Jul 01, 2021
Update Date: Jul 01, 2021

Summary: In our study, we identified a new transgenerational epigenetic mutant in rice, Epi-LF (Epi-LongFeng). The metastable phenotype of Epi-lf resembles that of rice Epi-d1, and the inheritance is largely similar to that of the agoutiAy mouse mutant. We found that in Epi-lf,mCHH methylation was present in a 155-bp sequence of DNA in close proximity to the 3' end of the LF gene, but the abundance of the siRNAs corresponding to this region was not significantly different from that of the wild type (WT), suggesting that this is a mCHH island like modification. In addition, ectopic mCHH modifications accumulated in the sporophytic tissues and embryos of the Epi-lf mutants. The majority of these ectopic CHH regions did not fit with the OsDRM2 (Domain Rearranged Methylase2)-mediated CHH methylation of genomic regions, suggesting that unknown genetic factors may co-ordinate DNA demethylation and methylation, especially for non-CG methylation.

Overall Design: We sequenced transcriptome, methylome and small RNA in different tissues of Eip-lf mutants and wild-type rice

GEN Datasets:
GEND000524
Strategy:
Species:
Tissue:
Healthy Condition:
Cell Type:
Cell Line:
Development Stage:
Protocol
Growth Protocol: The Epi-lf mutant was identified in a screen of an anther-cultured population of Japonica rice (Oryza sativa ssp. japonica; variety ZhongHua11; ZH11) in 2005. The mutant phenotype of Epi-lf was stably inherited over more than 20 generations. The mutant and WT rice plants were planted in paddy fields under natural conditions or in glasshouses with a day temperature of 30°C and a night temperature of 24°C.
Treatment Protocol: The ambient temperature above 35 ℃ was about 6 h a day and the highest temperature was about 40 ℃ during almost the whole growth period of the natural heat stressed plants, the ambient temperature above 35 ℃ was only 3 days and the mean daytime temperature was about 30 ℃ during the growth period of the normal cultivation condition plants.
Extract Protocol: Total RNA was extracted from different tissues using TRIzol reagent (Thermo Fisher Scientific). The first-strand cDNA was synthesized using 1 μg RNA and ReverTra Ace qPCR RT Master Mix with gDNA Remover (Toyobo), according to the manufacturer’s instructions, using 29 PCR cycles to amplify the LF transcripts and 23 PCR cycles for Actin1. The RT-PCR analysis was repeated three times with similar results. Genomic DNA was isolated from the flag leaves of WT and tillers with different phenotypes of Epi-lf using the CTAB method. Briefly, 1 μg of genomic DNA was treated with sodium bisulphite using the Qiagen EpiTect Bisulfite kit, according to the manufacturer’s instructions. The treated DNA was dissolved in 15 mL distilled water, and 3 μL of this solution was used as the template in a 50-μL PCR (94°C for 4 min; followed by 40 cycles of 94°C for 30 s , 55°C for 30 s and 60°C for 30 s; with a final 60°C for 10 min).The PCR products were purified and cloned into pEASY-T5-Blunt Zero Cloning Kit vectors.
Library Construction Protocol: RNA libraries were prepared for sequencing using standard Illumina protocols.
Sequencing
Molecule Type: rRNA- RNA
Library Source:
Library Layout: SINGLE
Library Strand: Forward; -
Platform: ILLUMINA
Instrument Model: Illumina HiSeq 4000; Illumina HiSeq X Ten
Strand-Specific: Specific; Unspecific
Samples
Basic Information:
Sample Characteristic:
Biological Condition:
Experimental Variables:
Protocol:
Sequencing:
Assessing Quality:
Data Resource GEN Sample ID GEN Dataset ID Project ID BioProject ID Sample ID Sample Name BioSample ID Sample Accession Experiment Accession Release Date Submission Date Update Date Species Race Ethnicity Age Age Unit Gender Source Name Tissue Cell Type Cell Subtype Cell Line Disease Disease State Development Stage Mutation Phenotype Condition Detail Growth Protocol Treatment Protocol Extract Protocol Library Construction Protocol Molecule Type Library Layout Strand-Specific Library Strand Spike-In Strategy Platform Instrument Model Cell Number Reads Number Gbases AvgSpotLen1 AvgSpotLen2 Uniq Mapping Rate Multiple Mapping Rate Coverage Rate