PRJNA253163: Mutation of a major CG methylase in rice causes genome-wide hypomethylation, dysregulated genome expression, and seedling lethality
概要: Cytosine methylation at CG sites (mCG) plays critical roles in development, epigenetic inheritance, and genome stability in mammals and plants. In the dicot model plant Arabidopsis thaliana, methyltransferase 1 (MET1), a principal CG methylase, functions to maintain mCG during DNA replication, with its null mutation resulting in global hypomethylation and pleiotropic developmental defects. Null mutation of a critical CG methylase has not been characterized at a whole-genome level in other higher eukaryotes, leaving the generality of the Arabidopsis findings largely speculative. Rice is a model plant of monocots, to which many of our important crops belong. Here we have characterized a null mutant of OsMet1-2, the major CG methylase in rice. We found that seeds homozygous for OsMet1-2 gene mutation (OsMET1-2-/-), which directly segregated from normal heterozygote plants (OsMET1-2+/-), were seriously maldeveloped, and all germinated seedlings underwent swift necrotic death. Compared with wild type, genome-wide loss of mCG occurred in the mutant methylome, which was accompanied by a plethora of quantitative molecular phenotypes including dysregulated expression of diverse protein-coding genes, activation and repression of transposable elements, and altered small RNA profiles. Our results have revealed conservation but also distinct functional differences in CG methylases between rice and Arabidopsis.
项目整体设计: mRNA-seq of homozygote, heterozygote and WT
| 生长方案: |
The three genotypes of the OsMET1-2 gene in rice (cv. Nipponbare), wild-type (WT, +/+), heterozygote (+/-) and homozygous mutant (-/-) were germinated and grown on the hormone-free, half-strength Murashige & Skoog (MS) medium under 16/8 hrs light/dark at 26/24℃. |
| 处理方案: |
- |
| 提取方案: |
Total RNA from the 11-d-old shoots of the three OsMET1-2 genotypes was isolated for RNA-seq. |
| 建库方案: |
For each genotype, two independent libraries were constructed and sequenced following the standard Illumina sequencing procedure on the HiSeq 2000. |
| 分子类型: |
poly(A)+ RNA |
| 库的片段类型: |
PAIRED |
| 库的链类型: |
- |
| 测序平台: |
ILLUMINA |
| 测序仪型号: |
Illumina HiSeq 2000 |
| 链特异性: |
Unspecific |
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Condition Detail |
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加标（Spike-In） |
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Mutation of a major CG methylase in rice causes genome-wide hypomethylation, dysregulated genome expression, and seedling lethality.
Proceedings of the National Academy of Sciences of the United States of America . 2014-07-07 [PMID:
25002488]
