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Published on: May 14, 2020
N6 -Methyladenosine mRNA modification regulates transcripts stability associated with cotton fiber elongation
Abstract:
N6 -Methyladenosine (m6 A) is the most abundant methylation modification in eukaryotic mRNA. The discovery of the dynamic and reversible regulatory mechanism of m6 A has greatly promoted the development of m6 A-led epitranscriptomics. However, the characterization of m6 A in cotton fiber is still unknown. Here, we reveal the potential link between m6 A modification and cotton fiber elongation by parallel m6 A-immunoprecipitation-sequencing (m6 A-seq) and RNA-seq analysis of fibers from the short fiber mutants Ligonliness-2 (Li2 ) and wild-type (WT). This study demonstrated a higher level of m6 A in the Li2 mutant, with the enrichment of m6 A modifications in the stop codon, 3'-untranslated region and coding sequence regions than in WT cotton. In the correlation analysis between genes containing differential m6 A modifications and differentially expressed genes, we identified several genes that could potentially regulate fiber elongation, including cytoskeleton, microtubule binding, cell wall and transcription factors (TFs). We further confirmed that the methylation of m6 A affected the mRNA stability of these fiber elongation-related genes including the TF GhMYB44, which showed the highest expression level in the RNA-seq data and m6 A methylation in the m6 A-seq data. Next, the overexpression of GhMYB44 reduces fiber elongation, whereas the silencing of GhMYB44 produces longer fibers. In summary, these results uncover that m6 A methylation regulated the expression of genes related to fiber development by affecting mRNA's stability, ultimately affecting cotton fiber elongation.
Insights
N6-Methyladenosine (m6A) RNA modification regulates cotton fiber length by impacting gene mRNA stability. This study links m6A levels to fiber elongation, identifying key genes like GhMYB44 involved in this process.
Area of Science:
- Plant molecular biology
- Epitranscriptomics
- Cotton genetics
Background:
- N6-Methyladenosine (m6A) is the most prevalent mRNA modification in eukaryotes, crucial for gene expression regulation.
- The role of m6A in cotton fiber development, particularly elongation, remains largely uncharacterized.
- Understanding m6A dynamics is key to advancing epitranscriptomics and crop improvement.
Purpose of the Study:
- To investigate the role and patterns of m6A modification in cotton fiber elongation.
- To identify genes and regulatory pathways influenced by m6A during fiber development.
- To elucidate the functional impact of m6A on key genes controlling cotton fiber length.
Main Methods:
- Parallel m6A-immunoprecipitation-sequencing (m6A-seq) and RNA-sequencing (RNA-seq) were performed on cotton fibers from wild-type and short fiber mutant (Li2) plants.
- Differential m6A modification and gene expression analyses were conducted to identify affected genes.
- Functional validation of candidate genes, including GhMYB44, using overexpression and gene silencing techniques.
Main Results:
- Cotton fibers exhibited distinct m6A modification patterns, with higher levels observed in the Li2 mutant.
- m6A modifications were enriched in specific mRNA regions (stop codon, 3'-UTR, CDS) in the mutant.
- Correlation analysis identified genes related to cytoskeleton, cell wall, and transcription factors (TFs) with differential m6A and expression.
- GhMYB44, a TF, showed significant m6A methylation and altered expression, functionally impacting fiber elongation upon manipulation.
Conclusions:
- m6A RNA methylation plays a significant role in regulating cotton fiber elongation.
- The study reveals that m6A affects mRNA stability of key genes, including GhMYB44, influencing fiber development.
- These findings provide novel insights into the epitranscriptomic regulation of cotton fiber growth and offer potential targets for crop improvement.
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