Investigating the role of DNA methyltransferases in the mantled somaclonal variation of oil palm
DNA methyltransferase (DMTases) genes have emerged as targets of interest in the exploration of epigenetic mechanisms underlying the mantled variant phenotype in oil palm. Indeed, reduction in global DNA methylation rates and perturbations of floral phenotype could be due to underexpresion of these genes. Therefore, our research efforts focused on the isolation of members of all three families of DNA-methyltranferases identified in higher plants, namely MET, CMT (chromomethylase) and DRM (domain-rearranged). Using both a library screening and a PCR-based approach involving degenerated primers, we successfully cloned partial cDNA sequences belonging to all three families. The respective full-length cDNAs were subsequently obtained through the RACE (Rapid Amplification of cDNA Ends) method. For each DMTase type, we then determined the size of the family in the oil palm genome, and we examined the overall transcription level of each family member individually. The comparative transcription pattern of each class of DMTase was studied in normal and variant tissues through semi-quantitative Reverse-Transcription-PCR (sqRT-PCR) and Real Time quantitative PCR. Our work shows for the first time the characterization of the three DNA methyltransferase gene families in oil palm, and addresses the hypothesis of their role in the determinism of the mantled variant phenotype.
| Main Authors: | , , , |
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| Format: | conference_item biblioteca |
| Language: | eng |
| Published: |
s.n.
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| Subjects: | F60 - Physiologie et biochimie végétale, F30 - Génétique et amélioration des plantes, |
| Online Access: | http://agritrop.cirad.fr/545488/ http://agritrop.cirad.fr/545488/1/document_545488.pdf |
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| Summary: | DNA methyltransferase (DMTases) genes have emerged as targets of interest in the exploration of epigenetic mechanisms underlying the mantled variant phenotype in oil palm. Indeed, reduction in global DNA methylation rates and perturbations of floral phenotype could be due to underexpresion of these genes. Therefore, our research efforts focused on the isolation of members of all three families of DNA-methyltranferases identified in higher plants, namely MET, CMT (chromomethylase) and DRM (domain-rearranged). Using both a library screening and a PCR-based approach involving degenerated primers, we successfully cloned partial cDNA sequences belonging to all three families. The respective full-length cDNAs were subsequently obtained through the RACE (Rapid Amplification of cDNA Ends) method. For each DMTase type, we then determined the size of the family in the oil palm genome, and we examined the overall transcription level of each family member individually. The comparative transcription pattern of each class of DMTase was studied in normal and variant tissues through semi-quantitative Reverse-Transcription-PCR (sqRT-PCR) and Real Time quantitative PCR. Our work shows for the first time the characterization of the three DNA methyltransferase gene families in oil palm, and addresses the hypothesis of their role in the determinism of the mantled variant phenotype. |
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