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Molecular phenology regulating bud dormancy of the Yoshino-cherry tree

ABSTRACT: Molecular phenology regulating bud dormancy of the Yoshino-cherry tree

PROVIDER: PRJNA1032135 | ENA |

REPOSITORIES: ENA

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Molecular phenology regulating bud dormancy of the Yoshino-cherry tree

Project description:We explored seasonal gene expression profiles in the Yoshino cherry trees (Cerasus × yedoensis ‘Somei-yoshino’) at three distinct sites, each situated at varying latitudes in Japan. Our goal was to gather molecular phenology data that could provide insights into mechanism of bud dormancy. The molecular phenology approach, which monitors the seasonal dynamics of global gene expression profiles in leaves and buds under natural conditions, enables the study of physiological responses to seasonal environmental changes at the molecular level.

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Project description:Chromatin immunoprecipitation-sequencing (ChIP-seq) is a robust technique to study interactions between proteins, such as histones or transcription factors, and DNA. This technique in combination with RNA-sequencing (RNA-seq) is a powerful tool to better understand biological processes in eukaryotes. We developed a combined ChIP-seq and RNA-seq protocol for tree buds (Prunus avium L., Prunus persica L Batch, Malus x domestica Borkh.) that has also been successfully tested on Arabidopsis thaliana and Saccharomyces cerevisiae. Tree buds contain phenolic compounds that negatively interfere with ChIP and RNA extraction. In addition to solving this problem, our protocol is optimised to work on small amounts of material. Furthermore, one of the advantages of this protocol is that samples for ChIP-seq are cross-linked after flash freezing, making it possible to work on trees growing in the field and to perform ChIP-seq and RNA-seq on the same starting material. Focusing on dormant buds in sweet cherry, we explored the link between expression level and H3K4me3 enrichment for all genes, including a strong correlation between H3K4me3 enrichment at the DORMANCY-ASSOCIATED MADS-box 5 (PavDAM5) loci and its expression pattern. This protocol will allow analysis of chromatin and transcriptomic dynamics in tree buds, notably during its development and response to the environment.

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