Plants live in constantly changing environments and the increased frequency of extreme weather, as a result of climate change, will exacerbate such stresses. The MemoryRNA project aims to expand the fundamental knowledge of non-genetically-based regulations of the plant transcriptome during heat stress (HS). Post-transcriptional modifications of RNA create an additional layer to reversibly control RNA fate. The sophisticated and poorly understood thermomemory phenomenon represents a powerful system to study the role of the RNA modifications and its high impact on RNA metabolism to reprogram the transcriptome, thus leading plants to adapt and survive. To unravel the role of epitranscriptomic-based reprogramming of the transcriptome of thermomemory RNAs at the shoot apical meristem, the project aims to: i) Reveal the m5C and m6A modifications transcriptome-wide that actively contribute to the global cellular response to HS; ii) Characterize the m5C and m6A marks on thermomemomery mRNAs; and iii) Determine how a m6A reader protein ECT2 is regulated upon HS, and its functional role on transcriptional regulation. Data and biological resources obtained by the team support the MemoryRNA project and incorporate a comprehensive set of cutting-edge technologies and approaches, including: i) Direct sequencing of long-read fragments and computational analysis for accurate mapping of RNA marks; ii) Antibody-based techniques and mutant plants to validate and characterise the function of m5C- and m6A-containing mRNAs; and iii) RNA and fluorescence in situ hybridization, and laser confocal microscopy to investigate the role of ECT2 during thermomemory. Understanding the mechanisms that coordinate transcriptional activity of HS-memory genes in response to environmental stressors is of utmost importance to modern agriculture, and throughout the the funding period, opportunities for translation of our results to biotechnological application will be monitored and explored.