AbstractPrevious studies suggest that the senescence and death of the replaceable bud of the Chinese chestnut cultivar (cv.) “Tima Zhenzhu” involves programmed cell death (PCD). However, the molecular network regulating replaceable bud PCD is poorly characterized. Here, we performed transcriptomic profiling on the chestnut cv. “Tima Zhenzhu” replaceable bud before (S20), during (S25), and after (S30) PCD to unravel the molecular mechanism underlying the PCD process. A total of 5,779, 9,867, and 2,674 differentially expressed genes (DEGs) were discovered upon comparison of S20 vs S25, S20 vs S30, and S25 vs S30, respectively. Approximately 6,137 DEGs common to at least two comparisons were selected for gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses to interrogate the main corresponding biological functions and pathways. GO analysis showed that these common DEGs could be divided into three functional categories, including 15 cellular components, 14 molecular functions, and 19 biological processes. KEGG analysis found that “plant hormone signal transduction” included 93 DEGs. Overall, 441 DEGs were identified as related to the process of PCD. Most of these were found to be genes associated with ethylene signaling, as well as the initiation and execution of various PCD processes.