To investigate proteomic changes during desiccation in recalcitrant Quercus acutissima seeds, TMT labeling technology was used to analyze protein profiles at initial moisture content, semi-lethal moisture content, and lethal moisture content stages. A total of 400,166 secondary spectra were obtained, identifying 39,777 peptides, among which 6,682 proteins were qualitatively identified and 6,398 were reliable proteins.