Update publication information. Strawberry is an important economic crop in the world, and strawberry stigmata are vital to promote pollen germination and allow pollen tubes to enter into stigma tissue. Post-translational modification (PTM) of proteins can significantly affect protein structure and function, and plays a very important regulatory role in plant development. So the functional study of PTM during the pollen-stigma recognition process will improve understanding of the molecular mechanism of the pollen-stigma recognition reactions in strawberry plants. However, to date, little is known about the functions of PTM in strawberry stigmata. In this study, we first analyzed various protein PTMs that likely exist in strawberry stigmata, and found lysine succinylation is highly abundant among six types of PTM. Then a highly efficient enrichment method for succinylation peptides was used to identify 209 succinylation modification sites in 120 proteins in the stigmata. Our bioinformatics analysis further revealed that the succinylated proteins are involved in many important biological processes including resistance and stress responses, vesicular transport, as well as processes related to energy and metabolism. Mass spectrometry (MS)-based proteomics, combining with immunoprecipitation (IP) and western blotting (WB), showed that there was anobvious increase in assembly polypeptide 2 (AP2)/clathrin succinylation abundance at 0.5 h or 2 h after pollination, revealing the significance of succinylation in the processes involved in the recognition of pollen-stigma signal substances and vesicular transport. In conclusion, our results suggest that lysine succinylation is involved in many important biological processes, and also reveal that AP2/clathrin-mediated vesicular transport process is strongly regulated by lysine succinylation during pollen recognition. These results suggest novel research directions for studying the molecular mechanism underlying the interaction and recognition between pollen and stigmata in plants, as well as the signal regulatory mechanism.