BackgroundPanax Ginsengis a perennial and semi-shady herb with tremendous medicinal value. Due to its unique botanical characteristics, ginseng is vulnerable to various abiotic factors during its growth and development, especially in high temperatures. Proteins encoded by14-3-3genes form a highly conserved protein family that widely exists in eukaryotes. The14-3-3family regulates the vital movement of cells and plays an essential role in the response of plants to abiotic stresses, including high temperatures. Currently, there is no relevant research on the14-3-3genes of ginseng.MethodsThe identification of the ginseng14-3-3gene family was mainly based on ginseng genomic data and Hidden Markov Models (HMM). We used bioinformatics-related databases and tools to analyze the gene structure, physicochemical properties,cis-acting elements, gene ontology (GO), phylogenetic tree, interacting proteins, and transcription factor regulatory networks. We analyzed the transcriptome data of different ginseng tissues to clarify the expression pattern of the14-3-3gene family in ginseng. The expression level and modes of14-3-3genes under heat stress were analyzed by quantitative real-time PCR (qRT-PCR) technology to determine the genes in the14-3-3gene family responding to high-temperature stress.ResultsIn this study, 4214-3-3genes were identified from the ginseng genome and renamedPgGF14-1toPgGF14-42. Gene structure and evolutionary relationship research dividedPgGF14sinto epsilon (ε) and non-epsilon (non-ε) groups, mainly located in four evolutionary branches. The gene structure and motif remained highly consistent within a subgroup. The physicochemical properties and structure of the predictedPgGF14proteins conformed to the essential characteristics of14-3-3proteins. RNA-seq results indicated that the detectedPgGF14sexisted in different organs and tissues but differed in abundance; their expression was higher in roots, stems, leaves, and fruits but lower in seeds. The analysis of GO,cis-acting elements, interacting proteins, and regulatory networks of transcription factors indicated thatPgGF14smight participate in physiological processes, such as response to stress, signal transduction, material synthesis-metabolism, and cell development. The qRT-PCR results indicatedPgGF14shad multiple expression patterns under high-temperature stress with different change trends in several treatment times, and 38 of them had an apparent response to high-temperature stress. Furthermore,PgGF14-5was significantly upregulated, andPgGF14-4was significantly downregulated in all treatment times. This research lays a foundation for further study on the function of14-3-3genes and provides theoretical guidance for investigating abiotic stresses in ginseng.