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MYB transcription factors play important roles in plant growth and development,metabolism,responses to biotic and abiotic stresses. In order to explore the structure and function of R2R3-MYB genes in Pinus massoniana, we identified 17 R2R3-MYB genes based on the transcriptome dataset of P. massoniana, and then investigated their physical and chemical properties, phylogenetic relations, gene tissue specificity with qRT-PCR, and expression patterns of these genes in the development process of flower organs and under different abiotic stresses. The results were as follows:(1)Seventeen PmMYBs subcellular were predicted to locate in cell nucleus, without transmembrane structure. They all contained Motif1 and Motif2 conserved motifs. These 17 PmMYBs genes could be divided into nine subgroups according to phylogenetic analysis, which were closely related to gymnosperms such as Pinus taeda and Picea glauca.(2)The expression patterns of PmMYBs genes in different tissues and under salt stress, drought stress, lead stress and low temperature stress were validated by qRT-PCR. As a result, a total of 17 genes were constitutive expressed, but the expression levels were different in different tissues. All genes appeared to be involved in flower development. Seven MYB genes seemed to be involved in the transformation of male and female traits as they were differently expressed in different stages of flower development. Moreover, those genes may also be involved in abiotic stress responses, as most of the genes were up-regulated under abiotic stress. Their unique expression patterns may suggest that they function at different stages of stress. A few genes were down-regulated under stress, especially PmMYB11 gene. This study systematically analyze the characteristics, evolutionary and expression patterns of R2R3-MYB genes in Pinus massoniana during flower development and abiotic stress, and provides a reference for further study on the function and stress response mechanism of R2R3-MYB gene in P. massoniana. [ABSTRACT FROM AUTHOR]