An artificial crossing was made between four Viola ×wittrockiana (2n = 48) lines and one Viola cornuta inbred (2n = 26) line as well as among V. ×wittrockiana lines to investigate the cytologic characteristics, fertility, and heteroses of hybrids. The sizes of flowers and stomata and the leaf areas of the V. ×wittrockiana plants were larger than those of the V. cornuta plants. The karyologic stability of V. cornuta was greater than that of V. ×wittrockiana, and ≈20% of individuals in the latter deviated from the given 2n = 48. Capsule setting after interspecies hand pollination ranged from 28% to 94%, and F1 seed germination ranged from 16% to 88%, revealing no obvious pre- or postzygotic selection mechanisms. The chromosome numbers in V. ×wittrockiana × V. cornuta F1 seedlings varied (2n = 34–38), with about half the cells exhibiting the intermediate (2n = 36) value. These interspecific hybrids exhibited positive heterosis in flower size (range, 11% to 66%) and negative heterosis in plant height (range, –12% to –57%). Both of these traits are desirable in pansy breeding programs.