Abstract The objective of this work was to characterize the expression of gametophytic self-incompatibility in a Coffea canephora breeding population, to assist in the management and development of new cultivars. For that purpose, 550 in vitro pollinations were carried out among 62 parent plants, of which 27 were from the conilon botanical variety and 35 from the robusta. Thirty-two genotypes compatible with all previously known testers were identified, suggesting the existence of new compatibility groups. From these results, hybridizations were carried out in a complete diallel design with reciprocal crosses to characterize new test plants. Based on the compatibility response with the test plants, the genotypes were clustered into the six following groups: group I, 11 (17.74%) genotypes; group II, 13 (20.97%); group III, 6 (9.68%); group IV, 9 (14.52%); group V, 8 (12.90%); and group VI, 15 (24.19%). The genotypes of the botanical variety robusta show a higher frequency of plants in compatibility group VI and a greater genetic variability, whereas those of the conilon variety have a higher frequency of plants in compatibility group II. The identification of new compatibility groups assists in new management practices that seek to increase the efficiency of pollination by favoring, through natural means, fully compatible crosses.