Coffee is a very important crop for the world economy. The commercial coffee production is based on two species, Coffea arabica L. (70%) and C. canephora Pierre (30%). C. arabica is the only allotetraploid (2n=4x=44) species, which has its primary center of diversity in the southwestern Ethiopia highlands. C. arabica is the only species cultivated in Colombia and represents a very important economic and social value. However, C. arabica has a very narrow genetic diversity and therefore, the use of diploid species on coffee breeding is very important because they allow to broad its diversity. In this work, the genetic segregation patterns were evaluated on a population of 101 F1 hybrid plants from a cross between the diploid species C. liberica and C. eugenioides using 618 molecular markers, of which 168 SSRs and 2 ESTs exhibited polymorphic patterns that allowed segregation analysis. Approximately 24% of the loci were null alleles, and the segregation distortion reached 23.5% at a < 0.01. A total of four segregant types were observed, from up to seven possibilities. The origins of null alleles, possible causes of segregation distortion, different segregation patterns obtained and recombination frequencies are discussed. The knowledge derived from this study allowed to better understand the genetic segregation behavior of these markers with the aim of developing genetic maps that have important applications for coffee breeding programs as well as other commercial crops.