A genetic linkage analysis involving 60 random decamer primers in Aleppo pine (Pinus halepensis MILL.), one of the most important Mediterranean conifers, is reported. Five trees originating from five natural Spanish populations and 40 haploid megagametophytes per tree were investigated based on joint segregation and independent assortment. Twenty-two decamers were selected for their stable and repeatable banding patterns and 10 of these produced 24 polymorphic loci that presented Mendelian inheritance. Some degree of segregation distortion was evident in 16% of the loci tested. A total of 155 linkage tests were executed based on LOD-scores and χ2 contingency tables. Six linkage groups that include 13 loci were detected OPA01750 OPP041200 OPP04930, OPA112500 OPA11950, OPA191150 OPA191090, OPN06690 OPN06420, OPN12450 OPN12300 and OPP10640 : OPP10600. Recombination frequencies were homogeneous across trees and chromosomal interference was found to be negative. Total consensus genetic map length ranged from 175 cM to 225 cM depending on the mapping function used. This is the first linkage study in this species using RAPD markers and single tree megagametophytes