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All cycads are strictly dioecious with a long juvenile stage. Currently, there is no method available to determine the sexuality of seedlings prior to the onset of cone formation. This study aimed to develop a sex specific Random Amplified Polymorphic DNA (RAPD) marker for Encephalartos natalensis. Initially, 140 primers were used to amplify the bulk DNA of five individuals each of known male and female sexuality. While a high degree of polymorphism was observed in the amplification profiles of male and female plants, only primer OPD-20 generated a specific band (∼850 bp) in female DNA. To validate this observation, this primer was re-tested with 69 individuals of E. natalensis. The 850 bp DNA band was present in all 38 female individuals tested and it was consistently absent in all 31 male plants tested. The result offers a rapid and simple test to determine sexuality of E. natalensis seedlings at early stages of development, before the onset of reproductive maturity thereby saving time and economic resources when cultivating these specimens.Electronic Supplementary Material Supplementary material is available to authorised users in the online version of this article at . 相似文献