JOJOBA (Simmondsia chinensis L.) is a dioecious perennial evergreen shrub that is native to the south-western deserts of North America, and has now been introduced to the Middle East. Currently, the global production of jojoba is low; this is mainly due to the high male-to-female ratio in plantations, since the plants are mainly established from seed. Hence, a proper male:female population ratio is almost impossible to maintain in the field, leading to reduced production. We report a breakthrough in the methodology for determining the sex of jojoba plants. Our research revealed that the jojoba genome contains a sequence of the SRY gene, which encodes a protein that is similar to that found in the human sex-determining region (SRY) gene. This region was amplified and matched with sequences that are found in papaya and humans. The jojoba-SRY (SRY gene) was amplified, and we closely matched its sequences with those in the papaya and human. The complete sequence of the DNA was deposited into the GenBank (MK991776, 360pb) database. Our newly developed method is rapid and straightforward, representing a breakthrough for sex determination strategies of jojoba plants at early developmental stages. The uniqueness of this approach is that it targets specific sequences in the SRY, a region that has been extensively studied in human genetics but is also apparently recognizable in plants. This useful molecular diagnostic tool is vital to agronomical breeding programs for sex determination in long-lived jojoba crops.