The date palm fruit (Phoenix dactylifera L.) is a valuable fruit crop that requires low inputs and has high nutritional value. Propagating date palms through suckers is slow and can be led to disease transfer. The extended juvenility phase and life cycle of date palms present challenges in breeding. Seed propagation is quicker, but determining the sex early is difficult, taking 7–8 years. To aid in date palm breeding, studies have been conducted to identify morphological, physiological, biochemical, and genetic markers for sex identification. Physical traits like plant height, leaf length, and leaf width have been considered, but they are influenced by the environment and may not be reliable for sex identification. It has been noted that morphological traits are not ideal for sex identification in date palms. Male plants exhibit higher stomatal conductance and transpiration rates compared to females. Biochemical analysis has shown variations in nitrogen, phosphorus, and potassium levels, with females having higher (NPK) concentrations due to increased energy needs for embryo growth. The TPD1 gene is more expressed in males, while the SHP1 gene, related to ovule development, is more expressed in females. Gene-based identification can enhance breeding efforts. Additionally, tissue culture techniques can introduce variation among plantlets, aiding breeders. These techniques can help establish date palm orchards with verified female plants early in the sex identification process, potentially, increasing yield. Early sex determination is crucial for a breeding program. This chapter focuses on the large-scale early sex determination of date palm plants using biotechnology techniques and enhancing yield through tissue culture and micropropagation.