Sterols produced by bacteria and all eukaryotic organisms are essential for membrane functionality and stability. They play a vital role in growth, development and in abiotic stress tolerance. They are involved in diverse responses to biotic and abiotic stresses that lead to providing resistance against multiple diseases. Additionally, sterols serve as defensive compounds against herbivorous insects and animals. Phytosterols derived from plants, improve human nutrition and health and cure different ailments. The biosynthetic pathways for sterols and triterpenes exhibit similarities until the synthesis of 2,3-oxidosqualene. The complexity of sterol pathways increases during the advanced stages of polycyclic structure synthesis, and remain poorly comprehended in plants. This review explores the various omics techniques used to unveil the functions of genes associated with the phytosterol pathways. The study investigates the biosynthetic gene clusters to clarify the structural arrangements of genes linked to metabolic pathways. Both the upstream and downstream genes associated with these pathways, as well as their evolutionary connections and interrelations within the pathways were brought to the forefront. Moreover, developing strategies to unravel the biosynthesis completely and their multi-layered regulation are crucial to comprehend the global roles that sterols play in plant growth, development, stress tolerance and in imparting defence against pathogens.