Cannabis sativa L., also called hemp, is a plant of the Cannabaceae family that originates from central and southwest Asia. Over time, the genetic diversity of cannabis has expanded, leading to the emergence of numerous distinct genotypes characterized by varied phenotypes and a wide range of secondary metabolites. These metabolites exhibit diverse biological properties and are utilized in human medicine and the pharmaceutical industry. Cannabis sativa L. includes different genetically distinct biotypes, notably industrial hemp and recreational cannabis. It has been used for its fiber, oil, food, and medicinal properties, as well as for recreational and religious purposes. It contains various bioactive substances such as alkaloids, flavonoids, terpenoids, and cannabinoids. The concentrations of these secondary metabolites vary significantly depending on the plant genotype and environmental factors, whether abiotic or biotic. Microorganisms associated with Cannabis sativa L., whether residing on the surface, within tissues, or in cultivated soils, exhibit significant genetic diversity. They provide benefits, such as promoting plant growth, protecting against pathogens, improving mineral absorption, and influencing the production of secondary metabolites. The use of beneficial microorganisms, such as PGPR bacteria, endophytic fungi, or their bioactive molecules, could play a key role in enhancing cannabinoid production in cannabis plants. Although the chemistry of cannabis has been extensively studied, further research is needed to understand the influence of biotic and abiotic factors on the variation in synthesized cannabinoid levels. This review provides a comprehensive overview of Cannabis sativa L., specifically focusing on its associated microorganisms and their interactions with the plant. It also explores opportunities for the optimal utilization of this plant, particularly focusing on its microorganisms, to enhance growth, protect against bio-aggressors, and promote cannabinoid biosynthesis. This paper also reviews current investigations and suggests various opportunities for further research perspectives.