In order to achieve a desirable ecological and sustainable agriculture a thorough understanding of the plant-soil mycobiome is imperative. Commercial industrial agriculture alters greenhouse gas emissions, promotes loss of plant and soil biodiversity, increases pollution by raising atmospheric CO₂, and releases pesticides, thus affecting both terrestrial and aquatic ecosystems. Diversified farming systems, including perennial cultivated pastures, are among worldwide strategies that aim to reduce terrestrial greenhouse gas emissions and deal with threats to global sustainability. Additionally, stimulation of soil microbes and appropriate soil management can influence soil interactions as well as the rates of organic matter decomposition and the release of gases. Agricultural soil microbial communities play a central role in ecosystem processes and are affected by biocontrol agents, biofertilizers, and exposure to pesticides, the extent to which is yet to be fully elucidated. Intercropping different plant species is beneficial, as this can increase carbon fixation by plants, transferring carbon to the soil, especially via mycorrhizas, thus modifying interplant interactions. This review focuses on agro-ecosystems, showing the latest advances in the plant-soil interface (the mycobiome) for an eco-efficient agricultural production.