From the beginning of this century, non-Saccharomyces yeasts have taken increased relevance in wine processing. Several biotechnological companies now produce non-Saccharomyces yeasts at an industrial level to improve aroma or flavor, stabilize wine, produce biological acidification, or conversely metabolize malic acid. Species like Torulaspora delbrueckii, Metschnikowia pulcherrima, Kloeckera apiculata, Lachancea thermotolerans, Schizosaccharomyces pombe, and several others are common due to the technological applications they have in sensory quality but also in wine ageing and stabilization. Moreover, spoilage non-Saccharomyces yeasts like Brettanomyces bruxellensis, Saccharomycodes ludwigii, and Zygosacharomyces bailii are becoming important because of the alterations they are able to produce in high-quality wines. New strategies to control these defective yeasts have been developed to control them without affecting sensory quality. The knowledge of the physiology, ecology, biochemistry, and metabolomics of these yeasts can help to better use them in controlling traditional problems such as low fermentative power, excessive volatile acidity, low implantation under enological conditions, and sensibility to antimicrobial compounds like sulfites traditionally used in wine processing. This Special Issue intends to compile current research and revised information on non-Saccharomyces yeasts with enological applications to facilitate the use and the understanding of this biotechnological tool. In 1 year this SI has globally more than 15kdownloads and produced more than 30 citations.