The use of turfgrasses might provide an additional solution for establishing green roofs in urban environments. The aim of the present study was to determine Manilagrass [Zoysia matrella (L.) Merr. ‘Zeon’] drought tolerance when grown under green roof conditions and under two different irrigation regimes. Treatments included: 1) two extensive green roof substrates {locally produced substrate [3 sandy loam soil:8 pumice:4 perlite:4 compost:1 zeolite (by volume)] and a commercially available substrate based on crushed tiles}; 2) two substrate depths (7.5 cm or 15 cm); and 3) two irrigation regimes (3 mm or 6 mm of irrigation every 3 days). Substrate characteristics (particle size distribution, saturated and dry bulk density, total porosity, water potential curves, in situ substrate moisture, pH, electrical conductivity, and nutrient analysis), turfgrass growth, and physiological status [green turf cover (GTC), normalized difference vegetation index (NDVI), and leaf relative water content (RWC)] were determined. During moisture deficit periods, GTC, NDVI, and RWC were most affected by substrate depth; moderately affected by irrigation regime; and, to a lesser extent, by substrate type. Turfgrass growth and physiological status were best during moisture deficit conditions in the deeper profile (15 cm) using the higher amount of irrigation (6 mm) and the locally mixed substrate.