Native warm-season grasses (NWSG) are gaining merit as biofuel feedstocks for ethanol production with potential for concomitant production of cattle forage and wildlife habitat provision. However, uncertainty continues regarding optimal production approaches for biofuel yield and forage quality within landscapes of competing wildlife conservation objectives. We used a randomized complete block design of 4 treatments to compare vegetation structure, forage and biomass nutrients, and biomass yield between Panicum virgatum (Switchgrass) monocultures and NWSG polycultures harvested once or multiple times near West Point, MS, 2011–2013. Despite taller vegetation and greater biomass in Switchgrass monocultures, NWSG polycultures had greater vegetation structure heterogeneity and plant diversity that could benefit wildlife. However, nutritional content from harvest timings optimal for wildlife conservation (i.e., late dormant season-collected biomass and mid-summer hay samples) demonstrated greater support for biofuel production than quality cattle forage. Future research should consider testing various seed mixes for maximizing biofuel or forage production among multiple site conditions with parallel observations of wildlife use.