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Soil-less growing practices have opened up new possibilities for many specialty crops, including strawberries. US strawberry growers are facing a number of challenges that threaten the future of the industry. These include increasing labor and production costs, increased competition from imports, short production seasons, new emerging pests and diseases, extreme weather patterns and new government regulations. Soil-less production of greenhouse-grown strawberries has the potential to address some of those issues. While soil-less greenhouse strawberry production has been common for many years in a number of European and Asian countries, it has yet to become commonplace in the US. Reasons are high upfront cost as well as a general lack of technical knowledge, resulting in grower reluctance to make a transition into greenhouse strawberry production. One horticultural key for a successful soil-less strawberry production system is the choice of optimal substrate options. Therefore, the presented study evaluated the impact of six substrate blends on the growth and yield of strawberry (Fragaria x ananassa cv. Albion) in a commercial greenhouse in Eastern North Carolina. Following substrate blends were used: (1) 100% Coco Fiber; (2) 50% Canadian Peat / 50% Perlite; (3) 50% Canadian Peat / 50% Coco coir; (4) 50% Canadian Peat / 50% Wood Fiber; (5) 50% Canadian Peat / 50% Bark; (6) European Peat. Tray plants (250 cc) were grown in a modified tabletop system inside a poly-covered greenhouse with basic climate control infrastructure. The study was conducted in a randomized complete block design during the growing season 2020-2021 and repeated in 2021-2022. Our results show that strawberries grown in 50% Canadian Peat / 50 % Wood Fiber yielded similar high to 100% Coco Fiber and 100% European Peat, both grower standards. Based on the research, strawberry greenhouse production could use more cost effective, local available woodfiber and Canadian peat substrates instead of coconut coir or European peat that are sourced and shipped overseas. This option can be a more cost effective option for growers in the Southeast, considering making a shift to greenhouse strawberry production.