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Employing rooftops for the cultivation of crops in limited urban space has garnered interest in densely populated cities in the United States, where there is a growing demand for locally sourced vegetable products. Fertility management recommendations for rooftop farming, however, are scant. With insufficient research on nutrient cycling within rooftop farming systems, which tend to use soilless substrates with low organic matter content, the potential tradeoffs between the negative impacts (e.g., nutrient runoff) and the benefits (e.g., increased locally produced vegetables, stormwater retention, etc.) associated with rooftop farms are unclear. The objective of this study was to evaluate the effects of organic and inorganic nitrogen (N) inputs on the N dynamics within substrate typically used on rooftop farms. Substrate without added N inputs (control) was compared with substrates receiving N sources that are both realistic for and/or reflective of amendments currently applied on urban rooftop farms: a synthetic fertilizer (Osmocote® 14N–4.2P–11.6K), and three organic N inputs—composted poultry manure, municipal green waste (MGW) compost, and vermicompost. Aboveground crop biomass and yields of Beta vulgaris (swiss chard), along with inorganic N availability (ammonium: