Broader realization of both increasing biodiversity loss and pressures on ecosystems worldwide has highlighted the importance of public perceptions of species and the subsequent motivations towards improving the status of natural systems. Several new proposals have arisen in reference to environmental learning, including mimicking popular gaming media. Inspired by the popular game Pokémon, the Phylo Trading Card Game (Phylo game) is one such emerging possibility. It was invented as an open-source, competitive, and interactive game to inform players' knowledge of species, ecosystems, and negative environmental events (e.g., climate change, oil spills, wildfires). The game has now achieved global reach, yet the impact of this game on conservation behavior has never been tested. This study used a randomized control trial to evaluate the Phylo game's impact on conservation behavior (i.e., Phylo condition). This was compared to an information control condition with a more traditional learning method using a slideshow (i.e., Slideshow condition). A second card game was used to control for the act of playing a game (i.e., Projects condition). We found that ecological perceptions (i.e., the perceived relationship of species to their ecosystems) and species knowledge increased after both the game and the slideshow, but the Phylo Game had the added benefit of promoting more positive affect and more species name recall. It also motivated donation behavior in the direction of preventing negative environmental events instead of directly aiding an individual species or ecosystem. Our findings highlight the potential value of this game as a novel engagement tool for enhancing ecological literacy, motivations, and actions necessary to meet ecological challenges.