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The learning objectives of environmental education emphasize investigation in real life to enhance students’ skills and experiences in solving practical problems. This study used the virtual reality (VR) technology to develop a virtual ecological environment for learning about the Taipei tree frog, supported by situated learning and game-based learning design to enhance students’ learning interest and motivation. Users can wear the head-mounted display (HMD) to explore the virtual environment for learning the Taipei tree frog’s ecological behavior, such as foraging and mating as well as its habitats and predators. A teaching experiment was conducted to investigate students’ learning effectiveness and the senses of presence and anxiety after using the virtual ecological environment. The experimental group (wearing the HMD) contained 40 students, the control group (using the desktop VR) contained 40 students, and both groups were used as samples to learn about the Taipei tree frog. The results indicated that using HMD VR and desktop VR could both enhance learning achievements, but the learning effectiveness of the former was significantly higher than that of the latter. The levels of anxiety for both groups were about the same, but the level of presence for the experimental group was higher than that of the control group because the HMD VR provided a more immersive experience than the desktop VR. The virtual ecological environment can save the time and effort of travelling to the natural habitat for observing the Taipei tree frog, and the design of role-playing game (RPG) can enhance learners’ interest and motivation. Therefore, it is a useful tool for promoting environmental education.