The objectives of this research were to 1) develop a learning unit to promote biodiversity use in agricultural ecosystems, 2) assess student learning outcomes related to their knowledge of biodiversity in agricultural ecosystems and to their skills of knowledge management, and 3) evaluate the impacts of learning to manage biodiversity in agricultural ecosystems on cultivators. Agriculture is a major occupation in Thailand. Knowledge about utilizing biodiversity in an agricultural ecosystem is essential for safe food consumption. Teachers play a vital role in developing and managing learning opportunities for cultivators, leading to participatory learning. Important components of learning management are teachers, learning sources in the local community, learning units about managing agricultural ecosystem biodiversity, and collaborative learning assessments for developing the learning units. This study develops learning outcomes at secondary schools to address the use of biodiversity in agricultural ecosystems along with agricultural methods and teaching skills related to sustainable agricultural learning management. Teaching skills have been improved through the guideline Encouraging Learning Outcomes for Biodiversity in Agricultural Ecosystems, in accordance with the sustainable development goals of the United Nations.