Rural ecological landscape biodiversity is an important part of global biodiversity, which is an essential guarantee for rural landscape construction, evaluation and rural livable development providing important ecosystem services for sustainable human development. Currently, the research system of rural ecological landscape biodiversity is still ambiguous, and the relevant research results cannot effectively instruct the practice of rural ecological landscape construction. In this paper, we used 522 relevant literature in Web of Science from 1990 to 2020 to make a knowledge map of literature co-citation and keyword co-occurrence using the bibliometric tool CiteSpace, focusing on turning points, high emergence and high citation literature to clarify the development of rural ecological landscape biodiversity. The study found that (1) the research on rural ecological landscape biodiversity could be divided into three phases, namely, the beginning stage (1993—2002), which guided the direction of rural ecological landscape biodiversity research and proposed the impact of urbanization on rural biodiversity; the countdown stage (2003—2010), which was based on Hansen’s 2005 article exploring the response mechanisms of biodiversity to landscape change, focusing on how to sustainably restore and preserve rural ecological landscape biodiversity; and the development stage (2011—2020), whose emergent literature was Beilin’s 2014 article, which shifted the attention to social-ecological systems, that is to say the study of rural ecological landscape biodiversity from ecological, economic and social multi-dimensions; (2) The main line of research on rural ecological landscape biodiversity changed from a large number of case studies to the theoretical generalization of the impact of urbanization on rural ecological landscape biodiversity and to the application of theory to practice and sustainable conservation of rural ecological landscape biodiversity; (3) The rural ecological landscape biodiversity research hotspots changed from the impact of urbanization on rural ecological landscape biodiversity to sustainable rural development and to ensuring sustainable human livelihoods. The spirit of questioning and seeking is the most important enlightenments we have achieved for future rural ecological landscape biodiversity research. Solid research will be an important driver of the success of rural ecological landscape biodiversity in practice. Accordingly, the following studies should be strengthened in the future: (1) Research and establish a planning system applicable to the construction of different rural ecological landscapes; (2) Research and innovate techniques for maintaining rural ecological landscapes ecosystem diversity; (3) Research and construct a monitoring and early warning mechanism for rural ecological landscapes biodiversity. The ultimate research goals all point to how rural ecological landscape biodiversity can better serve human society.