The study of rumen microbiota is of great significance for understanding the health, growth and development, as well as digestion and absorption processes of ruminants. In recent years, with the development of high-throughput sequencing technology and the rise of microbiome research, significant progress has been made in the study of rumen microbiota in ruminants. This article mainly reviews the composition and influencing factors of rumen microbiota in ruminants, as well as the technical methods for studying rumen microbiota, including 16S rRNA gene sequencing, functional genomics analysis, metabolomics research, and fluorescence in situ hybridization (FISH). Meanwhile, the application and development prospects of these methods in the study of rumen microbiota in ruminants will also be explored, aiming to provide reference for the structural and functional analysis of rumen microbiota.