Abstract: 【Aim】 To analyze the phylogeny of the genus Apis, and to investigate the ancestral state and the evolutionary process of dance direction, dance sound, nesting environment and comb structure in Apis based on the phylogenetic relationships. 【Methods】 The COI gene fragments of A. cerana, A. mellifera, A. dorsata, A. laboriosa, A. florea and A. andreniformis distributed inChinawere amplified by PCR and sequenced. The homologous sequences of COI gene of the above six species in other states or regions and those of A. nulunsis, A. nigrocinta, A. koschevnikovi and A. dorsata binghami were downloaded from NCBI databases. The phylogenetic relationships were established using maximum parsimony (MP), maximum likelihood (ML) and Bayesian inference (BI) based on these sequence data. The dance language and nesting behavior of each Apis species mentioned above were coded and treated as trait markers, and added correspondingly into the phylogenetic trees. The ancestral state was traced with the ML model. 【Results】 The results of phylogenetic analysis demonstrated that Apis species can be divided into three clusters, i.e., cavity-nesting honeybees (A. cerana, A. mellifera, A. nigrocinta, A. nulunsis, and A. koschevnikovi), giant honeybees (A. dorsata, and A. laboriosa) and dwarf honeybees (A. florea, and A. andreniformis). The dwarf honeybees are more ancestral, and the giant and cavity-nesting species are monophyletic. Our results also suggest that the giant bee A. dorsata binghami is a separate species from A. dorsata and A. laboriosa and closer to A. laboriosa. Prototype inference indicated that ancestral honeybee species nested in the open with a vertical single comb. They disseminate information about food sources and nesting sites using a silent horizontal waggle dance. The behavior of dancing with buzz and nesting with multi-combs appeared subsequently. 【Conclusion】 As a molecular marker, COI gene can be used to analyze the ancestral state and the evolution of dancing and nesting of Apis species. The buzzing dance language and multi-comb nesting behavior could be an outcome of adaptive evolution during the later stage.

Key words: Apis, COI gene, phylogeny, dance language, nesting behavior