Using the combined gene approach and multiple analytical methods to improve the phylogeny and classification of Bombus (Hymenoptera, Apidae) in China

Bumble bees are vital to our agro-ecological system, with approximately 250 species reported around the world in the single genus Bombus. However, the health of bumble bees is threatened by multiple factors: habitat loss, climate change, pesticide use, and disease caused by pathogens and parasites. It is therefore vitally important to have a fully developed phylogeny for bumble bee species as part of our conservation efforts. The purpose of this study was to explore the phylogenetic relationships of the dominant bumble bees on the Tibetan plateau and in northern China as well as their placement and classification within the genus Bombus. The study used combined gene analysis consisting of sequence fragments from six genes, 16S rRNA, COI, EF-1α, Argk, Opsin and PEPCK, and the phylogenetic relationships of 209 Bombus species were explored. Twenty-six species, including 152 gene sequences, were collected from different regions throughout China, and 1037 gene sequences representing 183 species were obtained from GenBank or BOLD. The results suggest that the 209 analyzed species belong to fifteen subgenera and that most of the subgenera in Bombus are monophyletic, which is in accordance with conventional morphology-based classifications. The phylogenetic trees also show that nearly all subgenera easily fall into two distinct clades: short-faced and long-faced. The study is the first to investigate the phylogenetic placement of Bombus turneri (Richards), Bombus opulentus Smith, Bombus pyrosoma Morawitz, Bombus longipennis Friese, Bombus minshanensis Bischoff, and Bombus lantschouensis Vogt, all of which are widely distributed throughout different regions of China. The knowledge and understanding gained from the findings can provide a molecular basis to accurately classify Bombus in China and to define strategies to conserve biodiversity and promote pollinator populations.