Discovery of Rickettsia and Rickettsiella Intracellular Bacteria in Emerald Ash Borer Agrilus planipennis by Metagenomic Study of Larval Gut Microbiome in European Russia

Emerald ash borer Agrilus planipennis (Coleoptera: Buprestidae) is a quarantine pest posing a threat to ash trees all over Europe. This wood-boring beetle native to Asia is quickly spreading in North America and European Russia, and approaching the European Union and the Middle East. It is important to study microorganisms associated with this pest, because the knowledge of its “natural enemies” and “natural allies” could be potentially used for the control of the pest. All previously published information about the A. planipennis microbiome was obtained in North America and China. We present the first study on procaryotes associated with A. planipennis in Europe. Alive larvae were sampled from under the bark of Fraxinus pennsylvanica in the Moscow Oblast and the gut microbiome was studied using metagenomic methods. Next-generation Illumina-based amplicon sequencing of the v3-v4 region 16S-RNA gene was performed. In total, 439 operational taxonomic units from 39 families and five phyla were detected. The dominant families in our samples were Pseudomonadaceae, Erwiniaceae and Enterobacteriaceae, in accordance with the published information on the larval gut microbiome in North America and China. We detected intracellular bacteria in A. planipennis for the first time, namely Rickettsia (Rickettsiaceae) and Rickettsiella (Diplorickettsiaceae). Representatives of the genus Rickettsia are known to be in mutualistic symbiosis with some phytophagous insects, while Rickettsiella bacteria are pathogenic to many arthropods. The finding of Rickettsia and Rickettsiella opens perspectives for future research on the interactions between these bacteria and A. planipennis and the possible use of these interactions for the control of the pest.