Bacterial community associated with the ambrosia beetle Platypus cylindrus on declining Quercus suber trees in the Alentejo region of Portugal

In Portugal, the oak pinhole borer Platypus cylindrus and its mycobiota have been associated with cork oak (Quercus suber) death, but no knowledge exists regarding the associated bacterial community. However, it is known that some bacteria are important for ambrosia beetle symbiosis and play a role in oak tree health. To explore the bacteria associated with this beetle and its host, with the ultimate goal of highlighting potential roles in oak decline, this study used a culture-dependent approach for strain isolation and phylogenetic identification using 16S rRNA gene sequencing and multilocus sequence analysis (MLSA). The bored galleries of different cork oak trees from a cork stand in Alentejo, together with the body and mycangia of adult beetles, were investigated. The samples revealed a diverse community comprising 500 isolates with 64 distinct types of bacterial colonies. Sixty-eight strains were selected for sequencing and used for phylogenetic analysis, 40 from wood galleries and 28 from beetles. Thirty-two genera of bacteria were identified, 18 of which were described for the first time within oak–beetle interactions. Major taxonomic groups were Actinobacteria in beetles and Enterobacterales in wood galleries. Although specific oak bacterial pathogens were not detected, a group of distinct strains detected in wood galleries, potentially belonging to a new Pectobacteriaceae species, were able to produce mild symptoms on cork oak plantlets. This study reports for the first time the biodiversity of culturable bacteria associated with the Q. suber–P. cylindrus interaction, their relevance to both organisms and the possible contribution to oak decline.