Partnerships Between Ambrosia Beetles and Fungi: Lineage-Specific Promiscuity Among Vectors of the Laurel Wilt Pathogen, Raffaelea lauricola

Nutritional mutualisms that ambrosia beetles have with fungi are poorly understood. Although these interactions were initially thought to be specific associations with a primary symbiont, there is increasing evidence that some of these fungi are associated with, and move among, multiple beetle partners. We examined culturable fungi recovered from mycangia of ambrosia beetles associated with trees of Persea humilis (silk bay, one site) and P. americana (avocado, six commercial orchards) that were affected by laurel wilt, an invasive disease caused by a symbiont, Raffaelea lauricola , of an Asian ambrosia beetle, Xyleborus glabratus . Fungi were isolated from 20 adult females of X. glabratus from silk bay and 70 each of Xyleborus affinis , Xyleborus bispinatus , Xyleborus volvulus , Xyleborinus saxesenii , and Xylosandrus crassiusculus from avocado. With partial sequences of ribosomal (LSU and SSU) and nuclear (β-tubulin) genes, one to several operational taxonomic units (OTUs) of fungi were identified in assayed individuals. Distinct populations of fungi were recovered from each of the examined beetle species. Raffaelea lauricola was present in all beetles except X . saxesenii and X . crassiusculus , and Raffaelea spp. predominated in Xyleborus spp. Raffaelea arxii , R. subalba , and R. subfusca were present in more than a single species of Xyleborus , and R. arxii was the most abundant symbiont in both X . affinis and X. volvulus. Raffaelea aguacate was detected for the first time in an ambrosia beetle ( X. bispinatus ). Yeasts (Ascomycota, Saccharomycotina) were found consistently in the mycangia of the examined beetles, and distinct, putatively co-adapted populations of these fungi were associated with each beetle species. Greater understandings are needed for how mycangia in ambrosia beetles interact with fungi, including yeasts which play currently underresearched roles in these insects.