Identification of the Achilles heels of the laurel wilt pathogen and its beetle vector.

Ambrosia beetles harbor fungal symbionts that serve as food sources for larvae and adults. These beetles lay their eggs along tunnels in xylem sapwood, which is the substrate for fungal growth. Symbiotic fungi of the genus Raffaelea found in invasive and indigenous ambrosia beetles include the highly virulent plant pathogen Raffaelea lauricola affecting members of the Lauraceae family. R. lauricola is responsible for the deaths of > 500 million trees since 2005. Infection by as few as 100 spores can kill a healthy tree within months. Our data show that R. lauricola is cold-adapted with optimal growth between 16 and 26 °C, with little to no growth at temperatures ≥ 30 °C. The fungus is halophilic and shows a dramatic decrease in growth at pH ≥ 6.8. Fungicide resistance profiling revealed sensitivity of R. lauricola to prochloraz, dichlorofluanid, most conazoles, dithiocarbamates, and zineb (zinc fungicide), whereas the related species Raffaelea arxii showed more limited fungicide sensitivity. Entomopathogenic fungi potentially useful for beetle control were generally highly resistant to most fungicides tested. Coupling pH decreased the concentration for 95% inhibition of fungal growth (IC95 ) of the most potent R. lauricola fungicides by 3-4-fold. Use of avocado bark plug insect bioassays revealed that commercially available Beauveria bassiana can be used as a biological control agent capable of effectively killing the beetle vectors. These data provide simple and practical recommendations to specifically target R. lauricola while having minimal effects on other symbiotic and entomopathogenic fungi, the latter of which can be used to manage the beetle vectors.