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Ambrosia beetles Xylosandrus crassiusculus and X. germanus are among the most important exotic pests of orchards and nurseries in the US and are difficult to control using conventional insecticides because of their cryptic habits. The use of biological control agents may prove effective by targeting both beetles and fungal symbionts inside tree galleries: entomopathogenic fungi could be used to target beetle foundresses and their brood, or mycoparasitic fungi, e.g., Trichoderma harzianum, could be used to target their associated fungal symbionts. We used a combination of in vitro assays and beetle bioassays to examine competition between symbionts and biological control fungi and the impact of biological control fungi on beetle brood production. The in vitro assays showed T. harzianum outcompeted different strains of Ambrosiella roeperi and A. grosmanniae associated with X. crassiusculus and X. germanus, respectively, whether in primary or secondary resource capture assays. In contrast, entomopathogenic fungi Beauveria bassiana and Metarhizium brunneum blocked the spread of symbionts only in primary competition assays. Complementary beetle bioassays showed that beetle galleries in T. harzianum-treated beech stems had sparse symbiont growth, many with no or only a small number of eggs present. Brood numbers produced by foundresses in T. harzianum-treated stems were comparable to those in stems treated with either entomopathogen at the higher dosages, in which brood reduction was likely due to foundress mortality prior to laying eggs or after laying only a small number of eggs. These results show the potential of using biological control fungi in targeting ambrosia beetle populations either directly by killing foundresses and reducing brood production or indirectly by suppressing symbiont growth in their galleries.

Little is known about the genetic architecture of antifungal immunity in natural populations. Using two population genetic approaches, Quantitative Trait Locus (QTL) Mapping and Evolve and Resequence (E&R), we explored D. melanogaster immune defense against infection with the fungus Beauveria bassiana. Immune defense was highly variable both in the recombinant inbred lines from the Drosophila Synthetic Population Resource used for our QTL mapping and in the synthetic outbred populations used in our E&R study. Survivorship of infection improved dramatically over just 10 generations in the E&R study, and continued to increase for an additional 9 generations, revealing a trade-off with uninfected longevity. Populations selected for increased defense against B. bassiana evolved cross resistance to a second, distinct B. bassiana strain but not to bacterial pathogens. The QTL mapping study revealed that sexual dimorphism in defense depends on host genotype, and the E&R study indicated that sexual dimorphism also depends on the specific pathogen to which the host is exposed. Both the QTL mapping and E&R experiments generated lists of potentially causal candidate genes, although these lists were non-overlapping.

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