Proteomic Analyses Detect Higher Expression of C-Type Lectins in Imidacloprid-Resistant Colorado Potato Beetle Leptinotarsa decemlineata Say

SIMPLE SUMMARY: Surveillance and determining the mechanisms of pesticide resistance are key components of resistance management. Mechanisms can be investigated using biochemical, genomic, proteomic and other modern analytical techniques. In the present study, proteomic analyses of Colorado potato beetle (CPB), one of the most adaptable insect pests to both plant toxins and synthetic insecticides, were applied to identify protein differences in insecticide-susceptible and resistant strains. Proteins identified in abdominal and midgut tissues based on separating by 2-dimensional (2-D) gels and mass spectrometry were associated with insect innate immunity. A database search found that the highest match was a C-type lectin (CTL), which is a component in the insect’s innate immune system. The 2-D gel spot identified as a CTL was greater in the insecticide-resistant CPB strain, but the CTL spot size was increased by exposure to imidacloprid in the susceptible strain. This is a novel finding, which suggests that CTLs and insect immunity may respond to certain toxins as well as to pathogens. There may also be a potential application for pest management if insect immunity is targeted. ABSTRACT: The Colorado potato beetle (CPB) is one of the most adaptable insect pests to both plant toxins and synthetic insecticides. Resistance in CPB is reported for over 50 classes of insecticides, and mechanisms of insecticide-resistance include enhanced detoxification enzymes, ABC transporters and target site mutations. Adaptation to insecticides is also associated with changes in behaviour, energy metabolism and other physiological processes seemingly unrelated to resistance but partially explained through genomic analyses. In the present study, in place of genomics, we applied 2-dimensional (2-D) gel and mass spectrometry to investigate protein differences in abdominal and midgut tissue of insecticide-susceptible (S) and -resistant (R) CPB. The proteomic analyses measured constitutive differences in several proteins, but the highest match was identified as a C-type lectin (CTL), a component of innate immunity in insects. The constitutive expression of the CTL was greater in the multi-resistant (LI) strain, and the same spot was measured in both midgut and abdominal tissue. Exposure to the neonicotinoid insecticide, imidacloprid, increased the CTL spot found in the midgut but not in the abdominal tissue of the laboratory (Lab) strain. No increase in protein levels in the midgut tissue was observed in the LI or a field strain (NB) tolerant to neonicotinoids. With the exception of biopesticides, such as Bacillus thuringiensis (Bt), no previous studies have documented differences in the immune response by CTLs in insects exposed to synthetic insecticides or the fitness costs associated with expression levels of immune-related genes in insecticide-resistant strains. This study demonstrates again how CPB has been successful at adapting to insecticides, plant defenses as well as pathogens.