This study characterizes the chemical defense system of the invasive longhorn beetle Aromia bungii, a destructive pest of Prunus trees, addressing the limited understanding of chemical defensive mechanisms in Cerambycidae. High-speed cameras, environmental scanning electron microscopy (ESEM), dissection, and micro-CT imaging were used to investigate defensive behavior, and the structure of the defense system, in this beetle. Both sexes of A. bungii possess a pair of triangular, sac-like defensive glands symmetrically located in the metathorax, attached to the metasternum. Upon mechanical stimulation, white liquid defensive substances are rapidly ejected through a pair of slit-shaped openings (~200 µm) at the metasternum corners, without gland eversion, reaching over 50 cm. The average weight of substances ejected in first sprays was 7.95 ± 0.79 mg for females and 8.62 ± 2.13 mg for males (mean ± se), with no significant difference between sexes. However, the weight in second sprays after 10 days was significantly lower, at 2.93 ± 0.54 mg for females and 2.22 ± 0.40 mg for males (mean ± se), suggesting that the beetles cannot re-synthesize the substances soon after spray. The weight of ejected substances had no correlation with beetle body weight. Our findings represent the first detailed morphological and functional description of a chemical defense system in Cerambycidae, revealing a specialized metasternal gland and spray mechanism. The substantial but likely non-renewable defensive substances reflect an adaptive trade-off in energy allocation between reproduction and defense in this species that exhibits high fecundity but a short lifespan at the adult stage.

Host alternation is a common behavioral strategy among many herbivorous insects. The mulberry longhorn beetle, Apriona germari is a destructive wood-boring pest. The adults exhibit rhythmic host alternation between feeding and oviposition hosts. However, the temporal rhythm and regulatory mechanisms underlying this behavior remain unclear. In the present study, by observing the dynamics of the numbers of A. germari on mulberry (Morus alba, feeding host) and willow (Salix babylonica, oviposition host) trees in a cage, we first found that both females and males began to aggregate on mulberry trees at dawn. Following 18:00 at dusk, the number of females on mulberry declined sharply, whereas the number of males decreased slightly, with a greater proportion of males staying on mulberry. To investigate the role of host volatiles in regulating the host alternation in A. germari, we then carried out two-choice olfactory assays to test whether the behavioral responses of A. germari to the host volatiles from mulberry and willow twigs differed between 04:00 and 08:00 (dawn, aggregated on mulberry) and 18:00 to 22:00 (dusk, dispersed from mulberry). Males were consistently attracted to the volatiles from healthy mulberry at both dawn and dusk, but not to those from willow. Females showed no attraction to the volatiles from either of two hosts at dawn; however, at dusk, they were significantly attracted to willow volatiles. Furthermore, volatiles from feeding-damaged mulberry trees were repellent to both sexes, particularly to males at dawn. These results indicate that the rhythmic host alternation in A. germari is partly mediated by host volatiles and an unidentified male-produced pheromone may be present in this species. Our findings can deepen the understanding of the host alternation in longhorn beetles, and offer a theoretical foundation for developing semiochemical-based, eco-friendly strategies for controlling this pest.

The Asian longhorn beetle (ALB), Anoplophora glabripennis, and citrus longhorn beetle (CLB), Anoplophora chinensis, are two destructive invasive wood-boring pests, with high similarities in morphology, geographical distribution, host range, life cycle, adult behaviors and male-produced pheromone, implying a potential existence of interspecific interactions. Matings have been found to occur across females and males of the two species when manually paired in confined spaces. However, interspecific mating and its regulating factors are unclear between sympatric populations on hosts. Herein, by observing mountings and tracking the beetles that freely coexisted on host branches in cages, we found that the majority of mountings appeared within species; however, interspecific mountings occasionally occurred between male CLBs and female ALBs. The CLB was more active than the ALB at night. It seems that males actively searched for female ALBs, while the inverse was the case with CLBs. The main release periods of shared pheromone components overlapped between the two species, while compound ratios had significant differences. Our results unveil a trivial and asymmetrical interspecific mating between ALBs and CLBs, implying a risk of co-outbreaks of the two species in either native or invaded areas.