Factors to Male-Female Sex Approaches and the Identification of Volatiles and Compounds from the Terminalia of Proholopterus chilensis (Blanchard) (Coleoptera: Cerambycidae) Females in Nothofagus obliqua (Mirb.) Oerst. (Nothofagaceae) Forests in Chile

Abstract

During the spring–summer seasons between 2019 and 2023, in the localities of Maquehue (La Araucanía Region) and Llifén (Los Ríos Region), we collected 262 virgin Proholopterus chilensis (1 female/2.3 males) specimens emerging from the live trunks of N. obliqua trees, an atypical sex ratio in Cerambycidae, suggesting high male competition for females. Virgin specimens of both sexes were individually placed in panel traps in the field, capturing only males (n = 184) over the field study seasons and only in traps baited with females. This fact preliminarily suggests the P. chilensis females emit possible “volatile or airborne” pheromones (eventually being a long-range sex pheromone), something unusual in Cerambycinae, the subfamily to which it currently belongs, although the taxonomic status of the species is under debate. In Llifén and Santiago (Metropolitan Region), behavioral observations were conducted, which allowed us to define the conditions necessary for male–female encounters that were replicated when carrying out volatile captures (Head Space Dynamic = HSD) and collections of compounds from terminalias excised from females. In field trials, virgin females less than ten days old were more attractive than older ones and attracted males during the night, i.e., between 23:00 and 5:00 AM, when the ambient temperature exceeded 11.6 °C. The aeration of females under the conditions described above and subsequent analysis of extracts by GC-MS allowed the identification of compounds absent in males and the control, including two oxygenated sesquiterpenes, a nitrogenous compound (C20), and a long-chain hydrocarbon (C26). From the terminalia extracts, hentriacontane, heptacosane, and heneicosyl, heptacosyl and docosyl acetates were identified by GC-MS, and their roles are unknown in the development of short-distance sexual behaviors, but they could serve to mark a trail leading the male towards the female in the final stage of approach for courtship/mating. Thus, we proposed the hypothesis that P. chilensis females emit both a long-range and a trail-pheromone, which, if confirmed, would be a rare case in this family. The specific identity of the compounds obtained by HSD, as well as the activities of these chemicals and those obtained from the terminalias, should be evaluated in future behavioral studies, as well as regarding their potential to attract males under field conditions. The current document is the first report on volatiles obtained from aerations and compounds extracted from female terminalia in P. chilensis.