Pheromone emission patterns and courtship sequences across distinct populations within Anastrepha fraterculus (Diptera-Tephritidae) cryptic species complex

Sexual behavioural isolation can result from sexual selection and represents a relevant factor associated with the speciation process. We analysed the pheromone emission pattern and the courtship of males of five different populations of the Anastrepha fraterculus cryptic complex: Brazil (Vacaria, Tucuman and Piracicaba), Colombia and Peru. The time of pheromone emission was recorded in each population every 30 min during the day. The behavioural sequences of courting were video recorded and analysed using EthoSeq software. Males from different populations have showed different period of pheromone emission - Vacaria, Piracicaba and Tucuman executed calling only during the morning, Colombia only in the afternoon and Peru during both periods. The general frequencies of the courtship units of the males were distinct among the populations. Three groups were formed in the classification from the function of 14 behavioural routines: Vacaria, Piracicaba and Tucuman formed a single group (Brazil-1), while Colombia and Peru formed two distinct groups. In the probabilistic trees generated, the behavioural units that most contributed to the occurrence of copulation were distinct among the three groups formed: Brazil-1 (Contact, Alignment and Arrowhead-1); Colombia (Flying, Mobile, Contact and Alignment); Peru (Flying, Arrowhead-1 and Calling). Our results indicated differences in sexual behaviour that may explain the behavioural isolation found between the distinct groups in addition with the temporal isolation found between the Brazil-1 and Colombia populations. The evolutionary implications for the A. fraterculus cryptic species complex are discussed.