The corn- and rice-strains of Spodoptera frugiperda exhibit several genetic and behavioral differences and appear to be undergoing ecological speciation in sympatry. Previous studies reported conflicting results when investigating male attraction to pheromone lures in different regions, but this could have been due to inter-strain and/or geographic differences. Therefore, we investigated whether corn- and rice-strain males differed in their response to different synthetic pheromone blends in different regions in North America, the Caribbean and South America. All trapped males were strain-typed by two strain-specific mitochondrial DNA markers. In the first experiment, we found a nearly similar response of corn- and rice-strain males to two different 4-component blends, resembling the corn- and rice-strain female blend we previously described from females in Florida. This response showed some geographic variation in fields in Canada, North Carolina, Florida, Puerto Rico, and South America (Peru, Argentina). In dose-response experiments with the critical secondary sex pheromone component (Z)-7-dodecenyl acetate (Z7-12:OAc), we found some strain-specific differences in male attraction. While the response to Z7-12:OAc varied geographically in the corn-strain, rice-strain males showed almost no variation. We also found that the minor compound (Z)-11-hexadecenyl acetate (Z11-16:OAc) did not increase attraction of both strains in Florida and of corn-strain males in Peru. In a fourth experiment, where we added the stereo-isomer of the critical sex pheromone component, (E)-7-dodecenyl acetate, to the major pheromone component (Z)-9-tetradecenyl acetate (Z9-14:OAc), we found that this compound was attractive to males in North Carolina, but not to males in Peru. Overall, our results suggest that both strains show rather geographic than strain-specific differences in their response to pheromone lures, and that regional sexual communication differences might cause geographic differentiation between populations.

Very little is known on how changes in circadian rhythms evolve. The noctuid moth Spodoptera frugiperda (Lepidoptera: Noctuidae) consists of two strains that exhibit allochronic differentiation in their mating time, which acts as a premating isolation barrier between the strains. We investigated the genetic basis of the strain-specific timing differences to identify the molecular mechanisms of differentiation in circadian rhythms. Through QTL analyses we identified one major Quantitative trait chromosome (QTC) underlying differentiation in circadian timing of mating activity. Using RADtags, we identified this QTC to be homologous to Bombyx mori C27, on which the clock gene vrille is located, which thus became the major candidate gene. In S. frugiperda, vrille showed strain-specific polymorphisms. Also, vrille expression differed significantly between the strains, with the rice-strain showing higher expression levels than the corn-strain. In addition, RT-qPCR experiments with the other main clock genes showed that pdp1, antagonist of vrille in the modulatory feedback loop of the circadian clock, showed higher expression levels in the rice-strain than in the corn-strain. Together, our results indicate that the allochronic differentiation in the two strains of S. frugiperda is associated with differential transcription of vrille or a cis-acting gene close to vrille, which contributes to the evolution of prezygotic isolation in S. frugiperda.

The pest species Spodoptera frugiperda, which is native to North and South America, has invaded Africa in 2016. The species consists of two strains, the corn-strain and rice-strain, which differ in their sexual communication. When we investigated populations from Benin and Nigeria, consisting of corn-strain and rice-corn-hybrid descendants, we found no strain-specific sexual communication differences. Both genotypes exhibited the same pheromone composition, consisting of around 97% (Z)-9-tetradecenyl acetate (Z9–14:Ac), 2% (Z)-7-dodecenyl acetate (Z7–12:Ac), and 1% (Z)-9-dodecenyl acetate (Z9–12:Ac), they had similar electrophysiological responses, and all mated around three hours into scotophase. However, we found geographic variation between African and American populations. The sex pheromone of African corn-strain and hybrid descendant females was similar to American rice-strain females and showed higher percentages of the male-attracting minor component Z7–12:Ac. In addition, African males exhibited the highest antennal sensitivity towards Z7–12:Ac, while American males showed highest sensitivity towards the major pheromone component Z9–14:Ac. Increasing the production of and response to the critical minor component Z7–12:Ac may reduce communication interference with other African Spodoptera species that share the same major pheromone component. The implications of our results on pheromone-based pest management strategies are discussed.