Super Cooling Point Phenotypes and Cold Resistance in Hyles euphorbiae Hawk Moths from Different Climate Zones

The spurge hawkmoth Hyles euphorbiae L. (Sphingidae) comprises a remarkable species complex with still not fully resolved taxonomy. Its extensive natural distribution range covers diverse climatic zones. This predestinates particular populations to cope with different local seasonally unfavorable environmental conditions. The ability of the pupae to overcome outer frosty conditions is well known. However, the differences between two main ecotypes (‘euphorbiae’ and ‘tithymali’) in terms of the inherent degree of frost tolerance, its corresponding survival strategy, and underlying mechanism have not been studied in detail so far. The main aim of our study was to test the phenotypic exhibition of pupae (as the relevant life cycle stadia to outlast unfavorable conditions) in response to combined effects of exogenous stimuli, such as daylight length and cooling regime. Namely, we tested the turnout of subitan (with fast development, unadapted to unfavorable conditions) or diapause (paused development, adapted to unfavorable external influences and increased resistance) pupae under different conditions, as well as their mortality, and we measured the super cooling point (SCP) of whole pupae (in vivo) and pupal hemolymph (in vitro) as phenotypic indicators of cold acclimation. Our results show higher cold sensitivity in ‘tithymali’ populations, exhibiting rather opportunistic and short-termed cold hardiness, while ‘euphorbiae’ produces a phenotype of seasonal cold-hardy diapause pupae under a combined effect of short daylight length and continuous cold treatment. Further differences include the variability in duration and mortality of diapause pupae. This suggests different pre-adaptations to seasonal environmental conditions in each ecotype and may indicate a state of incipient speciation within the H. euphorbiae complex.

Morphological evolution in Hyles Hübner, 1819 hawkmoths (Lepidoptera, Sphingidae): reconstructing the ancestral Hyles habitus

Molecular phylogenetic studies suggest that similar wing and body patterns in the hawkmoth genus Hyles Hübner, [1819] do not necessarily reflect a close phylogenetic relationship. To improve our understanding of morphological evolution in these organisms, 75 characters derived from the external adult morphology are explicitly coded and analysed in a maximum parsimony cladistic framework. The results corroborate the hypothesis that wing and body patterns have indeed reappeared in different parts of the phylogeny but the underlying genetic mechanism remains to be determined. By reconstructing the suite of ancestral states of the morphological characters using Bayesian inference, we derived an approximation of the appearance of the proto-Hyles species. The overall habitus of this moth does not display a combination of characters found in any extant Hyles species. Rather, the forewings are most like those of members of the Hyles euphorbiae-complex but with better developed antemedial and postmedial lines, the hindwings are typical Hyles, and the abdominal pattern most closely resembles that of Hyles euphorbiarum (Guérin-Méneville & Percheron, 1835), but with one fewer pairs of black subdorsal patches. Within the context of the subtribe Choerocampina and Sphingidae more generally, the proto-Hyles reconstruction does not resemble any other species apart from Rhodafra opheltes (Cramer, 1780), but this appears to be another instance of convergent pattern expression.

Genome-wide SNP Data Reveal an Overestimation of Species Diversity in a Group of Hawkmoths

The interface between populations and evolving young species continues to generate much contemporary debate in systematics depending on the species concept(s) applied but which ultimately reduces to the fundamental question of “when do nondiscrete entities become distinct, mutually exclusive evolutionary units”? Species are perceived as critical biological entities, and the discovery and naming of new species is perceived by many authors as a major research aim for assessing current biodiversity before much of it becomes extinct. However, less attention is given to determining whether these names represent valid biological entities because this is perceived as both a laborious chore and an undesirable research outcome. The charismatic spurge hawkmoths (Hyles euphorbiae complex, HEC) offer an opportunity to study this less fashionable aspect of systematics. To elucidate this intriguing systematic challenge, we analyzed over 10,000 ddRAD single nucleotide polymorphisms from 62 individuals using coalescent-based and population genomic methodology. These genome-wide data reveal a clear overestimation of (sub)species-level diversity and demonstrate that the HEC taxonomy has been seriously oversplit. We conclude that only one valid species name should be retained for the entire HEC, namely Hyles euphorbiae, and we do not recognize any formal subspecies or other taxonomic subdivisions within it. Although the adoption of genetic tools has frequently revealed morphologically cryptic diversity, the converse, taxonomic oversplitting of species, is generally (and wrongly in our opinion) accepted as rare. Furthermore, taxonomic oversplitting is most likely to have taken place in intensively studied popular and charismatic organisms such as the HEC.

Larval pattern morphotypes in the Western Palaearctic Hyles euphorbiae complex (Lepidoptera: Sphingidae: Macroglossinae)

External morphological variation in larval pattern elements in the Hyles euphorbiae complex is documented and described from independent samples of numerous populations across Europe and North Africa. Variability in the distribution of black cuticle and other colours was found to be much higher within populations than previously believed and it proved difficult to characterize any species, subspecies and even local population unequivocally using this feature alone. A representative sample of larvae from the Canary Islands, Madeira, North Africa, Iberia, Italy, Germany, Eastern Europe, Malta, Greece and Yemen, is illustrated, supplemented by observations from photographs of larvae from the Middle East and Western Asia. Taxonomy-independent definitions of larval morphotypes demonstrate significant overlap between species. We discuss the results in the context of Western Palaearctic biogeography, postulating several areas of hybridisation between the more northern Hyles euphorbiae and more southern H. tithymali that have led to the mosaic distribution patterns observed, and consider the potential of these moths for tracking the future eff ects of climate change in the Mediterranean Basin.