Speciation problems in the Papilio machaon group of butterflies (Lepidoptera: Papilionidae)

Abstract. The taxonomic and genetic relationships between P.machaon rathjensi Warnecke from the west side of southern Arabia and P.machaon muetingi Seyer from the east are discussed, together with suggestions of their relationship to P.machaon saharae Oberthiir from the northern Hejaz, Sinai and desert North Africa. It is concluded that though P.m.rathjensi and P.m.saharae are subspecifically distinct, they are very closely related to each other, and both should be regarded as specifically distinct from the rest of the machaon complex.
The remainder of the machaon complex in its turn presents a somewhat similar problem. Thus P.machaon gorganus Fruhstorfer and P.hospiton Géné are normally regarded as good species, but Fl caterpillars have been found in the wild and the cross can be readily produced in captivity with no upset in the sex ratio. Furthermore, recently we have obtained sparse F2 offspring between the two species.
We therefore suggest that the entire P.machaon complex is in a labile state as regards speciation throughout its range and the relationships between the various forms are not easily expressed through the traditional species and subspecies concepts. Our tentative conclusions are that the status of P.machaon and P.hospiton as distinct species probably remains valid. The saharaelrathjensi complex might be considered distinct from P.machaon using the biological species concept, while their genetics indicate a more conservative approach. However, more important than the exact status of the taxa in question is the fact that the P.machaon complex illustrates evoluton in action, the end result of which cannot be predicted.     

Towards resolving the identities of the Graphium butterflies (Lepidoptera: Papilionidae) of Peninsular Malaysia

Graphium butterflies are famous in Peninsular Malaysia for their colourful wings, yet their taxonomy remains unresolved. The popular guides to Malaysian butterflies, place the species in one, two or three genera and identification to species using obscure morphological characters can be difficult, especially for the closely related species, G. bathycles bathycloides and G. chironides malayanum. We sequenced the COI mtDNA barcode for Graphium specimens in the Museum of Zoology, University of Malaya to test the utility of DNA barcoding for the identification of Graphium species. Additionally, we sequenced 28S rRNA to examine, in conjunction with COI, the phylogenetic relationships of these species and investigate the validity of Pathysa and Paranticopsis as distinct genera. We found that all species of Graphium possessed a distinctive cluster of DNA barcodes with the exception of specimens originally identified as G. bathycles bathycloides and G. chironides malayanum which shared DNA barcodes. On further examination we found that the morphological determinations were ambiguous as the specimens overlapped for diagnostic characters reported for each species. The COI and 28S rRNA phylogenetic trees showed a similar topology with Paranticopsis species forming a clade nested within a larger clade also comprising Pathysa species. Based on this topology, in order for Pathysa to be a valid genus, at least three other clades within Graphium s.l. would also have to be raised as genera.     

Molecular phylogeny of swallowtail butterflies of the tribe Papilionini (Papilionidae, Lepidoptera).

Swallowtail butterflies of the tribe Papilionini number about 225 species and are currently used as model organisms in several research areas, including genetics, chemical ecology and phylogenetics of host plant utilization and mimicry, mechanisms of speciation, and conservation. We have inferred phylogenetic relationships for a sample of 18 species of the genus Papilio (sensu lato) and five outgroup taxa by sequencing two stretches of mitochondrial DNA that correspond to segments 12886-13370 and 12083-12545 of Drosophila melanogaster mitochondrial DNA and consist of sections of the genes for the large ribosomal RNA and subunit 1 of NADH-dehydrogenase. Our data support the monophyly of Papilio and, within it, of several traditionally recognized subgroups. Species belonging to groups that utilize primarily Rutaceae as larval foodplants form two clusters, corresponding to Old World and American taxa, respectively, while two previously recognized clades-of American and South Asian-Austronesian origin-whose members were known to feed mostly on Lauraceae and Magnoliaceae, are observed to form a clade. The sister group of Papilio is found to be the South Asian genus Meandrusa, which also happens to feed on Lauraceae. The latter plant family is therefore the probable larval host of the ancestor Papilio and the shift to Rutaceae (which four-fifths of extant Papilio species use as foodplants) is more likely to have occurred only after the initial diversification of the genus.     

Molecular phylogeny, historical biogeography, and divergence time estimates for swallowtail butterflies of the genus Papilio (Lepidoptera: Papilionidae)

Swallowtail butterflies are recognized as model organisms in ecology, evolutionary biology, genetics, and conservation biology but present numerous unresolved phylogenetic problems. We inferred phylogenetic relationships for 51 of about 205 species of the genus Papilio (sensu lato) from 3.3-Kilobase (kb) sequences of mitochondrial and nuclear DNA (2.3 kb of cytochrome oxidases I and II and 1.0 kb of elongation factor 1 alpha). Congruent phylogenetic trees were recovered within Papilio from analyses of combined data using maximum likelihood, Bayesian analysis, and maximum parsimony bootstrap consensus. Several disagreements with the traditional classification of Papilio were found. Five major previously hypothesized subdivisions within Papilio were well supported: Heraclides, Pterourus, Chilasa, Papilio (sensu stricto), and Eleppone. Further studies are required to clarify relationships within traditional "Princeps," which was paraphyletic. Several biologically interesting characteristics of Papilio appear to have polyphyletic origins, including mimetic adults, larval host associations, and larval morphology. Early diversification within Papilio is estimated at 55-65 million years ago based on a combination of biogeographic time constraints rather than fossils. This divergence time suggests that Papilio has slower apparent substitution rates than do Drosophila and fig-pollinating wasps and/or divergences corrected using best-fit substitution models are still being consistently underestimated. The amount of sequence divergence between Papilio subdivisions is equivalent to divergences between genera in other tribes of the Papilionidae, and between genera of moths of the noctuid subfamily Heliothinae.     

The proximate control of pupal color in swallowtail butterflies: implications for the evolution of environmentally cued pupal color in butterflies (Lepidoptera: Papilionidae)

The environmentally cued production of cryptic green/yellow or brown/melanized pupae is widespread in butterflies, occurring in the Nymphalidae, Pieridae, and the Papilionidae subfamily Papilioninae. The dimorphism is controlled by the hormone pupal melanization reducing factor (PMRF). In the nymphalid Inachis io dibutryl cAMP mimics PMRF, and inhibits pupal melanization. However, in the papilionid Papilio polyxenes PMRF stimulates browning, suggesting that the control of pupal color by PMRF has evolved independently in the swallowtail and nymphalid-pierid lineages. We examined this hypothesis by using ligatures to prevent hormone release in five species representing three Papilioninae tribes. One species, Papilio glaucus, produces only brown pupae. Ligatures resulted in green cuticle posterior to the ligature in all five swallowtail species, including P. glaucus, suggesting that the mode of action of PMRF is the same in the three tribes. We also found that in P. polyxenes injections of dibutryl cAMP into prepupal larvae mimic the effect of PMRF, by causing dose-dependent pupal browning. Our results support the hypothesis that the control of pupal color by PMRF has evolved independently in the two lineages. The observation that green pupal color can be induced in P. glaucus by ligature indicates that environmentally cued pupal color could evolve by facultative inhibition of PMRF release.     

Population dynamics ofLuehdorfia japonica Leech (Lepidoptera: Papilionidae)

1. An adult population of a papilionid butterfly, Luehdorfia japonicaLeech , was studied by marking, release and recapture procedures in a hilly region in the suburbs of Kanazawa City, Japan.
2. Age of butterflies was estimated from the wing wear conditions, rated as winage categories 0 to 6.
3. Jolly (1965) andSeber's (1973) method was applied to the marking-recapture results for estimating the population parameters (sampling ratio, population size and survival rate).
4. Sampling ratio of males was consistently higher (around 50%) than that of females.
5. Newly emerged females were especially inactive, so that few of them were captured. From day 6.5 to day 10.0 they began to oviposit and became more active and more catchable.
6. An approximate sex ratio of 1∶1 was confirmed from the specimens collected in the field and by rearing experiments.
7. Daily survival rate was about 0.75–0.80 and mean longevity was about 4 days for both sexes. The maximum longevity observed was 17 days, for males and 21 days for females.
8. Dispersal by both sexes of the butterfly was more than 1 km.

     

Population dynamics ofLuehdorfia japonica Leech (Lepidoptera: Papilionidae)

Mortality processes from egg to final instar larval stage are examined in the papilionid butterfly Luehdorfia japonica for two generations in a natural populations. Special attention is given to the effects of initial egg cluster size on the survival rate by the 3rd instar. Mean egg cluster sizes for the two generations were 11.3 and 10.7. The hatching rate was not affected by the egg cluster size. Up to 3rd instar, larvae in a group were more likely to die en masse rather than to die individually, whereas most larvae in later instars died independently. The egg clusters of the average (and the most frequent) size class (11±1) had the second highest survival rate until the 3rd instar. A slightly higher survival rate occurred in clusters a little larger (14±1). The average size clusters were least likely to be exterminated and about 70% of them produced at least one individual surviving to the 3rd instar. Factors affecting the observed mortality patterns were discussed.     

Pair-clinging behavior by males ofAtrophaneura alcinous (Lepidoptera: Papilionidae)

Females ofAtrophaneura alcinous usually mate soon after eclosion. Their ostium bursae becomes plugged with male secretion which reduces chances of remating. Males frequently cling to a copulating pair and wait for completion of copulation. This was observed in 66% of 198 copulating pairs, with a maximum of 5 males clinging at one time during the course of a copulation. Males clinging for longer periods were more successful in copulation with the freshly mated female than those clinging for shorter periods. Despite the plugging effect, females may mate more than once. Clinging males were responsible for 61% of re-copulations and 53% of re-inseminations. Clinging behavior may be regarded as an effective male mating strategy to exploit freshly mated females, and an alternative to finding virgin females.     

Comparative processing of allelochemicals in the Papilionidae (Lepidoptera)

Within the family Papilionidae (Lepidoptera), species display a broad range of feeding patterns, from oligophagy on a single hostplant family to polyphagy on over a dozen families. Accompanying this diversity of feeding strategies is a diversity of physiological mechanisms for processing hostplant allelochemicals. Studies on members of this family as well as other Lepidoptera suggest that oligophagy is associated with high activity, in addition to high substrate specificity, of detoxicative enzymes.     

Phylogenetic relationships of three African swallowtail butterflies,Papilio dardanus,P.phorcas and P.constantinus: A cladistic analysis (Lepidoptera: Papilionidae)

Abstract. The well-known polymorphic mimetic African papilionid butterfly, Papilio dardanus Brown, is demonstrated by cladistic analysis to form a monophyletic group (the phorcas-group) with two other African swallowtails, P.phorcas Cramer and P.constantinus Ward. Within this group, dardanus and phorcas are shown to be sister-species, with constantinus as the stem. The supposed relationship of this group of three Rutaceae-feeding species to the five members of the Canellaceae/Lauraceae-feeding Papilio hesperus-group (including P.nobilis Rogenhofer) is questioned. The significance of this analysis is discussed with special reference to Trimen's original hypothesis on the evolution of mimetic patterns in female Papilio dardanus, the unique male patterns of phorcas and dardanus, and natural and laboratory-produced hybrids. A pathway modelling technique is developed to represent the possible course of evolution of the phorcas-group, in terms of speciation events and the accumulation and decay of the pattern polymorphisms.     

Biogeography and systematics of Aricia butterflies (Lepidoptera,Lycaenidae)

Butterflies of the Aricia species group represent a paradigm of unresolved taxonomy, both at the genus and species levels. We studied phylogenetic relationships, biogeography, and systematics based on genetic – nuclear and mitochondrial – and morphometric – external (wings) and internal (genitalia) – data. We show that Aricia is a monophyletic genus comprising the taxa Pseudoaricia, Ultraaricia and Umpria, which are here considered junior synonyms of Aricia. The taxa allous, inhonora, issekutzi, mandzhuriana, myrmecias and transalaica, which have often been raised to species rank, are shown to probably represent subspecies or synonyms. We show that montensis is likely a good species that is sister to all A. artaxerxes populations across the Palearctic region. The species A. anteros and A. morronensis are shown to display deep intraspecific divergences and they may harbor cryptic species. We also discovered that A. cramera and A. agestis exhibit a pattern of mutual exclusion on islands, and a parapatric distribution in mainland with a narrow contact zone where potential hybrids were detected. The lack of a prezygotic barrier that prevents their coexistence could explain this phenomenon. This study will hopefully contribute to the stability of the systematics of Aricia, a group with potential for the study of the link between speciation and biogeography.     

Phylogenetic relationships of three African swallowtail butterflies,Papilio dardanus,P.phorcas and P.constantinus: New data from hybrids (Lepidoptera: Papilionidae)

Abstract. Various laboratory-bred hybrids between Papilio dardanus, P.phorcas and P.constantinus are reported. The principal object of the research was to obtain evidence regarding the evolutionary relationships of the three species, with particular reference to understanding the evolution of the mimetic patterns of P.dardanus. With regard to the latter, little progress was made but other findings were interesting, notably those concerning sex ratio, pupal coloration, and differences in male genitalia.     

Mobility and oviposition site-selection in Zerynthia cassandra (Lepidoptera,Papilionidae): implications for its conservation

The adults’ mobility and oviposition preferences of Zerynthia cassandra have been studied for the first time, with the aim of integrating auto-ecological information into recommendations for the habitat’s management of this species. Results of our mark-release-recapture study have highlighted that Z. cassandra is a strictly sedentary species, since detected movements only occurred over very short distances (≤200 m) and mainly within the species’ reproductive habitat (i.e. around Aristolochia rotunda stands), with males moving further than females. Our study shows that the main oviposition habitat of Z. cassandra is found where A. rotunda plants are growing in large stands; sites where plants growing in half to full sun and mostly oriented to the south are preferred. The distance of deposited eggs from the plants’ roots was narrowly correlated with the plants’ length. Eggs were deposited singly, mainly on the underside of leaflets. Management strategies necessary for improving the most important habitat features for the conservation of this species are suggested.     

Nectar sugar and amino acid preferences of Battus philenor (Lepidoptera, Papilionidae)

Abstract.

• 1 Butterflies of Battus philenor were tested for their preferences for nectar sugars and amino acids in an outdoor cage experiment.
• 2 The butterflies clearly preferred both sucrose and fructose over glucose. They also preferred sucrose over fructose.
• 3 No other preferences were found to be statistically significant, although male butterflies tended to prefer a plain sugar solution over a sugar solution containing a mixture of amino acids: females consumed both of these solutions in almost equal proportions.
• 4 The results are discussed with respect to nectar composition of butterfly pollinated flowers, flower preferences, physiological and reproductive aspects of butterflies.

     

Annotated checklist of Albanian butterflies (Lepidoptera,Papilionoidea and Hesperioidea)

The Republic of Albania has a rich diversity of flora and fauna. However, due to its political isolation, it has never been studied in great depth, and consequently, the existing list of butterfly species is outdated and in need of radical amendment. In addition to our personal data, we have studied the available literature, and can report a total of 196 butterfly species recorded from the country. For some of the species in the list we have given explanations for their inclusion and made other annotations. Doubtful records have been removed from the list, and changes in taxonomy have been updated and discussed separately. The purpose of our paper is to remove confusion and conflict regarding published records. However, the revised checklist should not be considered complete: it represents a starting point for further research.     

A partitioned likelihood analysis of swallowtail butterfly phylogeny (Lepidoptera:Papilionidae)

Although it is widely agreed that data from multiple sources are necessary to confidently resolve phylogenetic relationships, procedures for accommodating and incorporating heterogeneity in such data remain underdeveloped. We explored the use of partitioned, model-based analyses of heterogeneous molecular data in the context of a phylogenetic study of swallowtail butterflies (Lepidoptera: Papilionidae). Despite substantial basic and applied study, phylogenetic relationships among the major lineages of this prominent group remain contentious. We sequenced 3.3 kb of mitochondrial and nuclear DNA (2.3 kb of cytochrome oxidase I and II and 1.0 kb of elongation factor-1 alpha, respectively) from 22 swallowtails, including representatives of Baroniinae, Parnassiinae, and Papilioninae, and from several moth and butterfly outgroups. Using parsimony, we encountered considerable difficulty in resolving the deepest splits among these taxa. We therefore chose two outgroups with undisputed relationships to each other and to Papilionidae and undertook detailed likelihood analyses of alternative topologies. Following from previous studies that have demonstrated substantial heterogeneity in the evolutionary dynamics among process partitions of these genes, we estimated evolutionary parameters separately for gene-based and codon-based partitions. These values were then used as the basis for examining the likelihoods of possible resolutions and rootings under several partitioned and unpartitioned likelihood models. Partitioned models gave markedly better fits to the data than did unpartitioned models and supported different topologies. However, the most likely topology varied from model to model. The most likely ingroup topology under the best-fitting, six-partition GTR + gamma model favors a paraphyletic Parnassiinae. However, when examining the likelihoods of alternative rootings of this tree relative to rootings of the classical hypothesis, two rootings of the latter emerge as most likely. Of these two, the most likely rooting is within the Papilioninae, although a rooting between Baronia and the remaining Papilionidae is only nonsignificantly less likely.