Evolution of larval food plant associations in Delias Hübner butterflies (Lepidoptera: Pieridae)

A review of larval food plants of the genus Delias is presented. Larvae specialize primarily on aerial‐stem and root hemiparasites (“mistletoes”) in the order Santalales. Although butterfly food plant associations have been recorded for only a small proportion of the genus (28 species or ～11%, representing 12/24 species‐groups), available data suggest that the family Loranthaceae is used most frequently (77%), followed by the Santalaceae sensu stricto (14%) and Viscaceae (8%). With the possible exception of Euphorbiaceae (1%), almost all non‐mistletoe records are considered to be erroneous and, in most cases, probably represent the mistletoe host tree on which the larvae sometimes pupate. Of the eight major clades recognized in Delias, food plants have been recorded for six of these, although the majority of records (89%) are for three clades (hyparete, belladonna, nigrina). Optimization of the larval food plant data in the context of recent phylogenetic hypotheses for both butterflies and plants revealed little evidence of cospeciation at the higher systematic levels. The most parsimonious reconstruction was an origin of larval feeding on Loranthaceae, followed by at least six independent colonizations to Santalaceae + Viscaceae. In contrast to related pierids in the Aporiina associated with mistletoes in which further shifts from aerial‐stem mistletoes to distantly related plants (e.g. host trees parasitized by mistletoes) have facilitated differentiation at the generic level, there is no firm evidence to indicate that such secondary, monomorphic shifts have evolved in Delias. However, larvae of D. henningia (pasithoe group of belladonna clade) from Palawan and Luzon, the Philippines, appear to be polymorphic, feeding on both Loranthaceae and Euphorbiaceae.     

Seasonal development and variation in abundance among four annual flight periods in a butterfly: a 20‐year study of the speckled wood (Pararge aegeria)

Insects typically spend the winter in a species‐specific diapause stage. The speckled wood butterfly, Pararge aegeria, is unique in having two alternative diapause stages, hibernating as larvae or pupae. In southern Sweden this creates a seasonal flight pattern with four annual adult flight periods: the first in May (pupal diapause), the second in June (larval diapause), and the third and fourth directly developing offspring generations in July and August, respectively. We address the raison d'être of the two diapause pathways by (1) outdoor rearing of cohorts, and (2) performing transect censuses throughout the season for 20 years. We contend that an early start of next season provides a benefit accruing to pupal diapause; conversely, a large proportion of the offspring from adults of the fourth flight peak are unable to reach the pupal stage before winter, providing a benefit accruing to larval winter diapause. The results obtained show that the two hibernation pathways are unlikely to be genetically distinct because of a strong overlap between the two offspring generations, and because sibling offspring from the third and fourth flight periods are likely to choose either of the two hibernation pathways, thereby resulting in a genetic mixing of the pathways. © 2011 The Linnean Society of London, Biological Journal of the Linnean Society, 2011, 102 , 635–649.     

Systematics, biogeography and diversification of the Indo-Australian genus Delias Hübner (Lepidoptera: Pieridae): phylogenetic evidence supports an 'out-of-Australia' origin

Abstract Two alternative hypotheses for the origin of butterflies in the Australian Region, that elements dispersed relatively recently from the Oriental Region into Australia (northern dispersal hypothesis) or descended from ancient stocks in Gondwana (southern vicariance hypothesis), were tested using methods of cladistic vicariance biogeography for the Delias group, a diverse and widespread clade in the Indo‐Australian Region. A phylogenetic hypothesis of the twenty‐four species‐groups recognized currently in Delias and its sister genus Leuciacria is inferred from molecular characters generated from the nuclear gene elongation factor‐1 alpha (EF‐1α) and the mitochondrial genes cytochrome oxidase subunits I and II (COI/COII) and NADH dehydrogenase 5 (ND5). Phylogenetic analyses based on maximum parsimony, maximum likelihood and Bayesian inference of the combined dataset (3888 bp, 1014 parsimony informative characters) confirmed the monophyly of Delias and recovered eight major lineages within the genus, informally designated the singhapura, belladonna, hyparete, chrysomelaena, eichhorni, cuningputi, belisama and nigrina clades. Species‐group relationships within these clades are, in general, concordant with current systematic arrangements based on morphology. The major discrepancies concern the placement of the aganippe, belisama and chrysomelaena groups, as well as several species‐groups endemic to mainland New Guinea. Two species (D. harpalyce (Donovan), D. messalina Arora) of uncertain group status are currently misplaced based on strong evidence of paraphyly, and are accordingly transferred to the nigrina and kummeri groups, respectively. Based on this phylogeny, a revised systematic classification is presented at the species‐group level. An historical biogeographical analysis of the Delias group revealed that the most parsimonious reconstruction is an origin in the Australian Region, with at least seven dispersal events across Wallacea to the Oriental Region. The eight major clades of Delias appear to have diverged rapidly following complete separation of the Australian plate from Gondwana and its collision with the Asian plate in the late Oligocene. Further diversification and dispersal of Delias in the Miocene–Pliocene are associated with major geological and climatic changes that occurred in Australia–New Guinea during the late Tertiary. The ‘out‐of‐Australia’ hypothesis for the Delias group supports an origin of the Aporiina in southern Gondwana (southern vicariance hypothesis), which proposes that the ancestor of Delias + Leuciacria differentiated vicariantly on the Australian plate.     

Asymmetric life-history decision-making in butterfly larvae

In temperate environments, insects appearing in several generations in the growth season typically have to decide during the larval period whether to develop into adulthood, or to postpone adult emergence until next season by entering a species-specific diapause stage. This decision is typically guided by environmental cues experienced during development. An early decision makes it possible to adjust growth rate, which would allow the growing larva to respond to time stress involved in direct development, whereas a last-minute decision would instead allow the larva to use up-to-date information about which developmental pathway is the most favourable under the current circumstances. We study the timing of the larval pathway decision-making between entering pupal winter diapause and direct development in three distantly related butterflies (Pieris napi, Araschnia levana and Pararge aegeria). We pinpoint the timing of the larval diapause decision by transferring larvae from first to last instars from long daylength (inducing direct development) to short daylength conditions (inducing diapause), and vice versa. Results show that the pathway decision is typically made in the late instars in all three species, and that the ability to switch developmental pathway late in juvenile life is conditional; larvae more freely switched from diapause to direct development than in the opposite direction. We contend that this asymmetry is influenced by the additional physiological preparations needed to survive the long and cold winter period, and that the reluctance to make a late decision to enter diapause has the potential to be a general trait among temperate insects.     

Extra molting,cannibalism and pupal diapause under unfavorable growth conditions in Atrophaneura alcinous (Lepidoptera: Papilionidae)

Atrophaneura alcinous adopt multiple strategies such as extra molting, cannibalism and pupal diapause under unfavorable growth conditions. The conditions under which these strategies are adopted have been separately verified, but their relationship has often been overlooked. We examined which strategy A. alcinous adopted and the relative advantages of strategies using four experimental groups under different food quantity and individual density conditions. Our results indicated that A. alcinous often extra‐molted, prolonged the larval period, and larval and pupal weights were lighter under food shortage. Cannibalism and disease often occurred under the high density and food shortage condition. We also showed that individuals cannibalized during the prepupal or pupal period were less likely to extra molt before they were killed. Extra molting tended to occur more frequently in females than in males under a shortage of food. In addition, we showed that, when food was insufficient, A. alcinous might initiate pupal diapause under low densities, but not under high densities. These results suggest that A. alcinous prolongs the larval period by engaging in extra molting during times of unfavorable food conditions. This strategy might decrease the risk of cannibalization because surrounding larvae who could potentially cannibalize others pupate before larvae who prolong the larval period pupate. Our results also suggest that diapausing pupae under a shortage of food can only survive when there are few surrounding conspecifics, due to a lower cannibalization risk. In conclusion, there is a complex interaction between extra molting, cannibalism and pupal diapause strategies.     

Distribution and phylogenetic relationships of Australian glow-worms Arachnocampa (Diptera, Keroplatidae)

Glow-worms are bioluminescent fly larvae (Order Diptera, genus Arachnocampa) found only in Australia and New Zealand. Their core habitat is rainforest gullies and wet caves. Eight species are present in Australia; five of them have been recently described. The geographic distribution of species in Australia encompasses the montane regions of the eastern Australian coastline from the Wet Tropics region of northern Queensland to the cool temperate and montane rainforests of southern Australia and Tasmania. Phylogenetic trees based upon partial sequences of the mitochondrial genes cytochrome oxidase II and 16S mtDNA show that populations tend to be clustered into allopatric geographic groups showing overall concordance with the known species distributions. The deepest division is between the cool-adapted southern subgenus, Lucifera, and the more widespread subgenus, Campara. Lucifera comprises the sister groups, A. tasmaniensis, from Tasmania and the newly described species, A. buffaloensis, found in a high-altitude cave at Mt Buffalo in the Australian Alps in Victoria. The remaining Australian glow-worms in subgenus Campara are distributed in a swathe of geographic clusters that extend from the Wet Tropics in northern Queensland to the temperate forests of southern Victoria. Samples from caves and rainforests within any one geographic location tended to cluster together within a clade. We suggest that the morphological differences between hypogean (cave) and epigean (surface) glow-worm larvae are facultative adaptations to local microclimatic conditions rather than due to the presence of cryptic species in caves.     

Preventing insect diapause with hormones and cholera toxin

The decision to enter pupal diapause can be reversed in the flesh fly $\text{Sarcophaga crassipalpis}$ by treatment with a juvenile hormone analogue, ecdysterone, or cholera toxin. A topical application of juvenile hormone analogue (10μg) is effective in preventing diapause if applied to newly ecdysed 3rd instar larvae. An ecdysterone injection (1μg) into post-fed 3rd instar larvae can avert diapause, and even lower dosages (0.01μg) are effective if injected into young pupae immediately before the onset of diapause. Cholera toxin, a stimulant of adenylate cyclase, can prevent diapause if 1μg is injected into larvae 24 hrs. prior to pupariation. This is the first report of an effect of cholera toxin on insects. A 2-day exposure to 33°C was the only physical manipulation shown to be capable of reversing the previous commitment to enter pupal diapause.     

Müllerian mimetic radiation of Delias butterflies (Lepidoptera: Pieridae) in Bali and Timor

Mimicry rings are present among Delias butterflies, and those butterflies are also considered to be mimetic models of other lepidopteran insects; however, experimental evidence for their unpalatability to predators is limited. In Bali and Timor, a total of three mimicry rings of Delias species are present; particularly, male and female D. lemoulti join different rings in Timor. The present study examined the unpalatability of Delias in Bali and Timor to the caged avian predator Pycnonotus aurigaster. The birds ate eight Delias species in similar numbers, and ate the palatable butterfly Mycalesis horsfieldii much more frequently than Delias butterflies. The result suggests that the three mimicry rings of Delias species in Bali and Timor are Müllerian rather than Batesian. Based on previous findings on their phylogenetic relationships, the Müllerian mimicry rings of Delias in Bali and Timor are suggested to have emerged through the convergent evolution and phylogenetic constraints of wing color patterns. In the D. hyparete species group, mimetic radiation may have occurred between Bali and Timor.     

Regulation of heat shock proteins in the apple maggot Rhagoletis pomonella during hot summer days and overwintering diapause

Abstract Developing larvae of the apple maggot Rhagoletis pomonella are frequently exposed to summertime apple temperatures that exceed 40 °C and, during their overwintering diapause, pupae are exposed to sub‐zero soil temperatures for prolonged periods. To investigate the potential involvement of heat shock proteins (Hsps) in response to these environmental extremes, the genes encoding Hsp70 and Hsp90 in R. pomonella are cloned and expression monitored during larval feeding within the apple and during overwintering pupal diapause. Larvae reared in the laboratory at constant temperatures of 25, 28 or 35 °C express Hsp90 but very little Hsp70. Larvae do not survive rearing at 40 °C. The temperature cycles to which larvae were exposed inside apples in the field, ranging 16–46.9 °C over a 24‐h period, elicit strong Hsp70 and Hsp90 expression, which begins at mid‐day and reaches a peak in late afternoon, coinciding with peak air and apple temperatures. Heat shock proteins are also expressed strongly by pupae during their overwintering diapause. Hsp70 is not expressed in nondiapausing pupae but is highly expressed throughout diapause. Hsp90 is constitutively expressed in both diapausing and nondiapausing pupae. Rhagoletis pomonella thus strongly expresses its Hsps during pupal diapause, presumably as a protection against low temperature injury, and during larval development to cope with natural temperature cycles prevailing in late summer.