Costs of Reproduction in Male Bicyclus anynana and Pieris napi Butterflies: Effects of Mating History and Food Limitation

Life history theory predicts trade‐offs arising from the costs of reproduction: individuals investing more into reproduction should have less energy available for their own somatic demands. Despite a wealth of research on such costs for females, very few studies have looked at the male side. We investigated the costs of reproduction in males of two butterfly species, Bicyclus anynana and Pieris napi. The males of these species differ tremendously in their allocation of nutrients to reproduction: virgin male spermatophores were 18.5 times larger in the latter compared with those in the former. Based on this striking difference, we expected to find a longevity cost of mating in P. napi, but not necessarily in B. anynana. In line with our predictions, we found no evidence for a survival cost of mating in B. anynana, while there was some evidence for such a cost in P. napi. Here, virgin males lived longer than once‐ or twice‐mated ones. However, this overall trend did not hold for all treatment groups, and there was no linear relationship between the number of copulations and longevity. Adult food limitation reduced longevity throughout, but had otherwise very little impact. Spermatophore dry mass decreased in later compared with first spermatophores, while water content increased, indicating that spermatophore production is indeed physiologically costly. Additionally, adult food limitation tended to reduce spermatophore fresh and dry mass, and tended to increase spermatophore water content. Based on our results we tentatively conclude that longevity and reproduction may not always be traded off against each other, as in our study organisms’ longevity seems to depend in first place on adult carbohydrate income, while male reproduction seems to be primarily (although not entirely) fuelled by larval storage reserves.     

Flight endurance in relation to adult age in the green‐veined white butterfly Pieris napi

• 1 The flight apparatus in butterflies, as well as in other insects, is costly to manufacture. Since most animals live in a world where resources are limited, trade‐offs are expected and available resources must thus be allocated between flight and other functions, such as reproduction.
• 2 To mitigate this trade‐off, previous studies have shown that butterflies can break down flight muscles in the thorax as they age in order to use muscle nutrients for reproduction.
• 3 Although breakdown of flight muscles is expected to reduce flight ability, relative flight muscle mass (thorax mass/body mass) in many butterfly species does not decrease with age. The aim of the present study was to test the relationship between flight endurance and adult age in the green‐veined white butterfly Pieris napi (L.). The tests were performed in the laboratory at five different temperatures.
• 4 The results showed that age has a significant influence on butterfly flight endurance; older butterflies showed reduced flight endurance. Male butterflies fly for a longer time than females and flight endurance increases with temperature in both sexes.

     

Host preference variation cannot explain microhabitat differentiation among sympatric Pieris napi and Pieris rapae butterflies

1. Often, closely related insect species feed on different host plant species, and the tremendous diversity of phytophagous insects is therefore attributed to host plant‐driven speciation. However, for most taxa, host use information comes from field observations of egg‐laying females or feeding caterpillars, which means that the underlying reason for a particular host‐affiliation is not easily determined. 2. Therefore, it is often unclear whether an insect feeds on a certain host because it prefers that plant to alternative hosts, or because the host distribution overlaps with the habitat requirements of the insect. 3. We ask to what extent a divergent host use in the field mirrors the host plant preferences of two closely related butterflies, Pieris napi and Pieris rapae (Pieridae). In nature, P. napi typically occurs in moister habitats than P. rapae. 4. We scanned several microhabitats at a field site in Southern Sweden during multiple years, and collected Pieris eggs from three different plants, Cardamine pratensis (wet meadows), Barbarea vulgaris (drier micro‐habitats) and Alliaria petiolata (intermediate areas). 5. As predicted, P. rapae eggs were more common than P. napi eggs on B. vulgaris, whereas all of the 358 individuals collected from C. pratensis were P. napi, indicating a divergence in host use between the Pieris species. However, under controlled laboratory conditions, both species had virtually identical oviposition preferences, laying eggs on all three plants, notably P. rapae also laying eggs on C. pratensis, indicating that habitat use, not plant preference, drives host plant use in nature.     

Mating frequency influences nectar amino acid preference of Pieris napi

It is generally assumed that butterflies, as is the case with many holometabolous insects, rely primarily on nutrients gathered by larval feeding for somatic maintenance and fecundity. These reserves can be supplemented by adult feeding and in some cases by nuptial gifts passed from the males to the females during mating. Recent findings indicate that female butterflies detect and prefer nectar with high levels of amino acids, thus calling new attention to this nutritive source. Polyandrous species can further supplement their larval stores with additional nuptial gifts. This study examined how mating frequency of the polyandrous butterfly Pieris napi affects the female's preference for nectar amino acids. Females of this species generally detect and prefer nectar mimics containing amino acids. However, nectar amino acid preference is significantly lower in mated females. Furthermore, nectar amino acid preference increases when females are not allowed to remate, whereas the preference of twice-mated females remains constant at a lower level. These results indicate a versatile response of females to nectar amino acids, depending on their nutritional status; they may even switch their source of amino acids between adult feeding and nuptial gifts.     

Sex‐linked inheritance of diapause induction in the butterfly Pieris napi

Many temperate insects survive harsh environmental conditions, such as winter, by entering a state of developmental arrest. This diapause state is predominantly induced by photoperiod. The photoperiod varies with latitude and has led to local adaptation in the photoperiodic induction of diapause in many insects. To understand the rapid evolution of the photoperiodic threshold, it is important to investigate and understand the underlying genetic mechanisms. In the present study, the genetic basis of photoperiodic diapause induction is investigated in the green‐veined white butterfly Pieris napi (Lepidoptera, Pieridae) by assaying diapause induction in a range of conditions for a Swedish and Spanish population. Furthermore, the inheritance of diapause induction is assessed in reciprocal F1 hybrids and backcrosses between the two populations. The southern population shows a clear photoperiodic threshold determining diapause or direct development, whereas the northern populations show a high incidence of diapause, regardless of photoperiod. The hybrid crosses reveal that the inheritance of diapause induction is strongly sex‐linked, and that diapause incidence in the genetic crosses is highly dependent on photoperiod. This emphasizes the importance of assaying a range of conditions in diapause inheritance studies. The results indicate a strongly heritable diapause induction with a major component on the Z‐chromosome, as well as a minor effect of the autosomal background.     

Developmental and phenotypic responses to photoperiod in uni-and bivoltine Pieris napi in California

• 1 Coastal Californian Pieris napi are facultatively bivoltine, with two seasonal phenotypes, whereas inland populations from the Inner Coast Ranges and Sierra Nevada are univoltine and monophenic.
• 2 When reared under continuous light at 25d̀C both coastal and inland stocks produce about 50% diapause pupae, which give rise to vernal-phenotype adults. Non-diapause pupae of all stocks give rise to summer-phenotype adults, even though this phenotype does not exist in the wild in univoltine populations.
• 3 Univoltinism, which implies developmental suppression of the summer phenotype, is interpreted as a derivative from multivoltinism and an adaptation to host plant phenology.

     

Competition between larvae in a butterfly Pieris napi and maintenance of different life-history strategies

1. In scramble competition all individuals suffer equally from competition, whereas in contest competition some individuals outperform the others. Generally, larger individuals gain asymmetric advantage in competition over smaller ones. Given the positive correlation between age and size, asynchronous birth may result in asymmetric competition among juveniles. 2. In Pieris napi (Lepidoptera: Pieridae), reproductive rate is determined by the females' intrinsic mating tactic. The early reproductive rate is high in females with a low mating frequency and low in females with a high mating frequency, whereas lifetime fecundity shows the opposite pattern. Thus, offspring of monandrous females start to develop in relatively low densities and they are relatively large when the offspring of highly polyandrous females start to hatch. 3. The purpose of this study was to explore if asymmetry in larval competition could outweigh the late-life benefits of polyandry. In a laboratory experiment, P. napi larvae of different ages were reared together in different densities. 4. Increasing density decreased both larval survival and reachable pupal mass, but had no effect on duration of larval period. Younger larvae suffered from high mortality and reduced size compared with the older larvae. Mortality decreased in the older cohort with increasing age difference between the cohorts, and the reverse occurred in the younger cohort. Increasing age difference between the cohorts was associated with increase in pupal mass in both cohorts. All the variables showed a lot of variation between broods of different females. 5. The results suggest that polyandrous females, or more generally females with a low early reproductive rate, may lose a great proportion of their late-life benefits, which may partly explain the maintenance of polymorphism in reproductive strategies within species.     

Nuptial gifts and the use of body resources for reproduction in the green-veined white butterfly Pieris napi

Reproduction in butterflies, as in many holometabolous insects, is usually constrained by the amount of nutrients the animals can collect as juveniles. In polyandric species the females can also supplement their larval-derived reserves with protein-rich donations, so-called nuptial gifts, delivered by the males at mating. Recent findings also indicate that females have access to nitrogen from the histolysis of flight muscles in the thorax. This field study examined how butterflies of the polyandric gift-giving species Pieris napi (Lepidoptera: Pieridae) use body resources in their reproduction and how the male donations affect the females use of stored reserves. The results support earlier studies, indicating that females use resources from the breakdown of thorax muscles to increase their reproductive potential and the results also indicate that males also use thorax material in their reproduction. The study also supports recent findings that the male donation increases the breakdown of body resources and thereby boosts the reproductive output of the female.     

Do Reproductive Status and Lamb Gender Affect the Foraging Behavior of Bighorn Ewes?

In female ungulates lactation carries a high energetic cost and mothers often devote more care to sons than to daughters. The aims of this study were to determine whether lactating bighorn ewes have higher foraging time, bite rate or selectivity of forage than barren ewes and whether reproductive status affects migration patterns. Ewes with male lambs were predicted to spend more time foraging and to have a higher bite rate than ewes with female lambs. There were no differences in foraging behavior according to ewe reproductive status from April to August. In September-November lactating ewes had a higher bite rate and spent more time foraging than nonlactating ewes but lamb gender did not affect foraging behavior. Lactating ewes gained less weight than nonlactating ones until mid-August; from mid-August to late September mass gain was similar for both groups of ewes. Nonlactating ewes spent more time standing but reproductive status did not affect vigilance behavior or step rate while foraging. Ewes with lambs did not differ from nonlactating ewes in step rate. Pregnant ewes migrated earlier than barren ewes to alpine areas in spring. By so doing they abandoned areas with good quality forage presumably to give birth in areas safer from predation. All ewes spent most of the summer in the alpine range but nonlactating ewes returned to the winter range earlier than lactating ewes, probably to profit from the abundant forage there.     

Natural Plant Chemicals Acting as Oviposition Deterrents on Cabbage Butterflies (Pieris btassicae (L.), P. rapae (L.) and P. napi (L.))

In several growers' reports Solatium lycopersicum, Sambucus nigra. Thymus vulgaris, Salvia officinalis, Artemisia absinthium, A. abrotanum , and Allium cepa are said to decrease the oviposition of Pieris brassicae, P. rapae and P. napi. In the present study the butterflies were fed with honey automats and reared throughout the year in artificial light in an insec-tarium. In a dual-choice chamber with a slow throughflow of air two equally sized cabbage leaves were placed on opposite sides. Significantly fewer eggs were layed on the cabbage leaf on which extracts of the mentioned plants had been applied. Ten butterflies were used in each experiment. The chemoreceptors and the chemicals involved are not identified but the inhibitory substances are surely secondary plant substances. Acceptance or rejection of secondary plant metabolics determines the complicated food relationships between plants and insects. The use of secondary plant substances for ecological control of insect pests is proposed.     

Contact chemoreception of oviposition-stimulating glucosinolates and an oviposition-deterrent cardenolide in two subspecies of Pieris napi

Abstract. Electrophysiological responses to ten glucosinolates, oviposition stimulants isolated from their cruciferous host plants, were recorded from tarsal taste neurones of two subspecies of Pieris napi: P. napi napi L. and P. napi olerucea (Harris) (Lepidoptera: Pieridae). In the cluster of fourteen medial B-sensilla on the fifth tarsomere, strong responses to glucosinolates were recorded. Three receptors cells, identified on the basis of spike amplitude and shape, were typically active in these responses, with the cell producing the smallest spike predominating. The response intensity of the latter cell to a particular glucosinolate at 1 mm concentration depended both on the chemical structure of the stimulus and on the subspecies of insect. In P. napi napi , aromatic glucosinolates were just as effective as aliphatic glucosinolates, whereas in P. napi oleracea the long-chain aliphatic glucosinolates gluconapin and progoitrin were stronger stimuli. At 10 and 100 pg/ml concentrations, significant differences between the subspecies were found in the absolute response intensity to seven compounds. A second cell in P. napi olerucea , producing a medium-sized spike, was excited most by sinalbin, whereas in Pnupi nupi three aliphatic glucosinolates were equally effective stimulants for a similar cell. In both subspecies the cell producing the medium-sized spikes in response to glucosinolates was also highly sensitive to erychroside, an oviposition deterring cardenolide occurring in a host of P. napi olerncea. The evolutionary divergence between the two geographically separated subspecies is reflected by differences in chemosensory recognition mechanisms.     

Time-Varying Wing-Twist Improves Aerodynamic Efficiency of Forward Flight in Butterflies

Insect wings can undergo significant chordwise (camber) as well as spanwise (twist) deformation during flapping flight but the effect of these deformations is not well understood. The shape and size of butterfly wings leads to particularly large wing deformations, making them an ideal test case for investigation of these effects. Here we use computational models derived from experiments on free-flying butterflies to understand the effect of time-varying twist and camber on the aerodynamic performance of these insects. High-speed videogrammetry is used to capture the wing kinematics, including deformation, of a Painted Lady butterfly (Vanessa cardui) in untethered, forward flight. These experimental results are then analyzed computationally using a high-fidelity, three-dimensional, unsteady Navier-Stokes flow solver. For comparison to this case, a set of non-deforming, flat-plate wing (FPW) models of wing motion are synthesized and subjected to the same analysis along with a wing model that matches the time-varying wing-twist observed for the butterfly, but has no deformation in camber. The simulations show that the observed butterfly wing (OBW) outperforms all the flat-plate wings in terms of usable force production as well as the ratio of lift to power by at least 29% and 46%, respectively. This increase in efficiency of lift production is at least three-fold greater than reported for other insects. Interestingly, we also find that the twist-only-wing (TOW) model recovers much of the performance of the OBW, demonstrating that wing-twist, and not camber is key to forward flight in these insects. The implications of this on the design of flapping wing micro-aerial vehicles are discussed.     

Ecomorphological and genetic divergence between lowland and montane forms of the Pieris napi species complex (Pieridae, Lepidoptera)

The present study aimed to investigate the morphological and genetic differentiation of lowland and montane populations of the Pieris napi species complex in Norway and to make inferences about the subspecific status of Pieris napi adalwinda and Pieris napi napi . We measured 22 morphological characters on 228 individuals from eight populations in Central Norway (20–1100 m a.s.l.). In addition, lowland and mountain animals were reared at a high altitude locality. Half the animals from either locality were reared on mountain plant, and the other half on a lowland plant. Finally, a fragment of the mitochondrial cytochrome b gene was sequenced for individuals from Central and South-eastern Norway and Germany. Principal component analysis of morphological characters showed a zone of abrupt change from lowland to mountain morphological character states between populations at 506 m and 730 m a.s.l., respectively. The transplant experiment showed that the morphological differences have a genetic basis and that food plants have no impact on morphology. Limited, but significant, molecular genetic differentiation was found between lowland and mountain animals, but the phylogenetic analysis, however, showed that the lowland form ( P. n. napi ) is paraphyletic and the montane form ( P. n. adalwinda ) is monophyletic. Further study is required before taxonomic recognition can be applied.  © 2007 The Linnean Society of London, Biological Journal of the Linnean Society , 2007, 92 , 727–745.     

The effect of color preferences on the foraging behavior of the green-veined white butterfly (Pieris napi L.)

The color preferences of the green-veined white butterfly Pieris napi (Linnaeus, 1758) that are associated with its foraging behavior were studied in experiments with artificial flower models. It was found that newly emerged inexperienced butterflies that are searching for food prefer flower models that are blue or red and select yellow ones at a considerably lower rate. With repeated visits on artificial flowers, P. napi individuals display are highly consistent in their preference for the first chosen color. Males and females of this species display similar color preferences.     

Microevolutionary selection dynamics acting on immune genes of the green‐veined white butterfly,Pieris napi

Insects rely on their innate immune system to successfully mediate complex interactions with their microbiota, as well as the microbes present in the environment. Previous work has shown that components of the canonical immune gene repertoire evolve rapidly and have evolutionary characteristics originating from interactions with fast‐evolving microorganisms. Although these interactions are likely to vary among populations, there is a poor understanding of the microevolutionary dynamics of immune genes, especially in non‐Dipteran insects. Here, we use the full set of canonical insect immune genes to investigate microevolutionary dynamics acting on these genes between and among populations by comparing three allopatric populations of the green‐veined white butterfly, Pieris napi (Linné; Lepidoptera, Pieridae). Immune genes showed increased genetic diversity compared to genes from the rest of the genome and various functional categories exhibited different types of signatures of selection, at different evolutionary scales, presenting a complex pattern of selection dynamics. Signatures of balancing selection were identified in 10 genes, and 17 genes appear to be under positive selection. Genes involved with the cellular arm of the immune response as well as the Toll pathway appear to be enriched among our outlier loci, regardless of functional category. This suggests that the targets of selection might focus upon an entire pathway, rather than functional subsets across pathways. Our microevolutionary results are similar to previously observed macroevolutionary patterns from diverse taxa, suggesting that either the immune system is robust to dramatic differences in life history and microbial communities, or that diverse microbes exert similar selection pressures.     

Phenotypic induction in Pieris napi L.: role of temperature and photoperiod in a coastal California population

Abstract. 1. Californian Pieris napi have previously been reported as producing dark-veined and light-veined adults from diapausing and. non-diapausing pupae respectively; the converse has not been reported.
2. A population of the coastal subspecies venosa from Monterey County was examined to determine whether pupal temperature exposure was involved in proximate control of seasonal phenotypes.
3. When reared under continuous light at 25°C this stock produced seventeen diapause pupae, thirteen of which have eclosed yielding typical dark-veined adults, and thirty-eight non-diapause pupae.
4. Of fifteen non-diapause pupae held in a dark box at 25°C, fourteen produced typical light-veined adults and one produced a dark-veined individual of intermediate phenotype.
5. Of twenty-three non-diapause pupae held in a dark box at 10°C for 24 days, twenty-two produced dark-veined butterflies of intermediate phenotype and one produced a light-veined adult.
6. When the experiment was repeated with an inland, population of subspecies microstriata , which is normally univoltine and monophenic, the temperature effect on phenotype was still present but less pronounced.
7. The nature of phenotypic determination is reviewed with particular regard for the developmental environment of butterflies from diapaused pupae. The thermal environment of reactivating diapausers, rather than the diapause state itself, may determine adult phenotype.     

Life history tradeoffs in relation to the degree of polyandry and developmental pathway in Pieris napi (Lepidoptera, Pieridae)

Pieris napi females have different heritable reproductive tactics. Polyandrous females have higher lifetime fecundity, whereas monandrous ones start to reproduce at a faster rate. Butterfly larvae are time‐constrained in seasonal environments. Thus, polyandry is expected to be associated with fast juvenile development, which may result in biased mortality due to physiological costs. We compared how females with varying degrees of polyandry allocate between duration of larval period and achievable size in directly developing and over‐wintering generations. Offspring survival and growth were monitored under a high density and low quality diet. Polyandrous females developed at a faster rate than monandrous ones, regardless of developmental pathway. The growth rate of female offspring correlated with their mothers’ degree of polyandry, which underpins polyandry and monandry as distinct strategies with life history differences reaching beyond mating frequency. The high growth rate of polyandrous females resulted in a short larval period among directly developing females, and in large size within an over‐wintering cohort. A change in either the duration of the larval period or pupal mass had no significant effect on the other, emphasising that growth rate is not necessarily a simple outcome of the tradeoff between development time and size at maturity. The correlation between the degree of polyandry and juvenile growth rate diminished when larvae were exposed to environmental stress, which offers an explanation why juvenile mortality was decoupled from mating tactic. We conclude that polyandry is a strategy that allows larvae to utilise optimal conditions in a more effective way than monandry. As a consequence, polyandrous females either achieve larger size or they mature faster. This gives them a double advantage over monandrous ones within an over‐wintering generation or diminishes the effects of asynchronous hatching of offspring within a directly developing generation. Possible costs of high growth rate are discussed.     

Does a lack of mating opportunities explain monandry in the green‐veined white butterfly (Pieris napi)?

In the nuptial gift-giving butterfly, Pieris napi, multiple mating corresponds to higher lifetime fecundity than monandry does. Yet, female mating frequency, which is genetically determined and heritable, varies from strict monandry to a high degree of polyandry. Polyandrous females are known to suffer from reduced longevity when denied the opportunity to mate according to their intrinsic mating frequency. Here we test if monandry is likely to be maintained due to a lack of mating opportunities in the wild. We also explore longitudinal variation in female mating frequency (i.e. lifetime number of matings) and remating rate (i.e. time interval between successive matings). The latter was explored by comparing the mating patterns of females with varying origins in a laboratory, and the former by comparing the mating frequencies of wild and laboratory females. We were able to reveal spatial variation in female mating frequency, with monandry being a clearly more common mating tactic in the north than in the south. However, we did not find direct evidence that either the maintenance of monandry or spatial variation in female mating frequency is promoted by a lack of mating opportunities. Still, our findings may have profound implications, since decreasing mating frequency towards the north generates spatial variation in the strength of sexual selection.