High‐prevalence male‐killing Wolbachia in the butterfly Acraea encedana

Inherited bacteria which kill males during early development are widely distributed throughout the insects, but have been little studied outside of a single family of beetles, the Coccinellidae. We have investigated a male‐killing bacterium discovered in the butterfly Acraea encedana. This bacterium belongs to the genus Wolbachia and is identical in wsp gene sequence to a male‐killer in the closely related butterfly A. encedon, suggesting that it has either recently moved between host species or was inherited from a common ancestor of the butterflies. The prevalence of Wolbachia is remarkably high, 95% of females are infected and only 6% of wild caught butterflies are male. Measurements of the vertical transmission efficiency were used to calculate that this high prevalence is the result of infected females producing at least 1.79 times the number of surviving daughters as uninfected females (lower confidence limit is 1.25).     

All-female broods and mimetic polymorphism in Acraea encedon (L.) (Lepidoptera: Acraeidae) in Tanzania

In some populations of the African butterfly, Acraea encedon, there are two kinds of females, one producing offspring in a normal 1:1 sex ratio, the other producing females only; in other populations the sex ratio is apparently normal. All-female broods had hitherto been mainly associated with populations in which field sampling revealed an excess of females. The all-female brood trait is described from a population at Dar es Salaam which field sampling suggested was normal, and this indicates that the trait may be much more widespread and common than had previously been supposed. This discovery also extends the known distribution of the trait across Africa from Sierra Leone to eastern Tanzania. The butterfly is also a polymorphic Müllerian mimic of Danaus chrysippus, which is a highly unusual phenomenon as Müllerian mimicry is almost invariably monomorphic. The relative frequencies of two corresponding colour forms of the two species of butterflies at Dar es Salaam adds support to the hypothesis that they are indeed Mullerian mimics. The results of breeding experiments suggest that the polymorphic forms in Acraea encedon are allelic with dominance.     

Population biology of tropical African butterflies. Sex ratio and genetic variation in Acraea encedon

In Uganda the butterfly, Acraea encedon (L.), occurs in well-defined populations that are largely genetically and ecologically isolated from each other. During the course of a large-scale capture-recapture programme about 18,000 different individuals were marked and released, and from these and many recaptures aspects of the population structure were investigated. Some populations are composed of almost all females while in other populations the sex ratio is normal. In the predominantly female populations many females remain unmated and produce infertile eggs, laying them on plants that are not foodplants and on each other. In such populations there is intense aggregating behaviour. Males live longer than females and each male may mate with several different females. There is evidence for the existence of a self-regulating mechanism in the predominantly female populations in which population size and male frequency are reciprocally dependent. Acraea encedon exists in a large variety of sympatric colour forms, the frequency of which varies from population to population. Some colour forms survive better than others. The hindwing of Acraea encedon has 20 black spots each of which can be absent in any particular butterfly. The relative frequency of missing spots varies from population to population. The ecological genetics of Acraea encedon is discussed in relation to theories of mimicry and polymorphism.     

Polymorphic Müllerian mimicry in a group of African butterflies: a re-assessment of the relationship between Danaus chrysippus, Acraea encedon and Acraea encedana (Lepidoptera: Nymphalidae)

Müllerian mimicry, in which there is convergence in coloration between unrelated unpalatable species, should lead to uniformity in appearance, not polymorphism, and so the occurrence in tropical Africa of unrelated species of unpalatable butterflies with corresponding polymorphic colour forms suggested a problem of special evolutionary interest. Field work in Uganda and Sierra Leone in 1964 72 demonstrated a statistical association between the occurrence and relative frequencies of polymorphic forms in Danaus chrysippus (Danainae) and Acraea encedon (Acraeinae) which was deemed as confirmation of a Müllerian relationship between them. There were, however, certain anomalies which at the time remained unresolved. Later, in 1976, it was discovered that what had been called A. encedon is in reality two sibling species. A. encedon and a new one, named as A. encedana. The two differ in the structure of both male and female genitalia and in the coloration and the food-plants of the larvae. The recognition of the additional species has enabled a re-assessment of the polymorphie Müllerian association with D. chrysippus. It emerges that, although there is a close qualitative and quantitative reciprocal mimetic relationship between A. encedana and D. chrysippus, the relationship between A. encedon and D. chrysippus is much weaker, and in places non-exitent. The possible origin of the mimetic polymorphism is discussed in terms of hybridization of previously allopatric and monomorphic populations which have met as a consequence of recent expansions of geographical range in all three species resulting from forest clearance and the spread of savanna-like conditions in previously forested areas.     

The inheritance and population genetics of sex ratio in the butterfly Acraea encedon

In West and East Africa the butterfly, Acraea encedon , occurs in well-defined populations that are often predominantly female. Breeding the butterfly in the laboratory revealed the presence of an all-female strain, which is inherited directly through the female parent. It is probable that the inheritance is controlled by a Y-linked gene, causing meiotic drive in the Y chromosome, but the possibility of cytoplasmic inheritance has not been ruled out. A simple model for the population genetics of a predominantly female population indicates that such a population should rapidly become extinct due to the spreading of the all-female strain, but in most field populations studied extinction does not occur as quickly as predicted, if at all. Three factors which could enable populations to avoid extinction are investigated: suppressing systems, frequency-dependent mating preference and the sequence of emergence of the sexes in normal broods. No positive evidence has yet been found for the existence of a gene or genes capable of suppressing the sex ratio aberration, and no frequency-dependent mating preference was found, but an argument is presented which shows that the sequence of emergence in normal broods could be partly responsible for the maintenance of stable equilibria in predominantly female populations. Attempts to upset the sex ratio in the normal strain by making crosses between widely separated populations were not successful.     

Climatic warming increases voltinism in European butterflies and moths

Climate change is altering geographical ranges, population dynamics and phenologies of many organisms. For ectotherms, increased ambient temperatures frequently have direct consequences for metabolic rates, activity patterns and developmental rates. Consequently, in many insect species both an earlier beginning and prolongation of seasonal duration occurred in parallel with recent global warming. However, from an ecological and evolutionary perspective, the number of generations (voltinism) and investment into each generation may be even more important than seasonality, since an additional generation per unit time may accelerate population growth or adaptation. Using a dataset extending back to the mid-nineteenth century, I report changes in the voltinism of butterfly and moth species of Central Europe. A significant proportion of 263 multi-voltine species showed augmented frequency of second and subsequent generations relative to the first generation in a warm period since 1980, and 44 species even increased the number of generations after 1980. Expected ecological consequences are diverse. Since multi-voltinism has been linked to insect outbreaks they include an increase in the abundance of herbivorous pests of agriculture and forestry. However, disruption of the developmental synchrony associated with multi-voltinism and host plant phenology may also reduce fitness, potentially having unexpected consequences for species of conservation concern. The ability of species to adapt evolutionarily to a changing environment may be facilitated by increased voltinism.     

A novel method of behavioural thermoregulation in butterflies

Abstract The requirement for efficient thermoregulation has directed the coevolution of specialized morphological and behavioural traits in ectotherms. Adult butterflies exhibit three thermoregulatory mechanisms, termed dorsal, lateral and reflectance basking. In this study, we investigate a potential fourth mechanism whereby individuals perch with their wings fully spread and angled downwards such that the margins are appressed to the substrate. We find that mate‐locating male Hypolimnas bolina (L.) (Nymphalidae) adopt this posture when operational thoracic temperatures are lowest (less than approximately 34 °C). As thoracic temperature increases, males perch with wings increasingly closed and ultimately select shaded microhabitats. Using thermocouple‐implanted dead models, we show that appressed posture individuals warm faster than those adopting the conventional dorsal‐basking (horizontal wing) posture. This thermal advantage is not mitigated by shading of the outer 60–70% of the wing area, which suggests that – as with the conventional dorsal posture – only the basal wing surfaces contribute to heat gain via the absorption of solar irradiation. These investigations suggest that appression represents a novel extension of conventional dorsal basking behaviour in butterflies.     

Widespread infection and diverse infection patterns of Wolbachia in Chinese aphids

Wolbachia are intracellular symbionts that infect a wide range of arthropods and filarial nematodes. Aphids are engaged in diverse and complex relationships withtheir endosymbionts. Four supergroups （A, B, M and N） of Wolbachia were previously detected in aphids and supergroups M and N were only found in aphids. In this study, wedetected and described Wolbachia infections in natural populations of aphids in China. Three supergroups （A, B and M） were found in the examined aphid species. SupergroupM was preponderant, whereas supergroups A and B were only detected in certain species. Supergroup N was not found in this study. There were four infection patterns of Wolbachiain aphids, namely, infection with supergroup M alone, co-infection with supergroups A and M, co-infection with supergroups B and M, and co-infection with supergroups A, Band M. The pattern of infection only with supergroup M was universal and was tbund in all evaluated subfamilies. Only two subfamilies, Aphidinae and Lachninae, manifestedto present all four infection patterns. Three patterns were observed in Calaphidinae （M, A＆M, B＆M） and Eriosomatinae （M, B＆M, A＆B＆M）. Two patterns were observed inthe Anoeciinae （M, A＆M） and Greenideinae （M, B＆M）, and only one pattern （M） was observed in the remaining families and/or subfamilies of Aphidoidea. These results indi-cated that Wolbachia infections in Chinese aphids are widespread. Phylogenetic analyses suggest that Wolbachia supergroup M spread rapidly and recently among all host speciesof aphids in China. Reasons for this spread and its mechanisms are discussed along with the possible effects of Wolbachia on their aphid hosts.     

Cretaceous origin and repeated tertiary diversification of the redefined butterflies

Although the taxonomy of the ca 18 000 species of butterflies and skippers is well known, the family-level relationships are still debated. Here, we present, to our knowledge, the most comprehensive phylogenetic analysis of the superfamilies Papilionoidea, Hesperioidea and Hedyloidea to date based on morphological and molecular data. We reconstructed their phylogenetic relationships using parsimony and Bayesian approaches. We estimated times and rates of diversification along lineages in order to reconstruct their evolutionary history. Our results suggest that the butterflies, as traditionally understood, are paraphyletic, with Papilionidae being the sister-group to Hesperioidea, Hedyloidea and all other butterflies. Hence, the families in the current three superfamilies should be placed in a single superfamily Papilionoidea. In addition, we find that Hedylidae is sister to Hesperiidae, and this novel relationship is supported by two morphological characters. The families diverged in the Early Cretaceous but diversified after the Cretaceous-Palaeogene event. The diversification of butterflies is characterized by a slow speciation rate in the lineage leading to Baronia brevicornis, a period of stasis by the skippers after divergence and a burst of diversification in the lineages leading to Nymphalidae, Riodinidae and Lycaenidae.     

The parasitic bacterium Wolbachia and the origin of the eukaryotic cell

The acquisition of endosymbiotic alphaproteobacteria that gave rise to mitochondria was one of the key events in the origin of eukaryotic cell. To reconstruct this process, it is important to analyze relationships that developed later between eukaryotes and other alphaproteobacteria. Wolbachia pipientis, a bacterium that inhabits cells of numerous terrestrial invertebrates and exerts diverse effects on its hosts, is used as a model. Although Wolbachia is similar to mitochondria in many important features (basic metabolism, small molecule membrane transport, envelope structure, etc.), their relationships with the nucleocytoplasm are different. Mitochondria import most of their required proteins from the nucleocytoplasm and are controlled by the nucleocytoplasmic regulatory systems. On the contrary, Wolbachia exports its proteins into the host’s cytoplasm, thus causing dramatic aberrations in the ontogeny and reproduction of the host. This difference may be due to the fact that most of the protomitochondrial genes had been transferred into the central (nuclear) genome at the early stages of the development of the endosymbiotic system, while Wolbachia genes were not transferred into the nucleus. This fits well with the previously suggested hypothesis that there was a period of rapid lateral gene transfer in the evolution of proto-eukaryotes; the acquisition of mitochondria took place during this period. Later, eukaryotes, and especially metazoans, developed powerful mechanisms for prevention of lateral gene transfer. Therefore, the genes of the newly acquired endosymbionts cannot be transferred into the central genome, and the endosymbionts retain the capacity for selfish evolution.     

感染稻水象甲的Wolbachia基因组中插入序列的鉴定与分析

共生细菌Wolbachia对宿主的生殖起多种调控作用。以往研究表明, Wolbachia基因组中广泛存在插入序列(insertion sequence, IS), 它们对宿主基因组的可塑性、 多样性和进化起重要作用。稻水象甲Lissorhoptrus oryzophilus Kuschel在东亚是一种外来水稻害虫, 在原产地北美营两性生殖, 而在所有入侵地均营孤雌生殖。本研究采用PCR法从河北唐海孤雌生殖型稻水象甲体内克隆获得了Wolbachia的2条IS序列, 即ISWosp4和ISWosp6; 从美国德克萨斯州两性生殖型稻水象甲成虫体内克隆获得了Wolbachia的2条IS序列, 即ISWosp3和ISWosp5。碱基序列比对显示: ISWosp3和ISWosp4属于IS3家族IS3组成员, ISWosp5为IS4家族IS231组成员, ISWosp6为IS5家族IS1031组成员。对这些IS的ORF结构、 所编码氨基酸序列的结构等进行了分析, 推测ISWosp5具有潜在转座活性。所得结果增进了我们对Wolbachia IS3, IS4和IS5家族插入序列的认识, 同时为今后从IS的角度探讨Wolbachia与稻水象甲生殖的关系奠定了基础。     

A Survey of the bacteriophage WO in the endosymbiotic bacteria Wolbachia

Bacteriophages are common viruses infecting prokaryotes. In addition to their deadly effect, phages are also involved in several evolutionary processes of bacteria, such as coding functional proteins potentially beneficial to them, or favoring horizontal gene transfer through transduction. The particular lifestyle of obligatory intracellular bacteria usually protects them from phage infection. However, Wolbachia, an intracellular alpha-proteobacterium, infecting diverse arthropod and nematode species and best known for the reproductive alterations it induces, harbors a phage named WO, which has recently been proven to be lytic. Here, phage infection was checked in 31 Wolbachia strains, which induce 5 different effects in their hosts and infect 25 insect species and 3 nematodes. Only the Wolbachia infecting nematodes and Trichogramma were found devoid of phage infection. All the 25 detected phages were characterized by the DNA sequence of a minor capsid protein gene. Based on all data currently available, phylogenetic analyses show a lack of congruency between Wolbachia or insect and phage WO phylogenies, indicating numerous horizontal transfers of phage among the different Wolbachia strains. The absence of relation between phage phylogeny and the effects induced by Wolbachia suggests that WO is not directly involved in these effects. Implications on phage WO evolution are discussed.     

Yield and activity of Autographa californica multicapsid nucleopolyhedrovirus and Phthorimaea operculella granulosis virus in cloned and uncloned cell lines of P. operculella

Three selected uncloned Pop 2, Pop 3, Pop 4 and two cloned cell lines Pop cl1A and Pop cl2B were derived from the original cell line established from Phthorimaea operculella (ORS-Pop-93). Three new non-selected cell lines ORS-Pop-94A, ORS-Pop-94B and ORS-Pop-95 were also established from embryos of the same insect. Differences in morphology, growth rate and polypeptide profile were determined between these cell lines. All the cell lines were susceptible to the Autographa californica nucleopolyhedrovirus (AcMNPV). The cloned cell lines produced higher levels of AcMNPV (TCID-50 and PIB) than the parental cells and at the same rate as the Sf9 reference cell line. Substantial amounts of viral DNA were synthesized in the clone Pop cl 2B after infection with the granulosis virus of the potato tuber moth P. operculella (PTMGV) and a complete multiplication was obtained in the ORS-Pop-95 cell line. The comparison between Pop cell lines which support limited or complete replication of certain baculoviruses can offer insights into some of the molecular barriers which restrict the host range of these viruses. These cell lines with variable susceptibility to baculoviruses could also be used for in vitro recombinations, increasing their virus host range to be used for the control of this pest. This revised version was published online in August 2006 with corrections to the Cover Date.     

On the mechanics and energetics of nectar feeding in butterflies.

A mechanistic model describing the mechanics and energetics of nectar-feeding in butterflies is developed. The butterflies Collas eurytheme and Danaus plexippus are used to illustrate the model. Simulation results indicate that there are mechanical limitations upon the range of nectar sugar concentrations and nectar extraction times available to butterflies. There is a unique optimum for net rate of energy gain at 20–25% nectar sugar concentration which is independent of the metabolic rate and of proboscis shape and size over the ranges found in butterflies. The optimal nectar extraction rate depends upon the size and shape of the proboscis. These results are discussed in relation to the design of nectar feeding structures, optimal foraging strategy, and the evolution of insect pollination.     

Widespread occurrence of the micro-organism Wolbachia in ants.

For more than 20 years, sex allocation in hymenopteran societies has been a major topic in insect sociobiology. A recurring idea was that relatedness asymmetrics arising from their haplodiploid sex determination system would lead to various parent-offspring conflicts over optimal reproduction. A possible weakness of existing theory is that only interests of nuclear genes are properly accounted for. Yet, a diversity of maternally transmitted elements manipulate the reproduction of their host in many solitary arthropod groups. The bacterium Wolbachia is a striking example of such a selfish cytoplasmic element, with effects ranging from reproductive incompatibility between host strains, induction of parthenogenesis and feminization of males. This paper reports on a first PCR-based Wolbachia screening in ants. Out of 50 Indo-Australian species, 50% screened positive for an A-group strain. One of these species also harboured a B-group strain in a double infection. Various factors that might explain the unusually high incidence of Wolbachia in ants are discussed. In general, Wolbachia may represent a widespread and previously unrecognized party active in the conflicts of interest within social insect colonies.     

漳州市蝶类名录初报

１９９７－１９９９年在福建漳州调查蝶类,共计１０科、１１０属、１７６种。     

Loss rates of butterfly species with urban development. A test of atlas data and sampling artefacts at a fine scale

Data for the United Kingdom (UK) Manchester Butterfly Atlas produced conflicting species loss rates for increased urban development. In particular, a very low rate of loss was recorded (0.19 species for every 10% increase in urban cover) for the Mersey Valley mapped at a high resolution of 1 ha units. It was suggested that sampling artefacts (uneven recording) or failure to distinguish vagrant individuals from breeding populations cause this. Herein, results are reported for 30 sample squares, within the Mersey Valley, surveyed uniformly throughout 1999. It is shown that loss rates are as high as areas mapped at lower resolution over wider areas (0.67–0.68 species for every 10% increase in urban cover) and that increasingly stringent definitions of urban cover result in higher loss rates. Comparison with the data from the Atlas, but for the same 30 sample squares, indicate that the low rates at a fine scale for the complete Atlas data are more likely to be caused by uneven recording than from failure to record species status. However, progressive sampling of squares, despite uniform recording, will inevitably cause a reduction in loss rates of total species for increases in urban development.