Maintenance of a male-killing Wolbachia in Drosophila innubila by male-killing dependent and male-killing independent mechanisms

Many maternally inherited endosymbionts manipulate their host's reproduction in various ways to enhance their own fitness. One such mechanism is male killing (MK), in which sons of infected mothers are killed by the endosymbiont during development. Several hypotheses have been proposed to explain the advantages of MK, including resource reallocation from sons to daughters of infected females, avoidance of inbreeding by infected females, and, if transmission is not purely maternal, the facilitation of horizontal transmission to uninfected females. We tested these hypotheses in Drosophila innubila, a mycophagous species infected with MK Wolbachia. There was no evidence of horizontal transmission in the wild and no evidence Wolbachia reduced levels of inbreeding. Resource reallocation does appear to be operative, as Wolbachia-infected females are slightly larger, on average, than uninfected females, although the selective advantage of larger size is insufficient to account for the frequency of infection in natural populations. Wolbachia-infected females from the wild-although not those from the laboratory-were more fecund than uninfected females. Experimental studies revealed that Wolbachia can boost the fecundity of nutrient-deprived flies and reduce the adverse effect of RNA virus infection. Thus, this MK endosymbiont can provide direct, MK-independent fitness benefits to infected female hosts in addition to possible benefits mediated via MK.     

Extreme streams: species persistence and genomic change in montane insect populations across a flooding gradient

The ecological and evolutionary consequences of extreme events are poorly understood. Here, we tested predictions about species persistence and population genomic change in aquatic insects in 14 Colorado mountain streams across a hydrological disturbance gradient caused by a one in 500‐year rainfall event. Taxa persistence ranged from 39 to 77% across sites and declined with increasing disturbance in relation to species' resistance and resilience traits. For taxa with mobile larvae and terrestrial adult stages present at the time of the flood, average persistence was 84% compared to 25% for immobile taxa that lacked terrestrial adults. For two of six species analysed, genomic diversity (allelic richness) declined after the event. For one species it greatly expanded, suggesting resilience via re‐colonisation from upstream populations. Thus, while resistance and resilience traits can explain species persistence to extreme disturbance, population genomic change varies among species, challenging generalisations about evolutionary responses to extreme events at landscape scales.     

Asymmetrical interactions between Wolbachia and Spiroplasma endosymbionts coexisting in the same insect host

We investigated the interactions between the endosymbionts Wolbachia pipientis strain wMel and Spiroplasma sp. strain NSRO coinfecting the host insect Drosophila melanogaster. By making use of antibiotic therapy, temperature stress, and hemolymph microinjection, we established the following strains in the same host genetic background: the SW strain, infected with both Spiroplasma and Wolbachia; the S strain, infected with Spiroplasma only; and the W strain, infected with Wolbachia only. The infection dynamics of the symbionts in these strains were monitored by quantitative PCR during host development. The infection densities of Spiroplasma exhibited no significant differences between the SW and S strains throughout the developmental course. In contrast, the infection densities of Wolbachia were significantly lower in the SW strain than in the W strain at the pupal and young adult stages. These results indicated that the interactions between the coinfecting symbionts were asymmetrical, i.e., Spiroplasma organisms negatively affected the population of Wolbachia organisms, while Wolbachia organisms did not influence the population of Spiroplasma organisms. In the host body, the symbionts exhibited their own tissue tropisms: among the tissues examined, Spiroplasma was the most abundant in the ovaries, while Wolbachia showed the highest density in Malpighian tubules. Strikingly, basically no Wolbachia organisms were detected in hemolymph, the principal location of Spiroplasma. These results suggest that different host tissues act as distinct microhabitats for the symbionts and that the lytic process in host metamorphosis might be involved in the asymmetrical interactions between the coinfecting symbionts.     

A diversity of endosymbionts across Australian aphids and their persistence in aphid cultures

There is increasing interest in the use of endosymbionts in pest control, which will benefit from the identification of endosymbionts from potential donor species for transfer to pest species. Here, we screened for endosymbionts in 123 Australian aphid samples across 32 species using 16S DNA metabarcoding. We then developed a qPCR method to validate the metabarcoding data set and to monitor endosymbiont persistence in aphid cultures. Pea aphids (Acyrthosiphon pisum) were frequently coinfected with Rickettsiella and Serratia, and glasshouse potato aphids (Aulacorthum solani) were coinfected with Regiella and Spiroplasma; other secondary endosymbionts detected in samples occurred by themselves. Hamiltonella, Rickettsia and Wolbachia were restricted to a single aphid species, whereas Regiella was found in multiple species. Rickettsiella, Hamiltonella and Serratia were stably maintained in laboratory cultures, although others were lost rapidly. The overall incidence of secondary endosymbionts in Australian samples tended to be lower than recorded from aphids overseas. These results indicate that aphid endosymbionts probably exhibit different levels of infectivity and vertical transmission efficiency across hosts, which may contribute to natural infection patterns. The rapid loss of some endosymbionts in cultures raises questions about factors that maintain them under field conditions, while endosymbionts that persisted in laboratory culture provide candidates for interspecific transfers.     

昆虫内共生菌在昆虫防御中的作用

共生菌在昆虫体内的生长、繁殖过程中起着十分重要的作用,如营养功能、解毒作用、调控生殖及与寄主适应性等等。近些年,一些学者提出了共生菌在寄主抵御天敌等逆境的胁迫时也具有重要的作用。其在寄主防御中的作用可分为3类:(1)昆虫体内共生菌能产生具有抗真菌作用的代谢物,能够抵御病原真菌的入侵,提高寄主的适应性。(2)昆虫抵御寄生蜂的作用与其体内的共生细菌密切相关。(3)共生菌也能产生有毒物质保护寄主免于被捕食。昆虫共生菌在寄主防御中的作用机制除了产生抗菌物质和聚酮类等有毒物质外,还可能通过与水平传播的寄生物(horizontal transmission parasites,HTPs)竞争寄主资源来直接保护寄主。对共生菌的作用的深入研究将有可能进一步明确其功能,并加以利用。     

Prevalence and penetrance variation of male-killing Wolbachia across Indo-Pacific populations of the butterfly Hypolimnas bolina

Male-killing bacteria are generally thought to attain low to intermediate prevalence in natural populations, with only mild effects on the host population sex ratio. This view was recently challenged by reports of extremely high infection frequencies in three butterfly species, raising the prospect that male killers, by making males rare, might drive many features of host ecology and evolution. To assess this hypothesis, it is necessary to evaluate how often male killers actually produce a highly female-biased population sex ratio in nature, which requires both high prevalence of infection and high penetrance of action. To this end, we surveyed South Pacific and Southeast Asian populations of Hypolimnas bolina, a butterfly in which extreme prevalence of male-killing Wolbachia bacteria has recently been recorded. Our results indicate that highly female-biased populations are common in Polynesia, with 6 out of 12 populations studied having in excess of 70% of females infected with a fully efficient male killer. However, heterogeneity is extreme in Polynesia, with the male-killing Wolbachia absent from three populations. In contrast to the Polynesian situation, Wolbachia does not kill males in any of the three Southeast Asian populations studied, despite its very high prevalence there. We conclude that male killers are likely to have significant ongoing ecological and evolutionary impact in 6 of the 15 populations surveyed. The causes and consequences of the observed spatial variation are discussed with respect to host resistance evolution, host ecology and interference with additional symbionts.     

Propagule pressure and persistence in experimental populations

Average inoculum size and number of introductions are known to have positive effects on population persistence. However, whether these factors affect persistence independently or interact is unknown. We conducted a two-factor experiment in which 112 populations of parthenogenetic Daphnia magna were maintained for 41 days to study effects of inoculum size and introduction frequency on: (i) population growth, (ii) population persistence and (iii) time-to-extinction. We found that the interaction of inoculum size and introduction frequency-the immigration rate-affected all three dependent variables, while population growth was additionally affected by introduction frequency. We conclude that for this system the most important aspect of propagule pressure is immigration rate, with relatively minor additional effects of introduction frequency and negligible effects of inoculum size.     

Patch edges and insect populations

Responses of insect populations may be related to patch size and patch edge responses, but it is not clear how to identify these rapidly. We used a random-walk model to identify three qualitative responses to edges: no edge effect (the null model), reflecting edges and absorbing edges. Interestingly, no edge effect meant that abundance was lower at edges than in the center of patches, and reflecting edges have similar abundance at edges and centers. We then characterized several insect species’ response within maize plots to patch edges and patch size, using a simple, quick, qualitative experiment. Coleomegilla maculata and Trichogramma spp. were the only organisms that responded to patch size and edges as patch theory and the null edge model would predict. Ostrinia nubilalis larvae and possibly Rhopalosiphum maidis and eggs of Chrysopa spp. responded to patch size and edges as predicted by an attracting edge model. Estimation of predation rates suggested that the spatial distribution of these species might be determined by predators. Edge effects or the lack thereof relative to patch size may be rapidly determined for arthropod species, which could lead to understanding the mechanism(s) underlying these effects. This information may be useful in reaction diffusion models through a scaling-up approach to predict population structure of species among patches in a landscape. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Fluctuations in resource availability and insect populations

Summary Data are presented which show that fluctuations in the numbers of many insects are determined primarily by fluctuations in the carrying capacity of their habitats. Few studies exist, however, in which the resources available to insect populations have been quantified; due possibly to a too ready acceptance by many biologists of the concept of population equilibria and a concomitant dismissal of the likelihood of resource limitation.     

Macroevolutionary persistence of heritable endosymbionts: acquisition,retention and expression of adaptive phenotypes in Spiroplasma

The phylogenetic incongruence between insects and their facultative maternally transmitted endosymbionts indicates that these infections are generally short‐lived evolutionarily. Therefore, long‐term persistence of many endosymbionts must depend on their ability to colonize and spread within new host species. At least 17 species of Drosophila are infected with endosymbiotic Spiroplasma that have various phenotypic effects. We transinfected five strains of Spiroplasma from three divergent clades into Drosophila neotestacea to test their capacity to spread in a novel host. A strain that causes male killing in Drosophila melanogaster (its native host) also does so in D. neotestacea, even though these host species diverged 40–60 mya. A strain native to D. neotestacea (designated sNeo) and the two other strains of the poulsonii clade of Spiroplasma confer resistance to wasp parasitism, suggesting that this trait may be ancestral within this clade of Spiroplasma. Conversely, no strain other than sNeo conferred resistance to the sterilizing effects of nematode parasitism, suggesting that nematode resistance is a recently derived condition. The apparent addition of nematode resistance to a Spiroplasma lineage that already confers resistance to wasp parasitism suggests endosymbionts can increase the repertoire of traits conducive to their spread. The capacity of an endosymbiont to undergo maternal transmission and express adaptive phenotypes in novel hosts, without requiring a period of host–symbiont co‐evolution, enables the spread of such symbionts immediately after the colonization of a new host. This could be critical for the macroevolutionary persistence of facultative endosymbionts whose sojourn times within individual host species are relatively brief.     

Estimation of nektonic insect populations

SUMMARY 1. A water column sampler is described for obtaining absolute population estimates of nektonic insects in shallow waters.
2. Relative population estimates were obtained for Corixidae (Hemip-tera) and Buenoa (Hemiptera: Notonectidae) using a standardized sweep net procedure and compared to absolute estimates from column samples. These two types of estimates were highly correlated among lakes.
3. Simultaneous prediction and tolerance intervals for predictions from linear regressions indicated that reliable absolute population estimates generally cannot be expected from relative estimates     

Reduced flight capability in British Virgin Island populations of a wing-dimorphic insect: the role of habitat isolation, persistence, and structure

1. The effects of habitat isolation, persistence, and host‐plant structure on the incidence of dispersal capability (per cent macroptery) in populations of the delphacid planthopper Toya venilia were examined throughout the British Virgin Islands. The host plant of this delphacid is salt grass Sporobolus virginicus, which grows either in undisturbed habitats (large expanses on intertidal salt flats and around the margins of salt ponds, or small patches of sparse vegetation on sand dunes along the shore), or in less persistent, disturbed habitats (managed lawns). 2. Both sexes of T. venilia were significantly more macropterous in disturbed habitats (77.1% in males, 12.5% in females) than in more persistent, undisturbed habitats (19.2% in males, < 1% in females). 3. Males exhibited significantly higher levels of macroptery (26.9 ± 7.6%) than did females (2.0 ± 1.7%), and per cent macroptery was positively density dependent for both sexes in field populations. 4. There was no evidence that the low incidence of female macroptery in a subset of island populations inhabiting natural habitats (1.7 ± 1.2%) was attributable to the effects of isolation on oceanic islands. The incidence of macroptery in British Virgin Island populations of T. venilia was not different from that observed in mainland delphacid species existing in habitats of similar duration. 5. Rather, the persistence of most salt grass habitats throughout the British Virgin Islands best explains the evolution of flight reduction in females of this island‐inhabiting delphacid. 6. Males were significantly more macropterous in populations occupying dune vegetation (37.6 ± 9.8%) than in populations occupying salt flat–pond margin habitats (7.6 ± 5.6%). By contrast, females exhibited low levels of macroptery in both dune (0%) and salt flat–pond margin (< 1%) habitats. Variation in salt‐grass structure probably underlies this habitat‐related difference in macroptery because flight‐capable males of planthoppers are better able to locate females in the sparse‐structured grass growing on dunes. This habitat‐related difference in male macroptery accounted for the generally higher level of macroptery observed in males than in females throughout the islands. 7. The importance of habitat persistence and structure in explaining the incidence of dispersal capability in T. venilia is probably indicative of the key role these two factors play in shaping the dispersal strategies of many insects.     

Unpredicted impacts of insect endosymbionts on interactions between soil organisms,plants and aphids

Ecologically significant symbiotic associations are frequently studied in isolation, but such studies of two-way interactions cannot always predict the responses of organisms in a community setting. To explore this issue, we adopt a community approach to examine the role of plant–microbial and insect–microbial symbioses in modulating a plant–herbivore interaction. Potato plants were grown under glass in controlled conditions and subjected to feeding from the potato aphid Macrosiphum euphorbiae. By comparing plant growth in sterile, uncultivated and cultivated soils and the performance of M. euphorbiae clones with and without the facultative endosymbiont Hamiltonella defensa, we provide evidence for complex indirect interactions between insect– and plant–microbial systems. Plant biomass responded positively to the live soil treatments, on average increasing by 15% relative to sterile soil, while aphid feeding produced shifts (increases in stem biomass and reductions in stolon biomass) in plant resource allocation irrespective of soil treatment. Aphid fecundity also responded to soil treatment with aphids on sterile soil exhibiting higher fecundities than those in the uncultivated treatment. The relative allocation of biomass to roots was reduced in the presence of aphids harbouring H. defensa compared with plants inoculated with H. defensa-free aphids and aphid-free control plants. This study provides evidence for the potential of plant and insect symbionts to shift the dynamics of plant–herbivore interactions.     

Stability of environment and of insect populations

The abundance of moths was monitored with light-traps in two sites in southern Bohemia, České Budějovice for 22 years and in Černiš for 9 years. In these sites, that are vastly different in environmental stability and predictability, stability of insect populations was studied. The amplitude of fluctuations in abundance of the insect populations, as measured by the coefficient of variation (CV), varied a great deal between species so that there was a large overlap between the two sites. Nevertheless there was a highly significant tendency for species at Černiš, the more stable site, to have smaller values of CV, i.e., to be less fluctuating. Also in species co-occurring in the two sites, the CV at Černiš tended to be smaller. Trends in abundance of individual species over time, both increases and decreases, were common in both sites and did not differ between habitats. Environmental stability begets insect population stability in terms of the amplitude of the fluctuations, but trends in time occur irrespective of stability of the habitat.     

The endosymbionts Wolbachia and Cardinium and their effects in three populations of the predatory mite Neoseiulus paspalivorus

Whereas endosymbiont-induced incompatibility is known to occur in various arthropod taxa, such as spider mites, insects and isopods, it has been rarely reported in plant-inhabiting predatory mites (Acari: Phytoseiidae). Recent cross-breeding studies with the phytoseiid mite Neoseiulus paspalivorus De Leon revealed a complete post-mating reproductive isolation between specimens collected from three geographic origins—Northeast Brazil (South America), Benin and Ghana (West Africa)—even though they are morphologically similar. We carried out a study to assess to what extent these populations exhibit genetic differences and whether endosymbionts are involved in the incompatibility. First, we used the mitochondrial cytochrome oxidase I (COI) gene to assess genetic diversity among the three populations. Second, we used a PCR-based method to check for the presence of Wolbachia and/or Cardinium in these populations, and we determined their phylogenetic relationships using specific primers for Wolbachia and Cardinium 16S rDNA genes. Third, we also conducted a test using an antibiotic (tetracycline) in an attempt to eliminate the symbionts and evaluate their effects on the reproductive compatibility of their host. Based on the DNA sequences of their COI genes, specimens of the three populations appear to be genetically similar. However, the 16S rDNA gene sequences of their associated endosymbionts differed among the three populations: the Benin and Brazil populations harbour different strains of Wolbachia symbionts, whereas the Ghana population harbours Cardinium symbionts. In response to antibiotic treatment females of each of the three populations became incompatible with untreated males of their own population, similar to that observed in crossings between females from one geographic population and males from another. Compatibility was restored in crosses involving uninfected Brazil females and uninfected Benin males, whereas the reciprocal crosses remained incompatible. Cardinium symbionts seem to be essential for oviposition in the Ghana population. It is concluded that their associated bacterial symbionts are the cause of the post-mating reproductive isolation previously observed among the three geographic populations. This insight is relevant to biological control of coconut mites for which N. paspalivorus is an effective predator, because introducing one geographic strain into the population of another (e.g. in field releases or mass cultures) may cause population growth depression.     

Metapopulation processes and persistence in remnant water vole populations

We examined the spatial distribution of water vole populations in four consecutive years and investigated whether the regional population processes of extinction, recolonisation and migration influence distribution and persistence. We examined how such regional processes are influenced by spatial variation in habitat quality. In addition, we assessed the relevance of metapopulation concepts for understanding the dynamics of species that deviate from classical metapopulation assumptions and developing conservation measures for them. Populations were patchy and discrete, and the patchy distribution was not static between years. Population turnover occurred even in the absence of predatory mink, which only influenced the network of populations at the end of the study. Most populations were clustered close together in the upper tributaries. Local population persistence was predominantly influenced by population size: large populations were more persistent. Recolonisation rates were influenced by isolation and habitat quality. The isolation estimates which best explained the distribution of water vole populations incorporated straight‐line distances, suggesting water voles disperse overland. The distribution of recolonised sites indicated that dispersing voles actively selected habitat on the basis of its quality. Water voles depart from some of the assumptions made by frequently used metapopulation models. In particular there is no clear binary distinction between suitable and non‐suitable habitat. Accounting for variation in habitat quality before investigating temporal changes in population distribution allowed us to demonstrate that the key metapopulation processes were important. The significance of regional population processes relative to local population processes may have increased in declining, fragmented populations compared to pristine regional populations. We hypothesise that although mink predation is likely to eventually cause regional extinction in many areas, metapopulation processes have delayed this decline. Consequently, conservation measures should take into account mink predation rates and regional population processes, before considering aspects of habitat quality.