Comparisons of host mitochondrial, nuclear and endosymbiont bacterial genes reveal cryptic fig wasp species and the effects of Wolbachia on host mtDNA evolution and diversity

Background Figs and fig-pollinating wasp species usually display a highly specific one-to-one association. However, more and more studies have revealed that the "one-to-one" rule has been broken. Co-pollinators have been reported, but we do not yet know how they evolve. They may evolve from insect speciation induced or facilitated by Wolbachia which can manipulate host reproduction and induce reproductive isolation. In addition, Wolbachia can affect host mitochondrial DNA evolution, because of the linkage between Wolbachia and associated mitochondrial haplotypes, and thus confound host phylogeny based on mtDNA. Previous research has shown that fig wasps have the highest incidence of Wolbachia infection in all insect taxa, and Wolbachia may have great influence on fig wasp biology. Therefore, we look forward to understanding the influence of Wolbachia on mitochondrial DNA evolution and speciation in fig wasps. Results We surveyed 76 pollinator wasp specimens from nine Ficus microcarpa trees each growing at a different location in Hainan and Fujian Provinces, China. We found that all wasps were morphologically identified as Eupristina verticillata, but diverged into three clades with 4.22-5.28% mtDNA divergence and 2.29-20.72% nuclear gene divergence. We also found very strong concordance between E. verticillata clades and Wolbachia infection status, and the predicted effects of Wolbachia on both mtDNA diversity and evolution by decreasing mitochondrial haplotypes. Conclusions Our study reveals that the pollinating wasp E. verticillata on F. microcarpa has diverged into three cryptic species, and Wolbachia may have a role in this divergence. The results also indicate that Wolbachia strains infecting E. verticillata have likely resulted in selective sweeps on host mitochondrial DNA.

Measuring the discrepancy between fecundity and lifetime reproductive success in a pollinating fig wasp

Lifetime reproductive success in female insects is often egg- or time-limited. For instance in pro-ovigenic species, when oviposition sites are abundant, females may quickly become devoid of eggs. Conversely, in the absence of suitable oviposition sites, females may die before laying all of their eggs. In pollinating fig wasps (Hymenoptera: Agaonidae), each species has an obligate mutualism with its host fig tree species [Ficus spp. (Moraceae)]. These pro-ovigenic wasps oviposit in individual ovaries within the inflorescences of monoecious Ficus (syconia, or ‘figs’), which contain many flowers. Each female flower can thus become a seed or be converted into a wasp gall. The mystery is that the wasps never oviposit in all fig ovaries, even when a fig contains enough wasp females with enough eggs to do so. The failure of all wasps to translate all of their eggs into offspring clearly contributes to mutualism persistence, but the underlying causal mechanisms are unclear. We found in an undescribed Brazilian Pegoscapus wasp population that the lifetime reproductive success of lone foundresses was relatively unaffected by constraints on oviposition. The number of offspring produced by lone foundresses experimentally introduced into receptive figs was generally lower than the numbers of eggs carried, despite the fact that the wasps were able to lay all or most of their eggs. Because we excluded any effects of intraspecific competitors and parasitic non-pollinating wasps, our data suggest that some pollinators produce few offspring because some of their eggs or larvae are unviable or are victims of plant defences.

Convergent structure of multitrophic communities over three continents

Ecological theory predicts that communities using the same resources should have similar structure, but evolutionary constraints on colonization and niche shifts may hamper such convergence. Multitrophic communities of wasps exploiting fig fruits, which first evolved about 75MYA, do not show long-term “inheritance” of taxonomic (lineage) composition or species diversity. However, communities on three continents have converged ecologically in the presence and relative abundance of five insect guilds that we define. Some taxa fill the same niches in each community (phylogenetic niche conservatism). However, we show that overall convergence in ecological community structure depends also on a combination of niche shifts by resident lineages and local colonizations of figs by other insect lineages. Our study explores new ground, and develops new heuristic tools, in combining ecology and phylogeny to address patterns in the complex multitrophic communities of insect on plants, which comprise a large part of terrestrial biodiversity.

Biased oviposition and biased survival together help resolve a fig-wasp conflict

We report evidence that helps resolve two competing explanations for stability in the mutualism between Ficus racemosa fig trees and the Ceratosolen fusciceps wasps that pollinate them. The wasps lay eggs in the tree's ovules, with each wasp larva developing at the expense of a fig seed. Upon maturity, the female wasps collect pollen and disperse to a new tree, continuing the cycle. Fig fitness is increased by producing both seeds and female wasps, whereas short-term wasp fitness increases only with more wasps, thereby resulting in a conflict of interests. We show experimentally that wasps exploit the inner layers of ovules first (the biased oviposition explanation), which is consistent with optimal-foraging theory. As oviposition increases, seeds in the middle layer are replaced on a one-to-one basis by pollinator offspring, which is also consistent with biased oviposition. Finally, in the outer layer of ovules, seeds disappear but are only partially replaced by pollinator offspring, which suggests high wasp mortality (the biased survival or ‘unbeatable seeds’ explanation). Our results therefore suggest that both biased oviposition and biased survival ensure seed production, thereby stabilizing the mutualism. We further argue that biased oviposition can maintain biased survival by selecting against wasp traits to overcome fig defenses. Finally, we report evidence suggesting that F. racemosa balances seed and wasp production at the level of the tree. Because figs are probably selected to allocate equally to male and female function, a 1:1 seed:wasp ratio suggests that fig trees are in control of the mutualism.

Chaos of Wolbachia Sequences Inside the Compact Fig Syconia of Ficus benjamina (Ficus: Moraceae)

Figs and fig wasps form a peculiar closed community in which the Ficus tree provides a compact syconium (inflorescence) habitat for the lives of a complex assemblage of Chalcidoid insects. These diverse fig wasp species have intimate ecological relationships within the closed world of the fig syconia. Previous surveys of Wolbachia, maternally inherited endosymbiotic bacteria that infect vast numbers of arthropod hosts, showed that fig wasps have some of the highest known incidences of Wolbachia amongst all insects. We ask whether the evolutionary patterns of Wolbachia sequences in this closed syconium community are different from those in the outside world. In the present study, we sampled all 17 fig wasp species living on Ficus benjamina, covering 4 families, 6 subfamilies, and 8 genera of wasps. We made a thorough survey of Wolbachia infection patterns and studied evolutionary patterns in wsp (Wolbachia Surface Protein) sequences. We find evidence for high infection incidences, frequent recombination between Wolbachia strains, and considerable horizontal transfer, suggesting rapid evolution of Wolbachia sequences within the syconium community. Though the fig wasps have relatively limited contact with outside world, Wolbachia may be introduced to the syconium community via horizontal transmission by fig wasps species that have winged males and visit the syconia earlier.

Molecular species delimitation of a symbiotic fig-pollinating wasp species complex reveals extreme deviation from reciprocal partner specificity

Background: Symbiotic relationships have contributed to major evolutionary innovations, the maintenance of fundamental ecosystem functions, and the generation and maintenance of biodiversity. However, the exact nature of host/symbiont associations, which has important consequences for their dynamics, is often poorly known due to limited understanding of symbiont taxonomy and species diversity. Among classical symbioses, figs and their pollinating wasps constitute a highly diverse keystone resource in tropical forest and savannah environments. Historically, they were considered to exemplify extreme reciprocal partner specificity (one-to-one host-symbiont species relationships), but recent work has revealed several more complex cases. However, there is a striking lack of studies with the specific aims of assessing symbiont diversity and how this varies across the geographic range of the host. Results: Here, we use molecular methods to investigate cryptic diversity in the pollinating wasps of a widespread Australian fig species. Standard barcoding genes and methods were not conclusive, but incorporation of phylogenetic analyses and a recently developed nuclear barcoding gene (ITS2), gave strong support for five pollinator species. Each pollinator species was most common in a different geographic region, emphasising the importance of wide geographic sampling to uncover diversity, and the scope for divergence in coevolutionary trajectories across the host plant range. In addition, most regions had multiple coexisting pollinators, raising the question of how they coexist in apparently similar or identical resource niches. Conclusion: Our study offers a striking example of extreme deviation from reciprocal partner specificity over the full geographical range of a fig-wasp system. It also suggests that superficially identical species may be able to co-exist in a mutualistic setting albeit at different frequencies in relation to their fig host’s range. We show that comprehensive sampling and molecular taxonomic techniques may be required to uncover the true structure of cryptic biodiversity underpinning intimate ecological interactions.

A trophic cascade induced by predatory ants in a fig-fig wasp mutualism

Summary 1. A trophic cascade occurs when predators directly decrease the densities, or change the behaviour, of herbivores and thus indirectly increase plant productivity. The predator–herbivore– plant context is well known, but some predators attack species beneficial to plants (e.g. pollinators) and/or enemies of herbivores (e.g. parasites), and their role in the dynamics of mutualisms remains largely unexplored. 2. We surveyed the predatory ant species and studied predation by the dominant ant species, the weaver ant Oecophylla smaragdina, associated with the fig tree Ficus racemosa in southwest China. We then tested the effects of weaver ants on the oviposition behaviour of pollinating and non-pollinating fig wasps in an ant-exclusion experiment. The effects of weaver ants on fig wasp community structure and fig seed production were then compared between trees with and without O. smaragdina. 3. Oecophylla smaragdina captured more non-pollinating wasps (Platyneura mayri) than pollinators as the insects arrived to lay eggs. When ants were excluded, more non-pollinators laid eggs into figs and fewer pollinators entered figs. Furthermore, trees with O. smaragdina produced more pollinator offspring and fewer non-pollinator offspring, shifting the community structure significantly. In addition, F. racemosa produced significantly more seeds on trees inhabited by weaver ants. 4. Oecophylla smaragdina predation reverses the dominance of the two commonest wasp species at the egg-laying stage and favours the pollinators. This behavioural pattern is mirrored by wasp offspring production, with pollinators’ offspring dominating figs produced by trees inhabited by weaver ants, and offspring of the non-pollinator P. mayri most abundant in figs on trees inhabited by other ants. 5. Overall, our results suggest that predation by weaver ants limits the success of the non-pollinating P. mayri and therefore indirectly benefits the mutualism by increasing the reproductive success of both the pollinators and the plant. Predation is thus a key functional factor that can shape the community structure of a pollinator-plant mutualistic system. Key-words: competitive release, fig wasp, mutualism, predation, predator-exclusion experiment, trophic cascade

How to be a fig wasp down under: the diversity and structure of an Australian fig wasp community

Endophytic insects and their parasitoids provide valuable models for community ecology. The wasp communities in inflorescences of fig trees have great potential for comparative studies, but we must first describe individual communities. Here, we add to the few detailed studies of such communities by describing the one associated with Ficus rubiginosa in Australia. First, we describe community composition, using two different sampling procedures. Overall, we identified 14 species of non-pollinating fig wasp (NPFW) that fall into two size classes. Small wasps, including pollinators, gallers and their parasitoids, were more abundant than large wasps (both galler and parasitoid species). We show that in figs where wasps emerge naturally, the presence of large wasps may partly explain the low emergence of small wasps. During fig development, large gallers oviposit first, before and around the time of pollination, while parasitoids lay eggs after pollination. We further show that parasitoids in the subfamily Sycoryctinae, which comprise the majority of all individual NPFWs, segregate temporally by laying eggs at different stages of fig development. We discuss our results in terms of species co-existence and community structure and compare our findings to those from fig wasp communities on other continents.

Fighting in fig wasps: do males avoid killing brothers or do they never meet them?

1. In many fig wasp species, armoured wingless males regularly engage in lethal fights for access to females inside figs, which act as discrete mating patches. 2. Kin selection generally opposes killing brothers, because their reproductive success provides indirect genetic benefits (inclusive fitness). However, siblicide may be avoided if (i) brothers do not occur in the same figs, or (ii) males avoid fighting brothers in the same fig. Alternatively, (iii) siblicide may occur because intense mate competition between brothers at the local scale overcomes kin selection effects, or (iv) males do not recognise kin. 3. A fig may also contain wasps from other closely related species and it is not known if males also fight with these individuals. 4. Nine microsatellite loci were used in the first genetic analysis of fighting in fig wasps. We assigned species and sibling identities to males and tested alternative fighting scenarios for three Sycoscapter wasp species in figs of Ficus rubiginosa. 5. Approximately 60% of figs contained males frommore than one Sycoscapter species and approximately 80% of fights were between conspecifics, but a surprising 20% were between heterospecific males. 6.Within species, fewfigs contained brothers, suggesting that females typically lay one son per fig. Overall, most males do not compete with brothers and all fights observed were between unrelated males. Key words:Competition, fighting, genetics, kin selection, microsatellites, relatedness.

Polietileno de baixa densidade (PEBD) na conservação pós-colheita de figos cv. Roxo de Valinhos

Estudou-se o efeito de diferentes embalagens de polietileno em figos cv. Roxo de Valinhos, sob condições de frigoconservação. Os frutos colhidos no início do estádio de maturação, foram limpos e selecionados, sendo após embalados em filmes plásticos de polietileno de diferentes espessuras, constituindo assim os tratamentos: 1(controle), 2 - PEBD com 6 µm de espessura, 3 - PEBD com 10 µm de espessura, 4 - PEBD com 15 µm de espessura, 5 - PEBD com 22 µm de espessura. Os frutos foram então armazenados em câmara fria com temperatura de -- 0,5 °C e 85-90 % de UR, por oito dias. As análises foram realizadas diariamente, avaliando-se os seguintes parâmetros: perda de massa fresca, aspecto visual, firmeza de polpa, sólidos solúveis totais (SST), acidez total titulável (ATT) e relação SST/ATT. Ao final, verificou-se que os frutos embalados em PEBD de 22 µm apresentaram maior firmeza de polpa, melhor aspecto visual, menores teores de sólidos solúveis totais, maiores níveis de acidez total titulável e menores valores na relação SST/ATT, quando comparados aos demais tratamentos. No parâmetro perda de massa fresca, todos os tratamentos foram estatisticamente superiores ao controle.

Épocas de poda na produção de figos verdes 'Roxo de Valinhos' em sistema orgânico na região oeste do Paraná

O objetivo deste trabalho foi verificar o efeito da época de poda sobre o crescimento e a produção da figueira 'Roxo de Valinhos', cultivada na região oeste do Paraná, sob sistema orgânico, para a produção de figos-verdes. O delineamento utilizado foi em blocos ao acaso, com quatro blocos e seis tratamentos - épocas de poda: abril, maio, junho, julho, agosto e setembro. em cada parcela, constituída de três plantas úteis, foram coletados dados no ciclo de produção de 2007/2008. As variáveis fenológicas, comprimento final médio dos ramos, produção por mês (número e massa média de frutos por planta e produtividade estimada, entre os meses de outubro a fevereiro), produção acumulada, produtividade estimada acumulada e massa fresca média dos frutos. Houve diferenças na produção entre as épocas de poda; plantas podadas em julho e agosto apresentaram as maiores produções, escalonadas entre os meses de dezembro a fevereiro, com pico de produção em janeiro.

Sistema desponte na produção de figos verdes 'Roxo de Valinhos'

O objetivo deste trabalho foi estudar o efeito do sistema desponte sobre o desenvolvimento e a produção de figos verdes 'Roxo de Valinhos'. O experimento utilizando plantas de quatro anos de idade, com espaçamento 3 x 2m, foi conduzido de julho de 2007 a março de 2008, em Quatro Pontes, Paraná (PR). O delineamento utilizado foi em blocos ao acaso, com quatro blocos, e os tratamentos foram arranjados em fatorial 2 x 5, tomando por fatores o número de ramos produtivos (plantas conduzidas com seis ou 12 ramos) e o número de despontes (um, dois, três ou quatro, além do controle sem desponte). No sistema desponte, após a emissão da 16a folha, o ramo foi despontado (gema apical removida), selecionando-se duas brotações por ramo produtivo. Novos despontes foram realizados posteriormente, sempre após a emissão da sexta folha. em cada parcela, constituída de três plantas úteis, foram coletados dados no ciclo de produção 2007/08. A maior produção (2.208,87g planta-1) e produtividade estimada (3.681,19kg ha-1) observada de figos verdes foram obtidas quando as plantas foram conduzidas com 12 ramos produtivos, efetuando-se três despontes.

Seleção de clones de figueira cv. roxo de Valinhos formados por gemas irradiadas

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Conservação pós-colheita de figos verdes (Ficus carica L.) cv. roxo de Valinhos tratados com hipoclorito de sódio e armazenados sob refrigeração em atmosfera modificada passiva

Os principais causadores de perdas da qualidade de figos são: colheita e embalagens inadequadas, falta de padronização do produto na classificação e péssimas condições de transporte e armazenamento. Com este trabalho objetivou-se avaliar o efeito da imersão em hipoclorito, tipo de embalagem e refrigeração na conservação pós-colheita de figos verdes, cv. Roxo de Valinhos, mediante as características físicas, físico-químicas e químicas, durante o armazenamento. Após seleção, os figos foram imersos ou não em solução de hipoclorito de sódio a 40 ppm conforme os tratamentos, secos ao ar e embalados em filme de PVC de 50 m ou sacos plásticos. Após os tratamentos, os frutos foram submetidos a armazenamento refrigerado (1ºC e 70% de UR em BOD), por um período de 35 dias, sendo avaliados a cada 7 dias. O uso da embalagem reduziu drasticamente a perda de massa dos figos. Frutos não embalados apresentaram-se mais ácidos e com menores teores de açúcares. Com o uso da embalagem, os figos podem ser comercializados até os 35 dias de armazenamento desde que armazenados a 1ºC e 70% de UR em BOD. Já os figos não embalados, estes podem ser comercializados somente até os 2 dias de armazenamento sob refrigeração.

Mapeamento do contorno estrutural do embasamento da bacia sedimentar de Curitiba - PR

The sedimentary Curitiba basin is located in the Central-Southern part of the first Parananense plateau, and comprises Curitiba (PR), and part of the neighbour Municipalities (fig.1). It is supposed to be of Plio-Pleistocene age. It has a shallow sedimentary fulfillment, represented by the Guabirotuba formation (BIGARELLA and SALAMUNI, 1962) which is dristributed over a large area of about 3.000km2. The internal geometry, not entirely known yet, is actually object of detailed research, that shows its geological evolution to Cenozoic tectonic movements. For the purpose of this study the definition of the structural contour of the basement and their depo-centers is fundamental. This paper presents the results of the integration of surface and subsurface data, processed by statistical methods, which allowed a more precise definition of the morphostructural framework of the basement. For the analysis of the geological spacial data, specific softwares were used for statistical processing for trend surfaces analysis. The data used in this study are of following types: a) drilling logs for ground water; b) description of surface points of geological maps (CRPM, 1977); c) description of points of geotechnical drillings and down geological survey. The data of 223 drilling logs for ground water were selected out of 770 wells. The description files of 700 outcrops, as well as planialtimetric field data, were used for the localization of the basement outcrop. Thus, a matrix with five columns was set up: utm E-W (x) and utm N-S (y); surface altitude (z); altimetric cote of the contact between sedimentary rocks and the basement (k); isopachs (l). For the study of the basement limits, the analysis of surface trends of 2(nd) and 3(rd) degree polinomial for the altimetric data (figs. 2 and 3) were used. For the residuals the method of the inverse of the square of the distance (fig.4) was used. The adjustments and the explanations of the surfaces were made with the aid of multiple linear regressions. The analysis of 3rd degree polinomial trend surface (fig.3) confirmed that the basement tends to be more exposed towards NNW-SSE explaining better the data trend through an ellipse, which striking NE-SW and dipping SW axis coincides with the trough of the basin observed in the trending surface of the basement. The performed analysis and the respective images offer a good degree of certainty of the geometric model of the Curitiba Basin and of the morphostructure of its basement. The surface trend allows to sketch with a greater degree of confidence the structural contour of the topgraphic surface (figs. 5 and 6) and of the basement (figs. 7 and 8), as well as the delimitation of intermediate structural heights, which were responsible for isolated and assymmetric depocenters. These details are shown in the map of figures 9 and 10. Thus, the Curitiba Basin is made up by a structural trough stretching NE-SW, with maximum preserved depths of about 80m, which are separated by heights and depocenters striking NW-SE (fig. 11). These structural features seems to have been controlled by tectonic reactivation during the Tertiary (HASUI, 1990) and which younger dissection was conditioned by neotectonic processes (SALAMUNI and EBERT, 1994).