Cytoplasmic Incompatibility Research Articles

Cardinium Localization During Its Parasitoid Wasp Host’s Development Provides Insights Into Cytoplasmic Incompatibility

Arthropods harbor heritable intracellular symbionts that may manipulate host reproduction to favor symbiont transmission. In cytoplasmic incompatibility (CI), the symbiont sabotages the reproduction of infected males such that high levels of offspring mortality result when they mate with uninfected females. In crosses with infected males and infected females, however (the “rescue” cross), normal numbers of offspring are produced. A common CI-inducing symbiont, Cardinium hertigii, causes variable levels of CI mortality in the parasitoid wasp, Encarsia suzannae. Previous work correlated CI-induced mortality with male development time in this system, although the timing of Cardinium CI-induction and the relationship between development time and CI mortality was not well understood. Here, using a combination of crosses, manipulation of development time, and fluorescence microscopy, we identify the localization and the timing of the CI-induction step in the Cardinium-E. suzannae system. Antibiotic treatment of adult Cardinium-infected males did not reduce the mortality associated with the CI phenotype, suggesting that CI-alteration occurs prior to adulthood. Our results suggest that the alteration step occurs during the pupal period, and is limited by the duration of pupal development: 1) Encarsia produces most sperm prior to adulthood, 2) FISH localization of Cardinium in testes showed an association with sperm nuclei throughout spermatogenesis but not with mature sperm, and 3) two methods of prolonging the pupal period (cool temperatures and the juvenile hormone analog methoprene) both caused greater CI mortality, suggesting the degree of alteration is limited by the duration of the pupal stage. Based on these results, we compare two models for potential mechanisms of Cardinium sperm modification in the context of what is known about analogous mechanisms of Wolbachia, a more extensively studied CI-inducing symbiont.

The impacts of cytoplasmic incompatibility factor (cifA and cifB) genetic variation on phenotypes.

Wolbachia are maternally transmitted, intracellular bacteria that can often selfishly spread through arthropod populations via cytoplasmic incompatibility (CI). CI manifests as embryonic death when males expressing prophage WO genes cifA and cifB mate with uninfected females or females harboring an incompatible Wolbachia strain. Females with a compatible cifA-expressing strain rescue CI. Thus, cif-mediated CI confers a relative fitness advantage to females transmitting Wolbachia. However, whether cif sequence variation underpins incompatibilities between Wolbachia strains and variation in CI penetrance remains unknown. Here, we engineer Drosophila melanogaster to transgenically express cognate and non-cognate cif homologs and assess their CI and rescue capability. Cognate expression revealed that cifA;B native to D. melanogaster causes strong CI, and cognate cifA;B homologs from two other Drosophila-associated Wolbachia cause weak transgenic CI, including the first demonstration of phylogenetic type 2 cifA;B CI. Intriguingly, non-cognate expression of cifA and cifB alleles from different strains revealed that cifA homologs generally contribute to strong transgenic CI and interchangeable rescue despite their evolutionary divergence, and cifB genetic divergence contributes to weak or no transgenic CI. Finally, we find that a type 1 cifA can rescue CI caused by a genetically divergent type 2 cifA;B in a manner consistent with unidirectional incompatibility. By genetically dissecting individual CI functions for type 1 and 2 cifA and cifB, this work illuminates new relationships between cif genotype and CI phenotype. We discuss the relevance of these findings to CI’s genetic basis, phenotypic variation patterns, and mechanism.

Managing and monitoring genetic isolation and local adaptation of endemic and introduced Cotesia sesamiae for the biological control of the cereal stemborer Busseola fusca in Cameroon

The success of biological bontrol (BC) introductions can be enhanced by considering theory and knowledge of biological systems. The gregarious braconid parasitoid Cotesia sesamiae (Cameroon) is one of the best studied biological control agent from the perspective of molecular ecology. Its evolutionary adaptation to the target host involves symbiotic partners. Polydnaviruses are responsible for immune and developmental adaptations whereas Wolbachia bacteria may reinforce this local adaptation though genetic isolation mechanisms. The noctuid Busseola fusca is a major stemborer pest of maize in sub-Saharan Africa. In contrast to eastern Africa, C. sesamiae is rarely found on B. fusca in western Africa. It is however often obtained from other stemborer species feeding on wild grasses. A biological control project was launched in 2006–2007 by introducing to Cameroon seven crosses of Kenyan populations of C. sesamiae collected in different ecozones. They included populations adapted to B. fusca that develop on maize as well as populations adapted to other hosts feeding on wild plants to allow carryover between cropping seasons. Wolbachia strains responsible for cytoplasmic reproductive incompatibilities with endemic strains were included in the crosses to limit genetic exchanges between introduced and endemic C. sesamiae and preserve genetic adaptation to B. fusca of the introduced populations, while at the same time preserving their ability to survive on wild plants. Six post release surveys were carried out on maize from 2007 to 2013, and on wild grasses in 2013. A total of 393 C. sesamiae individuals, each from one cocoon mass, were genotyped for 11 microsatellite loci. Multidimensional scaling analysis, STRUCTURE and GENECLASS analyses assigned almost all the parasitoids recovered from maize to those introduced from Kenya. The introduced strains were also recovered from wild host plants with little genetic exchanges with endemics. Each population remained strongly associated with its original Wolbachia component, suggesting that Wolbachia may contribute to genetic isolation between endemics and introduced populations in wild host plants when maize is absent, thereby conciliating biological control success and safety.

Supplemental Material for Shropshire, Rosenberg, and Bordenstein, 2020

Wolbachia endosymbionts in arthropods often cause cytoplasmic incompatibility (CI) that results in embryonic death when infected males mate with uninfected females or females harboring different Wolbachia strains. Here, we use Drosophila transgenics and a variety of CI gene (cifA and cifB) homologs to test if sequence variation causes phenotypic variation in the strength of CI and incompatibilities between strains. We report that cif homologs cause weak and rescuable CI, closely related cifA homologs are often interchangeable and compatible, cifB sequence variation associates with CI levels, and divergent homologs can exhibit unidirectional CI against each other.

A Role for Maternal Factors in Suppressing Cytoplasmic Incompatibility.

Wolbachia are maternally transmitted bacterial endosymbionts, carried by approximately half of all insect species. Wolbachia prevalence in nature stems from manipulation of host reproduction to favor the success of infected females. The best known reproductive modification induced by Wolbachia is referred to as sperm-egg Cytoplasmic Incompatibility (CI). In CI, the sperm of Wolbachia-infected males cause embryonic lethality, attributed to paternal chromatin segregation defects during early mitotic divisions. Remarkably, the embryos of Wolbachia-infected females “rescue” CI lethality, yielding egg hatch rates equivalent to uninfected female crosses. Several models have been discussed as the basis for Rescue, and functional evidence indicates a major contribution by Wolbachia CI factors. A role for host contributions to Rescue remains largely untested. In this study, we used a chemical feeding approach to test for CI suppression capabilities by Drosophila simulans. We found that uninfected females exhibited significantly higher CI egg hatch rates in response to seven chemical treatments that affect DNA integrity, cell cycle control, and protein turnover. Three of these treatments suppressed CI induced by endogenous wRi Wolbachia, as well as an ectopic wMel Wolbachia infection. The results implicate DNA integrity as a focal aspect of CI suppression for different Wolbachia strains. The framework presented here, applied to diverse CI models, will further enrich our understanding of host reproductive manipulation by insect endosymbionts.

Large scale genome reconstructions illuminate Wolbachia evolution

Wolbachia is an iconic example of a successful intracellular bacterium. Despite its importance as a manipulator of invertebrate biology, its evolutionary dynamics have been poorly studied from a genomic viewpoint. To expand the number of Wolbachia genomes, we screen over 30,000 publicly available shotgun DNA sequencing samples from 500 hosts. By assembling over 1000 Wolbachia genomes, we provide a substantial increase in host representation. Our phylogenies based on both core-genome and gene content provide a robust reference for future studies, support new strains in model organisms, and reveal recent horizontal transfers amongst distantly related hosts. We find various instances of gene function gains and losses in different super-groups and in cytoplasmic incompatibility inducing strains. Our Wolbachia-host co-phylogenies indicate that horizontal transmission is widespread at the host intraspecific level and that there is no support for a general Wolbachia-mitochondrial synchronous divergence.

Genes associated with Wolbachia-induced cytoplasmic incompatibility in natural populations of Culex pipiens pallens: a preliminary study

To investigate the genes involved in Wolbachia-induced cytoplasmic incompatibility among three natural populations of Culex pipiens pallens in eastern China, so as to provide insights into the development of preventive and control measures for mosquito-borne diseases based on Wolbachia. The cytoplasmic incompatibility was tested among three natural populations of C. pipiens pallens collected from Nanjing and Wuxi of Jiangsu Province and Tangkou of Shandong Province using reciprocal crosses. Wolbachia infection was detected in C. pipiens pallens using a PCR assay, and the expression of Wolbachia wsp and WD0513 genes was quantified using a fluorescent quantitative real-time PCR (qPCR) assay. Bidirectional compatibility was found between the natural populations of C. pipiens pallens collected from Nanjing and Wuxi of Jiangsu Province (t = 0.57 and 0.15, both P values > 0.05), while bidirectional incompatibility was seen between the natural populations of C. pipiens pallens collected from Tangkou of Shandong Province and Wuxi of Jiangsu Province (t = 63.81 and 43.51, both P values < 0.01), and between the natural populations of C. pipiens pallens collected from Nanjing of Jiangsu Province and Tangkou of Shandong Province (t = 39.62 and 43.12, both P values < 0.01). Wolbachia wsp gene was amplified in all three natural populations of C. pipiens pallens, and qPCR assay detected no significant difference in the Wolbachia wsp gene expression among the three natural populations of C. pipiens pallens (F = 2.15, P > 0.05). In addition, there was no significant difference in the WD0513 gene expression between the natural populations of C. pipiens pallens collected from Tangkou of Shandong Province and Nanjing of Jiangsu Province (q = 8.42, P < 0.05) or between the natural populations of C. pipiens pallens collected from Tangkou of Shandong Province and Wuxi of Jiangsu Province (q = 7.84, P < 0.05); however, there was a significant difference detected in the WD0513 gene expression between the natural populations of C. pipiens pallens collected from Nanjing and Wuxi of Jiangsu Province (q = 0.40, P > 0.05). Different Wolbachia numbers are detected in natural populations of C. pipiens pallens collected from Nanjing and Wuxi of Jiangsu Province and Tangkou of Shandong Province, and WD0513 gene may be involved in the Wolbachia-induced cytoplasmic incompatibility among three natural populations of C. pipiens pallens.

Stable Introduction of Plant-Virus-Inhibiting Wolbachia into Planthoppers for Rice Protection

Progress has been made in developing the maternally inherited endosymbiotic bacterium Wolbachia as a tool for protecting humans from mosquito-borne diseases. In contrast, Wolbachia-based approaches have not yet been developed for the protection of plants from insect pests and their associated diseases, with a major challenge being the establishment of artificial Wolbachia infections expressing desired characteristics in the hemipterans that transmit the majority of plant viruses. Here, we report stable introduction of Wolbachia into the brown planthopper, Nilaparvata lugens, the most destructive rice pest that annually destroys millions of hectares of staple crops. The Wolbachia strain wStri from the small brown planthopper, Laodelphax striatellus, was transferred to this new host, where it showed high levels of cytoplasmic incompatibility, enabling rapid invasion of laboratory populations. Furthermore, wStri inhibited infection and transmission of Rice ragged stunt virus and mitigated virus-induced symptoms in rice plants, opening up the development of Wolbachia-based strategies against major agricultural pests and their transmitted pathogens. VIDEO ABSTRACT.

Wolbachia-Mitochondrial DNA Associations in Transitional Populations of Rhagoletis cerasi.

Simple SummaryWolbachia is an endosymbiotic bacterium that infects numerous insects and crustaceans. Its ability to alter the reproduction of hosts results in incompatibilities of differentially infected individuals. Therefore, Wolbachia has been applied to suppress agricultural and medical insect pests. The European cherry fruit fly, Rhagoletis cerasi, is mainly distributed throughout Europe and Western Asia, and is infected with at least five different Wolbachia strains. The strain wCer2 causes incompatibilities between infected males and uninfected females, making it a potential candidate to control R. cerasi. Thus, the prediction of its spread is of practical importance. Like mitochondria, Wolbachia is inherited from mother to offspring, causing associations between mitochondrial DNA and endosymbiont infection. Misassociations, however, can be the result of imperfect maternal transmission, the loss of Wolbachia, or its horizontal transmission from infected to uninfected individuals. These are important parameters influencing the spread of infection. Here, we studied Wolbachia-mitochondrial haplotype associations in R. cerasi in two transition zones in the Czech Republic and Hungary, where wCer2 is currently spreading. Our results suggest imperfect maternal transmission only in the early phases of wCer2 invasion and no evidence of horizontal transmission of wCer2 in R. cerasi.The endosymbiont Wolbachia can manipulate arthropod host reproduction by inducing cytoplasmic incompatibility (CI), which results in embryonic mortality when infected males mate with uninfected females. A CI-driven invasion of Wolbachia can result in a selective sweep of associated mitochondrial haplotype. The co-inheritance of Wolbachia and host mitochondrial DNA can therefore provide significant information on the dynamics of an ongoing Wolbachia invasion. Therefore, transition zones (i.e., regions where a Wolbachia strain is currently spreading from infected to uninfected populations) represent an ideal area to investigate the relationship between Wolbachia and host mitochondrial haplotype. Here, we studied Wolbachia-mitochondrial haplotype associations in the European cherry fruit fly, Rhagoletis cerasi, in two transition zones in the Czech Republic and Hungary, where the CI-inducing strain wCer2 is currently spreading. The wCer2-infection status of 881 individuals was compared with the two known R. cerasi mitochondrial haplotypes, HT1 and HT2. In accordance with previous studies, wCer2-uninfected individuals were associated with HT1, and wCer2-infected individuals were mainly associated with HT2. We found misassociations only within the transition zones, where HT2 flies were wCer2-uninfected, suggesting the occurrence of imperfect maternal transmission. We did not find any HT1 flies that were wCer2-infected, suggesting that Wolbachia was not acquired horizontally. Our study provides new insights into the dynamics of the early phase of a Wolbachia invasion.

Wolbachia

Ary Hoffman introduces Wolbachia.

Wolbachia affects reproduction in the spider mite Tetranychus truncatus (Acari: Tetranychidae) by regulating chorion protein S38‐like and Rop

Wolbachia-induced reproductive regulation in hosts has been used to control pest populations, but little is known about the molecular mechanism underlying Wolbachia regulation of host genes. Here, reproductive regulation by Wolbachia in the spider mite Tetranychus truncatus was studied at the molecular level. Infection with Wolbachia resulted in decreasing oviposition and cytoplasmic incompatibility in T. truncatus. Further RNA-seq revealed genes regulated by Wolbachia in T. truncatus. Real-time quantitative polymerase chain reaction (qPCR) showed that genes, including chorion protein S38-like and Rop were down-regulated by Wolbachia. RNA interference (RNAi) of chorion protein S38-like and Rop in Wolbachia-uninfected T. truncatus decreased oviposition, which was consistent with Wolbachia-induced oviposition decrease. Interestingly, suppressing Rop in Wolbachia-infected T. truncatus led to increased Wolbachia titres in eggs; however, this did not occur after RNAi of chorion protein S38-like. This is the first study to show that chorion protein S38-like and Rop facilitate Wolbachia-mediated changes in T. truncatus fertility. In addition, RNAi of Rop turned the body colour of Wolbachia-uninfected T. truncatus black, which indicates that the role of Rop is not limited to the reproductive regulation of T. truncatus.

Symbiont-mediated cytoplasmic incompatibility: what have we learned in 50 years?

Cytoplasmic incompatibility (CI) is the most common symbiont-induced reproductive manipulation. Specifically, symbiont-induced sperm modifications cause catastrophic mitotic defects in the fertilized embryo and ensuing lethality in crosses between symbiotic males and either aposymbiotic females or females harboring a different symbiont strain. However, if the female carries the same symbiont strain, then embryos develop properly, thereby imparting a relative fitness benefit to symbiont-transmitting mothers. Thus, CI drives maternally-transmitted bacteria to high frequencies in arthropods worldwide. In the past two decades, CI experienced a boom in interest due to its (i) deployment in worldwide efforts to curb mosquito-borne diseases, (ii) causation by bacteriophage genes, cifA and cifB, that modify sexual reproduction, and (iii) important impacts on arthropod speciation. This review serves as a gateway to experimental, conceptual, and quantitative themes of CI and outlines significant gaps in understanding CI’s mechanism that are ripe for investigation from diverse subdisciplines in the life sciences.

Wolbachia modulates prevalence and viral load of Culex pipiens densoviruses in natural populations.

The inadequacy of standard mosquito control strategies calls for ecologically safe novel approaches, for example the use of biological agents such as the endosymbiotic α-proteobacteria Wolbachia or insect-specific viruses (ISVs). Understanding the ecological interactions between these "biocontrol endosymbionts" is thus a fundamental step. Wolbachia are transmitted vertically from mother to offspring and modify their hosts' phenotypes, including reproduction (e.g., cytoplasmic incompatibility) and survival (e.g., viral interference). In nature, Culex pipiens (sensu lato) mosquitoes are always found infected with genetically diverse Wolbachia called wPip that belong to five phylogenetic groups. In recent years, ISVs have also been discovered in these mosquito species, although their interactions with Wolbachia in nature are unknown. Here, we studied the interactions between a widely prevalent ISV, the Culex pipiens densovirus (CpDV, Densovirinae), and Wolbachia in northern Tunisian C. pipiens populations. We showed an influence of different Wolbachia groups on CpDV prevalence and a general positive correlation between Wolbachia and CpDV loads. By investigating the putative relationship between CpDV diversification and wPip groups in the different sites, we detected a signal linked to wPip groups in CpDV phylogeny in sites where all larvae were infected by the same wPip group. However, no such signal was detected where the wPip groups coexisted, suggesting CpDV horizontal transfer between hosts. Overall, our results provide good evidence for an ecological influence of Wolbachia on an ISV, CpDV, in natural populations and highlight the importance of integrating Wolbachia in our understanding of ISV ecology in nature.

Evolution-guided mutagenesis of the cytoplasmic incompatibility proteins: Identifying CifA's complex functional repertoire and new essential regions in CifB.

Wolbachia are the world's most common, maternally-inherited, arthropod endosymbionts. Their worldwide distribution is due, in part, to a selfish drive system termed cytoplasmic incompatibility (CI) that confers a relative fitness advantage to females that transmit Wolbachia to their offspring. CI results in embryonic death when infected males mate with uninfected females but not infected females. Under the Two-by-One genetic model of CI, males expressing the two phage WO proteins CifA and CifB cause CI, and females expressing CifA rescue CI. While each protein is predicted to harbor three functional domains, there is no knowledge on how sites across these Cif domains, rather than in any one particular domain, contribute to CI and rescue. Here, we use evolution-guided, substitution mutagenesis of conserved amino acids across the Cif proteins, coupled with transgenic expression in uninfected Drosophila melanogaster, to determine the functional impacts of conserved residues evolving mostly under purifying selection. We report that amino acids in CifA's N-terminal unannotated region and annotated catalase-related domain are important for both complete CI and rescue, whereas C-terminal residues in CifA's putative domain of unknown function are solely important for CI. Moreover, conserved CifB amino acids in the predicted nucleases, peptidase, and unannotated regions are essential for CI. Taken together, these findings indicate that (i) all CifA amino acids determined to be crucial in rescue are correspondingly crucial in CI, (ii) an additional set of CifA amino acids are uniquely important in CI, and (iii) CifB amino acids across the protein, rather than in one particular domain, are all crucial for CI. We discuss how these findings advance an expanded view of Cif protein evolution and function, inform the mechanistic and biochemical bases of Cif-induced CI/rescue, and continue to substantiate the Two-by-One genetic model of CI.

Antagonistic interaction between male-killing and cytoplasmic incompatibility induced by Cardinium and Wolbachia in the whitefly, Bemisia tabaci.

Cardinium and Wolbachia are maternally inherited bacterial symbionts of arthropods that can manipulate host reproduction by increasing the fitness of infected females. Here, we report that Cardinium and Wolbachia coinfection induced male-killing and cytoplasmic incompatibility (CI) when they coexisted in a cryptic species of whitefly, Bemisia tabaci Asia II7. Cardinium and Wolbachia symbionts were either singly or simultaneously localized in the bacteriocytes placed in the abdomen of B. tabaci nymphs and adults. Cardinium-Wolbachia coinfection induced male-killing and resulted in a higher female sex ratio in the intraspecific amphigenetic progeny of Asia II7 ICWH and ICWL lines; interestingly, male-killing induction was enhanced with increased Cardinium titer. Moreover, single infection of Wolbachia induced partial CI in the Asia II7 IW line and resulted in reduced fecundity, higher embryonic mortality, and lower female sex ratio. The uninfected Asia II7 IU line had significantly higher fecundity, lower embryonic and nymphal mortalities, and a lower level of CI than both the Wolbachia-infected Asia II7 IW line and the Cardinium-Wolbachia-coinfected Asia II7 ICWH line. Our findings indicate that Cardinium-Wolbachia coinfection induced male-killing, which may have had antagonistic effects on Wolbachia-induced CI in the Asia II7 whiteflies. For the first time, our study revealed that B. tabaci Asia II7 reproduction is co-manipulated by Cardinium and Wolbachia endosymbionts.

Life and Death of Selfish Genes: Comparative Genomics Reveals the Dynamic Evolution of Cytoplasmic Incompatibility.

Cytoplasmic incompatibility is a selfish reproductive manipulation induced by the endosymbiont Wolbachia in arthropods. In males Wolbachia modifies sperm, leading to embryonic mortality in crosses with Wolbachia-free females. In females, Wolbachia rescues the cross and allows development to proceed normally. This provides a reproductive advantage to infected females, allowing the maternally transmitted symbiont to spread rapidly through host populations. We identified homologs of the genes underlying this phenotype, cifA and cifB, in 52 of 71 new and published Wolbachia genome sequences. They are strongly associated with cytoplasmic incompatibility. There are up to seven copies of the genes in each genome, and phylogenetic analysis shows that Wolbachia frequently acquires new copies due to pervasive horizontal transfer between strains. In many cases, the genes have subsequently acquired loss-of-function mutations to become pseudogenes. As predicted by theory, this tends to occur first in cifB, whose sole function is to modify sperm, and then in cifA, which is required to rescue the cross in females. Although cif genes recombine, recombination is largely restricted to closely related homologs. This is predicted under a model of coevolution between sperm modification and embryonic rescue, where recombination between distantly related pairs of genes would create a self-incompatible strain. Together, these patterns of gene gain, loss, and recombination support evolutionary models of cytoplasmic incompatibility.

Cellular Localization of Two Rickettsia Symbionts in the Digestive System and within the Ovaries of the Mirid Bug, Macrolophous pygmaeus.

Simple SummaryLike most insects, those that feed on both prey and plant materials harbor symbiotic bacteria in their body. Yet the involvement of bacteria in the feeding habits of these omnivorous consumers has yet to be investigated. In the present study, we took the first step toward testing the hypothesis that bacterial symbionts are involved in the feeding habits of the omnivorous bug Macrolophus pygmaeus. We (I) characterized the microbiome (the assembly of bacteria and fungi) of M. pygmaeus, and (II) determined the identity and location of the most dominant bacteria species within the host body. We found that M. pygmaeus microbiome is dominated by two Rickettsia species, R. belli and R. limoniae. These bacteria are found in high numbers in the digestive system of the bug, each exhibiting a unique distribution pattern, and for the most part, do not share the same cells in the gut. These results strongly suggest that the host bug may gain some nutritional benefits by hosting the two dominant symbiotic bacteria in its gut.Bacterial symbionts in arthropods are common, vary in their effects, and can dramatically influence the outcome of biological control efforts. Macrolophus pygmaeus (Heteroptera: Miridae), a key component of biological control programs, is mainly predaceous but may also display phytophagy. M. pygmaeus hosts symbiotic Wolbachia, which induce cytoplasmic incompatibility, and two Rickettsia species, R. bellii and R. limoniae, which are found in all individuals tested. To test possible involvement of the two Rickettsia species in the feeding habits of M. pygmaeus, we first showed that the microbiome of the insect is dominated by these three symbionts, and later described the distribution pattern of the two Rickettsia species in its digestive system. Although both Rickettsia species were located in certain gut bacteriocyes, in caeca and in Malpighian tubules of both sexes, each species has a unique cellular occupancy pattern and specific distribution along digestive system compartments. Infrequently, both species were found in a cell. In females, both Rickettsia species were detected in the germarium, the apical end of the ovarioles within the ovaries, but not in oocytes. Although the cause for these Rickettsia distribution patterns is yet unknown, it is likely linked to host nutrition while feeding on prey or plants.

Environmental and Genetic Contributions to Imperfect wMel-Like Wolbachia Transmission and Frequency Variation.

Maternally transmitted Wolbachia bacteria infect about half of all insect species. They usually show imperfect maternal transmission and often produce cytoplasmic incompatibility (CI). Irrespective of CI, Wolbachia frequencies tend to increase when rare only if they benefit host fitness. Several Wolbachia, including wMel that infects Drosophila melanogaster, cause weak or no CI and persist at intermediate frequencies. On the island of São Tomé off West Africa, the frequencies of wMel-like Wolbachia infecting Drosophila yakuba (wYak) and Drosophila santomea (wSan) fluctuate, and the contributions of imperfect maternal transmission, fitness effects, and CI to these fluctuations are unknown. We demonstrate spatial variation in wYak frequency and transmission on São Tomé. Concurrent field estimates of imperfect maternal transmission do not predict spatial variation in wYak frequencies, which are highest at high altitudes where maternal transmission is the most imperfect. Genomic and genetic analyses provide little support for D. yakuba effects on wYak transmission. Instead, rearing at cool temperatures reduces wYak titer and increases imperfect transmission to levels observed on São Tomé. Using mathematical models of Wolbachia frequency dynamics and equilibria, we infer that temporally variable imperfect transmission or spatially variable effects on host fitness and reproduction are required to explain wYak frequencies. In contrast, spatially stable wSan frequencies are plausibly explained by imperfect transmission, modest fitness effects, and weak CI. Our results provide insight into causes of wMel-like frequency variation in divergent hosts. Understanding this variation is crucial to explain Wolbachia spread and to improve wMel biocontrol of human disease in transinfected mosquito systems.

The Biochemistry of Cytoplasmic Incompatibility Caused by Endosymbiotic Bacteria

Many species of arthropods carry maternally inherited bacterial endosymbionts that can influence host sexual reproduction to benefit the bacterium. The most well-known of such reproductive parasites is Wolbachia pipientis. Wolbachia are obligate intracellular α-proteobacteria found in nearly half of all arthropod species. This success has been attributed in part to their ability to manipulate host reproduction to favor infected females. Cytoplasmic incompatibility (CI), a phenomenon wherein Wolbachia infection renders males sterile when they mate with uninfected females, but not infected females (the rescue mating), appears to be the most common. CI provides a reproductive advantage to infected females in the presence of a threshold level of infected males. The molecular mechanisms of CI and other reproductive manipulations, such as male killing, parthenogenesis, and feminization, have remained mysterious for many decades. It had been proposed by Werren more than two decades ago that CI is caused by a Wolbachia-mediated sperm modification and that rescue is achieved by a Wolbachia-encoded rescue factor in the infected egg. In the past few years, new research has highlighted a set of syntenic Wolbachia gene pairs encoding CI-inducing factors (Cifs) as the key players for the induction of CI and its rescue. Within each Cif pair, the protein encoded by the upstream gene is denoted A and the downstream gene B. To date, two types of Cifs have been characterized based on the enzymatic activity identified in the B protein of each protein pair; one type encodes a deubiquitylase (thus named CI-inducing deubiquitylase or cid), and a second type encodes a nuclease (named CI-inducing nuclease or cin). The CidA and CinA proteins bind tightly and specifically to their respective CidB and CinB partners. In transgenic Drosophila melanogaster, the expression of either the Cid or Cin protein pair in the male germline induces CI and the expression of the cognate A protein in females is sufficient for rescue. With the identity of the Wolbachia CI induction and rescue factors now known, research in the field has turned to directed studies on the molecular mechanisms of CI, which we review here.

Dynamics of a two-sex model for the population ecology of dengue mosquitoes in the presence of Wolbachia

The release of Wolbachia-infected mosquitoes into the population of wild mosquitoes is one of the promising biological control method for combating the population abundance of mosquitoes that cause deadly diseases, such as dengue. In this study, a new two-sex mathematical model for the population ecology of dengue mosquitoes and disease is designed and used to assess the population-level impact of the periodic release of Wolbachia-infected mosquitoes. Rigorous analysis of the model, which incorporates many of the lifecycle features of dengue disease and the cytoplasmic incompatibility property of Wolbachia bacterium in mosquitoes, reveal that the disease-free equilibrium of the model is locally-asymptotically stable whenever a certain epidemiological threshold, known as the reproduction number of the model (denoted by R0W), is less than unity. The model is shown, using centre manifold theory, to undergo the phenomenon of backward bifurcation at R0W=1. The consequence of this bifurcation is that Wolbachia may not persist, or dengue disease may not be effectively-controlled, when R0W is less than unity. Such persistence and elimination will depend on the initial sizes of the sub-populations of the model. Two mechanisms were identified for which the backward bifurcation phenomenon can be removed. When backward bifurcation does not occur, the associated non-trivial disease-free equilibrium is shown to be globally-asymptotically stable when the reproduction number of the model is less than unity. Numerical simulations, using data relevant to dengue transmission dynamics in northern Queensland, Australia, shows that releasing Wolbachia-infected mosquitoes every three weeks, for a one-year duration, can lead to the effective control of the population abundance of the local wild mosquitoes, and that such effective control increases with increasing number of Wolbachia-infected mosquitoes released (resulting in the reduction of over 90% of the wild mosquito population from their baseline values). Furthermore, simulations show that releasing only adult male Wolbachia-infected mosquitoes provide more beneficial population-level impact (in terms of reducing the population abundance of the wild mosquitoes), in comparison to releasing adult female Wolbachia-infected mosquitoes. Increasing the frequency of Wolbachia release (e.g., from the default release frequency of every three weeks to weekly) does not significantly affect the effectiveness of the Wolbachia-based control program in curtailing the local abundance of the wild mosquitoes. Finally, it was shown that the cytoplasmic incompatibility property of Wolbachia bacterium does not significantly affect the effectiveness of the Wolbachia-based mosquito control strategy implemented in the community.