Inactivation of Wolbachia reveals its biological roles in whitefly host

BACKGROUND: The whitefly Bemisia tabaci is cryptic species complex composed of numerous species. Individual species from the complex harbor a diversity of bacterial endosymbionts including Wolbachia. However, while Wolbachia is known to have a number of different roles, its role in B. tabaci is unclear. Here, the antibiotic rifampicin is used to selectively eliminate Wolbachia from B. tabaci so as to enable its roles in whitefly development and reproduction to be explored. The indirect effects of Wolbachia elimination on the biology of Encarsia bimaculata, a dominant parasitoid of B. tabaci in South China, were also investigated. METHODOLOGY/PRINCIPAL FINDING: qRT-PCR and FISH were used to show that after 48 h exposure to 1.0 mg/ml rifampicin, Wolbachia was completely inactivated from B. tabaci Mediterranean (MED) without any significant impact on either the primary symbiont, Portiera aleyrodidarum or any of the other secondary endosymbionts present. For B. tabaci MED, Wolbachia was shown to be associated with decreased juvenile development time, increased likelihood that nymphs completed development, increased adult life span and increased percentage of female progeny. Inactivation was associated with a significant decrease in the body size of the 4th instar which leads us to speculate as to whether Wolbachia may have a nutrient supplementation role. The reduction in nymph body size has consequences for its parasitoid, E. bimaculata. The elimination of Wolbachia lead to a marked increase in the proportion of parasitoid eggs that completed their development, but the reduced size of the whitefly host was also associated with a significant reduction in the size of the emerging parasitoid adult and this was in turn associated with a marked reduction in adult parasitoid longevity. CONCLUSIONS/SIGNIFICANCE: Wolbachia increases the fitness of the whitefly host and provides some protection against parasitization. These observations add to our understanding of the roles played by bacterial endosymbionts.This research was funded by the National Basic Research Program of China (2009CB119203), the Program for New Century Excellent Talents in University (NCET-0917–2011) and the China National Natural Science Foundation (31071732).http://www.plosone.or

Evidence for horizontal transmission of secondary Endosymbionts in the Bemisia tabaci cryptic species complex

Bemisia tabaci (Hemiptera: Aleyrodidae) is a globally distributed pest composed of at least 34 morphologically indistinguishable cryptic species. At least seven species of endosymbiont have been found infecting some or all members of the complex. The origin(s) of the associations between specific endosymbionts and their whitefly hosts is unknown. Infection is normally vertical, but horizontal transmission does occur and is one way for new infections to be introduced into individuals. The relationships between the different members of the cryptic species complex and the endosymbionts have not been well explored. In this study, the phylogenies of different cryptic species of the host with those of their endosymbionts were compared. Of particular interest was whether there was evidence for both coevolution and horizontal transmission. Congruence was observed for the primary endosymbiont, Portiera aleyrodidarum, and partial incongruence in the case of two secondary endosymbionts, Arsenophonus and Cardinium and incongruence for a third, Wolbachia. The patterns observed for the primary endosymbiont supported cospeciation with the host while the patterns for the secondary endosymbionts, and especially Wolbachia showed evidence of host shifts and extinctions through horizontal transmission rather than cospeciation. Of particular note is the observation of several very recent host shift events in China between exotic invader and indigenous members of the complex. These shifts were from indigenous members of the complex to the invader as well as from the invader to indigenous relatives.The Program for New Century Excellent Talents in University 2011, National Basic Research Program of China (973 Project, 2009CB119203) and the China National Natural Science Foundation (31071732).http://www.plosone.orgam201

Genetic networking of the Bemisia tabaci cryptic species complex reveals pattern of biological invasions

BACKGROUND: A challenge within the context of cryptic species is the delimitation of individual species within the complex. Statistical parsimony network analytics offers the opportunity to explore limits in situations where there are insufficient species-specific morphological characters to separate taxa. The results also enable us to explore the spread in taxa that have invaded globally. METHODOLOGY/PRINCIPAL FINDINGS: Using a 657 bp portion of mitochondrial cytochrome oxidase 1 from 352 unique haplotypes belonging to the Bemisia tabaci cryptic species complex, the analysis revealed 28 networks plus 7 unconnected individual haplotypes. Of the networks, 24 corresponded to the putative species identified using the rule set devised by Dinsdale et al. (2010). Only two species proposed in Dinsdale et al. (2010) departed substantially from the structure suggested by the analysis. The analysis of the two invasive members of the complex, Mediterranean (MED) and Middle East – Asia Minor 1 (MEAM1), showed that in both cases only a small number of haplotypes represent the majority that have spread beyond the home range; one MEAM1 and three MED haplotypes account for .80% of the GenBank records. Israel is a possible source of the globally invasive MEAM1 whereas MED has two possible sources. The first is the eastern Mediterranean which has invaded only the USA, primarily Florida and to a lesser extent California. The second are western Mediterranean haplotypes that have spread to the USA, Asia and South America. The structure for MED supports two home range distributions, a Sub-Saharan range and a Mediterranean range. The MEAM1 network supports the Middle East - Asia Minor region. CONCLUSION/SIGNIFICANCE: The network analyses show a high level of congruence with the species identified in a previous phylogenetic analysis. The analysis of the two globally invasive members of the complex support the view that global invasion often involve very small portions of the available genetic diversity.M.Z. Ahmed thanks University of Pretoria for awarding him a Commonwealth scholarship to support his doctoral studies.http://www.plosone.or

Structure of genetic diversity of Bemisia tabaci (Genn.) (Hemiptera: Aleyrodidae) populations in Brazilian crops and locations

Bemisia tabaci (Genn.) represents a complex of cryptic species that causes losses of many valuable crops. Even though there are differences in their ability to colonize hosts, transmit phytovirus and develop resistance to insecticides, the genetic differentiation of the insect populations is important for the adoption of control measures. Therefore, the genetic diversity of B. tabaci populations in economically important crops in Brazilian locations was characterized through a microsatellite analysis. Eight microsatellite markers were used for the analysis of eight populations, three in Solanum tuberosum (States of São Paulo, Minas Gerais and Bahia), two in Glycine max (States of São Paulo and Mato Grosso), one in Phaseolus vulgaris and Brassica oleracea var. acephala (States of São Paulo and Distrito Federal, respectively) and a cabbage population from Florida (USA). The number of alleles varied between two and 13 and the average value of F ST was 0.13. The population occurring in beans was genetically different, suggesting that the excessive use of insecticide or the host itself may have caused the modification of its allele frequency. The American population presented a large diversity and small differentiation compared to the Brazilian populations, especially from the Southeast, supporting the hypothesis that the B biotype was probably introduced into Brazil by the trade of plant material between the USA and the State of São Paulo. The genetic diversity found within and among the populations is geographically structured, and the insects from the central region of Brazil had superior genetic divergence when compared to the others Brazilian locations

Endophytic fungi decrease available resources for the aphid Rhopalosiphum padi and impair their ability to induce defences against predators

1. The production of winged morphs is a well known mechanism of induced defence in aphids to escape from natural enemies, and is also a reaction to poor resource quality. 2. Host plants of aphids often associate with endophytic fungi that have been shown to reduce the fitness of some species of aphids. 3. It was hypothesised that endophyte infection of host plants that represent a low quality plant resource should increase the aphid’s induced response to a predator because both low plant quality and predator presence represent a stronger cue for wing production than predator presence alone. 4. In a laboratory experiment, bird cherry-oat aphids Rhopalosiphum padi L. were exposed to the factors predator threat and endophyte infection and the effects of these factors on the proportion of winged morphs produced by the aphid colonies was analysed. 5. The presence of endophytic fungi strongly decreased aphid colony sizes. When a predator threat was present, all colonies on endophyte-free grasses produced winged morphs whereas only a few colonies were able to produce winged morphs on endophyteinfected grasses. However, these few colonies produced larger proportions of winged morphs than colonies on endophyte-free grasses. Without a predator threat, no colonies on endophyte-infected grasses produced any winged morphs. 6. These results show that aphids in stressed conditions and with reduced fitness will only invest in inducible defences when predators are present but are unable to produce winged morphs in response to endophyte presence