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Simplifi ed map of Europe showing the locations of the sites where Crioceris beetles (CP-C. paracenthesis, CA-C. asparagi, C12-C. quatuordecimpunctata, C14-C. duodecimpunctata, C5-C. quinquepunctata) and their Asparagus host plants were sampled. Open circles-sites without Wolbachia, solid circles-sites with infected beetles. Numbers (I-IX) correspond to site numbers described in Table 1.
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Intracellular bacteria of the genus Wolbachia (α-Proteobacteria) are the most widespread endosymbionts of insects.
Host infection is usually associated with alterations in reproduction, such as cytoplasmic incompatibility, the induction of parthenogenesis and offspring sex ratio bias: all phenomena that may infl uence host speciation. In the presen...
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The intracellular symbiont bacterium, Wolbachia is known as a reproductive parasite in different species of Trichogramma. Regard to importance of this symbiont in biological control programms, the present study addressed the efficiency of wsp gene to typing this symbiont from some populations of Trichogramma and also its prevalence. In this study w...
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... In such a study, bacterial strains appeared distantly related in the phylogenetic tree, not comprising a monophyletic clade and supporting the occurrence of horizontal transfer. Multiple mechanisms of Wolbachia horizontal transmission have been proposed, including predators, parasitoids, hemolymph transfer, cohabitation, and foraging on the same host plants (Ahmed et al. 2015;Huigens et al. 2004;Kolasa et al. 2017;Le Clec'h et al. 2013). So far, in could be detrimental for wNau1, as incompatibility among microorganisms could be a potential barrier to transmission of heritable symbionts . ...
The intracellular bacteria Wolbachia pipientis infects arthropods and filarial nematodes and is able to manipulate host reproduction. It has been reported an association between parthenogenesis and Wolbachia infection in weevils from the tribe Naupactini. A curing experiment suggested that a threshold density of Wolbachia is required for parthenogenetic reproduction to occur. The aim of this study was to analyze Wolbachia infection status and density in two sexually reproducing species from the tribe Naupactini, Naupactus xanthographus and Naupactus dissimulator. Wolbachia infection was detected in individuals from both species in several geographic locations, not being fixed. Quantification through real time PCR confirmed that Wolbachia loads in sexual species were significantly lower than in parthenogenetic ones; these results support the hypothesis of a threshold level for parthenogenetic reproduction to occur in Naupactini weevils. Strain typing showed that both sexual species carry wNau1, the most frequent strain in parthenogenetic Naupactini weevils. In addition, the presence of the WO phage, which might be an important factor regulating infection density in some hosts, was detected in this strain. Finally, Wolbachia wNau1 was located throughout the whole insect body, which is in agreement with the idea of a recent acquisition by horizontal transfer of wNau1 across the tribe Naupactini.
... Horizontal transmission may be a key factor in the infection dynamic and maintenance of heritable microbial symbionts, especially if they cause weak reproductive manipulations [143,147]. Acquisition and subsequent transmission of bacterial symbionts through feeding on plants may complement vertical transmission, favoring the spread and persistence of symbionts within the insect host populations [148][149][150]. In P. spumarius, intra-and inter-specific horizontal transmission via plant may be a rampant process contributing to the maintenance of Wolbachia within the infected populations of northern Italy, but probably not sufficient to compensate for the loss of infections, due to imperfect maternal transmission and environmental curing. ...
Philaenus spumarius is a cosmopolitan species that has become a major threat to European agriculture being recognized as the main vector of the introduced plant pathogen Xylella fastidiosa , the agent of the “olive quick decline syndrome”, a disease which is devastating olive orchards in southern Italy. Wolbachia are bacterial symbionts of many insects, frequently as reproductive parasites, sometime by establishing mutualistic relationships, able to spread within host populations. Philaenus spumarius harbors Wolbachia , but the role played by this symbiont is unknown and data on the infection prevalence within host populations are limited. Here, the Wolbachia infection rate was analyzed in relation to the geographic distribution and the genetic diversity of the Italian populations of P . spumarius . Analysis of the COI gene sequences revealed a geographically structured distribution of the three main mitochondrial lineages of P . spumarius . Wolbachia was detected in half of the populations sampled in northern Italy where most individuals belonged to the western-Mediterranean lineage. All populations sampled in southern and central Italy, where the individuals of the eastern-Mediterranean lineage were largely prevalent, were uninfected. Individuals of the north-eastern lineage were found only in populations from the Alps in the northernmost part of Italy, at high altitudes. In this area, Wolbachia infection reached the highest prevalence, with no difference between north-eastern and western-Mediterranean lineage. Analysis of molecular diversity of COI sequences suggested no significant effect of Wolbachia on population genetics of P . spumarius . Using the MLST approach, six new Wolbachia sequence types were identified. Using FISH, Wolbachia were observed within the host’s reproductive tissues and salivary glands. Results obtained led us to discuss the role of Wolbachia in P . spumarius , the factors influencing the geographic distribution of the infection, and the exploitation of Wolbachia for the control of the vector insect to reduce the spread of X . fastidiosa .
... Proposals for candidate genes to discriminate closely related Wolbachia strains vary. The most frequently analyzed gene set includes coxA, coding for cytochrome c oxidase, gatB, coding for glutamyltRNA (Gln) amidotransferase, hcpA, coding for conserved hypothetical protein, ftsZ, fbpA, coding for fructose-bisphosphate aldolase (Kolasa et al., 2017;Martins et al., 2021;Ren et al., 2020), and additionally the wsp gene (Baldo et al., 2006;Hu et al., 2020;Jeffries et al., 2021;Prezotto et al., 2017). An example of another set of genes in addition to the ftsZ gene is gltA, coding for citrate synthase and groEL, coding for 60-kDa heat-shock protein (Casiraghi et al., 2005). ...
Wolbachia is the most abundant intracellular symbiont among terrestrial Arthropoda. This bacterium together with other microorganisms, i.e., Cardinium, gained fame mainly as the causative agent of host sex-ratio distortion. Across the impressive diversity of oribatid mites (Acari: Oribatida), the microbes have been found in both parthenogenetic (Oppiella nova, Ceratozetes thienemanni, Hypochthonius rufulus) as well as sexually-reproducing (Gustavia microcephala, Achipteria coleoptrata, Microzetorchestes emeryi, Damaeus onustus) species. Wolbachia found in Oribatida represents supergroup E and is related to bacterial endosymbionts of springtails (Hexapoda: Collembola). Cardinium identified in O. nova and M. emeryi belongs to phylogenetic group A. In turn, Cardinium from A. coleoptrata constitutes a new separate group E. The occurrence of these bacterial endosymbionts in parthenogenetic and sexual oribatid mites species may suggests a different function other than manipulating host reproduction. Indeed, endosymbionts may have various “shades” of functions in invertebrate hosts, some of which cannot be excluded in the oribatid mites, e.g., enriching a nutrient-limited diet with B vitamins or contributing to host adaptation to colder and harsher climates. Nevertheless, the mystery behind the roles of bacteria in Oribatida still needs required to be unraveled.
... The collection of P. variipennis on cabbage represents the first record of this species as pest. P. erysimi was observed causing high damage on radish, as reported also in previous studies (Aslan & Özbek, 1998 (Kolasa et al., 2017;Magoga et al., 2016Magoga et al., , 2018Montagna, Sassi, & Giorgi, 2013). The values of the mean nucleotide intraspecific distance calculated for Chaetocnema and Phyllotreta genera separately are different, viz. ...
Flea beetles (Chrysomelidae: Galerucinae: Alticini), with ~8,000 species worldwide,
include pest species causing substantial economic damage to crops. The genera
Phyllotreta and Chaetocnema include both pest and non-pest
species. An accurate
and fast taxonomic identification approach is required for discriminating among taxa
for non-expert
taxonomists; moreover, the utility of this approach spans from biodiversity
conservation to the monitoring of pest species. DNA barcoding represents a
reliable and easy identification tool based on the use of short DNA sequences. In this
study, 45 new COI sequences of 13 Phyllotreta and five Chaetocnema species, representing
~30% and ~20% of the Turkish species belonging to these genera, were provided.
These sequences increased by ~18% and ~25% the number of species of these
genera whose sequences are available in BOLD. In order to test DNA barcoding efficiency
in Phyllotreta and Chaetocnema species identification, we created a data set
consisting of sequences belonging to species present in the Middle East and available
in BOLD plus the sequences developed in this study (36 species). The efficiency of
species identification, estimated using best close match analysis (with the ad hoc
calculated optimal distance threshold of 1.5%), was 99%. The overall intraspecific
and interspecific mean nucleotide divergences were 1.4% and 20%, respectively.
Interestingly, COI sequences of Phyllotreta nigripes clustered into two well-separated
groups with a high value of the between-group
nucleotide distance (11.4%), which
suggests the presence of cryptic species. In addition, information was provided on
the crops exploited by the collected organisms and the observed damage.
... This is consistent with the finding from Ahmed et al. (2013), who provided evidence for horizontal transmission of S-endosymbionts in the B. tabaci cryptic species complex based on phylogenies studies. There are substantial phylogenetic evidences showing that S-endosymbionts such as Wolbachia and Arsenophonus, undergoing horizontal transfer among host arthropod species (Ahmed et al., 2013;Chrostek et al., 2017;Kolasa et al., 2017;Li et al., 2017;Russell et al., 2003;Vavre et al., 1999). In some cases, the mechanisms for horizontal transmission of S-endosymbionts are already known, including transferring through parasitoid wasps (Ahmed et al., 2015;Gehrer & Vorburger, 2012), plants (Caspi-Fluger et al., 2012;Li et al., 2017) or even sexual transmission (Moran & Dunbar, 2006). ...
The whitefly Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) is a cryptic species complex, including members that are pests of global importance. This study presents a screening of B. tabaci species in China for infection by the primary endosymbiont, Portiera aleyrodidarum , and two secondary endosymbionts, Arsenophonus and Cardinium . The results showed that P. aleyrodidarum was detected in all B. tabaci individuals, while Arsenophonus was abundant in indigenous species of B. tabaci Asia II 1, Asia II 3, and China 1 but absent in the invasive species, Middle East-Asia Minor 1 (MEAM1); Cardinium presented in the Mediterranean (MED), Asia II 1 and Asia II 3 species but was rarely detected in the MEAM1 and China 1 species. Moreover, phylogenetic analyses revealed that the P. aleyrodidarum and mitochondrial cytochrome oxidase 1 ( mtCO1 ) phylograms were similar and corresponding with the five distinct cryptic species clades to some extent, probably indicating an ancient infection followed by vertical transmission and subsequent co-evolutionary diversification. In contrast, the phylogenetic trees of Arsenophonus and Cardinium were incongruent with the mtCO1 phylogram, potentially indicating horizontal transmission in B. tabaci cryptic species complex. Taken together, our study showed the distinct infection status of endosymbionts in invasive and indigenous whiteflies; we also most likely indicated the co-evolution of primary endosymbiont and its host as well as the potential horizontal transfer of secondary endosymbionts.
... The collection of P. variipennis on cabbage represents the first record of this species as pest. P. erysimi was observed causing high damage on radish, as reported also in previous studies (Aslan & Özbek, 1998 (Kolasa et al., 2017;Magoga et al., 2016Magoga et al., , 2018Montagna, Sassi, & Giorgi, 2013). The values of the mean nucleotide intraspecific distance calculated for Chaetocnema and Phyllotreta genera separately are different, viz. ...
Flea beetles (Chrysomelidae: Galerucinae: Alticini) include important economic pest species of the vegetable, industrial and other cultural crops. Due to their economic importance, an accurate and fast taxonomic identification is a crucial step and highly demanded in agricultural management particularly for morphologically hardly identifiable species that only some are regarded as economically important pests. DNA Barcoding offers a reliable and easy identification tool by using a short DNA sequences as a barcode in the case of a trustworthy database which was built on specimens identified by taxonomists. This study provides 45 new COI sequences belonging to 13 Phyllotreta and five Chaetocnema species, representing approximately the 30% and the 20% of the Turkey species, respectively. This provides an increase in terms of species numbers in BOLD of ~19% and 25%, respectively. Two dataset were constructed to test efficiency of the DNA barcodes in species identification of two genera and the nucleotide divergences within and between species were estimated. Best Closed Match analysis on datasets by using estimated optimal distance threshold was performed. The estimated barcoding efficiency in species identification (100% and 99%) for the two datasets confirmed the usefulness of this database for identification. Therefore, the database will be useful for fast and accurate discrimination among pests-not pests Phyllotreta and Chaetocnema species of Turkey, which will be helpful for making decision in establishing effective and eco-compatible pest management strategies.
A subject of particular interest concerns the understanding of which biotic and/or abiotic factors shape the insect’s microbiota composition and diversity. In this study, we focus our attention on the microbiota associated with leaf beetles (Coleoptera: Chrysomelidae) inhabiting a freshwater wetland in Northern Italy for investigating which factors shape it in this peculiar environment. A DNA metabarcoding approach targeting the 16S rRNA has been used to characterize the bacteria associated with seven leaf beetles species living in different zones of the wetland (trees/shrubs, meadow and emergent vegetation zones). The obtained results suggest that three factors potentially affecting microbiota diversity and composition are species membership in the first place, then living zone in the wetland and sex. Within the studied species, males were found possibly more prone to acquire bacteria from the surrounding environment, while females’ microbiota of most of the species resulted dominated by symbiotic bacteria (reproductive manipulators, e.g. Wolbachia, or other symbionts with functional roles in the species, e.g. Candidatus Macropleicola in Donacia genus). The present study, even if based on a limited sample size, contributed to exploring the factors affecting the composition and diversity of bacteria associated with Chrysomelidae.
Intracellular bacteria of the genus Wolbachia are the most abundant endosymbionts infecting many arthropods, with Coleoptera being the most diverse hosts in terms of taxonomy and ecology. There has been great progress in studies on the relations between Wolbachia and beetles, however, only some of the research details the consequences of infection. In this review, I summarise the knowledge on the evolutionary relations or ecological associations between Wolbachia and its beetle hosts. These bacteria often cause cytoplasmic incompatibility in the infected hosts and are responsible for a selective sweep of the mitochondrial genomes in some beetles. Wolbachia can manipulate the sex ratio or reproduction of some species of beetles, however, it does not induce parthenogenesis, with the possible rare exception of some Naupactini. Proof of the co-evolution of Wolbachia with beetles is missing, but some aquatic groups seem to be prone to co-speciation, unlike terrestrial taxa. On the other hand, there is a growing number of studies indicating or proving horizontal transmission of Wolbachia among beetle hosts, mostly via common host plants or the foraging substrate (such as dung). Wolbachia is not alone in infecting beetles as other endosymbiotic bacteria occur in beetles (Rickettsia, Spiroplasma, Cardinium, Arsenophorus), which have often been reported as interchangeable, suggesting the infection by various bacteria is dynamic. Nonetheless, there are still many issues associated with Wolbachia that are not yet been described in beetles (like the provision of nutrition or protection against pathogens) and high-throughput sequencing should be used to improve our understanding of Wolbachia-Coleoptera relations.
Chrysomelidae is a family of phytophagous insects with a highly variable degree of trophic specialization. The aim of this study is to test whether species feeding on different plants (generalists) harbour more complex microbiotas than those feeding on a few or a single plant species (specialists). The microbiota of representative leaf beetle species was characterized with a metabarcoding approach targeting V1–V2 and V4 regions of the bacterial 16S rRNA. Almost all the analysed species harbour at least one reproductive manipulator bacteria (e.g., Wolbachia, Rickettsia). Two putative primary symbionts, previously isolated only from a single species (Bromius obscurus), have been detected in two species of the same subfamily, suggesting a widespread symbiosis in Eumolpinae. Surprisingly, the well-known aphid symbiont Buchnera is well represented in the microbiota of Orsodacne humeralis. Moreover, in this study, using Hill numbers to dissect the components of the microbiota diversity (abundant and rare bacteria), it has been demonstrated that generalist insect species harbour a more diversified microbiota than specialists. The higher microbiota diversity associated with a wider host-plant spectrum could be seen as an adaptive trait, conferring new metabolic potential useful to expand the diet breath, or as a result of environmental stochastic acquisition conveyed by diet.
Myrmecophilous insects depend on their ant hosts during at least part of their life cycle. Lasius ants appear to be primarily involved as host in myrmecophilous interactions, especially with aphids and beetles, e.g. staphylinid. Pella laticollis is a rove beetle having a close ecological association with Lasius fuliginosus, which is correlated to its behavior adapted to avoid attack. Microorganisms can also play an important role in the maintenance of such relationships. We used 16S rRNA sequencing on the Illumina MiSeq platform to identify the bacterial communities associated with larvae and adults of both L. fuliginosus and P. laticollis. In addition, we determined the microbiota profiles of the nest-carton, an organic material lining the nest chambers. We obtained more than two million good quality reads. Taxonomic annotation showed that the profiles consisted of a total of 20 phyla, among which Proteobacteria was the most abundant. The samples were grouped according to the host's developmental stage or the nest material, and the differences between those groups were significant. Only the microbiota of L. fuliginosus larvae and adults did not differ significantly. Analyses at the genus level indicated a heightened abundance of Pseudomonas in the insects' profiles. The bacterial communities associated with the nest-carton included bacterial genera recorded from soil and dead wood. The profiles showed the presence of two well-known endosymbiotic bacteria, namely, Rickettsia and Wolbachia. According to our findings, the bacterial communities associated with larvae and adults of both L. fuliginosus and P. laticollis formed clusters according to the host's identity and its developmental stage. The profiles determined for the nest-carton formed a separate group. Our study is in line with a new research trend that is focusing on microbiota associated not only with ants, but also with myrmecophiles and ant nests inhabited by those species.
