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Fluorescent in situ hybridization of Wolbachia genes on the chromosomes of M. alternatus. (a) Male chromosomes in metaphase II arranged in the order of size. (b) Male chromosomes in the meiotic metaphase I. (c) Sperm nuclei. Wolbachia genes are visualized in yellow, and insect chromosomes are in blue.
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Monochamus alternatus is the longicorn beetle notorious as a vector of the pinewood nematode that causes the pine wilt disease. When two populations of M. alternatus were subjected to diagnostic polymerase chain reaction (PCR) detection of four Wolbachia genes, only the ftsZ gene was detected from one of the populations. The Wolbachia ftsZ gene per...
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... This is the early experimental evidence illustrating the HGT event between Wolbachia and the host insect. Subsequently, investigations through PCR detection and Southern blot confirmed that 57 genes derived from Wolbachia were integrated into the C. chinensis nuclear genome, albeit the transcriptional levels of ~50% genes were relatively low followed by being evidenced using FISH analysis [36] . A parallel work based on exhaustive PCR surveys and FISH analysis determined that 31 of 214 Wolbachia genes were transferred and located on an autosome of two populations of Cerambycidae Monochamus alternatus [37] . ...
Horizontal gene transfer (HGT) refers to the asexual exchange of genetic information between distantly related organisms. Although it is well acknowledged that HGT greatly contributes to the adaptive evolution in prokaryotes, its significance in shaping the orientation of eukaryote evolution remains obscure, especially in insect lineages. The massively expanded genomic data appears to be an excellent choice to uncover the mystery of HGTs in insects nowadays. Here we gather a body of evidence showing the HGT events from three broad donor origins, viruses, prokaryotes, and eukaryotes, in which most horizontally transferred genes are unlikely to be functional and will be eroded as a result of the difference of inheritable background between insect recipients and donor species. Nevertheless, particular interests in the prominent role of insect HGTs in maintaining and acquiring new functionalities have still been raised to underpin their adaptations. Among these, the previously investigated properties including reproduction regulation, detoxification of plant metabolites, formation of body color, and antimicrobic immunity are mainly included in different insect lineages. Albeit such cases are just the tip of the iceberg, we demonstrate that HGT drives insect evolution, especially in coevolution with host plants, and additional explorations into its functions should be given attention in order to access the complex evolutionary history of insects in the near future.
... Its usage has gained acceptance, especially in the last decades; however, some factors limit its usage (Collins & Cruickshank 2013). Recent speciation, the prevalence of paraphyly, hybridisation, taxonomic uncertainties, infections caused by endosymbiotic bacteria such as Wolbachia, and the availability of very little of the rich biodiversity on earth have been evaluated as factors that may adversely affect DNA barcoding performance (Funk & Omland 2003;Moritz et al. 2004;Cameron et al. 2006;Whitworth et al. 2007;Meier 2008;Smith et al. 2008;Aikawa et al. 2009). ...
Spondylidinae is a minor subfamily of Cerambycidae with around 100 species in 20 genera. The tribes Asemini and Spondylidini, both salient owing to vectors and invaders into the limelight of phytosanitary authorities, constitute the “spondylidine branch”. Although a few species of the branch were being included in some higher taxonomic level studies, the phylogenetic relationships within the taxa have never been evaluated from a molecular viewpoint. The present study identifies the phylogenetic relationships within the branch inferring from a single locus ( COI ) global dataset and two multi-locus local datasets, binary combinations of mitochondrial COI , 16S rRNA and nuclear 28S rRNA D1-D2 , that sequences were obtained from the samples of the East of Marmara Region, Turkey. [( Spondylis+Neospondylis ) + Megasemum ] and [ Tetropium+Asemum ] groups were separated from the same node in the phylogenetic trees. The genus Cephalallus was stated on the basalmost of these two groups. Arhopalus split from all other genera and divided into two subgroups. There were discrepancies between morphology and COI barcode sequences, which might be a sign of hybridisation between A. rusticus and A. syriacus . Intraspecific distances of COI gene regions of S. buprestoides , As. striatum and T. cinnamopterum were up to 4.26%, 8.51%, and 6%, respectively, which might point out to cryptic speciation.
... The relationship between Wolbachia and this insect is unique. The beetle is not infected with Wolbachia as a bacterium, but carries many Wolbachia genes on an autosome, indicating that lateral gene transfer has occurred from Wolbachia to M. alternatus [28,29]. Very few reports of the detection of Wolbachia in the cerambycid insects have been published, compared to those related to other insect taxa, and there has been no research into the effect of Wolbachia on the insects. ...
... Two artificial diets established for rearing M. alternatus [28,32] were used for rearing A. fraudatrix larvae and adults. In diet I, a mixture of 4.0 g of milled inner bark of P. densiflora, 6.6 g of artificial silkworm diet (Silkmate 2M, Norsan Co., Kanagawa, Japan), and 0.8 g of dried yeast (EBIOS, Asahi Group Foods, Ltd., Tokyo, Japan) was kneaded with 8.6 ml of distilled water, an approach modified from that described in [28]. ...
... Two artificial diets established for rearing M. alternatus [28,32] were used for rearing A. fraudatrix larvae and adults. In diet I, a mixture of 4.0 g of milled inner bark of P. densiflora, 6.6 g of artificial silkworm diet (Silkmate 2M, Norsan Co., Kanagawa, Japan), and 0.8 g of dried yeast (EBIOS, Asahi Group Foods, Ltd., Tokyo, Japan) was kneaded with 8.6 ml of distilled water, an approach modified from that described in [28]. Diet II was composed of 1.6 g of current and one-year-old needles of P. densiflora dried at 70˚C for one day and milled into powder, 5.4 g of Silkmate 2M, 0.6 g of EBIOS, and 12.4 ml of distilled water [32]. ...
Wolbachia are obligatory endosymbiotic α-proteobacteria found in many arthropods. They are maternally inherited, and can induce reproductive alterations in the hosts. Despite considerable recent progress in studies on the associations between Wolbachia and various taxonomic groups of insects, none of the researches have revealed the effects of Wolbachia on longicorn beetles as the host insect. Acalolepta fraudatrix is a forest longicorn beetle that is distributed in East Asia. In this study, the relationship between Wolbachia and A . fraudatrix was investigated. Out of two populations of A . fraudatrix screened for Wolbachia using the genes ftsZ , wsp , and 16S rRNA, only one of the populations showed detection of all three genes indicating the presence of Wolbachia . Electron microscopy and fluorescent in situ hybridization also confirmed that the A . fraudatrix population was infected with Wolbachia . Sequencing the wsp genes derived from single insects revealed that two strains of Wolbachia coexisted in the insects based on the detection of two different sequences of the wsp gene. We designated these strains as w Fra1 and w Fra2. The bacterial titers of w Fra1 were nearly 2-fold and 3-fold higher than w Fra2 in the testes and ovaries, respectively. The two strains of Wolbachia in the insects were completely eliminated by rearing the insects on artificial diets containing 1% concentration of tetracycline for 1 generation. Reciprocal crosses between Wolbachia -infected and Wolbachia -uninfected A . fraudatrix demonstrated that only eggs produced by the crosses between Wolbachia -infected males and Wolbachia -uninfected females did not hatch, indicating that Wolbachia infecting A . fraudatrix causes cytoplasmic incompatibility in the host insect. This is the first report showing the effect of Wolbachia on reproductive function in a longicorn beetle, A . fraudatrix .
... Alternatively, positive amplifications might also indicate a bacterial DNA integrated into the host genome after an event of LGT (Lateral Gene Transfer), in which case one can erroneously assume an ongoing infection. Lateral Gene Transfer of Wolbachia DNA into the host genome is a well-documented phenomenon in several insect taxa (Dunning Hotopp et al. 2007, Aikawa et al. 2009, Klasson et al. 2009, Morrow et al. 2015. ...
... The unsuccessful detection of the wsp gene in some of the positive species for 16S could be attributed to false negative results, as previously assumed in other studies (e.g., Jeyaprakash andHoy 2000, Li et al. 2007). On the other hand, the successful amplification of only one or few genes has been interpreted as an evidence for LGT of Wolbachia DNA into host genomes (e.g., Aikawa et al. 2009) that ultimately could lead to pseudogenization (e.g., Morrow et al. 2015). Multiple sources of information, such as multiloci approaches, genomics, and immunofluorescent assays, could unveil if these sand fly species carry live endosymbionts or just genomic vestiges of Wolbachia. ...
Wolbachia pipientis (Hertig) is an endosymbiotic microorganism widespread among arthropods and other invertebrate hosts, and employed in strategies to reduce the incidence of arthropod-borne diseases. Here, we used a PCR-based approach for 16S RNA and wsp genes to investigate the prevalence, geographical distribution, and strains of Wolbachia in sand flies (Diptera: Psychodidae: Phlebotominae), the main vectors of the causative agents of leishmaniasis, from three biomes in Brazil: Amazon, Cerrado, and Caatinga. We found that: 1) Wolbachia DNA is present in most (66.7%) of the sampled sand fly species, including vectors of Leishmania spp. (Ross, Trypanosomatida: Trypanosomatidae), 2) the prevalence of Wolbachia DNA varies among species and populations, 3) some strains of Wolbachia may have wider geographical and host range in South America, and 4) two phylogenetic distinct wsp sequences might represent two novel strains for Wolbachia in South America sand flies. Those findings increase the basic knowledge about Wolbachia in South American sand flies and might foster further researches on its use to reduce the transmission of sand fly-borne parasites.
... Recent horizontal transfers of genes or mobile elements can indicate ecological relationships, because HGT events occur between closely associated organisms. Well-known examples include HGT from Wolbachia symbionts to their host insect 50,51 and transfer of BovB retrotransposons between ruminants and snakes via parasitic ticks 52,53 . We found that RTE/BovB repeats are abundant in the S. proliferum genome, but were likely acquired by an ancestral pseudophyllidea, as indicated by their abundance in D. latus and S. solidus. ...
The cryptic parasite Sparganum proliferum proliferates in humans and invades tissues and organs. Only scattered cases have been reported, but S. proliferum infection is always fatal. However, S. proliferum's phylogeny and life cycle remain enigmatic. To investigate the phy-logenetic relationships between S. proliferum and other cestode species, and to examine the mechanisms underlying pathogenicity, we sequenced the entire genomes of S. proliferum and a closely related non-life-threatening tapeworm Spirometra erinaceieuropaei. Additionally, we performed larvae transcriptome analyses of S. proliferum plerocercoid to identify genes involved in asexual reproduction in the host. The genome sequences confirmed that the S. proliferum has experienced a clearly distinct evolutionary history from S. erinaceieuropaei. Moreover, we found that nonordinal extracellular matrix coordination allows asexual reproduction in the host, and loss of sexual maturity in S. proliferum are responsible for its fatal pathogenicity to humans. Our high-quality reference genome sequences should be valuable for future studies of pseudophyllidean tapeworm biology and parasitism.
... The intimate associations between host and intracellular Wolbachia frequently lead to lateral gene transfers (LGTs) between their genomes (Dunning Hotopp et al., 2007). In beetles, Wolbachia genomic fragments are transferred to the X chromosome and autosome of the host (Aikawa et al., 2009). The ll transferred genes are, however, pseudogenized and likely derived from a single LGT. ...
The most widespread intracellular bacteria in the animal kingdom are maternally inherited endosymbionts of the genus Wolbachia. Their prevalence in arthropods and nematodes worldwide and stunning arsenal of parasitic and mutualistic adaptations make these bacteria a biological archetype for basic studies of symbiosis and applied outcomes for curbing human and agricultural diseases. Here, we conduct a summative, centennial analysis of living in the Wolbachia world. We synthesize literature on Wolbachia’s host range, phylogenetic diversity, genomics, cell biology, and applications to filarial, arboviral, and agricultural diseases. We also review the mobilome of Wolbachia including phage WO and its essentiality to hallmark reproductive phenotypes in arthropods. Finally, the Wolbachia system is an exemplar for discovery-based science education using biodiversity, biotechnology, and bioinformatics lessons. As we approach a century of Wolbachia research, the interdisciplinary science of this symbiosis stands as a model for consolidating and teaching the integrative rules of endosymbiotic life.
... Monochamus alternatus is the main vector of the pinewood nematode, Burasphelenchus xylophilus (Steiner and Buhrer) (Nematode: Aphelenchoididae), which causes serious damage to pines 13 . At present, there are no effective strategies for controlling M. alternatus, such as physical, chemical, or biological methods 14 . ...
Abiotic stress factors can significantly affect insects. In particular, the stressful effects of exposure to ultrasound on insects are considered important. In the present study, we investigated the effects of ultrasound on the important global pest Monochamus alternatus (Coleoptera: Cerambycidae), which is the main vector of the pinewood nematode. We exposed M. alternatu s adults (aged 1 day, 3 days, and 5 days) to ultrasound at different frequencies (using two ultrasonic devices, i.e., LHC20 with a mixture of frequencies at 35 kHz, 70 kHz, and 105 kHz; and GFG-8016G at two separate frequencies of 30 kHz and 60 kHz) for different periods of time (1 h, 12 h, and 24 h), before evaluating the juvenile hormone III (JHIII) titers. All of the ultrasound treatments significantly decreased the JHIII titers in M. alternatus adults. The decreases in the JHIII titers due to ultrasound exposure did not differ according to sex, but the effects on beetles of different ages differed significantly depending on the duration of exposure. The decreases in the JHIII titers were highest in male and female beetles after exposure to ultrasound for 12 h. Following exposure to ultrasound for any time period, the decreases in the JHIII titers were lower in adults aged 3 days than those aged 1 day and 5 days. The different ultrasonic frequencies led to variable decreases in the JHIII titers in M. alternatus adults, where the greatest decreases occurred in beetles exposed to ultrasound at 60 kHz. Our results indicate that ultrasound can negatively affect the normal JHIII levels and it may further disrupt sexual maturation by M. alternatus adults.
... Our previous work identified CI-like toxin and antidote genes on scaffolds in several arthropod genome assemblies, although antidote genes were found more frequently possibly due to utility in suppressing microbial invaders implementing RP (Gillespie et al., 2018). Many of these genomes contain multiple divergent antidote genes, though none were found to harbor large-scale Wolbachia gene transfer that is known to occur in certain invertebrates (Leclercq et al., 2016;Hotopp et al., 2007;Aikawa et al., 2009;Kondo et al., 2002;Dhaygude et al., 2019;McNulty et al., 2010;Werren et al., 2010). In light of this observation, we originally speculated that CI-like genes are mobilized from wolbachiae to host through WO phage insertion into host nuclear genomes; however, the absence of other WO phage genes in these genomes made this assertion tenuous. ...
Wolbachiae are obligate intracellular bacteria that infect arthropods and certain nematodes. Usually maternally inherited, they may provision nutrients to (mutualism) or alter sexual biology of (reproductive parasitism) their invertebrate hosts. We report the assembly of closed genomes for two novel wolbachiae, w CfeT and w CfeJ, found co-infecting cat fleas ( Ctenocephalides felis ) of the Elward Laboratory colony (Soquel, CA, USA). w CfeT is basal to nearly all described Wolbachia supergroups, while w CfeJ is related to supergroups C, D and F. Both genomes contain laterally transferred genes that inform on the evolution of Wolbachia host associations. w CfeT carries the Biotin synthesis Operon of Obligate intracellular Microbes (BOOM); our analyses reveal five independent acquisitions of BOOM across the Wolbachia tree, indicating parallel evolution towards mutualism. Alternately, w CfeJ harbors a toxin-antidote operon analogous to the w Pip cinAB operon recently characterized as an inducer of cytoplasmic incompatibility (CI) in flies. w CfeJ cinB and three adjacent genes are collectively similar to large modular toxins encoded in CI-like operons of certain Wolbachia strains and Rickettsia species, signifying that CI toxins streamline by fission of large modular toxins. Remarkably, the C . felis genome itself contains two CI-like antidote genes, divergent from w CfeJ cinA , revealing episodic reproductive parasitism in cat fleas and evidencing mobility of CI loci independent of WO-phage. Additional screening revealed predominant co-infection ( w CfeT/ w CfeJ) amongst C . felis colonies, though fleas in wild populations mostly harbor w CfeT alone. Collectively, genomes of w CfeT, w CfeJ, and their cat flea host supply instances of lateral gene transfers that could drive transitions between parasitism and mutualism.
... Wolbachia bacteria are indeed able to colonize female germline cells, through which they are transovarially transmitted to the progeny. Wolbachia interspecific horizontal transmission was initially described as a rare phenomenon (O'Neill et al. 1992;Rousset et al. 1992;Turelli et al. 2018;Werren et al. 1995), but studies are increasingly showing that, in a variety of insect species, Wolbachia genes have been horizontally transferred to host chromosomes (Aikawa et al. 2009;Dunning Hotopp et al. 2007;Fenn et al. 2006;Klasson et al. 2009;Kondo et al. 2002;Nikoh and Nakabachi 2009;Nikoh et al. 2008;Woolfit et al. 2009). ...
Tsetse flies (Glossina spp.) act as the sole vectors of the African trypanosome species that cause Human African Trypanosomiasis (HAT or African Sleeping Sickness) and Nagana in animals. These flies have undergone a variety of specializations during their evolution including an exclusive diet consisting solely of vertebrate blood for both sexes as well as an obligate viviparous reproductive biology. Alongside these adaptations, Glossina species have developed intricate relationships with specific microbes ranging from mutualistic to parasitic. These relationships provide fundamental support required to sustain the specializations associated with tsetse’s biology. This chapter provides an overview on the knowledge to date regarding the biology behind these relationships and focuses primarily on four bacterial species that are consistently associated with Glossina species. Here their interactions with the host are reviewed at the morphological, biochemical and genetic levels. This includes: the obligate symbiont Wigglesworthia, which is found in all tsetse species and is essential for nutritional supplementation to the blood-specific diet, immune system maturation and facilitation of viviparous reproduction; the commensal symbiont Sodalis, which is a frequently associated symbiont optimized for survival within the fly via nutritional adaptation, vertical transmission through mating and may alter vectorial capacity of Glossina for trypanosomes; the parasitic symbiont Wolbachia, which can manipulate Glossina via cytoplasmic incompatibility and shows unique interactions at the genetic level via horizontal transmission of its genetic material into the genome in two Glossina species; finally, knowledge on recently observed relations between Spiroplasma and Glossina is explored and potential interactions are discussed based on knowledge of interactions between this bacterial Genera and other insect species. These flies have a simple microbiome relative to that of other insects. However, these relationships are deep, well-studied and provide a window into the complexity and function of host/symbiont interactions in an important disease vector.
... Horizontal gene transfer can occur in prokaryotic organisms and facilitates the acquisition of novel genes with functions such as pathogenicity, antibiotic resistance and regulation of metabolic pathways (Boucher et al., 2003). Although horizontal transfer between prokaryotic bacteria and multicellular eukaryotic organisms is generally rare, it has been widely reported in the case of Wolbachia and their hosts (Aikawa et al., 2009;Hotopp et al., 2007;Klasson et al., 2009;Kondo et al., 2002;Nikoh et al., 2008;Woolfit et al., 2009). A whole genome sequencing study showed multiple insertions of Wolbachia genes into the host (G. ...
Ectoparasites and livestock-associated insects are a major concern throughout the world because of their economic and welfare impacts. Effective control is challenging and relies mainly on the use of chemical insecticides and acaricides. Wolbachia, an arthropod and nematode-infecting, maternally-transmitted endosymbiont is currently of widespread interest for use in novel strategies for the control of a range of arthropod-vectored human diseases and plant pests but to date has received only limited consideration for use in the control of diseases of veterinary concern. Here, we review the currently available information on Wolbachia in veterinary ectoparasites and disease vectors, consider the feasibility for use of Wolbachia in the control of livestock pests and diseases and highlight critical issues which need further investigation.
