The Wolbachia bacterium, one of the most prevalent endosymbiotic bacteria, is known to induce reproductive anomalies in various invertebrate taxa. We investigated Wolbachia infection frequency in 203 Coleopteran insects collected from Korea. Among them 26 species (12.8%) across families proved to harbor Wolbachia. The phylogenetic trees of two Wolbachia specific genes imply that there have been complex horizontal gene transfers and recombination events within and between divergent Wolbachia subgroups. Interestingly, we also find that the infection frequency is noticeably low and incongruent at the family level when compared to other literature cases of Wolbachia infection in beetle species.

By definition, endosymbionts reside in the hosts' cells and are maternally inheritted to offspring as the mitochondriondoes. Recent progresses reveal that some endosymbionts have evolved their manipulabilities of hosts' reproduction. Inthis nature, once an endosymbiont shares its host with others, their association may last long unless their evolutionaryinterest or hosts' fitness is undermined.We tested whether the association existed among the endosymbionts; Wolbachia, Spiroplasma, Cardinium and Rickettsia.The results show that there are some multiple infection cases but there is no obvious pattern in their association.

Loop-mediated isothermal amplification (LAMP) is a rapid, specific, cost-effective detection method by amplifying nucleic acid under isothermal conditions. In this study, we used LAMP for detection of Hamiltonella defensa that lives as a facultive endosymbiont of whitefly ‘Bemisia tabaci’. We designed the Hamiltonella-specific primers by targeting 16S ribosomal RNA gene and validated the specificity of one primer set. To find the optimum temperature for our primer set, the LAMP reaction was held at the temperature, 60℃, 62℃ and 65℃. As a result, 62℃ was the optimum reation temperature for LAMP reaction. Specificity of primer set was tested by the reaction to both Trialeurodes vaporariorum and B. tabaci. After the whole procedure, the amplicons by LAMP were visualized by adding SYBR Green to the reaction tube.

The intracellular endosymbiont bacterium Wolbachia is currently considered the most abundant in arthropods and has also been isolated from nematodes, amphipods, isopods, mites and spiders. Recently, Wolbachia-based research was focused on the control of disease vector-population, such as several mosquitoes such as genus Aedes and Anopheles which cause dengue fever and malaria, respectively. For the analysis of regional difference between vector mosquito Aedes albopictus and Wolbachia, we selected different regions and collected Ae. albopictus which were distinguished with mountain chain and waterway. Whole genomic DNA were extracted from collected specimens with 9 regions. PCR analysis and sequencing were accomplished in each specimen for Wolbachia detection and identification using WSP gene. As a results, almost mosquitoes were infected with two strain of Wolbachia both wAlbA and wAlbB. However, regional separation of vector mosquitoes, wAlbA strain of Wolbachia were showed more than 98% sequence similarity. In this study, we first reported that Wolbachia infection and type of Wolbachia in Korea and endosymbiont Wolbachia was showed highly sequence homologies.

Wolbachia is one of the most common endosymbionts best known to induce several reproductive alterations in its insect hosts. In some cases, the insect hosts harbor more than two strains of the bacterium. The Vollenhovia emeryi ant lives in dead trees and is morphologically subdivided into the long-winged and the short-winged. Interestingly the short-winged morph is free of Wolbachia, but only the long-winged morph is multiple- infected with the Wolbachia bacterium. We sampled four populations of the long-winged morph in Korea and performed pyrosequencing in Multilocus Sequence Typing (MLST), to determine the bacterial strain diversity. Six different gene regions (coxA, fbpA, ftsZ, gatB, hcpA and wsp gene) were targeted and amplified. However, the result shows that diversity of haplotypes is very high. The pyrosequencing approach in MLST, a new method of discriminating Wolbachia strains, is promising to effectively detect multiple infections and rare haplotypes.

Sweetpotato whitefly, Bemisia tabaci, is a vector of more than 100 plant-diseased viruses, as well as a serious pest of various horticultural plants. This species harbors a primary endosymbiont Portiera along with several secondary endosymbionts such as Cardinium and Hamiltonella. We investigated whether or not TYLCV acquisition alters the densities of endosymbionts in the body of B. tabaci using quantitative real-time PCR. Our results showed that the densities of both Cardinium and Hamiltonella, but not Portiera, increased upon acquisition of TYLCV. In addition, expression of GroEL, a molecular chaperone produced by Hamiltonella, was significantly upregulated in TYLCV-infected whiteflies. Our results suggest that endosymbionts may play an important role in TYLCV transmission mechanism within the body of B. tabaci.

Many previous ecological studies on three major bacterial symbionts and a newly discovered symbiont PAXS (pea aphid X-type symbiont) in the pea aphid Acrythosiphon pisum have shown that these symbionts are associated with the expression of a variety of host phenotypes, including resistance to parasitoid Aphidius ervi and tolerance to heat stress. The principal role of all four symbionts “Candidatus Serratia symbiotica”, “Candidatus Hamiltonella defensa”, “Candidatus Regiella insecticola”, and PAXS is to protect aphids against abiotic stress by preserving the cells in which most of symbionts dwell and by reducing the rate of parasitism. In this experiment, we detected endosymbionts from four aphid clones by means of genomic DNA extraction, PCR with gene specific primer, and restriction enzyme cutting. The patterns of PCR and restriction enzyme cutting were all identical in the four aphid clones. In order to specifically identify the endosymbiont, we searched the sequences using BLAST. The BLAST search revealed that nucleotide sequences of the symbiont were 98% identical to Serratia entomophil. S. entomophil is also known to provide tolerance to heat stress, resistance to parasitoid wasps, and restoration of reproduction in aphids, suggesting its role in host protection.

The sweetpotato whitefly, Bemisia tabaci, acts as a vector of more than 100 plant viruses. B. tabaci is known to harbor a primary endosymbiont (Portiera) and 6 secondary endosymbionts (Arsenophonus, Cardinium, Fritschea, Hamiltonella, Rickettsia and Wolbachia). These endosymbionts play important roles in the acquisition and transmission of plant viruses. Using PCR analysis, we identified endosymbiotic bacteria in various B. tabaci populations collected from different places of Korea. Distribution of endosymbionts was different according to the biotype of B. tabaci. Subsequently, their relative densities of endosymbionts were compared between TYLCV-viruliferous and non-viruliferous populations of the Q biotype using quantitative realtime PCR. We found that the densities of Portiera, Cardinium and Hamiltonella are higher in viruliferous than non-viruliferous whiteflies. Our results suggest the role of endosymbiont for the TYLCV transmission of whiteflies.

evolution of multicellular organisms. Especially in insects, a large amount of bacterial symbionts are illustrated. Among the insect taxonomic group, sap feeding insects, mostly Heteroptera, that feeds on poor nutrient sources from plants, have developed special organs where symbiotic bacteria can reside. The bacteria mainly provide the host insects with some essential amino acids. This has built evolutionarily unbreakable tie between the symbionts and the their hosts. Here we present an obligate symbiont from Lycorma delicatula (Heteroptera: Fulgoridae) being thought to invade the Korean penninsula recently. Based on the partial 16s rRNA gene sequence, the symbiotic bacterium is identified as Cadidatus Sulcia mullleri. The Genbank data indicates that the bacterium is found in various heteropteran families. This may imply the acquisition of the bacterium precedes the hosts' divergence, though there is an alternative postulation. We discuss its distribution in the sap-feeding insects and potential role on survival of L. delicatula and paratransgenetic application of the bacterium in controlling L. delicatula.

The Wolbachia bacterium, one of the most prevalent endosymbiotic bacteria, is known to induce reproductive anomalies such as cytoplasmic incompatibility, feminization, male killing and parthenogenesis in various arthropod species. The bacterium is considered to have had huge impacts on hosts' reproductive biology, immunity, evolution, and molecular machineries. Infection surveys on the bacterium have rather been limited to specific taxa that are mainly of economical importance or conducted with randomly collected organisms. Here we investigated infection frequency of Wolbachia in 206 Coleopteran insects collected from Korea. Among them 28 species (13.59%) across families proved to harbor Wolbachia. The phylogenetic trees based on the partial 16s rRNA gene and the partial Wolbachia surface protein (wsp) gene of Wolbachia show that all the Wolbachia strains belong to either Supergroup A or B and Wolbachia evolved independently from its hosts. In addition, the cophylogenetic analysis of the 16s rRNA gene and wsp gene implies that there have been horizontal DNA transfers and recombination events within and between divergent Wolbachia supergroups.