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      • Detection of the Bacterial Symbiosis in the Pea Aphid Acrythosiphon pisum.

        Kyungjae Andrew Yoon,Young Ho Kim,Si Hyeock Lee 한국응용곤충학회 2012 한국응용곤충학회 학술대회논문집 Vol.2012 No.10

        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.

      • Comparison of Endogenous Cellulase Genes from Four Termite Species with Different Habitats

        Kyungjae Andrew Yoon,Young Ho Kim,Si Hyeock Lee 한국응용곤충학회 2013 한국응용곤충학회 학술대회논문집 Vol.2013 No.04

        To search for a variety of cellulase genes from termites with different habitats consuming different foods, we collected three species (Neotermes spp., Odontotermes spp., Macrotermes spp.) from the wood and one species (Nasutitermes spp.) from the cow dung. Total RNA was isolated both from alimentary track tissues containing paunch and from other tissues, and used for the suppression subtractive hybridization (SSH). The resulting EST libraries were sequenced and searched by BLAST to identify cellulase genes. A total of 16 cellulase genes were found from the wood-dwelling termites whereas 4 cellulase genes from the cow dung-dwelling termites. Endo-beta-1,4-glucanase and beta-glucosidase were identified as the most abundant cellulase from the wood-dwelling termites and cow dung-dwelling termites, respectively. This finding suggests that cellulase profiles are significantly different depending on the termite’s habitat and food. In addition, we analyzed phylogenetic relationships among the cellulase genes along with other cellulase genes reported to date. All cellulase genes appeared to be originated from endosymbioants without any hint of horizontal gene transfer. Functional expression of endo-beta-1,4-glucanase using a baculovirus expression system is in progress to characterize its enzymatic properties.

      • Comparative transcriptomic analysis of the venom gland of social wasps Vespa crabro and Vespa analis

        Kyungjae Andrew Yoon,Kyungmun Kim,Hyo-min Ahn,Ki-Gyoung Kim,Hong-Yul Seo,Young Ho Koh,Si Hyeock Lee 한국응용곤충학회 2015 한국응용곤충학회 학술대회논문집 Vol.2015 No.04

        Vespa crabro is a cosmopolitan social wasp species whereas Vespa analis is commonly found in Asia. Both species are widely distributed in Korea and known to be aggressive when disturbed, resulting in frequent sting accidents. Although major venom components of well known Vespa wasps have been reported, no comparative transcriptomic analysis of venom gland between V. crabro and V. analis has been conducted to date. To investigate the differences in venom properties between these two wasps, total RNA was extracted from each venom gland and used for RNA-sequencing. A total of 31 venom-specific genes were identified in both venom gland transcriptomes but their expression profiles were different between V. crabro and V. analis. Venom allergen 5, premastoparan A and phospholipase A were the top three genes that were most prevalently transcribed in the venom gland of V. crabro, and their transcription rates were 902-, 112- and 4164-fold higher compared with V. analis, respectively, as judged by FPKM values. Their differential transcription profiles were confirmed by quantitative real-time PCR. In the venom gland of V. analis, however, premastoparan A was most abundantly transcribed gene, followed by calponin and tropomysin. In general, most venom-specific genes were more abundantly expressed in V. crabro but some genes exhibited higher transcription rates in V. analis, including muscle LIM protein, troponin, paramyosin, calponin, etc. Our findings suggest that V. crabro produce venom with much more enriched venom components, thereby with higher toxicity compared with V. analis.

      • Comparative functional analysis of mastoparans from social hornets Vespa crabro and Vespa analis

        Kyungjae Andrew Yoon,Kyungmun Kim,Hyo-min Ahn,Ki-Gyoung Kim,Hong-Yul Seo,Young Ho Koh,Si Hyeock Lee 한국응용곤충학회 2015 한국응용곤충학회 학술대회논문집 Vol.2015 No.10

        Vespa crabro and V. analis are social hornet species commonly found in Asia, including Korea. Mastoparan is one of the major venom peptides of these two hornets but its amino acid sequence defers substantially. To examine the differences in the potential toxicity and bioactivity of mastoparans between these two social hornets, differential toxicological and pharmacological activities of synthesized mastoparan were investigated. V. analis mastoparan showed a 7-fold higher hemolytic activity, suggesting its higher cytotoxic potential compared with V. crabro mastoparan. Mastoparans from both hornet species exhibited similar levels of antimicrobial activities against Staphylococcus aureus and Botrytis cinerea, whereas the mastoparan from V. analis showed more potent antimicrobial activities against Escherichia coli and Candida albicans. Nevertheless, the antimicrobial activities of mastoparans of V. crabro and V. analis were relatively lower compared with those of other wasps. Both mastoparans also exhibited some levels of antitumor activity but the activity was significantly higher in V. analis mastoparan. In summary, the hemolytic, antimicrobial, and antitumor activities of synthesized V. analis mastoparan were higher than those of V. crabro mastoparan. These differential bioactivities are likely due to the amino acid sequence differences in the mature peptides. In particular, the additional Lys residue present in V. analis mastoparan may contribute to the higher levels of bioactivity as proposed by secondary structure prediction.

      • Characterization of biological activities of abaecin from bumblebee Bombus ussurensis

        Kyungjae Andrew Yoon,Young Ho Koh,Si Hyeock Lee 한국응용곤충학회 2017 한국응용곤충학회 학술대회논문집 Vol.2017 No.04

        Abaecin is a proline-rich anti-bacterial peptide which is known to have high activity against Gram-negative bacteria and it is originally isolated from honeybee Apis mellifera. Since the anti-bacterial peptides including abaecin were identified in the hemolymph of A. mellifera infected with Gram-negative bacteria Escherichia coli, this result implies that honeybees express abaecin for defense against natural infection. Anti-microbial peptides including abaecin were also identified in the adult hemolymph of bumblebees. To investigate pharmacological and toxicological properties of abaecin from Bombus ussurensis, biological activities were evaluated by conducting anti-tumor, anti-microbial and cytotoxic activity assays. Abaecin exhibited significantly high anti-tumor activity against ovarian tumor cells SK-OV-3 and NIH; OVCAR-3 at 100 μM after 72 h treatment. Interestingly, abaecin showed none of anti-microbial activity against Gram-negative and Gram-positive bacteria. Since abaecin exhibited extremely low level of hemolytic activity against human erythrocytes, it may serve as a good model peptide for studying its potential as a selective anti-tumor drug.

      • Bioactivity and molecular characterization of bombolitins from Bombus ardens, B. consobrinus, B. terrestris and B. ussurensis

        Kyungjae Andrew Yoon,Young Han Park,Young Ho Koh,Si Hyeock Lee 한국응용곤충학회 2017 한국응용곤충학회 학술대회논문집 Vol.2017 No.10

        Biological activities of bombolitins from Bombus ardens, B. consobrinus, B. terrestris and B. ussurensis (bombolitinsA, C, T and U, respectively) were examined using hemolytic, anti-microbial, anti-fungal and anti-tumor activity assays.Among the four bombolitins tested, bombolitin T showed the highest hemolytic and anti-tumor activities. All bombolitinsexhibited strong anti-microbial and anti-fungal activities, and bombolitin A specifically possessed the highest anti-microbialactivity against the Gram-negative bacteria Escherichia coli. Circular dichroism spectrometry analysis revealed that allfour bombolitins had over 61.7% and 45.5% of α-helicity in 30 mM sodium dodecyl sulfate and 50% trifluoroethanolbuffers, respectively, which form lipid-membrane-mimicking environments. Bombolitin T showed the lowest IC50 valuesof 8.5 μM and 8.8 μM against SK-OV-3 and NIH-OVCAR-3 cell lines, respectively, after 72 h of treatment, but itsrelative hemolytic activity at a concentration of 200 μM was 2.3-fold higher than that of 0.1% Triton X-100. Thisstudy provides new information on the biological and molecular properties of venom peptides of bumblebees. However,further studies on reducing cytotoxicity of bombolitins are needed for designing selective anti-tumor peptides.

      • Characterization of venom evolution via venom component phylogenetic analysis in Aculeata Hymenoptera

        Kyungjae Andrew Yoon,Do Eun Lee,Woo Young Bang,Chang Hwan Bae,Joo-Hong Yeo,Young Ho Koh,Si Hyeock Lee 한국응용곤충학회 2018 한국응용곤충학회 학술대회논문집 Vol.2018 No.10

        To identify the venom components and their expression patterns of some Aculeata bees/wasps, venom gland-specific transcriptome analysis was conducted. FPKM values were normalized with the average of the transcription level of reference gene (a-tubulin). Common components in both solitary and social wasp venoms include hyaluronidase, phospholipase A2, metalloendopeptidase, etc. Although it has been expected that more diverse bioactive components with the functions of prey inactivation and physiology manipulation are present in solitary wasps, the information on venom compositions of solitary wasps obtained in this study was not sufficient to generalizae this notion. Nevertheless, some neurotoxic peptides (e.g., pompilidotoxin and dendrotoxin-like peptide) and proteins (e.g., insuline-like peptide binding protein) appear to be specific to solitary wasp venom. In contrast, several proteins, such as venom allergen 5 protein, venom acid phosphatase, and various phospholipases, appear to be relatively more abundant in social wasp venom. In the venom gland trancsriptome of bumblebees, major allergens or pain producing factors were barely identified, implying that bumblebee venoms are relatively less toxic than those of social or solitary wasps.

      • Characterization of venom evolution via venom component phylogenetic analysis in Aculeata Hymenoptera

        Kyungjae Andrew Yoon,Do Eun Lee,Woo Young Bang,Chang Hwan Bae,Joo-Hong Yeo,Young Ho Koh,Si Hyeock Lee 한국응용곤충학회 2019 한국응용곤충학회 학술대회논문집 Vol.2019 No.04

        To identify and compare the venom components and expression patterns of some bees/wasps, venom gland-specific transcriptome analyses were conducted for 14 Aculeate bees/wasps. Most of the allergens and pain-producing factors showed extremely high expression levels in social wasps, implying that social wasps have evolved to use venom to defend the colony against intruders. Acid phosphatase and tachykinin, which are known as allergens and neurotoxic peptides, were found with high frequencies in the venom glands of solitary wasps. This suggests that solitary wasps might use their venom for catching and preserving prey. In the venom glands of bumblebees, little or no transcripts of major allergens or pain producing factors were identified, implying that bumblebees venoms are relatively less toxic than those of social or solitary wasps. Taken together, the differential expression patterns of venom genes in some Aculeate bees/wasps implies that bees/wasps have unique groups of highly expressed venom components, which appear to have evolved in response to both ecological and behavioral influences.

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