The effects of seed soaking treatment with the solutions of plant growth regulators IAA, GA3 and BAP on seed germination and shoot and bulb growth of Allium victorialis var. platyphyllum (Korean wild garlic) were determined. A significant variation in the seed germination rate was recorded at all treatments for various soaking periods. Maximum seed germination was obtained when seeds were soaked in IAA or GA3 solution at 200 mg L-1. The MAP treated seeds started to germinate after 3 months. Among treatments, IAA was found to be most effective in improving seed germination, but further seedling growth was not correlated to the soaking time. Seed soaking in IAA or GA3 solution enhanced further growth of seedlings compared with water control treatment. Shoot and bulb growth was highest in GA3 treatments.

The study was to examine the variation and pattern of colony components of Vollenhovia emeryi. The total of 38 colonies was collected from 10 locations, including 35 colonies from 9 locations in South Korea and 3 colonies from 1 location in USA. To know colony components variation, we studied the approximate colony size, the approximate number of reproductives: queens, gyne and male, the approximate number of brood during 7 months. The observation period was from September 2012 to March 2013 over the breeding season from July to August. As a result, the study has found that colony components of the 38 colonies are not constant. This also indicates that gynes and males appear during non-breeding season at least in the laboratory condition. There are 4 types of gynes and males emergence: both(31.6%), only gynes(2.6%), only males(28.9%), absence(28.9%). The number of gynes and males of each colony also shows variance. It indicates that Vollenhovia emeryi may show colony components continuum. In this species, fertilized eggs become worker females and males; worker females are diploid but males are haplid because of maternal genomic elimination. Unfertilized eggs become queens which are diploid by clonal reproduction. Therefore, each individual of the sex can exceptionally transfer its own genes. This unique haplodiploid sex determination may create the conflict between queen and male and cause colony components continuum.

The phylogenetic analysis the queen polymorphic Vollenhovia emeryi ant reveals derivation of the Wolbachia- free short-winged from the Wolbachia-infected long-winged. However, intriguingly, some Japanese short-winged colonies harbor Wolbachia. Wolbachia specific bacteriophage (WO) is also detected in more than half of the infected colonies with no clear distribution pattern across the host insect lineage. We hypothesized that 1) the infected Japanese short-winged is in the intermediate stage to complete loss of Wolbachia and 2) the phage invaded the host after the host insect diverged. To test the hypotheses, we studied the strain diversity using the multi-locus sequence typing (MLST) of five ant colonies; three longwinged colonies from Korea and one long-winged colony and one short-winged colony from Japan. Both Korean and Japanese V. emeryi colonies show unexpectedly high level of Wolbachia strain diversity. However, the diversity is not significantly different between the long-winged and the short-winged against our first hypothesis. Phylogenies of Wolbachia show Korean strains and Japanese strains are largely monophyletic indicating prior infection before the host divergence. The strain diversity of the phage is also surprisingly high. Phylogenies of orf2 and orf7 genes are incongruent to that of Wolbachia and geographically distinct. This indicates that the phage is spatially static and the current infection pattern may be the consequence of local repeated gain and loss of the phage.

The ant species, Vollenhovia emeryi Wheeler (Hymenoptera: Myrmicinae) is endemic in East Asia and has invaded into North America. In this species, the queen caste shows polymorphism in its wing morphology; long-winged queen and short-winged queen, and two morphs are thought not to coexist in nature. This research is conducted to 1) deduce the phylogeographical structure of the two wing morphs in South Korea and to trace the distribution pattern from East Asia to North America, and 2) investigate the Wolbachia and WO phage infection frequency of the species. Either individuals or colonies of V. emeryi were collected from 80 locations, encompassing 68 locations in South Korea, 11 in Japan, and one in USA. Among the collected samples in South Korea, the long-winged morph is dominant and considered as the ancestral type, while the short-winged morph is very rare and derived character. The origin of the US population is neither Korea nor Japan at least in this study. However, we do not exclude the possibility that its origin is the other parts of Japan or the other countries. All of the long-winged morph are infected with Wolbachia, while the short-winged seems to be geographically partially infected. It suggests the possibility that the short wing trait is linked with the evolution of resistance to Wolbachia infection. Bacteriophage WO infection status has no correlation with host insect lineage.

CD63, a member of tetraspanin membrane protein family, plays pivotal role in cell growth, motility, signal transduction, host-pathogen interactions and cancer. In this work, the cDNA encoding CD63 homologue (TmCD63) was cloned from larvae of coleopteran beetle, Tenebrio molitor. The cDNA is comprised of an open reading frame of 705 bp, encoding putative protein of 235 amino acid residues. In silico analysis shows that the protein has four putative transmembrane domains and one large extracellular loop. The characteristic ‘Cys-Cys-Gly’ motif and ‘Cys188’ residues are highly conserved in the large extracellular loop. Phylogenetic analysis of TmCD63 revealed that they belong to the insect cluster with 50-56% identity. Analysis of spatial expression patterns demonstrated that TmCD63 mRNA is mainly expressed in gut and Malphigian tubules of larvae and the testis of the adult. Developmental expression patterns of CD63 mRNA showed that TmCD63 transcripts are detected in late larval, pupal and adult stages. Interestingly, TmCD63 transcript was upregulated the maximum 4.5 fold in response to DAP-type peptidoglycan during the first 6 h, although other immune elicitors also made significant increase in the transcript level at later time-points. These results suggest that CD63 might contribute to T. molitor immune response against various microbial pathogens.

The little sized ant species, Vollenhovia emeryi Wheeler (Hymenoptera: Myrmicinae) is the only species belonging to genus Vollenhovia in South Korea, and it is endemic in East Asia encompassing Korea, Japan, China, Taiwan and has recently invaded into North America. In this species, the queen caste shows polymorphism in its wing form, the queen with normal wings called the long-winged (L) queen and short-winged (S) queen with aberrant small wings, and the two morphs are thought not to coexist in nature, however the morphology of workers and males of the two wing morphs are indistinguishable. We obtained a L gyne from a S colony that had been maintained in the laboratory. In addition, we compared the genome size of entire castes of the two wing morphs using flowcytometry. Our results confirm that the two wing morphs are obviously the same species, and moreover, the wing morph may be determinated by the epigenetical process.

In Korea, twelve species in genus Myrmica have been described. Some of them are morphologically similar and this makes their identification difficult. For this reason, we collected several Myrmica species in question and inferred their phylogenetic relationship using the 418bp partial COI (cytochrome C oxidase 1) region from a total of 33 individuals. We found that the CO1 haplotypes are effectively grouped into three clusters that match well to their external morphological characters. Although this three species could be distinguished by the only small part of the COI region, the two individual sample of the M. kotokuii and one sample of the M. carinata are included in the M. kurokii group. The results indicate that the morphological identification could be obscure in the three species and it requires a close examination for this phenomenon.

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.

In haplodiploid sex determination, females are sexually reproduced from fertilized diploid eggs, and males from unfertilized haploid eggs. Haplodiploid sex determination seems simple in that sex depends simply on the ploid level. However, the underlying genetic mechanisms are thought to be much more complicated than expected. Among them, a powerful proposed mechanism is genomic imprinting. All epigenetic on-off systems require target genes, unless the systems target histone proteins on chromosomes. For Hymenoptera, a good candidate target gene in terms of sex determination is known either as feminizer (fem) or transformer (tra) in many insects. These two genes are essential for expressing femaleness. In most Hymenopteran insects, the maternal tra seems to be methylated and consequently not expressed, while the paternally derived tra gene is not methylated. Therefore, a fertilized egg with the paternally derived active tra gene will develop into a functional female. Like all Hymenoptera, ants (Formicidae) have haplodiploid sex determination. In Vollenhovia emeryi, however, queens are produced clonally while workers derive from fertilized eggs. Males are haploid, likewise deriving from fertilized eggs, but only after selective elimination of their maternal genome. Under the conventional genomic imprinting model, we would have expected that the opposite pattern of what is observed in others. Here we present extraordinary sex determination and suggest our hypothesis about genomic imprinting pattern in V. emeryi

Vollenhovia emeryi ant is distinguished by its wing morphology; short winged (SW) and long winged (LW). Its reproduction shows a bizarre genetic caste system distinct from other social hymenopteran insects. Unfertilized eggs undergo genome duplication and develop into clonal gynes. Fertilized eggs develop either into workers or males. The fate of the fertilized eggs is determined whether maternal genome loss (MGL) takes place after fertilization. Eggs with MGL become haploid males with only paternal half of the genome. Without MGL, the eggs become workers with maternal and paternal half of the genome. In this research, we analysed 5 nuclear genes of SW and LW individual ants. Among them, two genes from an SW male are identical to those of LW, and one gene from the SW male seems a variant of LW. The result indicates that SW males are derived from LW colonies. From the genetic relatedness point of view individuals in the same castes are genetically identical. On the other hand, between workers and two reproductives, the relatedness is asymmetrical and there is even no gene sharing between gynes and males. The conventional genetic relatedness by Hamilton is revised under this condition.

The ant species, Vollenhovia emeryi, is distributed in Far East. The species can be divided into two major groups by their wing morphology of reproductives: short-winged and long-winged. A nationwide survey of the species was conducted for analyzing the mitochondrial haplotype diversity and genetic population structure. We collected 91 samples from 40 locations. A total of the 1239 bp partial COI (cytochrome C oxidase 1) region was used for the analyses. We found the total of 21 haplotypes. The mitochondrial haplotypes may correspond to the wing morphology. The genetic population structure examined potential geographic barriers of gene flow such as distance, mountains, rivers and plains which are non-mountain areas to prevent dispersal through mountain range. The result implied that no barriers considered in this study affected differently gene flow. Therefore, the behavioral characteristics of the ant may be the causal constraint of its genetic exchange.

Vollenhovia emeryi chosenica (Wheeler) (Hymenoptera: Myrmicinae) is an ant species frequently found in forests. In nature, two phenotypically distinct forms are found e.g. long winged and short winged. Unlike other hymenopteran insects, the ant is unique in its mode of reproduction. In this species, queens are clonally reproduced from unfertilized eggs. On the other hand, workers develop from fertilized eggs. Strikingly, haploid males are reproduced from fertilized eggs after destroying the maternal half of the genome e.g. maternal genome loss (MGL) consequently only with the paternal half of the genome. We collected the ant colonies nationwide in 2011. In this study, we demonstrate that the ant is infected with Wolbachia, the bacterial reproductive manipulator in various insects. Interestingly, only the long winged morphs seem to be infected. Furthermore, most colonies are mulitple-infected except two colonies collected from Chuncheon and Mt. Deogyu. We will discuss potential interactions among the Wolbachia infection polymorphism and wing morphology, and evolution of clonal reproduction and MGL.

The black soldier fly (Hermetia illucens) (Diptera: Stratiomyidae) is an invasive species that is thought to be endemic in Korea. The insect has been paid much attention thanks to its ability to decompose organic wastes. We collected the insect nation-wide in 2006 and 2010 to investigate its genetic structure based on the mitochondrial cytochrome oxidase 1 (mt CO1) haplotype. The results show that during the time period the insect seems to lose some rare haplotypes. We conclude that the insect may have experienced genetic bottleneck while its settlement in Korea. In our presentation, we show its updated haplotype network and genetic differentiation and propose potential explanations.

This study was aimed to develop a novel qualitative multiplex polymerase chain reaction (PCR) for simultaneous detection of genetically modified (GM) soy and maize within a single reaction. The specific primers designed to detect four respective GM events (A2704-12, MON88017, Bt11, and MON863) were included in the tetraplex PCR system. Each of PCR products for four GM events could be distinguished by agarose gel based on their different lengths. The specificity and reproducibility of this multiplex PCR were evaluated. This multiplex PCR consistently amplified only a fragment corresponding to a specific inserted gene in each of the four GM events and also amplified all four of the PCR products in the simulated GM mixture. These results indicate that this multiplex PCR method could be an effective qualitative detection method for screening GM soy and maize in a single reaction.

This study was conducted to investigate insect fauna in the four insect garden sites of Suwon, Yeongyang, Buyeo and Yecheon from 2005 to 2007. Seasonal population size of insects was largest from June to August in all the four sites. In the four sites, Coleopteran insects were dominant followed by Hemiptera and Orthoptera. Unique education/learning programs are successfully run at the insect gardens based on the three geographic types (i.e. urban, mountainous and rural). These activities will help preserve insect biodiversity in the area and visitors better understand life forms such as insects found in the areas.

The porcine reproductive and respiratory syndrome virus (PRRSV) has six structural proteins which encoded by ORFs 2 to 7 are designated as GP2, 3, 4, 5, M and N, repectively. In this study, we determined the expression of each protein using novel transfer vector, pBmKSK4 which has the polyhedrin promoter of BmNPV and 6xHis tag. The recombinant transfer vector was co-transfected into Bm5 cells along with bBpGOZA DNA. Recombinant virus was purified by plaque assay and amplified in Bm5 cells. Expression of each protein was identified by SDS-PAGE and Western blot analysis using anti-6xHis monoclonal antibody. The expression levels of the structural proteins in Bm5 cells were stronger than the expression system using pBacPAK9 transfer vector in Sf21 cells. As expected, GP5 was expressed at low levels from its structural properties and its toxicity for cells. In addition, each recombinant protein was purified using Ni-NTA spin columns. The ability to produce each protein in the baculovirus system indicates that these could be major candidates for the development of a vaccine against PRRSV.

As a continuation of a previous work by Park et al. (2006), we have developed a two-element radio interferometer that can measure both the phase and amplitude of a visibility function. Two small radio telescopes with diameters of 2.3 m are used as before, but this time an external reference oscillator is shared by the two telescopes so that the local oscillator frequencies are identical. We do not use a hardware correlator; instead we record signals from the two telescopes onto a PC and then perform software correlation. Complex visibilities are obtained toward the sun at λ=21cm for 24 baselines with the use of the earth rotation and positional changes of one element, where the maximum baseline length projected onto UV plane is ~90λ As expected, the visibility amplitude decreases with the baseline length, while the phase is almost constant. The image obtained by the Fourier transformation of the visibility function nicely delineates the sun, which is barely resolved due to the limited baseline length. The experiment demonstrates that this system can be used as a "toy" interferometer at least for the education of (under)graduate students.

This study identified and compared the volatile flavor components of two commercial rice wines: one fermented using the mycelium of Phellinus linteus and a regular commercial rice wine. The volatile flavor components were isolated from the infusions by Porapak Q (50-80 mesh) column adsorption. The concentrated aroma extracts were then analyzed and identified by GC and GC-MS. Thirty-four kinds of flavor components were identified in the mycelium-fermented rice wine, including 11 alcohols, 8 esters, 3 ketones, 6 acids, 3 hydrocarbones, and 4 others. In the regular commercial rice wine, 36 kindss of flavor compounds were identified, including 9 alcohols, 6 esters, 4 ketones, 6 acids, 9 hydrocarbones, and 2 others. Therefore, the data indicate that the primary flavor components in the rice wines were alcohols and esters.

Al though the changes in tooth morphology and hardness by hydrogen peroxide(H20 z) have been r‘epor‘.ted .‘ the pαr。o야t뻐ec야tive role of heme oxygenase-l(HO-l) against the cytotoxic effects of H202 has not been clarifïed i n human pulp cells ln this st udy. we investigated whether HO-l is involved in Hz0 2-induced cytoLox icity a nd examined the production 0 1' dentin sia lophosphoprotein(DSPP) and other mineralization markers‘ in hllman pu lp cells H202 decreased cell viabi lity, but increased HO-l and DSPP expression in a concentra tion and time dependent manner . Inhibitors of gllanylate cyclase. PI3K, ERK. and p38 MAP kinase blocked H202-indllced cytotoxicity and the expression of HO-l and DSPP mRNAs in pulp cells. These data suggest that the induction of HO-l by H202 in plllp cells plays a protective role against the cytotoxic effects 0 1' HzOz and stimulates DSPP expression‘ reslllting in prematllre odontoblast dilTerentiation throllgh pathways that involve cGMP‘ p38. and ERK.