에메리개미는 여왕개미와 수개미가 유전적으로 복제되어 번식한다고 알려져 있으며, 여왕개미의 날개형태가 장시형과 단시형으로 나타난다. 장시형은 정상적인 날개형태이고, 이보다 짧은 날개형태는 단시형이라고 한다. 장시형과 단시형 모두 한 종으로 취급되지만, 두 가지 점에서 종 지위에 대한 조사가 필요하다. 첫째, 자연 상태에서는 두 날개형이 함께 발견되지 않고, 둘째, 날개형이 육안으로 뚜렷하게 구분된다. 또한 복제되어 번식한 여왕개미가 단수체인지 배수체인지 조사가 필요하다. 따라서 우리는 본 연구에서 에메리개미 유전체 크기를 추정하여 두 날개형은 동종이며, 여왕개미는 배수체임을 확인하였다.

Vollenhovia emeryi (Formicidae: Myrmicinae) is divided into two morphs based on the wing length of the queen caste: the long-winged with a normal wing and the short-winged with the aberrant short wing. The phylogenetic analysis shows that the short-winged is derived from the long-winged. In Korea, only the long-winged morph is infected with Wolbachia while the short-winged is devoid of the bacterium suggesting that the short-winged evolved the resistance to the bacterial infection. Intriguingly, some Japanese short-winged colonies proved to still be infected with the bacterium. We hypothesized that the infected Japanese short-winged is the intermediate form in the process to complete cure. However, the data of the MLST and the measurement of Wolbachia density did not support our hypothesis. Our further experiment using microsatellite markers shows that the infected Japanese short-winged shows the similar genetic background to the long-winged. In this presentation we will discuss the potential resistance evolution in the Korean short-winged and future research direction at the genome level.

In every summer, cicadas emerge and become numerically and ecologically dominant in Korean penninsula. Especially, cicada emergence is affected by the environmental factors. In order to evaluate the effect of environmental factors in cicada species, we analyzed the temporal changes in cicada exuviae based on meteorological and non-meteorological factors such as artificial light intensity and habitat characters on urban park area surrounded by residential houses. Combined multivariate analyses with a cluster analysis and a principal component analysis (PCA) were conducted. Samples were classified into 3 different cluster based on differences of meteorological factors such as temperature and humidity. Moreover, Random Forest model (RF) showed a high predictability of daily mean temperature on species peak abundance. These results provide a strong evidence that meteorological factors have significant effects on cicada emergences. Regarding the non-meteorological factors, we found no relationship in cicada emergence.

Vollenhovia emeryi (Hymenoptera: Myrmicinae) is dimorphic in its wing morphology of alate females: the long-winged and the short-winged. In our previous study, we found that the long-winged is ancestral and the short-winged is derived. Intriguingly, the former is infected with the intracellular symbiotic Wolbachia bacterium and the derived is void of the bacterium indicating that the latter somehow evolved resistance to the bacterium. This may be one of few cases in which transition from susceptibility to the bacterium can be traceable via the divergence estimation. As a consequence, we inferred that the two morphs diverged approximately quarter million years ago; a remarkably recent event in evolutionary perspective. In this presentation, we will further discuss genetic orchestration in the host insect and future research directions.

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.

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 Hymenoptera, thanks to its haplodiploid sex determination, arrhenotokous parasitoid females are able to control offspring sex ratio by determining whether to use sperm or not at the time of oviposition. However, the offspring sex ratio may also be influenced by the mating behavior of a male. When a male searches females, he will have more chances to encounter with other males in a high male density population than low male density population. If the male is competitive, the male can find females faster and may have longer copulation time, and eventually transfer more sperm than other males. In this study, we investigated mate searching time, copulation duration and offspring sex ratio as a function of the male density. Although the results show difference between searching time and copulation duration, they are not statistically significant. We will discuss the statistical consequence, male density dependent offspring sex ratio and its evolutionary implications.

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.

The hologenome theory of evolution proposes that an organism is simply not an individual but the totality of numerous microbial symbionts to the host. In fact, the evidence of biochemical and physiological associations between the symbionts and the host has been growing fast in all major taxa. The Vollenhovia emeryi ant is tiny and found nationwide in Korea. The ant can be further categorized by its wing morphology, eg. long-winged and short-winged. Our initial screening process showed that the microbial reproductive manipulator, the Wolbachia bacterium, only infected the long-winged morph. This gave us a good opportunity to investigate the effect of the Wolbachia infection on the bacterial community diversity using the next generation sequencing technique. We find that there are about 180 bacterial symbionts in the short-winged morph. On the other hand, the long-winged morph has only about 20 bacterial symbionts. This implies that the bacterial community diversity may be subject to the existence of Wolbachia. Furthermore, the Wolbachia strain diversity is unexpectedly high. In addition, the bacterial structure difference among castes indicates that there may be labour division even between queens. The results and future research direction will be discussed from the hologenome theory perspective.

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 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.

Molecular phylogenetic analysis was conducted to evaluate the taxonomic relationships of genus Arisaema L. distributed in Korea and the molecular phylogenetic characteristics of three authentic Arisaema species for the herbal medicine Arisaematis Rhizoma (the rhizomes of A. amurense, A. heterophyllum, and A. erubescens). The sequences of three DNA barcodes (rDNA-ITS, matK, and rbcL) were analyzed using 50 samples of nine taxa consisted of eight Korean and one Chinese Arisaema with one outgroup (Dracunculus vulgaris). Both individual and combined phylogenetic analyses of three DNA barcode sequences revealed that the treated nine taxa are independently classified into six distinct clades (Clade I, A. amurense f. amurense and A. amurense f. serratum; Clade II, A. serratum and A. takesimense; Clade III, A. ringens; Clade IV, A. erubescens; Clade V, A. heterophyllum; Clade VI, A. thunbergii subsp. thunbergii and A. thunbergii subsp. geomundoense). These six clades were reasonably divided into three individual sections, Pedatisecta, Sinarisaema, and Tortuosa. Futhermore, the results of comparative DNA barcode sequences analyses provided a significant information for the taxonomic reconsideration of Arisaema L. at the specific and intraspecific level. However, we could not confirm the taxonomic characteristics or identity among the three authentic medicinal species through the molecular phylogenetic analyses of genus Arisaema L. for Arisaematis Rhizoma.