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.

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 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.

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.

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 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.