무당벌레(Harmonia axyridis)는 종내에서 초시색상패턴이 매우 다양하게 존재한다. 본 논문에서는 서로 다른 색상패턴의 무당벌레를 대상 으로 amplified fragment length polymorphism (AFLP)을 실시하여 무당벌레의 초시색상 패턴간 유전형질의 차이를 확인하고자 하였다. 총 28 개의 프라이머 조합으로 실험을 실시한 결과, 총 2,741 개의 밴드가 검출되었다. 그 중 20 개의 밴드(S1-S20)만이 특정 색상패턴에서 나타났다. 이들 가운데 9 개의 밴드를 색상에 연관된 AFLP 후보 지표로 선발하였다. 밴드 가운데 S1과 S2, S20은 Succinea 1, 2 변이형에 공통적으로 나타났으며, S3와 S5는 Conspicua 변이형에 특이적이었다. 또한 S13는 Spectabilis 변이형에, S15와 S18, S19는 Succinea 2 변이형에 특이적이었다. 특정 색상패턴에만 나타나는 9 개의 AFLP 지표들은 cloning을 통해 염기서열 분석을 실시하였고, GenBank를 이용해 다른 염기 서열과 비교를 해보았지만 아무런 상동성도 찾을 수가 없었다. 무당벌레 종 내 유전적 다양성을 평가한 결과, Spectabilis가 Conspicua보다 Succinea 변이형에 높은 유사성을 보였다. 색상에 연관된 AFLP 후보 지표를 기준으로 sequence characterized amplified region (SCAR) 지표로 변환하여 9 개의 AFLP 분자지표들 가운데에서 5 개만이 SCAR 지표로 전환될 수 있었으며, 이를 통해 AFLP 지표가 무당벌레의 색상과 연관되어 있는지 확인할 수 있었다.

There was different between two differential geographical and environmental condition areas on elytra color expression patterns of the multicolored Asian lady beetles (Harmonia axyridis). Especially, it was investigated that expression rates of melanic patterns (conspicua, spectabilis and axyridis) relatively increased in overwintering populations collected in highly mean temperature and longer cumulative daylength area. In addition, in the same collection site, the seasonal difference had influenced on color patterns of H. axyridis. Although these effects didn"t were not observed in the laboratory, environmental conditions such as temperature or cumulative daylength might be factors that gave an effect on color pattern formation.

To identify DNA markers linked to a elytra polymorphism, amplified fragment length polymorphism (AFLP) analysis was performed on DNA samples from four each colour pattern individuals (2 females and males), for example, succinea 1, succinea 2, conspicua, and spectabilis. As a result of performing AFLP analysis with the restriction endonuclease combination EcoRⅠ and Mse I, total of 2,269 AFLP fragments which were specific to succinea, conspicua and spectabilis was identified using 24 different AFLP primer combinations. Among these 2,269 fragments, 16 bands which were the most specific to one color patterns were isolated, cloned and sequenced. Subsequent UPGMA cluster analysis revealed that population of H. axyridis was divided four major group and these genetic tree showed that H. axyridis elytra colour diversity was affected by genetic polymorphism. It is considered that these genetic analyses may be facilitated the understanding of molecular genetic mechanism related with the wing colour pattern formation in this species.

As an effective generalist predator of aphids and other hemipteran pests, Harmonia axyridis has been a successful biological control agent. Interestingly, it was known that there were varied in color patterns on H. axyridis elytra. In fact, Seo & Youn (2007) reported that H. axyridis had five color patterns, for example, succinea 1, 2, conspicua, spectabilis, and axyridis. But there are uncertain that H. axyridis elytra colour patterns are regulated by genetic polymorphism. So we tried to what is the reason that color patterns are greatly variable. To identify DNA markers linked to a elytra polymorphism, amplified fragment length polymorphism (AFLP) analysis was performed on DNA samples from four female succinea, conspicua, spectabilis and Coccinella septempunctata which is another species in Coccinellidae. AFLP analysis with the restriction endonuclease combination EcoRⅠ and MseⅠwas performed. Using 12 AFLP primer pairs, nine AFLP fragments which is specific between succinea, conspicua, spectabilis was identified. These nine AFLP fragments were isolated, cloned and sequenced. Subsequent UPGMA cluster analysis revealed three major group of H. axyridis populations. These genetic tree showed that H. axyridis elytra colour diversity was affected by genetic polymorphism. For more genetically understanding elytra colour genes, different primer combinations may be need to be generate enough polymorphic markers. These genetic analyses may be facilitate the understanding of molecular mechanism behind wing colour pattern formation.

The surface of most coccinellids (particularly the elytra) has characteristic colour patterns, which show great variability within many species. Individuals of a species of ladybird often differ from one another in colour and pattern. In case of Harmonia axyridis, environmental factors, such as temperature or food, can probably influence colour and pattern. However, virtually no work has been carried out in the influence of such factor. Therefore, in this study, according to rearing temperature, photoperiod and diet, variation of elytra color patterns were investigated. First instar larva on each color pattern were reared in an incubator at one of two temperatures; 25 and 30℃, under the following photoperiod; L16: D8, L12: D12 and L8: D16 and were provided three species of aphids. And their color pattern of adults were estimated.

The multicolored Asian ladybird beetle, Harmonia axyridis, which demonstrates typical genetic polymorphism in its elytra color patterns. Early studies have color polymorphism in terms of geographical clines while a few investigated temporal populations in Coccinellidae. Nevertheless, note that geographical and temporal morph variation does not always correspond to what is expected from thermal and industrial adaption theories. A recent study of transformation and RNAi of the ladybird beetle, however, there is yet no evidence to indicate the variation is genetic or environmental factors. Here we describe a relatively new molecular fingerprinting technique, amplified fragment length polymorphism (AFLP). Because we think that color polymorphism in Coccinellidae is affected by genetic polymorphism. In total 38 markers were scored from which some markers were polymorphic. Supsequent UPGMA cluster analysis revealed 3 major group of Harmonia axyridis populations. But for strains that are more genetically similar, different primer combinations may be need to generate enough polymorphic marker.

The multicolored Asian lady beetles (Harmonia axyridis) has characteristic color patterns, which show great variability within species. Up to now, it has been well known that main factors affected on individual color pattern variations in the population of H. axyridis are external, physical, and environmental characteristics. Indeed, there is as yet no evidence to indicate whether the variation is genetic or environmental factors. Also the factors which produce this variation are unknown in this species, although it is suspected that much of the variation is under genetic control. However, the genetic relationships among many of color types were investigated by observing the progeny of each particular pairs. It is worth mentioning a few particular breeding cases to illustrate certain facets of variability, and to indicate examples suitable for genetic analysis of the color pattern variation.