The red-spotted apollo butterfly, Parnassius bremeri Bremer, 1864 (Lepidoptera: Papilionidae), has been listed as an endangered species in Korea. We developed microsatellite markers by nest-generation sequencing (NGS), selected 12 markers, and applied the markers to available South Korean populations to understand population genetic characteristics. The genotyping of 40 P. bremeri individuals from three localities in South Korea showed that at each locus, the observed number of alleles ranged from 17 to 43, the observed and expected heterozygosities were 0.84722-0.90556 and 0.76045-0.79208, respectively, and FIS was –0.155 to –0.121. STRUCTURE analysis supported the presence of two genetic pools in all three populations, although an immediate reason for this subdivision is not known. The population based FST, RST, and front wing length collectively suggest that at least Samchuk population in Gangwon Province has a significant distance. Further scrutinized analysis is undergoing.

The population size of Red-spotted Apollo Butterfly(Parnassius bremeri) has been reduced because of their habitats destruction and partly climate change. Estimation of metapopulation size and survival day of Red-spotted Apollo Butterfly was made in Samcheok where release was carried for 5 years, Korea, by using the mark-release-recapture method. 421(female: 188, male: 233) of Red-spotted Apollo Butterfly were captured and 177 individuals(female: 89, male: 88) were recaptured and rates of recapture was 42%. Average of survival day was 3.59 and max survival day was 11. The migration of Red-spotted Apollo Butterfly was occurred significantly between short patches. Their max distance of migration was 6.74km. Estimate of P. bremeri was from minimum 125 to maximum 1844.

This study was conducted to investigate exact life cycle and Parnassius bremeri’s response to heat shock stress from 2012 to 2016 in Holoce Ecosystem Conservation Research Institution through captive breeding. About 16 days(15.7±0.3days) later embryo in eggs were developed as pharate first instar and the eggs of Parnassius bremeri possessed the ability to undergo an extended aestivation hosting a fully developed pharate 1st instar larvae within the chorion. Pharate 1st instar in egg hatched from late in November to early in December about 194 days(194.3±1.1days) later. All pharate 1st instar in egg and out of egg survived at 25℃, 35℃. In case of 45℃ survival rate of pharate 1st instar in an egg (93.8±6.3%) was significantly higher than those (22.2±10%) of pharate 1st instar out of an egg. All was dead at 47.5, 50, 52.5 and 55℃.

Egg SCP ( –47.2 ±1.0℃) of Red-spotted Apollo butterfly, Parnassius bremeri, was lower than larva(–35.0 ±0.9℃). The difference (12℃) between egg and larva was revealed by chorion through scanning electron microscope (COXEM EM-30, Korea). But considering of about 6 months’ pharate 1st instar in egg from June to November, function of egg seems not to be overwintering mechanism but rather estivation. In order to identify estivation function of chorion heat stress was carried out. Experiment was conducted on the 4 conditions such as high temperatures of 25, 35, 45 and 55 °C for 2 hour, then allowed them to recover at 25 °C for 1 hour. Total 65 samples 0f 31 eggs and 34 pharate 1st instar out of an egg were used and replicated 3 times. All pharate 1st instar in egg and pharate 1st instar out of egg survived at 25℃ and 35℃. In case of 45℃ survival rate of pharate 1st instar in an egg (91.6±3.5%) was significantly higher than those (37.8±9.4%) of pharate 1st instar out of an egg. All was dead at 55℃.

The Red-spotted Apollo Butterfly (Parnassius bremeri ) is member of Snow Apollo Butterfly in Genus Parnassius which has been designated as the most endangered insect around the world and recorded in IUCN Red List of Threatened Animals and Appendix II in CITES. The lack of distributional information of Parnassius bremeri is one of the main problems when trying to assess the threats to and planning conservation strategies for this endangered species. The pheromone method made it easier to detect local populations of the species, not to disturb the habitat and can be used by providing a useful tool for conservation research. Sampling by breeding captivity in Holoce Ecosystem Conservation Research Institution (HECRI) was used. Each of 3 individuals male, mated and unmated female, end of the abdomen tissue and abdominal tissue containing the sphragis were cut. The pheromone was extracted as method presented by Hernández-Roldán et al. (2014). Among pheromone fatty alcohol to the (Z, Z, Z) -9,12,15-octadecatrien-1-ol and Eicosane classified as hydrocarbon of unmated female was detected higher rate than mated female. Two pheromone can be helpful to attract males in potential population.

SCPs on larvae and eggs of red-spotted apollo butterfly, P. brimeri were measured according to the method of Kim and Kim (1977) with a thermocouple, BTM-4208SD (LT Lutron, Taipei, Taiwan), to detect the release of the latent heat of fusion as body water freezed. SCP of larvae during March goes below –27.4±1.7℃ and egg scp during November goes –47.2±1.0℃. In order to identify the reason of the difference(-17℃) between egg and larva we took photographed egg through scanning electron microscope (COXEM EM-30, Korea). Chorion of P. bremeri were 100.1㎛, Papilio machaon and Sericinus montela in same family was 10.8㎛, 5.5㎛ respectively. P. bremeri was 10 times, 20times thicker than another species within the same family .

The early spring swallowtail, Luehdorfia puziloi (Lepidoptera: Papilionidae) is univoltine with adult emergence in early spring and the larvae feed on the genus Asarum (Aristolochiaceae). This study was conducted to investigate temperature effect on adult emergence of overwintering L. puziloi pupae at five different constant temperatures (15, 20, 25, 27.5 and 30℃) with photoperiod 10:14(L:D) and relative humidity 60% by using 150 pupae, 2013 in Holoce Ecosystem Conservation Research Institution (HECRI). The lower developmental thresholds of female, male, and both sexes combined were 6.131, 8.64, and 8.088℃, respectively. Developmental period of overwintering pupae to adults decreased with increasing temperature from 15 to 30℃. Mortality of five different constant temperatures (15, 20, 25, 27.5, and 30℃) was 13.3, 3.3, 16.67, 13.3 and 13.3%. Especially mortality of constant temperature 25℃ was higher than other conditions and survival rate 20℃ was the highest. Thermal constants of female, male and both sexes combined were 207.11, 125.97 and 150.18DD, respectively. The relationship between the developmental rate and temperature was fitted by linear and five nonlinear developmental rate models (Logan 6, Lactin 1, 2 and Briere 1, 2). The development variation of overwintering pupae was described by the two-parameter Weibull distribution model. The adult emergence of overwintering L. puziloi pupae in spring can be explained and predicted by using developmental rate models and developmental distribution model.

Luehdorfia puziloi (Erschoff, 1872) is a monophagous butterfly species that occurs only once during spring in a year and distributed nationwide in Korea. From 2012 to 2013 the adult emergence has been monitored to assess how the species responds to temperature rise in the country. Monitoring sites were located in four different provinces, two in the northern part of the country and the others the southern part. In each site a small emergence cage was set up with 60 to 70 pupae and a HOBO data logger to record the soil temperature of the site from February to April. The earliest record from literatures and specimen collection data was 28th of March in Geojae-gun, one of far south parts of the country, during the late 90’s. In 2013, the butterfly started emerging in a southern site, Namhae-gun, from 20th of March, and it was the earliest record so far. The emergence from all of the sites occurred on average 7.75 days faster in 2013 than 2012. The emergence patters during the two years clearly showed that the adult emergence reached to 50% emergence rate faster as the soil temperature rose.

Temperature is one of important factors to determine insect phenology. Based on the bioclimatic law, the relationship between climate change and ecosystem change was studied from 2008 to 2013 in HECRI by monitoring the spring emergence patterns of three Papilionidae species (Papilio xuthus, P. machaon, and Sericinus montela). The overwintering pupae were set on the wood plate and adult emergence were monitored and recorded in every morning. The first spring emergence of P. xuthus, P. machaon and S. montela in 2013 were Apr 19th, May 1st and Apr 22th, respectively. And peak time of three species were May 7th, May 11th and May 9th, respectively. Study on temperature-dependent development was conducted to investigate the temperature effect on adult emergence of overwintering S. montela pupae at four different constant temperatures (15, 20, 25 and 30°C) with photoperiod 10:14(L:D). The low temperature threshold of female, male and both sexes combined were 12.39, 12.16, and 12.37°C, respectively. Developmental period of overwintering pupae to adults decreased with increasing temperature from 15 to 30°C. Thermal constant of female, male and both sexes combined were 220.26, 192.31, and 200.18DD, respectively. The relationship between thermal constant and cumulative adult emergence was predicted by temperature-dependent development. Estimate through 7 times on the highest temperature was equal and results were distinctively divided into two pattern(2008～2010 and 2011～2013). The relationship between observed and estimated values was presented by linear regression (r2=0.97)

Papilio xuthus is known as the Asian swallowtail, Chinese yellow swallowtail, or as the Xuthus swallowtail. It overwinters in pupae stage. The study was conducted to investigate the temperature effect on adult emergence of overwintering P. xuthus pupae at four different constant temperatures (15, 20, 25 and 30°C) with photoperiod 10:14(L:D). The lower developmental thresholds of female, male and both sexes combined were 9.71, 11.03, and 10.49°C, respectively. Developmental period of overwintering pupae to adults decreased with increasing temperature between 15 and 30°C. Degree-days for adult emergence to female, male and both sexes combined were 227.27, 175.44, and 200.80DD, respectively. The relationship between the developmental rate and temperature was fitted by linear and five nonlinear developmental rate models (Logan 6, Lactin 1, 2 and Briere 1, 2). The variation of overwintering pupae development was described by the two-parameter Weibull distribution model. The adult emergence of overwintering P. xuthus pupae in spring can be explained and predicted by using developmental rate models and developmental distribution model. The effect of climate change on phenology and distribution of P. xuthus may be presented by the model.

Papilio macilentus is the longest tail butterfly of Papilionidae found in Japan, China and Korea and has distinctive morphological differences between male and female. P .macilentus feeds on Zanthoxylum schinifolium and has three generations per year. The lower developmental threshold and thermal constant for adult emergence of P. macilentus overwintering pupae were estimated at four different constant temperatures (15, 20, 25 and 30°C) with photoperiod 10:14 (L:D). The lower developmental threshold of both sexes combined was 7.95°C. The development of overwintering pupae decreased with increasing temperature between 15 and 30°C. Thermal constant of both sexes combined was 251.26DD. The relationship between the developmental rate and temperature was fitted by linear and five nonlinear developmental rate models (Logan 6, Lactin 1, 2 and Briere 1, 2). The development of overwintering P. macilentus pupae was described by the two-parameter Weibull distribution model. The temperature-dependent adult emergence model of P. macilentus developed in this study could be used to predict the full-cycle phenology model of P. macilentus and applied to understand the effects of climate change on the emergence distribution of overwintering pupae.

 ,  , Understanding the ecological complexity and habitat of a species are crucially important to conserve an endangered species. This study evaluated the patch network ecology of the endangered species Parnassius bremeri. The results indicated that 188 individuals were captured and 220 were recaptured, respectively. The sex ratio of female: male was 42:146, males were four times more abundant than females. The average longevity of an adult was 3.93 ± 3.93 days (male, 4.0 ± 3.9, female, 2.5 ± 1.0 days), the maximum longevity was 14 days for males and 13 days for females, respectively. Therefore, the expected longevity of males was longer than that of females. The average emigration distance for the species was 377 m, and the maximum emigration distance was 1550 m. The analysis of patch connectivity and individual colonization revealed that the ideal distance between patches was about 300 m. Moreover, a >, 600 m patch distance decreased the colonization rate severely. We also observed higher immigration and emigration between patches that were clustered in close proximity. This leads us to conclude that a higher number of patches at a close distance is best suited for P.bremeri. We find these results to be crucial to determine a policy to protect and conserve this endangered species.

Understanding the metapopulation structure and movement of a species are required for conserving the species. In this paper, migration patterns and connectivity of patches of a threatened butterfly, Pamassius bremeri Bremer, were postulated using the mark-release-recapture (MRR) technique in a habitat located in the mid-southern region of the Korean peninsula. A total of 194 individuals were captured (137 males and 57 females) and, of them, 93 individuals (73 males and 20 females) were recaptured during the MRR experiment. The migration analysis showed 23-150% immigration and 28-53% emigration. There were high correlations between the migrating individuals and the distance between patches, but there was no correlation between migrating individuals and patch size or between migrating individuals and the number of host plants. Consequently, the migration of butterflies occurred frequently between closer patches, while patch size and quantity of the food plant had minor effects on migration behavior. Additionally, males migrated more frequently than females. Analysis of the migration patterns of P. bremeri showed that the central patch played an important role on linking patch groups and more frequent migrations were monitored between nearby patches than between the remote patches. This study suggested that active migrations take place between the neighboring multiple patches and these are accelerated if there is a stepping-stone patch between them.

The loss of natural habitats and their fragmentation caused by human activities pose a great threat to biodiversity worldwide, reducing animal population to small, fragmented patches of natural habitat. In this paper, a metapopulation dynamics of Sericinus montela koreanus, a species of butterfly, was studied for two years by mark-release-recapture (MRR) techiques in the Musimcheon, Cheongju Korea. 2,749 individuals (males: 2,295, females: 454) were marked and released, with 343 individuals (12.5%) recaptured during the MRR experiment. Capture sex ratio and recapture sex ratios was both consistently male-biased. The Jolly-Seber model was used to estimate daily metapopulation size, survival rates, addition rate, and accidental deaths. We urge conservation biologists to consider the local population dynamics of species for the management of metapopulations in fragmented landscapes. In the case of the S. montela koreanus, continuing site protection is essential.

In order to establish an indoor-rearing system for A. Alcinous, the effect of temperature and pupal diapause was investigated. Temperature has been suggested as an important factor in determining the developmental rate, Lifespan and mortality in invertebrates. As temperature increase, the developmental period was gradually reduced. The developmental period of A. alcinous larvae was 12.9 days and 25.8 days at 30℃ and 20℃, respectively. And we investigated the effect of different chilling days on diapause termination. Under longday condition(LD 16:8h, 20℃) oxygen consumption gradually increase. But oxygen consumption on the first day of pupa was the highest and rapidly fell down the next day and attained lowest point after 7days, with average value of 1.92mol/min/g under shortday condition(LD 8:16h). Diapause pupae were chilled for 60, 75, 90, 105 days at 8℃ and thereafter subjected to LD 16:8h at 25℃. And emergence rate at 60~90days was 57.1~80.7%.

The 15,389-bp long complete mitogenome of the endangered red-spotted apollo butterfly, Parnassius bremeri (Lepidoptera: Papilionidae) was determined. This genome has a gene arrangement identical to those of all other sequenced lepidopteran insects, which have the gene order of tRNAMet, tRNAIle, and tRNAGln at the beginning. Due to the uncertainty the start codon for COI gene in insect has been discussed extensively. We propose the CGA sequence as the start codon for COI gene in lepidopteran insects, based on complete mitogenome sequences of lepidopteran insects including our P. bremerii and additional sequences of the COI start region from a diverse taxonomic range of lepidopteran species (a total of 51 species belonging to 15 families). As has been suggested in other sequenced lepidopteran insects the 18 bp-long poly-T stretch and the downstream conserved motif ATAGA that were previously suggested to serve as a structural signal for minor-strand mtDNA replication also was found at the 3’-end region of the P. bremerii A+T-rich region. In an extensive search to find out tRNA-like structure in the A+T-rich region, each one tRNATrp-like sequence and tRNALeu (UUR)-like sequence were found in the P. bremeri A+T-rich region, and most of other sequenced lepidopteran insects were shown to have tRNA-like structure within the A+T-rich region, thereby indicating that such feature is frequent in the lepidopteran A+T-rich region. Phylogenetic analysis using the concatenated 13 amino acid sequences and nucleotide sequences of PCGs of the four macrolepidopteran suferfamilies together with Tortricoidea and Pyraloidea well recovered a monophyly of Papilionoidea and a monophyly of Bombycoidea. However, Geometroidea and Noctuoidea were unexpectedly clustered as one group and placed this group to the sister group to Bombycoidea, instead of Papilionoidea in most analyses.

We analyzed a portion of mitochondrial COI gene sequences (658 bp) to investigate the genetic diversity and geographic variation of the swallowtail butterfly, Papilioxuthus L., and the cabbage butterfly, Pieris rapae (Lepidoptera: Papilionidae). P. xuthus showed a moderate level of sequence divergence (0.91% at maximum) in 15 haplotypes, whereas P. rapae showed a moderate to high level of sequence divergence (1.67% at maximum) in 30 haplotypes, compared with other relevant studies. Analyses of population genetic structure showed that most populations are not genetically differentiated in both species. The distribution pattern of both species appears to be consistent with category IV of the phylogeographic pattern sensu Avise (Avise et al. 1987): a phylogenetic continuity, an absence of regional isolation of mtDNA clones, and extensive distribution of close clones. The observed pattern of genetic diversity and geographic variation of the two butterfly species seems to reflect the abundant habitats, abundant host plants, and flying abilities in connection with the lack of historical biogeographic barriers.

The 15,389-bp long complete mitochondrial genome (mitogenome) of the redspotted apollo butterfly, Parnassius bremeri (Lepidoptera: Papilionidae) was determined. This genome has a gene arrangement identical to those of all other sequenced lepidopteran insects, but differs from the most common type, as the result of the movement of tRNAMet to a position 5’-upstream of tRNAIle. No typical start codon of the P. bremeri COI gene is available. Instead, a tetranucleotide, TTAG, which is found at the beginning context of all sequenced lepidopteran insects was tentatively designated as the start codon for P. bremeri COI gene. All protein-coding genes (PCGs), but COII (T) have complete termination codon TAA or TAG. One of the most unusual feature of the P. bremeri mitochondrial genome is the presence of two tRNA-like structures, such as the tRNATrp-like sequence and tRNALeu (UUR)-like sequence with proper anticodon and clover-leaf structures in the 514-bp long A+T-rich region. Furthermore, the A+T-rich region possesses three sequences that have the potential to form stem-and-loop structures, flanked by the conserved sequences, "TA(A)TA" at the 5’ end and "G(A)nT’ at the 3’ end. After more genomic and phylogenetic analyses are performed, further detailed information will be available.

본 연구는 꼬리명주나비(Sericinus montela Grey)의 유충을 재료로 하여 투과전자현미쟁과 주사전자현미경을 이용하여 혈구의 미세구조를 관찰하였다. 꼬리명주나비 종령유충의 혈구는 구조와 특징에 따라 5종류로 구분되었다. 원시혈구(Prohemocyte)는 세포내에 하나의 큰 핵과 미분화된 세포내 소기관을 가지고 있었다. 부정형혈구(Plasmatocyte)는 섬유다발을 가진 방추행 또는 난형의 비교적 큰 세포로, 특히 방추형의 세포는 길게 발달한 핵을 갖고 있었다. 과립혈구(Granular cell)는 세포질내에 다양한 과립과 잘 발달한 소기관을 함유하고 있었으며 원형질 돌기를 많이 가지고 있었다. 소구형혈구(Spherule cell)는 세포질내에 미세한 물질을 함유한 소구(spherule)를 가자고 있으며 성숙한 세포는 소구내의 물질을 혈림프내로 방출시켰다. 편도혈구(Oenocytoid)는 세포질내 전자밀도가 높은 물결로 채워져 있으며 소기관은 잘 발달되어 있지 않았다.