기후변화에 따른 서식지 감소는 생물다양성의 커다란 위협 요소 중 하나이고 생물종이 서식하는 공간적 분포에 대한 이해는 멸종위기종 관리, 생태계 복원 등 다양한 분야에서 매우 중요하다. 본 연구는 남한지역에서 서식하는 멸종위기종 Ⅰ급으로 지정된 붉은점모시나비를 대상으로 기후변화에 따른 서식분포변화를 분석하고자 한다. 이 를 위해서 최근 보전생물학, 동물생태학 등 다양한 분야에서 널리 활용되는 MaxEnt 모델을 현재시기와 미래시기 의 생물기후변수에 적용하여 잠재적 서식지 분포 변화를 평가하였다. 붉은점모시나비는 미래시기에 서식지가 감소하는 경향으로 예측되었고, 기온보다 강수량에 의한 영향이 크고, 특히 강수량 계절성에 영향이 가장 클 것으 로 분석되었다. 분석결과는 국내 생물다양성 증진에 필요한 기초자료로서 활용할 수 있을 것으로 기대된다.

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℃.

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.

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℃.

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 red-spotted apollo butterfly, Parnassius bremeri, immatures grow during winter and spring. Supercooling point of larvae during January goes much below -20℃. Morphologically, the larvae appear to be adapted to cold temperatures. Dark-colored body surface is useful to absorb solar energy and spiny integument may prevent any external ice formation on the body surface. Biochemically, P. bremeri larvae elevate glycerol as a cryoprotectant. This study reports two genes associated with glycerol biosynthesis in P. bremeri. Larval transcripts were analyzed using RNA-Seq technique. A total of 14 Gb transcripts were read by Illumina HiSeq and assembled to be 127,279 contigs. To specify the the genes associated with glycerol biosynthesis, a biosynthetic pathway to synthesize glycerol from dihydroxyacetone-3-phosphate was predicted with two genes of glycerol-3-phosphate dehydrogenase (GPDH) and glycerol kinase (GK). Both genes were annotated in the transcriptome of P. bremeri. Pb-GPDH encodes 166 amino acid residues containing NAD+-binding region, catalytic site, and calcium binding region. The predicted amino acid sequence was clustered with other lepidpopteran GPDH genes. Three Pb-GK genes were annotated from the transcriptome. Pb-GK1 encodes a full open reading frame of 514 amino acid residues. A ohylogenetic analysis showed that these three GKs were separately clustered. Interestingly, Pb-GK1 was clustered with other GKs that were known to be associated with rapid cold hardiness.

Upon freezing temperatures, most insects should avoid cellular freezing by migration to warm hibernating sites, by becoming cold-hardy or by undergoing diapause development. However, a highly threatened butterfly, Parnassius bremeri, terminates egg diapause at early winter season and grows during entire winter and spring. Thus, the cold hardiness of P. bremeri needs to be explored to understand its cold tolerance limit and physiological factors. Supercooling points (SCPs) of P. bremeri vary from -10℃ to -48℃ among season. Especially, the young larvae during Jan – Mar kept SCPs at below -20℃. Larval plasma contained high level of glycerol (39.7 mM) at March, but it decreased the level (2.4 mM) at May. Transcriptome analysis indicated high levels of gene expressions associated with glycerol synthesis. Temporal expression patterns of polyol synthesis genes supported the change of glycerol. This study suggests that glycerol is a major cryoprotectant of P. bremeri to be cold-handy against freezing temperatures during winter.

 ,  , 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 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.

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.

붉은점모시나비의 국내 서식지를 확인하기 위하여 기존에 알려진 채집지와 표본자료, 그리고 실재 잔존 발생지로 알려진 경남 남부지방을 조사한 결과 경남 고성군과 의령군 2개소에서 발생이 재확인되었으며, 이 외에 조사기간 중 강원도 삼척에서 대량 서식지가 발견되었다. 종의 보전을 위한 복원계획 수립을 위해 대량사육기술 개발이 필요하므로 생활사 등 생태적 특성을 위한 기초조사를 실행하였다. 성충의 출현 시기는 5월 중순에서 6월 말까지였다. 산란은 기주식물 외에 주변의 마른 나뭇잎 등에도 하였으며, 암컷 한 마리당 127개까지 산란하였다. 늦은 봄에 산란된 난은 220여 일의 난 기간을 보내고 1월 11일부터 난각을 깨고 부화하며, 부화된 1령유충은 10여 일이 지나 섭식을 시작한다. 항온조건(, 75% RH, l6L:8D)에서 전 유충기간은 65.7일로 각각 1령충 11.2일, 2령충 7.3일, 3령충 12.8일, 4령충 16.2일, 5령충 18.2일이었다. 용기간은 21.3일이었으며, 성충의 수명은 26.2일이었다. 산란조건을 알아보기 위하여 크기를 달리한 산란실을 만들고 각 실 당 접종수를 달리하여 시험해 본 결과 222m이상 크기의 산란실에 암컷 3+수컷 1의 비율로 넣은 경우가 가장 효과적이었다. 흡밀용 당원을 개발하기 위한 실험에서는 흑설탕구와 Fructose구에서 높은 선호도를 나타내었다.

충청북도 옥천군 지역의 붉은점모시나비에 대한 소멸원인과 복원방안을 조사하였다. 붉은점모시나비는 과거 중부지역인 옥천군에서 비교적 많은 개체가 발생하였으나, 그 수가 계속 감소하여 1997에는 두 마리의 성충만 확인되었고, 1998년에는 확인되지 않았다. 따라서 현재 옥천군지역에서 붉은점모시나비는 소멸된 것으로 추정된다. 붉은점모시나비의 소멸원인을 알아보기 위하여 1990∼1998년간 옥천군 7곳의 서식지에 대한 조사 기록의 분석과 여러기관에 소장된 표본