Sanghuangporus sanghuang (Hymenochaetaceae, Agaricomycetes) is a medicinal mushroom which has gained particular attention in Korea as a natural immune booster and a cancer suppressor. Recent molecular phylogenetic studies clarified the species concept of S. sanghuang extensively used in Asia including Korea, China, and Japan. In this study, we assembled its mitochondrial genome and identified the locations and functions of genes and coding regions using next-generation sequencing. Raw sequencing data was produced using PacBio approach, and assembled through hierarchical genome assembly process (HGAP). Determining the mitocondrial regions GeSeq and Artemis annotation tools were used. And the resulting gene map was visualized using OGDraw software. A total of 108 reads was aligned with a coverage of 11.87X. The entire mitochondrial genome was 111,734 bp in length, in which 24 proteins and 46 transfer RNAs were recognized. The overall base composition is 38.37% A, 38.42% T, 11.05% C and 12.14% G with an GC content of 23.2%. This mitochondrial genome data can be used to understand its phylogenetic relationship and taxonomic resolution and as a source of markers for population and species delimitation.

Despite of a substantial effort, the phylogeny of Lepidoptera still has unstable taxa and unresolved relationships particularly form Apoditrysia to Ditrysia (e.g. phylogenetic position and familial relationships of Gelechioidea, the monophylies of Tineoidea and Zygaenoidea and so on). We performed phylogenetic analysis using ~13,000-bp length of mitogenome sequences form 50 species (with seven species in two superfamilies form this study) representing 13 superfamilies to improve the lepidopteran phylogeny from Apoditrysia to Ditrysia. A close relationship of Gelechioidea to Obtectomera than to apoditrysian taxa was noteworthy, although the nodal support is very weak (ML, 23%; BI, BPP = 0.94). Monophyly of Zygaenoidea without Epipyropidae (Epipomponia nawai) was also noteworthy in spite of weak nodal support (ML, 64%; BI, 1.0). Scrutinized analysis is currently underway.

We have determined the mitochondrial genome of Reticulitermes speratus kyushuensis Morimoto, 1968. The total length of the R. speratus kyushuensis is 15,898 bp with 65.3% A + T content. It consists of 13 protein–coding, 22 tRNA, 2 rRNA genes and an A+T–rich control region. All the protein–coding genes used ATN as start codon. But the stop codons were TAA, TAG, and an incomplete termination codon (T) abutting an adjacent tRNA gene. The A+T–rich control region was 1,105 bp in length with 67.8% A + T content

The number of reported mitochondrial genomes (mitogenomes) from the monotypic Lasiocampoidea has been limited until recently. In this study, we sequenced the complete mitogenome of the lappet moth, Kunugia undans (Lepidoptera: Lasiocampidae), and compared it to those of other lasiocampid species and macroheteroceran superfamilies (59 species in six superfamilies). The 15,570-bp long K. undans genome had the typical set of genes found in animal mitogenomes, with the exception of one additional trnR that are located between trnA and trnN loci. Considering that the two trnR copies are located in tandem with proper secondary structures and identical anticodons, a gene duplication event might be responsible for the presence of the two tRNAs. In summary, the general mitogenome characteristics of Lasiocampoidea did not differ greatly from the remaining macroheteroceran superfamilies, but it did exhibit some unique features.

Currently, only limited number of mitochondrial genomes (mitogenome) is available from Odonata. In order to extend current mitogenome data for comparative biology and phylogeny we sequenced complete mitogenomes of two endangered dragonfly species, Libellula angelina and Nannophya pygmaea (Ododana: Libellulidae). The whole genomes were 15,233 bp in L. angelina and 15,112 bp in N. pygmaea and included a typical set of genes (13 protein-coding genes, two rRNA genes, and 22 tRNA genes) and one major non-coding A+T-rich region. The arrangement of the genomes was identical to typical one found in insects. Phylogenetic reconstruction using concatenated sequences of 13 PCGs and two rRNAs of Odonata (17 species in eight families in three suborders) using both Bayesian Inference (BI) and Maximum Likelihood (ML) methods have shown a strong support for monophyletic Zygoptera (BI, BPP = 1 and ML, 100%). Currently, further scrutinized analysis is under progress.

In order to understand evolutionary characteristics of gene rearrangement in Lepidoptera, we collected all available complete mitogenome (mitogenome) sequences registered in GenBank (274 mitogenomes from 44 families in 23 superfamilies as of August 6, 2015). It turned up six rearrangements that differ from the arrangement of ancestral insects, including that of the gelechioid Mesophleps albilinella that we sequenced in this study. The M. albilinella mitogenome has a unique gene arrangement among the Gelechioidea: ARNESF (the underline signifies an inverted gene) at the ND3 and ND5 junction, as opposed to the ARNSEF that is found in ancestral insects. Most of the rearrangements can be explained by the tandem duplication-random loss model, but inversion, which requires recombination, is also found in two cases, including M. albilinella. Excluding the MIQ rearrangement at the A+T-rich region and ND2 junction, which is found in nearly all Ditrysia, most of the remaining rearrangements found in Lepidoptera appear to be independently derived in that they are automorphic at several taxonomic scales. Current mitogenomic data are limited, particularly for congeneric data. Thus, future research focused on congenerics could clarify evolutionary independency at the generic level also.

The spotted-wing drosophila Drosophila suzukii (Diptera: Drosophilidae) is an Asian species introduced into North America and Europe. It damages a wide variety of thin-skinned fruits. We sequenced the complete mitochondrial genome (mitogenome) of D. suzukii to better understand the mitogenomic characteristics of this species and understand phylogentic relationships of Drosophila. The 16,230-bp complete mitogenome of the species consists of a typical set of genes, including 13 protein-coding genes (PCGs), two rRNA genes, and 22 tRNA genes, and one major non-coding A+T-rich region, with an arrangement typical of insects. Twelve PCGs began with the typical ATN codon, whereas the COI began with TCG, which has been designated as the start codon for other Drosophila species. The 1,525-bp A+T-rich region is the second longest in Drosophila species for which the whole mitogenome has been sequenced, after D. melanogaster. Phylogenetic analysis with the 13 PCGs of the Drosophila species using Bayesian Inference and Maximum likelihood methods both placed D. suzukii at the basal lineage of the previously defined Melanogaster group, with a strong support.

The Gelechioidea is the second most species-rich group of Lepidoptera, but only limited number of mitochondrial genome (mitogenome) sequences is available. Thus, we sequenced the complete mitogenome of a gelechioid Hieromantis kurokoi (Lepidoptera: Stathmopodidae) to use the data for future study for the higher phylogeny of Ditrysia in Lepidoptera. The arrangement of the genome was identical to typical one found in Ditrysia (trnM-trnI-trnQ) (underline for inverted gene). The COI began with CGA, which has been designated as the start codon for majority of lepidopteran species, whereas other protein-coding genes (PCGs) began with the typical ATN codon. The 360-bp long A+T-rich region harbored the conserved sequence blocks Phylogenetic analysis using the 13 PCGs both by Bayesian inference (BI) and Maximum-likelihood (ML) methods indicated that H. kurokoi belonging to the family Stathmopodidae grouped together with within-familial species Atrijuglans hetaohei with the highest nodal support (BI, 1.0; ML, 100%).

In this study, we tested the effect of a range of insect orders including Trichoptera as outgroups for lepidopteran phylogeny. Phylogenetic analyses performed with four different partitioning schemes using the maximum-likelihood method provided four different topologies (T1-T4) and topological test most supported T1 topology. When the means of first principle component for nucleotide frequency between A/T and G/C of PCGs was considered Trichoptera, Diptera, Coleoptera, and Orthoptera tended to result in T1 topology more frequently in the given ingroup taxa and outgroups tested. This result contradicts to the general view that the sister taxon might be the best outgroup. The T1 topology was largely consistent with a recent large molecular dataset-based lepidopteran phylogeney, presenting the relationships ((((((((((Noctuoidea + Geometroidea) + (Bombycoidea + Lasiocampoidea)) + Drepanoidea) + Mimallonoidea) + Pyraloidea) + Gelechioidea) + Papilionoidea) + Tortricoidea) + (Gracillaroidea + Yponomeutoidea)) + Hepialoidea).

We sequenced the complete mitochondrial (mt) genome of Camponotus atrox (Hymenoptera: Formicidae) that is distributed only in Korea. This genome is 16,540 bp in size, contains typical sets of genes (13 protein-coding genes, 22 tRNAs, and two rRNAs). The C. atrox A+T-rich region is the longest in the sequenced ants as 1,402 bp and is comprised of an identical tandem repeat consisting of six 100-bp copies and one 96-bp copy. A total of 315 bp of intergenic-spacer sequences were spread over 23 regions. An attempt to align spacer sequences in ants turned out that alignment was mostly feasible among congeneric species, with a substantial sequence divergence, indicating the potential of these sequences as congeneric molecular markers. The A/T content in first and second codon positions of PCGs are similar in ants including C. atrox (73.9 vs. 72.3% on average). Estimation of degree of genetic divergence (e.g. non-synonymous substitution rate) with an increased taxon sampling among hymenopteran superfamilies indicated the presence of different rates of divergence between the suborders Symphyta and Apocrita as has previously been reported. The C. atrox mt genome has a unique gene arrangement, trnI-trnM-trnQ at the A+T-rich region and ND2 junction (underline for inverted gene), possibly originated from tandem duplication of trnM-trnI, resulting in trnM-trnI-trnM-trnI-trnQ and loss of first trnM and second trnI, resulting in trnI-trnM-trnQ.

The lycanid butterfly, Shijimiaeoides divina (Lepidoptera: Lycaenidae) is listed as the second-degree endangered wild animal in Korea from 2012. The 15,259-bp long complete mitochondrial genome of the species consisted of a typical set of genes (13 protein-coding genes, two rRNA genes and 22 tRNA genes) and one major non-coding A+T-rich region, with the typical arrangement found in majority of Lepidoptera. The 15,259-bp long S. divina mitogenome is well within the range found in Lycaenidae and has typical sets of 37 genes and a major non-coding A+T-rich region as 379 bp. As other lycanid butterflies S. divina COI also started with CGA. The gene arrangement of S. divina is identical to that of the Ditrysia in Lepidoptera that has the order trnM-trnI-trnQ (underline for inverted gene) between the A+T-rich region and ND2. Comparison of the skewness between the PCGs encoded in major and minor strand indicates a substantial difference between them in GC skewness (0.261 ~ 0.340 in minor strand vs. -0.081 ~ -0.115 in major strand). The 151-bp intergenic spacer sequence of the S. divina mitogenome is spread over 16 regions ranging in size from 1-53 bp. The longest one (53 bp) located between trnQ and ND2 shows substantially high sequence homology to neighboring ND2 may indicating the origination of the region by a partial duplication of the ND2 gene. One of the unusual features of the S. divina mitogenome is the presence of a trnK-like sequence that is encoded at the major strand of the genome in the A+T-rich region.

노랑부리백로(Egretta eulophotes)는 천연기념물 제 361 호이며, 멸종위기 Ⅰ급으로 등록 되어 있다. 국제적 희귀종 으로 IUCN의 Redlist에 취약종(Vulnerable: VU)이며, 야생 에서 2,500개체 미만만이 남은 것으로 알려져 있다. 국내에 서 노랑부리백로 집단번식지는 1987년 신도에서 최초 발견 되어 천연기념물 360호로 지정받았고, 1991년 약 400여 둥 지가 관찰되었으나, 이후 인위적 방해요인으로 인접 섬으로 이동하여 신도에는 현재 번식하지 않는다. 최근 노랑부리백 로의 번식이 확인된 곳은 천연기념물로 지정되어 보호받는 영광군 칠산도(제 389호), 연평도, 구지도, 서만도, 황서도, 보령의 목도 등이 있다. 노랑부리백로는 서해안 무인도서에 서 집단으로 번식하고 있고, 전 세계 최대 번식집단을 형성 하고 있다. 한편, 노랑부리백로와 같이 무인도서에 서식하는 종은 외 부 위협 요인에 쉽게 취약하거나 교란될 수 있고, 괭이갈매 기와 같이 무인도서내 집단번식의 확산은 서식지 감소로 이어져 멸종위기로 이어지고 있다. 이러한 점 때문에 종의 개체군 구조, 유전적 다양성, 소수개체군의 유전자 관리에 대한 분자생물학적 연구 방법은 노랑부리백로와 같이 무인 도서에서 집단으로 번식하는 멸종위기종의 보전에 중요한 정보와 기초자료를 제공할 수 있다. 본 논문은 노랑부리백로 서식지의 개체군 조사와 더불어 NGS(Next Generation Sequencing)의 빠른 분석기법을 이 용하여 노랑부리백로의 미토콘드리아 유전체 분석과 계통 분류를 수행하고 근연종과의 비교를 통해 종 다양성 및 개 체군 보전을 위한 기초자료로 이용하고자 한다. 노랑부리백로 번식 및 서식현황을 파악하기 위해 문화재 청의 허가를 받고, 번식 및 서식 피해를 최소화하면서 번식 지내 노랑부리백로 43개체의 혈액, 조직(사체), 알껍질 등에 서 유전자원을 확보하였다. 이후 DNA 추출 후 QC테스트 한 총 18개체의 유전자원 샘플로 2개의 primer set을 제작하 고 PCR 증폭을 진행한 후 100bp 이상 조각으로 해독하여, 약 1.2Gb의 미토콘드리아 유전체 데이터를 생산 및 분석하 였다. 노랑부리백로의 번식현황 조사결과 전남 영광군의 칠산 도, 납대기섬, 서만도, 황서도, 보령의 목도(나무섬) 등에서 번식이 확인되었다. 하지만 기존 번식지이었던 신도, 구지 도, 비도, 예도 등에서는 번식을 확인할 수 없었다. 최근 10년간 노랑부리백로의 둥지 수 변화를 보면 2004년 674개 의 둥지에서 2006년 461개, 2008년 524개의 둥지로 감소하 였다가 2010년 696개로 증가한 후, 본 조사인 2013년에는 367개로 감소하였는데, 이는 구지도의 번식 집단이 사라졌 고, 서만도와 황서도의 개체군이 감소한 결과로 판단된다. NGS로 생산된 데이터를 기 보고된 노랑부리백로의 reference sequence에 BWA 프로그램을 이용하여 align을 수행하였으며, samtools를 이용하여 각 샘플별 미토콘드리 아 서열을 생산하였다. 전체 미토콘드리아 서열을 비교해본 결과, 총 87개의 위치에서 다형성(polymorphism)이 발견되 었으며, 대부분이 D-loop(Control region)에 모여 있는 것을 확인할 수 있었다. 또한 기 보고된 중국의 노랑부리백로 데이터와 비교한 결과, 새로운 두 개의 Haplotype이 한국 노랑부리백로에 있음을 확인하였다. 미토콘드리아의 유전 자 다양성을 측정하기 위해 기 보고된 중국의 노랑부리백로 데이터의 미토콘드리아 서열 중 D-loop 영역의 일부를 DnaSP 프로그램을 이용하여 비교하였다. Haplotype/Nucleotide diversity를 계산한 결과 각각 0.956, 0.00908을 얻었고, 중국의 노랑부리백로(Haplotype diversity: 0.920, Nucleotide diversity: 0.00878)에 비해 높게 나타났다. Tajima's D test를 통해 한국에 번식하는 노랑부리백로의 미토콘드리아 control region에 선택압 (selection pressure)이 없다는 결과도 확인할 수 있었다.한국과 중국에 서식하는 노랑부리백로의 지리적 변이를 확인하기 위하여, Arlequin을 통해 AMOVA (Analysis of Molecular Variance)를 진행하였으며, Haplotype frequency를 이용하였을 때 약 2.27 %의 variation 차이가 있음을 확인할 수 있었으며, Genetic distance를 이용하였을 때는 유의한 값 을 얻을 수 없었다. 또한 번식지별로 개체군간 차이가 있는 지 확인해보았으나, 샘플의 수가 적어 통계적으로 유의한 값이 도출되지 않았다. 노랑부리백로의 번식지별 개체군간 의 비교와 새롭게 발견된 Haplotype의 분석을 위하여 RAxML을 이용하여 계통도 분석을 한 결과, 특별하게 번식 지별로 클러스터링(clustering) 되지 않아 번식지별 차이가 없었다. 한편, KHap1, KHap2는 이번 연구에서 새롭게 발견된 Haplotype으로서 기 보고된 중국의 노랑부리백로 Haplotype 과 클러스터링이 되는 것을 확인하였다. 이는 국내 번식하는 노랑부리백로 집단이 중국에서 번식하는 집단과 유전적 교 류가 있다고 판단되며, 번식시기보다는 월동지인 동남아시 아 등지에서 모여 집단 월동할 가능성이 있다. 향후, 노랑부 리백로 mtDNA와 STR genotyping의 분석을 추가적으로 수 행하여 한국에서 번식하는 노랑부리백로의 유전자 다양도 와 계통분석을 심층적으로 수행할 필요가 있다.