The sweetpotato whitefly Bemisia tabaci (Gennadius) is one of the most important sap sucking pests causing economic losses in a variety of vegetables in Bangladesh and as well as around the world. In the present study, the mtCOI sequence of B. tabaci was analysed using samples collected from different host plants (Potato, Brinjal, Tomato, Sweet potato, Bean) from district Gazipur, Patuakhali, Rajshahi and Nilphamari of Bangladesh. Phylogenetic analysis of our samples and relative sequences of B. tabaci in NCBI database was shown three independent clusters. Samples in Bangladesh were most similar with those of Thailand, Vietnam, Pakistan, China and India but did not show any B and Q aggressive biotypes.

NGS data was yielded by using Illumina Hiseq platform. The short reads were filtered by quality score and read length were mapped against the reference genome (KACC42780). Genome-wide reanalyzed data of Flammulina strains were compared against the reference genome to establish a genome-wide single nucleotide polymorphism (SNP). The rate of mapping differences between the strains reflected in the strain variation in its result. The genome-wide SNPs distribution divided into types of homozygous SNP and heterozygous SNP moreover all of the strains demonstrated a wide variation in all of the regions. In the further study of topological relationship between the collected strains, phylogenetic tree was separated into 3 major groups. Group I contained F. velutipes var. related strains of ASI 4062, 4148, 4195. Group Ⅱ contained strains that were different species of ASI 4188 F. elastica, ASI 4190 F. fennae, and ASI 4194 F. rossica. The other 19 strains F. velutipes were classified as a single group. Polymorphic SNPs of F. velutipes strains representing the phylogenetic segregation of whiteand brown-fruiting body forming groups were compared. As previously reported, white gene expression is recessive to brown in fruiting body color gene expression. The white strains produced 131,874 SNPs to be aa type and homozygous from of SNP. 407,947 SNPs were detected as AA, Aa type from the brown-fruiting body of SNP. We constructed a SNP matrix with 8 white strains and 12 brown strains. To develop the molecular marker related in to fruiting body color and geographical origin, we isolated 240 SNPs from the white-and brown-fruiting body forming. To determine the chromosome relationship on polymorphic SNP between Korea and Japan strains producing white-fruiting body, we analyzed that the Korea white strains detected 185,695 SNPs and the Japan white strains produced 263,811 SNPs. Using the constructed SNP matrix with 3 Korea white strains and 3 Japan white strains, the experiment generated 475 SNPs of phylogenetic SNPs fromKorea and Japan white-fruiting body. As a result, we regarded as they are potentially related to the white color. White and brown color and origin specific SNPs could be used as an identification marker for selection of F. veluipes strains in the breeding program.

Field surveys for the Bemisia tabaci complex were conducted from 2009 to 2013 in Korea, and the results were compared with published data of the B. tabaci complex. Three species, MED, MEAM1, and JpL, were collected from several provinces. The MED was mainly collected in greenhouses, displacing the earlier invasive species, MEAM1, and the JpL species was collected in the field. JpL is newly confirmed as a unique species of B. tabaci species complex in Korea and Japan.

The phylogenetic relationships of species and genera in the subfamily Nymphalinae from Myanmar were inferred using mtDNA sequence data from 608 bp of cytochrome oxidase subunit I (COI). A total of 20 species in 10 genera were sequenced and used to construct phylogenetic trees. The base composition of COI sequences was 38.1% T, 15.6% C, 31.6% A, 14.7% G, revealing strong AT bias (69.7%). The sequence distance of 20 species of Nymphalinae ranged from 1.5% to 15.5%. The transition of nucleotide substitution was more common than transversion. The transition between T and C were higher than transition between A and G, and the transversion between A and T was the highest amongst other types of transversion. The phylogenetic trees were constructed using the neighbor-joining (NJ) and maximum likelihood (ML) methods and showed almost identical topologies. The results indicated that the tribes Junoniini and Nymphalini (sensu Wahlberg et al., 2005) formed monophyletic groups but Kallimini was not monophyletic group. Rhinoplapa polynice formed sister group to Junoniini clade with moderate support in both trees. The relationship of species in Junoniini was ((Junonia + Yoma) + Hypolimnas) and the relationship in Nymphalini was (Symbrenthia + (Vanessa + (Kaniska + Polygonia))). The clustering results were almost identical to current morphological classification.

Field surveys for the genus Reticulitermes were conducted from 2013 to 2014 in Korea, and the results were compared with published data of the genus Reticulitermes. Totally, two species, R. speratus kyushuensis and R. kanmonensis were collected from several provinces, and R.kanmonensis is newly confirmed as an unrecorded species in Korea. The R. speratus kyushuensis was mainly collected in various trees, showing 0-2% genetic divergences; whereas, the R. kanmonensis was collected in pine woods of western regions, Gunsan, Wanju, Iksan (Jeollabuk-do), and Seocheon (Chungcheonnam-do). In the morphological comparison, R. kanmonensis is distinguished from R. speratus kyushuensis by more than 15 setae on pronotum and reveals 7.0% genetic difference (in a mitochondrial COII gene) from R. speratus kyushuensis in the molecular comparison.

Oyster mushrooms including of P. ostreatus, P. eryngii, P. pulmonarius and P. cornucopiae are one of the famous mushrooms for foods in Korea. RAPD were carried out using 14 of oligoprimers to analyze the phylogenetic relationship among 57 strains of 32 Pleurotus species. Most of species formed the minimum clade with strains within species and was divided respectively species. Therefore clade was separated well in accordance species. Pleurotus species formed again clade to be added in close related to other species, and were discriminated by sixteen clades with each representative species including high similarity groups. Sixteen clades were composed representative species according to each clade. There were clade I of P. pulmonarius(P. sajor-caju, P. opuntiae, P. sapidus), clade II of P. eryngii(P. fuscus var. ferulae, P. fossulatus), clade III of P. ostreatus(P. ostreatus var. columbinus, P. spodoleucus, P. floridanus), clade IV of P. florida, clade V of P. djamor(P. flabellatus, P. incarnates, P. salmoneo-stramineus), clade VII of P. populinus(P. subareolatus), clade VIII of P. cystidiosus(P. cystidiosus var. formosensis), clade X of P. dryinus(P. dryinus var. pometi), clade XIV of P. cornucopiae(P. citrinopilieatus, P. euosmus), and clade XV of P. australis. These species were representative species each clades. Five species, P. ulmarius(clade VI), P. griseus(clade IX), P. calyptratus(clade XI), P. lampas(clade XII), P. smithii(clade XIII)and P. serotinus (clade XVI) were used each one strain in analysis, so they were clustered other groups.

Pleurotus has increased rapidly production and consumption because of highly nutritional value, natural healthy food and so on. The basic studies for Pleurotus need for development of mushroom industry. This study was to investigate the cultural characteristics among 15 strains of 6 species and to analyze their phylogenetic relationships. The cultural characteristics were investigated by mycelial growth activity at different media, temperature and pH. The optimum media for mycelial growth were YM and MCM in most species. The optimum temperature for mycelial growth were 25˚C and 30˚C. The optimum pH for mycelial growth were widly range from pH 5.1 to 7.4. Through the RFLP (restriction fragment length polymorphism) of IGS (intergenic spacer) I region in ribosomal DNA, it was analyzed phylogeny of interspecies and intraspecies. Each species was discriminated well as isolates within each species formed clade to be distinguished other species. P. florida was highly similar to P. floridanus, and P. flabellatus was P. cornucopiae. P. fuscus var. ferulae was highly similar to P. eryngii but discriminated different species in analysis of RFLP of IGS I region and showed different characteristics in mycelial culture. RFLP of IGS I region was useful of studying phylogenetic relationships of species and population.

A rust on Asiatic dayflower plant (Commelina communis L.) was found in Geoje, Korea, in August 2010. Uredia are mostly produced on abaxial leaf surface or elongated on stem, early naked, surrounded by the ruptured epidermis, cinnamon-brown. Uredospores are globose, ellipsoid or ovate, echinulate, yellowish brown or brown, 20-30 × 20-25 ㎛. On the basis of mycological characteristics and molecular data, the fungus was identified as Uromyces commelinae Cooke. The phylogenetic position of U. commelinae is separate from the other rusts where the economically important rusts of the Poaceae are situated. Although host ranges of the rust caused by U. commelinae were previously recorded, full descriptions and illustrations, including symptoms and signs have not been described. This is the first description of rust disease on C. communis plant with molecular identification, symptoms, and signs.

The genetic relationships among five genera, seven species of Theaceae were examined through a random amplified polymorphic DNA (RAPD) analysis and internal transcribed spacer (ITS). In RAPD analysis, five of 15 arbitrary primers showed polymorphic bands, which were able to classify different genera and species of Theaceae. The genetic variations of Theaceae were from 0.031 to 0.484. In ITS analysis, the ITS sequences were analyzed using BLAST and showed high identities with sequences of Theaceae, seven species published in NCBI GenBank database, which ranged from 98 to 100%. Sequence alignment of seven species showed 34.9% identities for ITS 1 region and 43.7% for ITS 2 region. Pairwise sequence divergences among seven species ranged from 0 to 0.330%. In phylogenetic tree, they were divided into three groups. In conclusion, the molecular data generated in the present investigation will help to understand the genetic relationships of Theaceae and also might be useful for further studies in intra-species, inter-species, and molecular evolution researches.

Field surveys for the B. tabaci species complex were conducted from 2009 to 2013 in Korea, and the results were compared with published data of the B. tabaci complex. Three species, Mediterranean, Middle East-Asia Minor 1, and JpL, were determined from several regions based on mitochondrial COI sequences. The Mediterranean was mainly collected in greenhouses, displacing the earlier invasive species, Middle East-Asia Minor, and the JpL was collected in the field. The JpL is newly confirmed as a unique species of B. tabaci complex in Korea and Japan.

Part of a field survey to assess the extent of damage by termite (Reticulitermes speratus kyushuensis Morimoto), termite samples were collected from various locations of Korea from 2009 to 2011. During the study period, we firstly discovered Reticulitermes species on Jeju island, and partial sequences of mitochondrial COII gene, which is proved useful to study phylogenetic relationships of termites were investigated. Also, morphological characteristics of the collected samples were compared with previous reports. According to the result of taxonomic study and comparative sequence analysis, it was identified as Reticulitermes speratus kyushuensis Morimoto, and this is first report of R. s. kyushuensis from Jeju island in Korea which can be used as estimation of population structure and tool of control strategy.

한국 내 전국적으로 5곳의 담수에서 2010년 4월부터 2011년 6월에 걸쳐 담수시료를 채취한 후, 각 담수시료에서 Botryococcus braunii의 출현을 확인하고 microcapillary-pipetting method를 이용하여 5주를 분리하였다. 분리된 B. braunii의 18S rRNA sequencing 후, BLAST search를 통한 분석 결과는 B. braunii와 가장 높은 상동성을 나타낸 것으로 확인되어 형태적 동정결과와 일치하였다. 진화, 계통분류학적 분석결과, 분리된 5주의 B. braunii는 단일 계통인 Trebouxiophyceae으로 분류되었다. 18S rRNA sequence의 계통 분류학적 분석에서 B. braunii KNU6는 다른 분리된 B. braunii와 같은 분류군 내에서 거리의 차이가 있었고, 또한 세포크기, 점액질의 양 등의 형태적 특징도 차이를 보였다. 한국에서 분리된 5주의 B. braunii는 모두 race A type의 분류군에 포함되며, 탄소수 C23-C33의 지방산에서 유래된 alkadienes과 alkatrienes를 생산하는 것으로 사료되며, B. braunii를 이용한 바이오디젤을 생산하기 위한 최적의 배양조건을 찾는 연구가 필요하다.

본 시험은 국내외에서 수집한 꽃송이버섯균 22균주에 대하여 분자생물학적 유연관계를 분석하고자 하였다. 수집균주의 ribosomal DNA의 ITS 영역에 대한 cleaved amplified polymorphic sequence (CAPS) 분석 결과, KACC50866은 다른 균주들과 20%이하의 유연관계를 나타내었으며 나머지 균주들은 90% 이상의 유연관계를 보이면서 4그룹으로 구분되었다. 따라서 이들의 세분화된 분자생물학적 구분을 위하여 rDNA ITS 영역의 염기서열분석을 하여 구분하여 본 결과 KACC50866 균주는 다른 꽃송이버섯균과 유연관계가 매우 낮은 것으로 나타났다. 그리고 나머지 21개 균주는 같은 그룹으로 구분되어 있어 같은 종으로 생각할 수 있으나, 이들을 좀더 세분하기 위해서는 미토콘드리아의 유전자 서열 분석 등이 병행되어야 할 것으로 판단된다.

Up to now only 17 coleopteran mitogenomes have been fully or near fully sequenced, but this chart does not include any complete mitogenome sequence of the coleopteran infraorder Scarabaeiformia, in which the monotypic Scarabaeoidea is included. In this study, we firstly present three scarabaeoid mitogenomes, two of which are completed and one is near completed. The complete nucleotide sequences of the white-spotted flower chafer, Protaetia brevitarsis (Scarabaeiformia: Scarabaeidae), was determined to be 20,319 bp, the two-spotted stag beetle, Metopodontus blanchardi (Scarabaeiformia: Lucanidae), was 21,678 bp, and the incomplete garden chafer, Polyphylla laticollis (Scarabaeiformia: Scarabaeidae), which lacks sequence information from whole A+T-rich region and a partial srRNA gene was 14,473 bp. Thus, the two complete mitogenome sequences are longest among the completely sequenced insect mitogenomes, followed by the 19,517 bp-long dipteran Drosophila melanogaster. Phylogenetic analysis using the concatenated 13 amino acid sequences of PCGs of the 13 coleopteran insects including three newly sequenced scarabaeoid successfully recovered a monophyly of suborder Polyphaga, a monophyly of infraorder Cucujiformia, a monophyly of infraorder Elateriformia, and also a monophyly of infraorder Scarabaeiformia within Polyphga. However, the Scarabaeiformia represented by three newly sequenced species was unexpectedly placed as a sister group to the Cucujiformia, instead of the placement to the sister to the Cucujiformia and Elateriformia group.

These experiments were carried out to determine the optimal culture conditions for nine collected strains of Lyophyllum decastes(Fr.:Fr.)Sing. Bottle cultivation was used as a technique to characterize and develop new substrates as possible substitutes of sawdust. Nine kinds of Lyophyllum decastes were selected SPA 202 and SPA 205 fungus that mycelium grow fast and have high density. All cultures were incubated on Burke compost for 20 days. Growth of hyphae was optimal at temperatures between 20-25℃ and pH of 5.0-6.0. The best nutrient additives tested were lactose, yeast extract and MgSO4- 7H2O as carbon source, nitrogen source and mineral salt respectively. Collected strains of Lyophyllum decastes were incubated for 7 to 20 days after inoculation on agar medium followed by 45 to 75 days on sawdust substrate. The ideal optimal moisture percentage in sawdust media was 65%. The optimal species used was Quercus aliena , followed by Populus deltoides. Addition of 10% of Rice bran to Quercus aliena sawdust makes mycelial growth fast. Spawn running on the fermented sawdust media required 50 days at 20 to 25℃ and another 7 more days after covering the media with soil for promoting formation of fruitbodies. This was followed by cultivation for 10 to 11 days at 17 to 18℃ for initiation and 7 to 8 days for development of fruitbodies. The length of pilei and stipes which were harvested in optimal stage showed 60㎜ and 67㎜, respectively. Yields of strain SPA 202 grown on fermented sawdust medium could be up to 130g. The yields of strain Lyophyllum decastes used for fermented composition sawdust was the most appropriate recording 86% among them especially in SPA 202 artificial cultivation. Among 14 pleurotus eryngii , to identify the similarity ITS sequences and RAPD polymorphism are analyzed. ITS region sequences of 14 strains ,which have similar reqion sequences SPA 100, 101, 102 belong to Hypsizygus . 11strains belong to Lyophyllum decastes group. After classify 3 subgroup genus relationships among is analyzed. To analyzing diversity of artificial cultivated mushroom, after RAPD analysis excellent strains SPA 202 and SPA 203 show 75% similarity and pleurotus eryngii of Lyophyllum decastes SPA 103 and SPA 104 show 65%. As a result , SPA 202 which is cultivated in nature is estimated to Lyophyllum decastes by molecular biology method.

We have determined the complete mitochondrial genome of the yellow-spotted long horned beetle, Psacothea hilaris (Coleoptera: Cerambycidae), an endangered insect species in Korea. The 15,856-bp long P. hilaris mitogenome harbors gene content typical of the animal mitogenome and a gene arrangement identical to the most common type found in insect mitogenomes. As with all other sequenced coleopteran species, the 5-bp long TAGTA motif was also detected in the intergenic space sequence located between tRNASer (UCN) and ND1 of P. hilaris. The 1,190-bp long non-coding A+T-rich region harbors an unusual series of seven identical repeat sequences of 57-bp in length and several stretches of sequences with the potential to form stem-and-loop structures. Furthermore, it contains one tRNAArg-like sequence and one tRNALys-likes equence. Phylogenetic analysis among available coleopteran mitogenomes using the concatenated amino acid sequences of PCGs appear to support the sister group relationship of the suborder Polyphaga to all remaining suborders, including Adephaga, Myxophaga, and Archostemata. Among the two available infraorders in Polyphaga, a monophyletic Cucujiformia was confirmed, with the placement of Cleroidea as the basal lineage for Cucujiformia. On the other hand, the infraorder Elateriformia was not identified as monophyletic, thereby indicating that Scirtoidea and Buprestoidea are the basal lineages for Cucujiformia and the remaining Elateriformia.

Eumenis autonoe belonging to a lepidopteran family Nymphalidae (superfamily Papilionoidea) is an endangered species in Korea. Historically, the species was distributed in Europe and Asian region including a wide region in Korean peninsula. However, in Korean peninsula, the species is found only in two isolate dregions: South in a remote island Jeju, where altitude is higher than1, 400 meter on Halla Mt. and North in far northern Korean peninsula around Mt. Bekdu. In this study, we report the complete mitochondrial genome of the endangered E. autonoe collected from Mt. Halla. The 15,489-bp long E. autonoe genome has a typical gene content found in animal mitochondrial genomes and contains the gene arrangement identical to all other sequenced lepidopteran insects, which differs from the most common type found in insects, as the result of the movement of tRNAMet to a position 5’-upstream of tRNAIle. As seen in many other lepidopteran insects, no typical ATN codon for COI gene is available. Thus, we tentatively designated the CGA (arginine) found at the beginning of the COI gene, as has been suggested for lepidopteran COI starter. The intergenic spacer sequence located between tRNASer (UCN) and ND1 of E. autonoe mitogenome also contains the ATACTAA motif which is conserved in all sequenced lepidopteran species. The 678-bp long A+T-rich region, which is longest in sequenced lepidopteran insects contains ten identical tandem repeats composed of 27 bp plus one 13-bp long identical incomplete final repeat. Such repeat sequence is rare in the lepidopteran mitogenomes known so far. The E. autonoe A+T-rich region also contains a poly-T stretch located at the end of the region as 19 bp and also contains the downstream conserved motif ATAGA that were previously suggested to serve as a structural signal for minor-strand mtDNA replication. Phylogenetic analysis using the concatenated 13 amino acid sequences of PCGs among available six lepidopteran superfamilies (Tortricoidea, Pyraloidea, Papilionoidea, Bombycoidea, Geometroidea, and Noctuoidea) rooted with three dipteran species with BI and ML analyses supported the following topology: ((((Bombycoidea + Geometroidea +Noctuoidea) + Papilionoidea) + Pyraloidea) + Tortricoidea). Within Papilionoidea, a closer relationship between Lycaenidae and Pieridae, excluding Nymphalidae was observed. Further fruitful information will be available after more analysis is done.

Halacarid mites inhabit mainly in the bottom of the sea and their body lengths are approximately 0.5 mm. Few biological information about halacarids have been obtained. The phylogenetic study of halacarid mites has not been progressed and the phylogenetic relationship within the family Halacaridae is still unknown. Particularly there is no molecular study to infer phylogenetic relationship within the family. In the annual meeting of Acarological Society of Japan 2007, we developed the method for molecular analysis of 28S nuclear ribosomal gene in haracarid mites. Subsequently, we have further progressed the technique of the molecular analysis to infer the phylogenetic relationship of haracarids. The aim of the present study is to elucidate the phylogenetic position of the subfamily Rhombognathinae on the bases of molecular data. Halacarid specimens which were obtained from seaweeds and sandy segment at Japanese seacoasts, consist of eight genera in five subfamilies: Halacarinae (Halacarus sp., Agauopsis spp., Thalassarachna spp.), Halixodinae (Badyagaue sp., Agaue sp.), Copidgnathinae (Copidognathus spp.), Actacarinae (Actacarus spp.) and Rhombognathinae (Rhombognathus spp.). The template DNA was extracted and 28S nuclear ribosomal gene of the DNA was amplified by PCR reactions. And we determined approximately 400 base pair length of the gene. To construct phylogenetic trees by neighbor-joining method we calculated genetic distances of each operational taxonomic unit considering 1) transversions and 2) transitions and transversions based on Tamura - Nei model by using MEGA ver. 4 (Tamura et al. 2007). As a result, we got two trees both showing two main clusters. One of them includes only one subfamily Rhombognathinae and another consist of Halacarinae, Copidgnathnae, Actacarinae and Halixodinae. Rhombognathinae was greatly differentiated from the other subfamilies. Thus, Rhombognathinae possibly is a distinct taxon phylogenetically and this result supports the monophyletic nature obtained previously by morphological analysis.

In the preceding symposium, we presented the result of the phylogenetic analysis of Stigmaeopsis spider mites (Acari: Tetranychidae) in Japan using sequence data of the Cytochrome Oxidase subunit I (COI) region of mitochondria DNA. We analyzed haplotypes representing S. longus, S. celarius, S. takahashii and S. saharai, which inhabit bamboo species of the genera Sasa and Phyllostachys (Poaceae; Bambusoideae), and two forms of S. miscanthi (HG and LW) living on Miscanthus sinensis (Poaceae; Panicoideae). Both neighborjoining and maximum-likelihood methods revealed that Stigmaeopsis consists of seven monophyletic groups, in which two forms of S. miscanthi form two distinct monophyletic groups, whereas four species of bamboo-inhabiting species comprise a polyphyletic assembly. These results imply a mismatch between diagnostic morphological characters (i.e. the length of dorsal setae) and their phylogenetic lineages for bamboo-inhabiting species, requiring re-evaluation of the use of these characters. In addition, our results indicate that plural lineages coexist in a single habitat at various regions, suggesting that differentiated lineages secondarily encounter around southwestern Japan. Interestingly, the result of phylogenetic analysis on mitochondrial COI and nuclear ITS1 region was, in part, inconsistent in internal branches. In this symposium, I re-analyzed more these data by phylogenetic network analysis. Based on the results, I make some inference about the process of habitat expansion of these mites.

The 15,338-bp long complete mitochondrial genome (mitogenome) of the Japanese oak silkmoth, Antheraeayamamai (Lepidoptera: Saturniidae) 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 A. yamamai 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 A. yamamai COI gene. Three of the 13 protein-coding genes (PCGs) harbor the incomplete termination codon, T or TA. All tRNAs formed stable stem-and-loop structures, with the exception of tRNASer(AGN), the DHU arm of which formed a simple loop as has been observed in many other metazoan mt tRNASer(AGN). The 334-bp long A+T-rich region is noteworthy in that it harbors tRNA-likestructures, as has also been seen in the A+T-rich regions of other insect mitogenomes. Phylogenetic analyses of the available species of Bombycoidea, Pyraloidea, and Tortricidea bolstered the current morphology-based hypothesis that Bombycoidea and Pyraloidea are monophyletic (Obtectomera). As has been previously suggested, Bombycidae (Bombyxmori and B.mandarina) and Saturniidae (A.yamamai and Caligula boisduvalii) formed a reciprocal monophyletic group.