The striped fleas beetle (SFB), Phyllotreta striolata is a notorious and widely distributed pest in the world including Korea. We surveyed SFB insecticide resistance in Pyeongchang and Hoengseng population. Leaf dipping bioassay with recommended concentration of insecticides was performed using SFB adults. Two populations showed high level of resistance in various insecticides particularly neonicotinoids. Nicotinic acetylcholinesterase receptor (nAChR) is well known target site in neonicotinoid. Therefore, we tried to identify the nAChR from the SFB. Primary, partial contig sequence obtained by RNAseq from Hoengseng population and additional gap fillings were conducted. Total 12 nAChR subunits (10 alpha subunits and 2 beta subunits) were identified and phylogenetic relationships were predicted. Earlier studies reported that point mutations contribute to develop the imidacloprid resistance in aphids and brown planthopper, however we did not find it. Further studies relative to neonicotinoid insecticide resistance mechanism are demanded.

Liriomyza leafminer flies comprise a pest group that causes both considerable economic losses and serious quarantine problems. To investigate Liriomyza spp. abundance and distribution along with natural enemies, we surveyed of leafminer (LM) species and its natural enemies (NE) on potato crop from 146 farmers’ field in the potato growing provinces during 2015. Sweeping (10 sweeps/ field) and visual inspection were employed within 5 minute-walking distance, and damaged leaflets (10 leaflets/ field) were collected in each field. Leafminer species were identified by morphological, and its NE was identified by morphological and molecular techniques. L. huidobrensis was the most dominant Liriomyza species. Distribution of L. huidobrensis on potato fields was expanding. Highest potato fields were infested with leafminer in southern parts of country. Highest rate (100%) field damage was recorded in Miryang, Namwon, Henam, and Boseong. Overall, NE species collected constituted of 4 families and 18 and 20 species in sweeping, and damaged samples respectively. There was a rich complex of parasitoid species including 20 species (13 species of Eulophidae family). The endoparasitoid, Braconid sp., Ganaspidium sp., and ectoparasitoid, Diglyphus isaea were dominant species in sweeping. While Diglyphus isaea, Hemiptarsenus varicornis, and the Braconid sp. were most dominant species collected from damaged samples. The number and diversity of parasitoid species in potato crop in Korea provides insight into the development of integrated control strategies of invasive LM pests in the field, and glasshouse crops.

In the present study, Cydia kamijoi Oku is newly recognized in korean insect fauna. This tortricid moth was first found damaging the cones of Abies koreana in Jeju Island 2014. The moth can be a serious insect pest on A. koreana because of high damage rate on the cones, up to 71% average. The genus Cydia now was 11 korean species including C. kamijoi. Regarding this species, some basic information such as collection records, morphological characters, and ecology were provided

The habitat of the cockroach varies relative to species. The German cockroach (Blattella germanica) lives in human dwelling while B. nipponica lives in mountainous region. Even though, these two species have similar morphology, their habitats and walking speed were different. Underlying this fact, we hypothesized that habitats might influence on the walking speed via altering particularly nervous system including acetylcholinesterase (EC3.1.1.7, AChE). Full length ORFs were cloned from each species and sequenced. Sequence analyses showed that both genes in each species possess typical features of ace, and that Bgace1 and Bnace1. Those features are orthologous to the insect ace1 whereas Bgace2 and Bnace2 are to the insect ace2. Some SNPs were observed but Bgace1 and 2 showed high similarity (99%) with Bgace1 and 2, respectively. Multiple AChE bands were identified by native PAGE and IEF from three tissues (head, leg and other body) and their expression pattern was almost identical between two species. However, Esterase expression patterns were significantly different. Furthermore, length as well as detailed structure of antenna, leg and tail cerci was also significantly different. Taken together, various factors such as sensory organ detailed morphology and esterases are responsible for habitat and walking speed difference.

A new gregarine species (Eugregarinida: Gregarinidae) was previously reported from daikon leaf beetle, Phaedon brassicae at Daegwallyeong in Korea. Here, we found another novel gregarine species from the striped flea beetle, Phyllotreta striolata (Coleoptera: Chrysomelidae). This paper describes the morphological and molecular-biological characteristics of Gregarine species associated to P. brassicae and P. striolata. The size and body shape at mature trophozoite stage were the main diagnostic characteristics of those species. Gregarines from P. brassicae were bigger size (100um) with square shaped body while Gregarines from P. striolata were slightly smaller size (80um) with egg shaped body. Based on the partial 18S rDNA sequence, gregarine species from P. striolata was grouped in eugregarine and also this new gregarine showed 85% homology that of the reported Gregarina sp. from P. brassicae. However, gut dissection result showed that each species had own gregarine parasite species association. These results were identical in local populations from three surveyed locations. There was positive relationship between infection rates and elevation for gregarine related to P. striolata. Therefore, we suggested that the gregarines and their coleopteran hosts have species-specific interaction via co-evolution.

The pea leafminer fly, Liriomyza huidobrensis (Blanchard) (Diptera: Agromyzidae), is a polyphagous pest attacking wide range of field and greenhouse crops and ornamental plants. Surprisingly, localized outbreaks with high population and heavy unique damage of leafminers were observed from the field survey of potato fields in the southern parts (Miryand and Goryeong) of Korea in 2012. L. huidobrensis was the responsible for those potato damages based on the morphological and genetic characteristics. Then after, it added a one more emerging species in Korean Liriomyza account. Infestation of L. huidobrensis was expanding and reported in major potato growing areas of the country. Genetic evidence showed the close relationship with Chinese L. huidobrensis population, so it might be invaded from China. As a new emerging pest, very few informations relative to this pest are available in Korean prospective. There is also significant potential for it to widen its distribution, as the Korean Peninsula is a hot spot of climate change, particularly global warming. There are a lot of challenges uprooting infront of researchers and growers. It is demanded to explore new secrets and to be ready to tackle of this invasive pest, L. huidobrensis.

Pogonini Laporte, 1834 is one of the tribes in Trechinae Bonelli, 1810 (Coleoptera: Carabidae). This tribe is morphologically similar to Trechini, Benbidiini, Zolini and Patrobini of the subfamily. However, Pogonini can be distinguished from the other tribes by having a combination of the following characters: apical maxillary palpomere fusiform; umbilical series of setae on elytron more or less contiguous (though sparse in middle); each tarsomeres with a median sulcus on dorsal side. Pogonini includes about 85 species of 12 genera worldwide and is found in all zoogeographical regions of the world. Many species inhabit along the sea coasts or near salt lakes, and are recognized as halobiont. In Korea, Paik (1998) reported Pogonus (Pogonus) itoshimaensis Habu, 1954. However, he added only the species name in the checklist without description or illustration although few specimens were examined. After Paik’s study, any studies on taxonomic status and distribution of Korean Pogonini have not been carried out. The genus Pogonus Dejean is the largest group in Pogonini with two subgenera: P. (Pogonoidius Carret, 1903) and P. (Pogonus Dejean, 1821). These two subgenera share very similar external morphological characters; Pogonoidius has setae on anterior angles of pronotum and setae at 7th interval of elytra while Pogonus does not have them. In the present study, two species, Pogonus (Pogonus) japonicus Putzeys, 1875 and P. (Pogonoidius) punctulatus Dejean, 1828, are newly recognized from Korea. A key to the Korean Pogonini species, redescription, distribution, photos of diagnostic characters, and illustrations of male and female genitalia are provided.

Multiple starters consisting of two Bacillus amyloliquefaciens strains (MJ1-4 and EMD17), Pichiafarinosa SY80, and Rhizopus oryzae were used for Doenjang making. Bacillus strains were selected based on their abilities to inhibit toxinogenic fungi and Bacillus cereus, fibrinolytic activity, and their ability to confer good flavor to Cheonggukjang. P. farinosa SY80 and R. oryzae, previously isolated from soy sauce, were selected because they were not inhibited by two bacilli. Doenjang was prepared by inoculation of multiple starters (A1 Doenjang). Control Doenjang was prepared by inoculation of B. subtilis KACC 16750 (Natto strain) and Aspergillus oryzae KCCM 60166 (A2 Doenjang). Another control (A3 Doenjang) was prepared by inoculation of microorganisms present in rice straw. Doenjang samples were fermented for 70 days at 20℃. pH of 3 samples decreased from the initial value of 6.4 to 5.8~6.0 and titratable acidity (TA) increased from 0.6 to 1.1~1.3. The amount of amino-type nitrogen increased during fermentation. There were slight differences in moisture, crude-protein, and crude-fat contents after 70 days. Contamination of fungi was observed only in A3 Doenjang and B. cereus was not detected from all 3 samples. A1 Doenjang showed the highest fibrinolytic activity and A2 Doenjang the second. These results indicate that Doenjang made with carefully selected starters was functionally improved and microbially more safe.

A new gregarine parasite (Apicomplexa: Protozoa) was observed in the population of the striped cabbage flea-beetle, Phyllotreta striolata (Coleoptera: Chrysomelidae) in Hoengseong-gun. Gregarines are well known species-specific parasites of various Arthropoda. Based on 954bp of 18S rDNA sequence, this gregarine species was grouped in eugregarine and also this new gregarine showed 85% homology that of the reported Gregarina sp. from daikon leaf beetle, Phaedon brassicae (FJ481523). These two coleopteran hosts were generally found coexist in highland cabbage fields. However, after gut dissection (n>50), each species confirmed has their own gregarine parasite species. These results were identical in surveyed three local populations (Pyeongchang-gun, Hoengseong-gun and Inje-gun). Therefore, we suggested that the gregarines and their coleopteran hosts has species specific interaction via co-evolution.

PVY (Potyviridae: potyvirus) is one of the most important potato virus affecting seed potato production and also it is transmitted non-persistently via aphids. For healthy seed potato production, a virus detection system is highly important in addition to aphid monitoring and control. To achieve this detection method, it need to fast and easy to use. About two decades ago RT-PCR based PVY detection method was developed. However that was very time consuming and has low sensitivity. Here, we developed an advanced PVY detection method which a uses the boiling extraction of the viral RNA from aphid stylet and amplification by specific primers located in the viral capsid protein gene. Therefore, it could directly synthesize cDNA of PVY viral capsid gene from extracted RNA of PVY using one-step RT-PCR method in very short time compared to previous methods due to the omission of RNA extraction step. We confirmed this PVY detection method using the two aphid species (Macrosiphum euphorbiae and Aphis gossypii) that known as PVY vectors. The efficiency of this PVY detection method was 60% to 80% from two the aphid species. Hence, this method could be potentially applied to virus free seed potato production programs.

The genus Diadegma is a well known parasitoid group and some are known to have symbiotic virus, PDV. A novel IV was discovered from the calyx of D. fenestrale female. This virus was named as D. fenestrale Ichnovirus (DfIV). The encapsidated DfIV genome contains 65 circular DNA segments with an aggregate size of 247,191 bp. Based on BLAST analysis, a total of 120 ORFs were predicted as follows: rep; 48, cys-motif; 11, vinnexin; 10, vankyrin; 9, PRRP; 3 and other unassigned genes (39). These viral genes were expressed in lepidopteran hosts (Phthorimaea operculella and Plutella xylostella) after parasitization which means DfIV genome segments were integrated into lepidopteran hosts. This study was focused on this result. Based on gene expression profile, candidate promoter and integration motifs were selected and then, fused with eGFP as a reporter gene. Modified DfIV genome segment was ligated to a commercial containing f1 ori and Ampr gene to propagate in E. coli. We have named this fusion vector as pIN. The construction methodology of pIN and its application would be further discussed in detail.

Despite that porcine spermatogonial stem cells (pSSCs) have been regarded as a practical tool for preserving eternally genetic backgrounds derived from pigs with high performance in the economic traits or phenotypes of specific human diseases, there were no reports about precise definition of niche conditions promoting proliferation and maintenance of pSSCs. Accordingly, we tried to determine niche conditions supporting proliferation and maintenance of undifferentiated pSSCs for short-term. For these, undifferentiated pSSCs were progressively cultured in different composition of culture medium, seeding density of pSSCs, type of feeder cells and concentration of growth factors, and then total number of and alkaline phosphatase (AP) activity of pSSCs were investigated at post-6 day culture. As the results, the culture of 4x105 pSSCs on mitotically in activated 2x105 STO cells in the mouse embryonic stem cell culture medium (mESCCM) supplemented with 30 ng/ml glial cell line-derived neurotrophic factor (GDNF) was identified as the best niche condition supporting effectively the short-term maintenance of undifferentiated pSSCs. Moreover, the optimized short-term culture system will be a basis for developing long-term culture system of pSSCs in the following researches.

Conservation biological control (CBC) is one of the pest management tactics based on protecting and maintaining natural enemies that already exist in the crop environment. Among many appropriate practices to control pests, CBC has been considered as a sustainable means and a less expensive alternatives to chemicals. It contains an introduction of non-crop plants as shelter habitat for providing natural enemies with food sources, overwintering or refuges shelters. As a small scale experiment of CBC, we tried to intercrop six kinds of floral plants (buckwheat, red clover, Agastache rugosa, Chrysanthemum indicum, Allium tuberosum, Lythrum anceps) with napa cabbage in order to attract some natural enemies, especially parasitic wasps that could be fed on them in the cabbage fields, which is a novel way for conservation of natural enemies, but none showed significant result. For the development of integrated pest management system by harmonizing biological and chemical control, we carried out evaluating toxicity of 30 pesticides to parasitoids and investigating effectiveness of traps. With leaf dipping method for adult and body dipping method for cocoons, 12 pesticides showed under 30% of toxicity being selected as safer by IOBC. Using sex-pheromone traps for attracting lepidopteran pests showed that DBM, CAW and CL occurred less than untreated fields; 67.5%, 70.6% and 44.0%, respectively. Also, yellow sticky traps could reduce some pests; 52% for flea beetle, 62% for cabbage sawfly and 41% for Phaedon brassicae. These results are expected to give basic information to develop conservation biological control of DBM with indigenous parasitoids in the cabbage fields on a large scale in the future. Before developing and distributing a promising pest control method, we should consider whether it can be compatible with other agricultural practices or various situations around fields.

The cotton aphid, Aphis gossypii (Glover), is one of the main pests in various vegetable crops due to insecticide resistance in Korea. Some insect pests noticed neonicotinoid insecticide resistance such as Nilaparvata lugens, Bemisia tabaci and Myzus persicae etc. and the major player which contributed for developing the resistance was over-expression of P450, particularly CPY6 family. However, A. gossypii was a unique case that they developed non-P450 dependent resistant mechanism. Previously we reported that two point mutations (RtoT in nicotinic acetylcholine receptor, nAChR, beta 1 subunit and RtoT with LtoS in a transcript variant) contribute to develop the imidacolprid resistance in A. gossypii. Moreover we surveyed the mutation(s) in various local field populations. Based on the 3D modeling, we hypothesize that RtoT mutation can reduce the imidacloprid sensitivity. A stretch of 33 amino acid was deleted in the N-terminal region of original transcript of nAChR beta 1 subunit that contained RtoT with LtoS mutations in resistant strain. Among the two transcripts, only original transcript differently expressed between imidacloprid susceptible and resistant strain (resistant ratio = 3,800). Six alpha subunit (1∼5, 7) transcript levels were not significantly different between two strains. Therefore mutation and down-regulation of nAChR beta 1 subunit is also associated with imidacloprid resistance in the A. gossypii.

Highland agriculture is a kind of specified agricultural term based on altitude and meteorological characteristics and their main crops are seed potato and vegetables. These crops are only cultivated in summer season when insect pests are most dynamically attacking such as diamondback moth (DBM, Plutella xylostella) and aphids son on. Aphids, insect virus vector, are really annoying factor, particularly in seed potato. Furthermore, these insect pests formed small sub-population or colony based on small field area. For instance, green peach aphid (GPA, Myzus persicae) populations’ genetic backgrounds and insecticide susceptibilities were different even in the same Pyeongchang area. Therefore, we suggested that highland agriculture should adopted IRM strategy for the control of insecticide-resistant aphids. First. Monitoring of annual fluctuation of insect population using traps for more effective monitoring. Second. Performing the insecticide resistant monitoring patch by patch or field using high throughput molecular diagnosis for more accurate monitoring. Third. Establishing the insect pest management program based on these results. We will deeply discussed practical monitoring results and IRM strategy in highland agriculture.

The genus Diadegma is a well known parasitoid group and some are known to have symbiotic virus, PDV. A novel IV was discovered from the calyx of D. fenestrale female. D. fenestrale has more than two hosts, including PTM and DBM. The oviposition and survival rate results showed that D. fenestrale preferred PTM to DBM as hosts. Nevertheless, the developmental period and morphology of D. fenestrale were not significantly different between PTM and DBM. To identify these phenomena, DfIV genome expression patterens were compared between PTM and DBM under various conditions. DfIV genes were more widely expressed in PTM than in DBM after parasitized by D. fenestrale, particularly at the initial point. In addition, large numbers of DfIV genes were expressed only in PTM and they showed differential expression patterns between two lepidopteran hosts. This DfIV genome expression plasticity showed a dependency on the lepidopteran host species and parasitization time, suggesting that it may contribute to the parasitoid survival rate increase. This may be one of the key elements that determine the symbiotic relationship between PDV and parasitoid.