본 연구에서는 신선편이 양배추 내 Bacillus cereus의 최 적 증균 온도를 선정하고 증균배양액에 소수성필터를 적용하여 multiplex PCR의 검출률을 확인하였다. B. cereus 증균온도는 B. cereus 균주 5 개를 혼합하여 1 Log CFU/ mL이 되도록 증균배지에 접종하고 30oC, 37oC, 42oC에서 증균한 뒤 3시간 간격으로 MYP agar에 도말한 후 계수하여 선정하였다. 소수성필터 미적용 그룹은 B. cereus 균 주 5 개를 혼합하여 신선편이 양배추에 접종한 뒤 최적 증균온도에서 증균하였으며, 증균배양액을 가열하여 DNA 를 추출한 뒤 multiplex PCR을 진행하였다. 소수성필터 적용 그룹은 증균배양액을 소수성 필터에 적용하고 필터에 있는 균을 멸균증류수로 현탁한 뒤 가열하여 추출된 DNA 로 multiplex PCR을 진행하였다. 증균온도 확인 결과, 6시간 증균 시 42oC에서 증균된 샘플(5.4 ± 0.3 Log CFU/mL) 과 30oC에서 증균된 샘플(4.6 ± 0.6 Log CFU/mL) 간 유의 차가 확인되었다(p < 0.05). 소수성필터 적용 유무에 따른 multiplex PCR 결과, 1 Log CFU/g 접종된 샘플의 검출률이 소수성 필터 적용 전 60%(3/5)에서 100%(5/5)로 향상 되었다. 2 Log CFU/g 접종 샘플은 소수성필터 적용 전 80%(4/5)에서 소수성 필터 적용 후 100%(5/5)로 검출률이 증가하였으나, 3 Log CFU/g 접종 샘플은 소수성 필터 적용 전후 모두 100%(5/5)로 확인되었다. 이상의 결과를 통해 신선편이 양배추 내 B. cereus 검출 시 증균배양액에 소수성필터를 적용하고 multiplex PCR을 적용했을 때 신속하고 효율적인 검출이 가능할 것으로 판단된다.

We present the first results of the invariant point (IVP) coordinates of the KVN Ulsan and Tamna radio telescopes. To determine the IVP coordinates in the geocentric frame (ITRF2014), a coordinate transformation method from the local frame, in which it is possible to survey using the optical instrument, to the geocentric frame was adopted. The least-square circles are fitted in three dimensions using the Gauss-Newton method to determine the azimuth and elevation axes in the local frame. The IVP in the local frame is defined as the mean value of the intersection points of the azimuth axis and the orthogonal vector between the azimuth and elevation axes. The geocentric coordinates of the IVP are determined by obtaining the seven transformation parameters between the local frame and the east-north-up (ENU) geodetic frame. The axis-offset between the azimuth and elevation axes is also estimated. To validate the results, the variation of coordinates of the GNSS station installed at KVN Ulsan was compared to the movement of the IVP coordinates over 9 months, showing good agreement in both magnitude and direction. This result will provide an important basis for geodetic and astrometric applications.

Toll and IMD pathways play an important role in producing antimicrobial peptides (AMPs) through NF-κB in insects. The functions of IκB kinase (IKK) complex regulating the NF-κB signaling cascade have not yet been investigated in Tenebrio model. Here, we identified TmIKK-β (or TmIrd5) which contains 2,112 bp encoding 703 amino acid residues. Domain analysis shows that TmIKK-β contains one Serine/Threonine protein kinases catalytic domain. Developmental expression patterns indicate that TmIKK- β gene was highly expressed in early pupal (P1) and adult (A5) stages. Tissue specific profiles show that TmIKK-β was highly expressed in the integuments in last instar larvae, and fat body and hemocytes in 5 day-old adults. TmIKK-β1 transcripts were strongly induced at 3 and 12 h-post injection of E. coli, and 3 h-post injection of S. aureus or C. albicans in hemocytes. In gut, TmIKK-β transcripts were slightly induced by E. coli (at 6, 9 and 24 h) and C. albicans (at 24 h), while it was not induced by S. aureus challenge. Moreover, it was highly induced at 6 h-post injection of E. coli and then it was gradually decreased in the fat body. To understand the immunological role of TmIKK-β, gene specific RNAi and mortality assay was performed. Depletion of TmIKK-β mRNA leads to increase microbial susceptibility of larvae against E. coli, S. aureus and C. albicans. In addition, induction patterns of fourteen AMP genes in response to microbial challenge was tissue specifically investigated in TmIKK-β–silenced T. molitor larvae. The results suggest that expression of ten AMP genes out of fourteen genes were drastically decreased by TmIKK-β RNAi in fat body, suggesting that TmIKK-β plays an important role in antimicrobial innate immune responses.

It has been well known that IKK-β, -ε and –γ play a pivotal role in IMD pathway. In this study, TmIKK-ε was identified and their functions in countering pathogenic infections were investigated. We identified TmIKK-ε gene which including 2,196 bp nucleotides (encoding 731 amino acid residues). Domain analysis of TmIKK-ε indicates that there is one Serine/Threonine protein kinases catalytic domain. TmIKK-ε gene was highly expressed in 2 day-old pupal stage and the expression was gradually decreased until 1 day-old adults. Then the expression was slightly increased until 4 day-old adult stage. Tissue specific expression of TmIKK-ε mRNA was high in the gut, integuments and hemocytes in last instar larvae, and fat body, Malpighian tubules and testis in 5-daysold adult. In hemocytes, TmIKK-ε was drastically induced by E. coli injection after 3 h and by S. aureus at 3 and 12 h-post injection. In gut, expression level of TmIKK-ε was high at 6 h-post injection of microbial injection. Expression of TmIKK-ε in fat body was drastically induced by E. coli at 3 and 24 h-post injection while it was not significantly induced by S. aureus and C. albicans. To understand the immunological role of TmIKK-ε, gene specific RNAi and mortality assay were performed. TmIKK-ε RNAi caused increased larval mortality against E. coli, not S. aureus and C. albicans. Finally, to investigate the induction patterns of Tenebrio fourteen AMP genes in response TmIKK-ε RNAi, three microorganisms were treated into TmIKK-ε-silenced T. molitor larvae. Nine out of fourteen AMP genes were not induced by microbial challenge in TmIKK-β dsRNA-injected group. Taken together, our results indicate that TmIKK-ε may regulates nine antimicrobial peptide genes in response to microbial challenge in T. molitor fat body.

Autophagy is an important self-eating process to eliminate damaged or unused organelles. We identified nine autophagy-related genes (Atg) including AaAtg-1, -3, -4b, -4d, -5, -6, -8, -12 and -13 from the Asian tiger mosquito, Aedes albopictus. Developmental expression patterns indicate that mRNA levels of AaAtg-1, -3, -4b, -4d, -5, -6, -12 and -13 were highly expressed in egg, whereas expression of AaAtg8 was high in 1stand3rdinstarlarvalstages. TissuespecificexpressionofthesegenesindicatesthatAaAtg1 was highly expressed in thorax and midgut in blood-fed adult female mosquitoes (BF), and head and thorax in sugar fed adult female mosquitoes (SF). Transcript level of AaAtg3 was high in thorax in BF, but head, thorax and Malpighian tubules in SF. AaAtg4b, -4d mRNA levels were significantly high in Malpighian tubules in BF, and head in SF, respectively. AaAtg-5 and -6 transcripts were highly expressed in head in BF, and expression of AaAtg-8 was high in Malpighian tubules in BF. Levels of AaAtg-12 and -13 mRNAs were significantly high in head and midgut in BF. Induction patterns of AaAtg genes against pathogens showed that AaAtg-1, -3, -4b, -8, -12 and -13 were strongly induced at 6 h-post injection of S. aureus, and mRNA levels of AaAtg-1, -3 and -13 were significantly induced by E. coli challenge after 3 h-post injection in SF abdominal carcass. In SF midgut, AaAtg-1, -3, -4b, -4d, -5, -6, -12 and -13 transcripts were drastically induced at 9 h-injection of E. coli and S. aureus, while expression of AaAtg-8 was highly induced by S. aureus and C. albicans at 9 h-post injection. Each AaAtg gene was slightly induced by E. coli, S. aureus or C. albicans at different time points in abdominal carcass in BF. Interestingly, AaAtg-8 was not induced by microbial challenge. While eight other Atg genes except AaAtg-8 were highly influenced by S. aureus at 6 and 9 h-post injection, E. coli at 3 h-post-treatment, and 3, 6, and 9 h-post inoculation. In the future, we will characterize the functional roles of autophagy during mosquito-microbes interaction.

Host defense against pathogen invasion highly relies on immune defense machinery that is controlled by the nuclear factor-κB (NF-κB) of transcription factors. The Toll pathway are well known as an insect innate immune mechanism to protect host itself from invaded pathogens. Basically, in the edible insect, Tenebrio molitor, the Toll pathway is primarily activated by polymeric Lys-type peptidoglycans (PGNs), and components of fungal cell walls, β-1,3-glucan. Based on the current studies, the tremendous study has been focused on recognition and subsequent activation of spätzle in haemolymph, hence, there is a grave gap for intracellular event. Herein, in order to understand intracellular event of Toll signaling pathway, the Dorsal gene were identified. Moreover, domain analyses of TmDorsal2 gene indicate that there are two major domains such as Rel homology domain (RHD), ig-like, plexins, and transcription factors (IPT) domains. Based on the achieved results, TmDorsal2 mRNA was highly expressed in 1-day old pupa. Furthermore, TmDorsal2 was highly expressed in Malpighian tubules and fat body in last instar larvae (LL), and likewise mainly expressed in Malpighian tubules during adult 5-day old period, also the lowest expression of TmDorsal2 was observed in gonads. Moreover, TmDorsal2 mRNA levels after infection with E. coli appreciably went up at 6 and 9h time points. To investigate the effects of TmDorsal2 RNAi on larval susceptibility against various pathogens namely E. coli, S. aureus or C.albicans, dsRNA of TmDorsal2 has been synthesized the larvae dissected after 24h. As a result, TmAttacin1a, 1b and 2, TmDefencine1 and 2, TmTenecin1, 2, 3 and 4, TmCecropin2, TmColeoptericin1 and 2, Thaumatin-like protein 1 and 2 markedly reduced in the gut after injecting all mentioned microbes. In contrast, TmTenecin 2, Thaumatin-like protein 1 and 2 strikingly increased after microbe injection in the fat body. Interestingly, the most AMPs gene expression in whole body experimental case were upregulated. On the horizon, we will investigate effects of TmDorsal1 RNAi on larval susceptibility against various pathogens. Taken together, our studies may aid to understand insect innate immunity.

후각 신경 시스템은 일반 생활 환경에서 많은 다양한 화학 물질을 인식하고 구별한다. 곤충에서는 다양한 화학 물질을 기호적 또는 회피적인 특이성을 부여하고 이를 구분해 낼 수 있는 고도로 발달된 후각신경 수용체들로 구성된 냄새 맡는 (odorant-gated) 이온 채널 군을 진화시켰다. 최근에 후각 수용체와 단백질을 포함한 olfaction 관련한 진딧물 게놈 밝혀졌고, 초파리에서 다양하게 분화되어 있는 후각 수용체들이 보고되고 있다. 후각 신경 수용체의 유전체는 매우 높은 보전적인 염기 서열을 가지고 있으며, 체계적인 신호 전달 시스템을 갖추고 있다. 대표적 수용체인 odorant-gated ion channels comprised of a highly conserved co-receptor (Orco)는 중심 구멍 주위에 대칭 적으로 배열된 4 개의 서브 유닛을 갖는 homotetramer 채널 구조를 가지고 있다. 이는 인체 내에 존재하는 7-transmembrane receptor와 매우 유사한 구조를 형성하고 있고, 신경전달물질의 수용체와 매우 유사한 구조적 형태 및 gating mechanism을 가지고 있다. 본 연구에서는 초파리에서 분리한 후각 신경 수용체 하위 유형인 OR65 유전자 분리하여 세포 발현 시켜 Xenopus oocyte를 이용하여 Whole cell voltage clamp recording을 실시하였다. 본 수용체의 성공적인 발현 이후 유해 해충 유인제 개발 회사인 마이크로자임의 미생물 배양 추출물을 이용해서 후각 신경 수용체 활성 조절 여부를 연구하였다. 미생물 배양 추출물을 10,000배 희석한 recording media에서 수용체의 활성을 확인하였고, 이를 농도 별로 처리하여 농도 의존성 수용체 활성 작용을 확인하였다. 따라서 곤충의 후각 신경 수용체 활성 조절 시스템을 이용하여 유인물질 또는 기피 물질을 발굴할 수 있으며, 본 연구를 통해서 MZ01은 곤충 수용체 OR65를 활성 시킴으로써 유인 현상을 나타내며, 본 연구를 통해서 현장에서 검증된 미생물 배양 추출물의 성능을 과학적 분석으로 결과를 제시하였다.

The existing ethyl formate fumigant is carbon dioxide (CO2) mixed liquified gas in metal cylinder, but this product type costs a lot to manufacture, translate and maintain cylinder. To supplement these problems, we have developed a new ethyl formate fumigation technique with nitrogen (N2) carrier. We assessed the susceptibility of mealy bugs, the most frequently detected pests in imported banana, and phytotoxicity of banana fruits. Ethyl formate and nitrogen were concurrently treated on citrus mealybug, one of the most resistant mealybug to fumigant, and ethyl formate was treated with LC50 product of independent treatment dosage. Nitrogen was treated with 7 dosages from 79% to 95% concentration. Phytotoxicity of banana was assessed by treating EF 35 mg/L with N2 79% for 14 days, and color, sugar contents and loss of weight were measured. EF with N2 treatment showed more than 50% of mortality on every growth stages, and there was no significant difference between control and treatment banana fruits. These results indicate that concurrent treatment of EF and N2 can be used to control mealybug in banana fruits.

농업용 무인멀티콥터를 활용한 방제면적은 수도작은 물론 밭작물에서도 방제 범위가 확대되고 있다. 밭작물은 작물의 형태가 다양하여 비행속도, 살포높이 등을 최적화하는 연구가 필요하다. 본 연구에서는 배추, 대파, 고추에서 발생하는 나방류에 대하여 무인멀티콥터의 살포 높이에 따른 방제효과를 확인하였다. 살포된 약제가 작물에 부착되는 정도를 조사한 결과, 배추는 2m, 대파는 3~4m, 고추는 모든 높이에서 비슷한 부착정도를 보였지만, 전체적으로 부착량이 적고 불균일 하였다. 살포 높이별 방제효과는 작물별로 각각 배추(9~10엽기)는 2m높이, 고추(약 1.2m)는 2~3m, 대파(약 60cm)는 2, 3, 4m 각각 비등한 방제효과를 보였다. 각각의 작물모두 경엽처리 대비 약효가 미흡하였으며, 고추는 4m높이 살포시 방제효과가 현저히 낮았다.

최근 수목 병해충을 방제, 예방하기 위해 나무주사법이 많이 사용되고 있다. 나무주사법은 기상에 상관없이 약제처리가 가능하며, 약제사용량 감소와 천적등 비표적 생물에 대한 영향을 줄일 수 있다는 장점이 있다. 하지만 현재 국내에서 나무주사법은 산림 해충을 대상으로만 등록, 적용되어 있을 뿐, 과수에 등록된 제품은 없다. 과수에 나무주사법이 적용되기 위해서는 농약잔류와 약효에 대한 연구가 필요하다. 본 연구에서는 복숭아(천중도)에 Acetamiprid 10% SL을 나무주사 후, 잔류성과 약효를 조사하여 실용가능성을 확인하였다. 약제가 나무주사된 나무는 복숭아 벚나무깍지벌레(Pseudaulacaspis prunicola)에 대하여 방제효과가 있으나, 가지별로 불균일한 약효와 잔류성을 보여 향후 수형, 흉고직경 등을 고려한 나무주사제의 처리법 최적화에 대한 연구가 지속적으로 필요할 것으로 생각된다.

갈색날개매미충은 2010년 8월 국내에서는 처음으로 충남 공주, 예산의 사과와 불루베리를 가해하는 것이 발견되었으며, 미국선녀벌레는 2005년 8월 김해 한림의 단감원에서 처음으로 성충이 발견되어 2008년과 2009년에 김해, 수원 등지에서 확인된 후 최근까지 전국으로 확산되고 있다. 꽃매미는 2004년 천안에서 처음 발견되어 2006년부터 발생이 꾸준히 증가되다 2013년을 정점으로 다소 감소하는 추세를 나타내고 있다. 2018년도 돌발해충들의 국내 발생분포와 현황을 확인하고자 전국 농경지 및 농경지 주변을 대상으로 3월(3.5-3.23)에는 월동난, 7∼8월(7.31-8.10)에는 약·성충 발생을 조사하였다. ‘18년 갈색날개매미충 월동난 조사결과 89개 시군에서 4,628ha로 전년보다 발생지역 7개 시군이 증가 되었으나 발생면적은 8.1% 감소되었으며, 꽃매미는 80개 시군에서 701ha로 발생되어 갈색날개매미충과 유사하게 전년보다 발생지역은 3개 시군이 증가되었으나 발생면적은 10.5% 감소되었다. ‘18년 약·성충 조사에서는 꽃매미는 95개 시군, 1,171ha, 갈색날개매미충은 94개시군, 7,867ha, 미국선녀벌레는 123개시군, 14,779ha에서 발생을 확인하였다. 전체적으로 발생지역은 확대되는 경향이었으며, 발생면적은 전년대비 꽃매미는 1,047ha, 갈색날개매미충은 967ha 감소되었으나 미국선녀벌레는 4,475ha 증가되었다. 특히 미국선녀벌레의 경우 경기와 경남지역에서 급증하는 경향을 나타내었다.