The red imported fire ant (Solenopsis invicta) is a species of ant native to South America. The fire ant was inadvertently introduced into USA, Australia, New Zealand, and other Asian countries including China and Taiwan. Since the first report of the fire ant in port city of Busan, Korea in 2017, it was found in many other cities of Korea in following year. To obtain the molecular information of this invasive species, total RNA was extracted from the abdominal segment of the ants collected in Incheon, and subjected to transcriptome sequencing. By using Illumina sequencer platform, 101 base pared-end sequencing generated 2 × 50,064,081 of raw reads to obtain 2 × 45.95 Gbase of quality filtered nucleotide sequences. The in silico cDNA library was constructed by Trinity de novo assembler followed by TransDecoder ORF finder and CD-HIT clustering program to streamline the library. The final version of cDNA library contains 20,442 contigs with protein coding capability. To survey the virome of this ant, these contigs were searched against the viral reference sequences from NCBI RefSeq database with BLASTN program. As a result, contigs which showed high sequence identities with several RNA viruses including previously reported SINV-2 were found from the fire ant. This virome information might give an idea of a shift of virological environment of this newly found ant isolate or population in Korea.

Morphology of antennal sensilla and their distribution were investigated in male and female adults of Gymnosoma rotundatum, a parasitic fly to hemipteran species, using scanning electron microscopy. The overall length of antenna was not different between male and female. Antenna of G. rotundatum was composed of scape, pedicel and funiculus in both sexes. Three types of sensilla (sensillum basiconica, s. chaetica and s. coeloconica) were identified from both sexes, in varying numbers and distribution along the antennae. The two sensillum types were further divided into different subtypes; s. basiconica into three subtypes and s. chaetica into two subtypes. Among sensilla, s. basiconica subtype 1, 2 and 3 were multiporous, indicating that the ir primary function is olfactory, and others were not. The s. basiconica was most numerous on the antennae in both sexes. The number of subtype 1 of s. basiconica was different between male and female. The morphological information obtained in our study provides a basis for electrophysiological and behavioral studies of the olfactory sensory function of each morphological type of sensilla. (Supported by PJ011756022018, RDA)

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.

We assessed the susceptibility of three fumigants on Phthorimaea operculella, which is an important pest of stored potato worldwide. 5 to 6 initial dosage of each fumigants were treated on every growth stages of P. operculella. Methyl bromide showed 100% mortality at CT 33.40mg h/ L on egg, CT 14.41mg h/L on late larvae, CT 31.89mg h/L on pupae and CT 16.01mg h/L on adult, respectively. The LCT50 of methyl bromide was 19.115mg h/L on egg, 3.934mg h/L on late larvae, 13.810mg h/L on pupae and 6.260mg h/L on adult, respectively. In case of phosphine, 98% mortality was achieved at CT 16.77mg h/L on egg, and 100% mortality was achieved at CT 16.58mg h/L on late larvae, CT 18.54mg h/L on pupae and CT 12.28mg h/L on adult, respectively. The LCT50 of phosphine was 1.457mg h/L on egg, 2.236mg h/L on late larvae, 1.282 mg h/L on pupae and 0.253mg h/L on adult, respectively. In case of ethyl formate, 100% mortality was achieved at CT 96.21mg h/L on egg, CT 101.30mg h/L on late larvae, CT 120.66mg h/L on pupae and CT 148.30mg h/L on adult, respectively. The LCT50 of ethyl formate was 23.730mg h/L on egg, 13.706 mg h/L on late larvae, 29.578mg h/L on pupae and 19.235mg h/L on adult, respectively.

Carbonyl sulfide(COS) is a naturally generated gas from fermentation process of microbes and from plant root and stem. COS was firstly registered as a fumigant at 1993 to control stored product pests. To supplement environmental problems and toxicity of commercial fumigants and develop new fumigant, we have processed the susceptibility assessment of carbonyl sulfide on important agricultural pests, Myzus persicae and Tetranychus urticae. Every growth stages of two insect species were tested, and five dosages of carbonyl sulfide were treated for 4 hours, and the mortality was investigated after 24 hours of treatment. Nymphal stage of M. persicae was completely controlled at more than 20 mg/L dosage, and adult stage showed 95.8% mortality at 80 mg/L dosage. The LC50 of M. persicae was 7.314mg/L for nymph and 26.117mg/L for adult stage. Egg stage of T. urticae showed 91.2% mortality when treated with 100mg/L carbonyl sulfide, and nymph and adult stage showed 100% and 94.1% mortality at 8mg/L and 80mg/L, respectively. The LC50 of T. urticae was 73.110mg/L for egg, 2.818mg/L for nymph and 12.054mg/L for adult stage.

Stored grain pests can cause reduction of grain quantity, quality, commercial value and germination rate. Susceptibility of three fumigants, methyl bromide, ethyl formate and phosphine, were assessed on Tribolium castaneum, which is an important stored grain pest. On susceptible insects, LCT50 of phosphine was 0.654mg h/L for egg, 0.127mg h/L for late larvae, 0.105mg h/L for pupae and 0.048mg h/L for adult stage, respectively. LCT50 of methyl bromide was 33.193mg h/L for egg, 14.585mg h/L for late larvae, 8.616mg h/L for pupae and 11.967mg h/L for adult stage, respectively. LCT50 of ethyl formate were 25.165mg h/L for egg, 80.912mg h/L for late larvae, 176.326mg h/L for pupae and 68.578mg h/L for adult stage, respectively. On resistant insects, LCT50 of phosphine were 82.325mg h/L for egg, 33.315mg h/L for late larvae, 73.546mg h/L for pupae and 55.707mg h/L for adult stage, respectively. LCT50 of methyl bromide were 19.250mg h/L for egg, 43.413mg h/L for late larvae, 76.842mg h/L for pupae and 19.387mg h/L for adult stage, respectively. LCT50 of ethyl formate were 87.552mg h/L for egg, 113.457mg h/L for late larvae, 200.122mg h/L for pupae and 85.394mg h/L for adult stage, respectively.

Recently, phosphine resistance of Sitophilus oryzae has been reported from China, India, Brazil and Australia. In this study, susceptibility of three fumigants were assesses on phosphine resistant and susceptible S. oryzae to investigate domestic phosphine resistance level and to use base data for resistance control. On susceptible insects, LCT50 of phosphine was 0.440mg h/L for egg, 0.602mg h/L for early larvae, 3.901mg h/L for late larvae, 6.171mg h/L for pupae and 0.295mg h/L for adult stage, respectively. LCT50 of methyl bromide was 9.997mg h/ L for egg, 12.113mg h/L for early larvae, 18.952mg h/L for late larvae, 21.104mg h/L for pupae and 17.824mg h/L for adult stage, respectively. LCT50 of ethyl formate was 75.795mg h/L for egg, 60.110mg h/L for early larvae, 160.491mg h/L for late larvae, 255.797mg h/L for pupae and 77.711mg h/L for adult stage, respectively. On resistant insects, LCT50 of phosphine was 6.959mg h/L for egg, 28.456mg h/L for early larvae, 48.170mg h/L for late larvae, 29.106mg h/L for pupae and 16.550mg h/L for adult stage, respectively. LCT50 product of methyl bromide was 17.842mg h/L for egg, 14.900mg h/L for early larvae, 25.840mg h/L for late larvae, 43.520mg h/L for pupae and 16.397mg h/ L for adult stage, respectively. LCT50 of ethyl formate was 60.034mg h/L for egg, 64.450mg h/L for early larvae, 149.028mg h/L for late larvae, 140.408mg h/L for pupae and 66.043mg h/L for adult stage, respectively. Domestic resistant S. oryzae showed 4 to 56 times higher resistance rate than susceptible insects.

The susceptibility of three fumigants, methyl bromide, ethyl formate and phosphine, and concurrent treatment of ethyl formate and phosphine were tested on Lasioderma serricorne. Susceptibility assessment were performed by treating 5 to 6 initial dosage on every growth stages of L. serricorne. The LCT50 of methyl bromide was 13.896mg h/L for egg, 36.038mg h/L for late larvae, 25.172mg h/L for pupae and 21.758mg h/ L for adult, respectively. The LCT50 of phosphine was 0.317mg h/L for egg, 0.649mg h/L for late larvae, 3.748mg h/L for pupae and 0.703mg h/L for adult, respectively. In case of ethyl formate, the LCT50 was 43.657mg h/L for egg, 137.606mg h/L for late larvae, 72.676mg h/L for pupae and 52.951mg h/L for adult, respectively. Concurrent treatment of ethyl formate and phosphine was performed by treating 5 to 6 initial dosage of ethyl formate with 0.5 mg/L phosphine for 4 hours on every growth stages of L. serricorne. The LCT50 of ethyl formate concurrent treatment was 13.746mg h/L for egg, 8.156mg h/L for late larvae, 27.087mg h/L for pupae and 11.353mg h/L for adult, respectively, and these results indicate that concurrent treatment can control pest with lower dosage and shorter period. Sorption rates and ventilation periods of each fumigants were also calculated for efficacy and safety

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.

Spotted wing drosophila, Drosophila suzukii, is an important pest of grape and strawberry because it lays egg inside fruit by damaging fruit surface. Recently, D. suzukii has been widely spreaded and causes several problems, so many countries include Australia designated D. suzukii as an important quarantine pest. Because of this, Korean farmers have trouble to export strawberry. In this study, we tested ethyl formate and phosphine to control D. suzukii, and also tested concurrent treatment of ethyl formate and phosphine to reduce phytotoxicity and enhance efficacy. When treated 35g/m3 of ethyl formate for 4 hours, mortality of egg, larvae, pupae and adult stages of D. suzukii was 22.2%, 21.1%, 19.2% and 28.3%, respectively. When treated 1g/m3 of phosphine for 24 hours, all stages of D. suzukii was completely controlled, but caused phytotoxic effect on strawberry. When treated with 35g/m3 of ethyl formate and 1g/m3 of phosphine concurrently for 4 hours, efficacy has been increased with less phytotoxicity than separate treatment. This result indicates that concurrent treatment enhanced efficacy with less phytotoxicity.

우드펠렛은 가정용, 상업용, 산업용 등으로 사용되며 특히 화력발전소에서 석탄 대용으로 수입량이 급증하여 연간 약 2,400만톤 가량 수입되고 있는 상황이다. 우드펠렛 검역훈증제로 기존에는 메틸브로마이드(이하 MB)가 최근까지 사용되어져 왔으나, MB의 높은 흡착률과 우드펠렛의 과도한 수용비로 인해 최종가스농도 기준에 미치지 못하여 재투약하는 사례가 빈번히 발생해 문제가 되어져왔다. 이를 해결하기 위해 농림축산검역본부에서는 올해 초 MB와 포스핀을 병행으로 처리하는 방식의 소독처리기준을 신설하였다. 새로운 소독처리기준이 설정됨에 따라 배기시 안전한 배기시간 수준을 설정하기 위해 병행처리 후 수착과 탈착되는 가스농도를 조사하였다. 실내 훈증상에 우드펠렛을 채우고 약제 처리한 결과 각 훈증제의 TLV(Threshold Limit Value)-TWA(Time Weighted Average) 기준이하로 농도가 감소되는 배기시간은 25℃일 때 MB는 120시간, 포스핀은 6시간으로 측정되었고 15℃일 때 MB는 186시간, 포스핀은 19시간으로 측정되었다.

선박이나 컨테이너로 수출입되는 곡류 및 박류는 검역용 훈증제인 메틸브로마이드(이하 MB)와 인화수소(이하 PH3)를 사용하는데, 처리 온도/수용비/선적방식에 따라 충분한 배기시간을 필요로 한다. 현장에서 배기시간 미설정으로 검역관 및 방제기술자에게 훈증제 TLV(Threshold Limit Value)-TWA(Time Weighted Average) 기준 이상의 농도에 노출될 위험이 있으므로, 소독 전, 후 작업자 안전을 고려하여 각 훈증제 별 작업자 안전기준에 적합한 배기시간이 설정되어야 한다. 따라서, 실내 훈증상에서 곡류(쌀, 대두) 및 박류(대두박, 주정박)를 대상으로 현행 소독처리기준으로 소독 후 탈착되는 가스농도를 측정해 MB 1 ppm, PH3 0.3 ppm 수준 이하로 감소되는 배기시간을 조사하였다. 조사결과 TLV-TWA 기준이하의 수준으로 감소되는데 필요한 배기시간은 주정박/대두박/대두에 MB가 처리된 경우는 모두 30시간, 쌀에 PH3가 처리된 경우는 30분이 소요되는 것을 확인하였다.

This study investigated the fumigant activity of phosphine (PH3) on 2 kinds of mealybug (Pseudococcus longispinus and P. orchidicola (Hemiptera: Pseudococcidae)) adults and nymphs. All of the two mealybugs adults showed higher LCT99 values than nymphs, and P. longispinus showed higher tolerance than P. orchidicola in a 12 L desiccator. The absorption of phosphine on 13 nursery plants showed 12.2~41.5% difference depending on the plant. All of the mealybugs treated with phosphine 2 mg/L in 0.5 m3 fumigation chamber for 4 h showed 100% fumigant activity, except P. longispinus adult (approximately 90% at bottom part). However, when the exposure time was increased to 24 h, all of them showed 100% mortality. In the treatment of 10 m3 container, the 24 h treatment of phosphine showed 100% mortality to P. longispinus and P. orchidicola adults and nymphs. In all the experiments, no phytotoxicity of phosphine observed on 13 plants until 1 month after treatment.

For the setting of control timing, the number of overwintering eggs on host plants and population density of Metcalfa pruinosa nymphs and adults was surveyed in the three peach orchards. According to a survey of the overwintering eggs density (∅2~3㎝, 20㎝ branches) of M. pruinosa on host plants, 221.8 in Mugunghwa trees, 189.3 Oak trees, and 152.7 Acacia trees, and 57.0 Peach showed the number of eggs laid. Examined peach branches within the 50cm (∅1~5cm) was not found population densities of M. pruinosa nymphs and adults, and it was also very low in the sticky trap. The peak density of the adult was highest at 5.4 per trap on August 7 after being first investigated on July 16 in a string wrap survey, and then decreased. In the peach orchard, it is judged that the control of the M. pruinosa is due to be 20 days after the end of the insect hatching, and that distribution of host plants is more important in the outside region than inside region.

붉은불개미는 세계 100대 침입외래 해충 중의 하나로 우리나라에서는 관리 해충으로 1996년도에 지정되었다. 2017년 9월28일 부산항 감만 부두에서 국내 최초로 발견된 후, 2018년에 부산항, 평택항 과 인천항에서 발견이 되었고, 9월18일에 대구의 아파트 공사장에서 내륙에서는 처음으로 발견이 되었다. 대구에서 발견된 붉은불개미도 아파트 건설을 위하여 중국에서 수입한 석재에 혼합되어 수입된 토양과 식물체와 같이 혼입된 것으로 추측이 된다. 그러므로, 현재까지 국내에서 붉은불개미가 번식을 하여 토착화되었다는 증거는 없다. 현재까지 붉은불개미에 대한 종 판명은 전문가에 의한 형태적인 분류가 대부분이다. 그래서, 붉은불개미에 대한 확증을 위해서는 채집된 개체를 검역기관으로 이송하여 분자진단을 실시해야 하므로, 최소 이틀의 시간이 필요하다. 현재 항원항체 반응을 이용한 붉은불개미 진단 키트가 해외에서 개발되어 판매되고 있지만, 낮은 민감도로 최소 3~5개의 충체가 필요한 실정이다. 현재 판매되고 있는 진단키트가 가지는 문제점은 이를 대신할 새로운 키트의 개발이 필요함을 보여주고 있다. 신속한 붉은불개미의 검역현장에서 진단을 가능하게하기 위한 노력 중의 하나로 우리는 붉은불개미의 복부에서 발현되는 유전체에 대한 연구를 수행하였다. 본 발표에서는 그에 대한 현재까지의 연구 진행과 결과를 설명하도록 하겠다. (본 연구는 농림축산 검역본부의 연구비 지원으로 이루어졌음).

In order to identify the specific antigens for pine wood nematode (PWN), we confirmed that one of the genes commonly found in the transcriptome, proteome and secretory proteins of PWN belonged to the Aldose Reductase (AR) family protein. 36.5 kDa PWN-AR1 was expressed and purified using Baculovirus Expression System. Total 1,546 hybridoma fusion library was generated and screened for specificity to PWN-AR1 by Enzyme Linked Immunosorbent assay (ELISA). Nine clones showed strong immunoreactivity to PWN-AR1 were limited-diluted. Total 864 limited-diluted clones were further screened using PWN-AR1 by ELISA and 34 monoclonal antibody (Mab) clones were selected. 34 Mab clones were further screened using PWN extracts and a standard PWN-infected pine tree extract by ELISA. Finally nine clones were selected and their immunoreactivities to 4 different nematodes were examined by ELISA. Seven clones pecifically recognized PWN while two clones recognized 4 nematodes. Our data suggested that PWN-AR1 is a PWN secretory enzyme while PWN is invading pine trees, Thus, PWN-AR1-Mabs could be used to develop diagnosis tools for PWN and its infected pine trees. (This work was supported by National Institute of Forest Science)

소나무재선충(Pine wood nematode, PWN)은 아시아와 유럽의 소나무류들에 침입하여 고사시키는 심각해 병원성 선충이다. 가장 완벽한 방제 방법은 감염목을 소각/분쇄의 방법으로 PWN 감염목을 제거하는 것이다. 현재 PWN의 다른 종과 차이를 가지는 유전자서열을 이용한 진단 방법이 다양하게 개발되어져서 실험실에서의 종 판명은 가능한 실정이다. 하지만, 대부분의 분자진단방법은 소나무 목편에서부터의 시료 추출, 그리고 분자진단 시약과 혼합하고, PCR을 이용하여 증폭을 진행 한 후, 결과를 확인하는 과정이 필요하다. 그러므로, 분자진단법을 현장에 적용하기 위해서는 향 후 많은 노력이 필요하다. 이에 반하여 항원/항체 반응을 이용한 진단 방법의 경우는 항원 추출 후, 일정량을 신속진단키트에 떨어 뜨려서 반응을 확인하는 과정을 거치므로, 전문적인 지식이나 기술이 필요하지 않다. 하지만, PWN과 PWN 감염목에 특이적인 항원이 밝혀지지 않은 상황이다. 최근에 우리는 PWN단백질 중에서 분비되는 Aspartic peptidase 1 (ASP1)을 항원으로 선정을 하고, 전장단백질을 Baculovirus Expression System으로 발현을 하였다. PWN-ASP1을 항원으로 단클론항체를 분비하는 세포 주를 확립하였다. 이러한 단클론항체는 향후, PWN의 소나무 침입에 관한 연구와 신속진단키트의 개발에 활용될 수 있을 것이다. (본 연구는 국립산림과학원의 연구비 지원으로 진행되었음)

Bursaphelenchus xylophilus, commonly known as pine wood nematode (PWN) and pathogen of pine wilt disease, is transmitted to new host trees through Monochamus spp.. In this study, five points were set 1 km apart placing the interface of the damaged area in the middle to measure density of the beetles and confirm the presence of PWN in Chungcheongbuk-do, Republic of Korea. The vectors collected from five traps on each point were sorted on weekly basis until the 12th week. We confirmed the presence of PWN in each collected vector by PCR with species-specific ITS primers.

This study was conducted to surveyed the community structure of insects on thirty varieties of Hibiscus syriacus in Busan and Suwon, Korea. In addition, seasonal changes and damage rates of major insect pest species were investigated. Insect pests were visually observed from the area of H. syriacus plantations. As a result, 16 species belonging to 9 families of 4 orders were observed, while 13 species were classified to insect pests. Among insect pests, hemipterans (6 species) and lepidopterans (4 species) were most diverse groups. During summer season, Rehimena surusalis (Lepidoptera: Crambidae) and Haritalodes derogate (Lepidoptera: Crambidae) were damaged to flower buds and leaves, respectively. In September, nymphs and adults of Plautia stali (Hemiptera: Pentatomidae) were found from flower buds and leaves.

소나무재선충병의 매개충인 북방수염하늘소의 체내 선충보유량에 따른 비행능력의 차이를 구명하기 위하여 춘천시 지내리의 잣나무림에서 어리소나무재선충(Bursaphelenchus mucronatus)에 감염이 확인된 북방수염하늘소 암·수 각 20마리를 채집, 실내사육 후 우화-섭식 1 주일이 지난 시점부터 1주 간격으로 Flight mill을 이용하여 비행실험을 진행하였다. 동시에 사육통 바닥에 여과지와 섭식한 잣나무 신초를 3~4일 마다 교체 후 Baermman funnel법으로 소나무 재선충을 분리하여 그 수를 조사, 비행능력과 선충보유량의 관계를 조사하였다. 그 결과 약 50,000마리 이상 선충을 체내에 보유시 비행능력이 급감하는 것을 확인 할 수 있었으나, 전체적으로 선충의 체내 보유량에 따른 비행능력의 차이는 상관관계가 없는 것으로 확인 되었다.