Quarantine Pest Research Facility (QPRF) had been built in Animal and Plant Quarantine Agency for study on exotic pests at 2018. This facility was constructed to prevent effective containment for preventing accidental escape of insects. We tested a developmental and susceptibility of pesticides (Fluxametamide, FLX and Hydramethylnon, HMT) on beet armyworm, Spodoptera exigua for testing containment level of QPRF. S. exigua showed 22.5 days at 28±1℃ and 26.5 days at 25±1℃ with 60±5% RH for entire life cycle. LC50 of FLX and HMT to 5th instar larvae for 24h revealed as lower than 1ppm and 100ppm, respectively. Also, there is no release of adults in QPRF and confirmed by sex pheromone trap during winter season.

파밤나방은 배추과 채소 등 여러 농작물에 피해를 주고 있는 중요한 나방류 중의 하나이며, 돌발적으로 다발생한 경우 큰 피해를 주기도 한다. 제주지역에서 배추과 채소 주요 재배지역인 제주시 한림읍에 2007년부터 2011년까지 파밤나방 성페로몬 트랩을 이용하여 시기별 발생밀도를 조사하였다. 트랩에 최초 유인 시기는 4월 상순이었으나 3월 온도가 높은 해에는 3월 상순에 유인되기도 하였다. Karimi-Malati et al.(2014)의 발육온도 모델을 이용하여 제주지역에서의 연간 발생세대수를 추정한 결과 4∼5세대를 경과하는 것으로 추정되었다. 성페로몬 트랩의 발생 절정기별 다음 세대 성충 발생시기와 트랩 유인밀도와의 관련성을 2009년과 2010년 조사 자료에 대해 검토한 결과 연관성이 낮은 시기가 나타났다. 이 시기 NOAA의 HYSPLIT 모델(Rolph et al., 2017)을 이용하여 기류의 역방향과 순방향 궤적선을 추적한 결과 중국으로부터 장거리 이동이 가능할 것으로 추정되었다. 장거리 이동에 관여하는 기상요인과 비래원은 6∼7월에는 장마전선 등에 의한 중국의 남중부지역이, 8∼9월에는 중국의 북동부지역으로 나타났다.

The beet armywom, Spodoptera exigua Hübner, originated from Southeast Asia is one of important agricultural pests worldwide. The moth having no diapause mechanism has highly migratory capacity which contributes to population outbreaks and geographic expansion. Although the moth was first reported in 1926 from Korea, it has been important since the late 1980s. In July 2018, the species outbreaked in Gimje, Jeonbuk Province causing serious damage to soybean in several paddy fields. In this study, we investigate the genetic diversity of the outbreaking populations using a partial sequence of mitochondrial DNA COI gene. 37 COI sequences obtained from specimens collected at the fields of Gimje, Korea were compared with 249 COI sequences from GenBank. In total, 25 haplotypes were detected from 18 countries. Three haplotypes (H1, H2, and H5) found in Korea were all shared with those in China. Haplotypes H1 and H2 were prominent in Korea as well as in invasive regions worldwide. This suggest that outbreaking populations in Korea were invaded from China. The migration source was estimated using backward trajectory analysis, and found to be Zhejiang Province, China.

Prostaglandin E2(PGE2) is an autocrine and paracrine signal in insects and other animals. Its signal pathways in target cells are well understood in mammalian system but not in insects. Here, we assessed PGE2 signaling in hemocytes of Spodoptera exigua through knocking-down of signal component genes by RNA interference (RNAi) and knocking-out (KO) of PGE2 receptor using CRISPR-Cas9. From S. exigua transcriptome, we selected hemocyte signaling components and analyzed their functions in cellular immune responses through RNAi. KO mutant against PGE2 receptor exhibited severely hampered larval development and adult fecundity.

최근 파밤나방 등 다양한 나비목 해충에서 diamide 계통의 살충제 (IRAC group 28)의 저항성이 많이 보고가 되고 있다. 국내에서도 이미 배추좀나방과 파밤나방의 저항성이 보고되었는데, 본 연구에서 사용된 강릉에서 2018년 채집한 파밤나방의 경우 Chlorantraniliprole에 대하여 감수성 대비 2,500배 이상의 높은 저항성을 나타냈다. 추천 농도 처리 시 30% 이하의 방제가를 나타냈으며 Cyantraniliprole, Cyclaniliprole, Flubendiamide에서 높은 교차저항성을 나타냈다. 이 저항성 집단에서 diamide 계통 살충제의 작용점인 ryanodine receptor에 이미 보고된 G4946E 돌연변이는 발견되지 않았으나 새로운 I4760M 돌연변이를 발견하였는데, 2014년 배추좀나방에서 보고된 돌연변이와 위치가 동일하였다. 따라서 I4760M 돌연변이는 파밤나방에서 diamide 저항성에 관여 할 것으로 판단한다.

The present study evaluated the susceptibility of field populations of Plutella xylostella and Spodoptera exigua larvae to four diamide insecticides—chlorantraniliprole, cyantraniliprole, cyclaniliprole, and flubendiamide. All the four diamide insecticides induced 100% mortality in the populations from Seongju (SJ) and Geochang (GC) when treated at a concentration recommended for P. xylostella. However, a very low insecticidal activity was observed in the population from Pyeongchang (PC) with 42.3% 3 d after treatment with chlorantraniliprole. Further, the populations of S. exigua from Cheongju (CJ), Jindo (JD), and Yeonggwang (YG) were not completely controlled by the 4 diamide insecticides. A comparison of susceptibility of S. exigua larvae to chlorantraniliprole between 2014 and 2017 showed that chlorantraniliprole induced 100% mortality in all populations in 2014, whereas a very low insecticidal activity was observed among the populations in 2017. This study can serve as a basis to control pests effectively using diamide insecticides.

Phospholipase A2 (PLA2) hydrolyzes fatty acids from phospholipids at sn-2 position. Two immune-associated PLA2s are known in Spodoptera exigua. Both are calcium-independent cellular PLA2 (iPLA2A and iPLA2B). These iPLA2s are inducible to immune challenge using different epitopes such as virus, Gram+, Gram- bacteria, and fungi. However, their expression profiles are different between these two iPLA2s. Especially, immune challenge with Gram+ bacteria and fungi significantly up-regulated these iPLA2 expression, suggesting their expression under Toll signal pathway. RNA interferences of Toll or IMD signal components are being investigated.

A prostaglandin (PG), PGE2, mediates both cellular and humoral immune responses of insects. Spodoptera exigua (Lepidoptera: Noctuidae) biosynthesizes PGE2 by an unknown enzyme. This study predicted the enzyme, which is a novel membrane-bound prostaglandin E synthase-2 (mPGES-2). The transcriptional expression analysis showed the mPGES-2 is inducible to immune challenge. After RNA interference (RNAi), the hemocyte nodulation and expression of eleven antimicrobial peptides were significantly impaired. The RNAi effect was rescued by addition of PGE2 suggesting that mPGES-2 is crucial for immune responses mediated by PGE2.

4종의 diamide계열 살충제를 이용한 배추좀나방과 파밤나방의 지역계통별 감수성을 조사하였다. 배추좀나방의 경우 성주와 거창지역 집단 에서 4종의 약제 모두 추천농도에서 100%의 살충활성을 보인 반면, 평창지역 집단에서는 chlorantraniliprole에 대해 42.3%의 낮은 살충활성을 보였다. 감수성계통 배추좀나방과 저항성비를 비교한 결과 cyclaniliprole은 비교적 4지역에서 유사하거나 낮은 저항성비(0.1~6.3배)를 보인 반면, 평창지역 집단은 chlorantraniliprole (1,196.3배)와 cyantraniliprole (105.6배), flubendiamide (191.6배)은 매우 높은 저항성비를 보였다. 파밤나방의 경우 청주와 진도, 영광지역의 집단 모두 4종의 약제에 낮은 감수성을 나타났는데, 특히 청주와 진도지역의 집단은 flubendiamide에 대해 100,000배 이상의 저항성비를 보였다. 채집 연도(2014년과 2017년)에 따른 파밤나방에 대한 chlorantraniliprole의 감수성을 비교한 결과 2014년에 채집된 집단들은 모두 100%의 살충률을 보인 반면 2017년에 채집된 집단들은 살충활성이 매우 낮게 나타났다. 본 연구결과는 diamide계열 살충제가 빠르게 저항성이 발현되고 있으며 작용기작이 다른 약제와의 교호살포 등 종합적 방제전략을 수립하는데 기초자료가 될 수 있을 것이다.

Entomopathogenic fungi have been used to control pest as alternative to chemical pesticide. To kill the pest entomopathogenicfungi penetrate cuticle of pest, reach the hemocoel and utilize nutrient of host pest. Finally fungi kill the host by consumingthe host nutrient and physically damaging the tissues. But these process of fungi to control pest is needed so much timeand this point is a disadvantage for fungi. Therefore we studied other application method of fungi to control pest. Weconducted behavior test of beet armyworm to Isaria fumosoroseus which is high virulent against beet armyworm. Adultof the beet armyworm avoided oviposition at Chinese cabbage treated with I. fumosoroseus compare to control and otherhigh pathogenic isolate, Metarhizium anisopliae and this repellency of I. fumosoroseus lasted for 5days in greenhouse.Behavior of larvae to I. fumosoroseus also investigated with choice and non-choice test. Third to fifth instar larvae detectedand avoided fungi. Repellent behavior of larvae to fungi was more noticeable in younger larvae. This result may be usedto prevent the infestation of moth in crop production.

An immune signal pathway using Toll receptor in response to Gram positive bacterial or fungal challenge induces cellularimmunity and production of antimicrobial peptides (AMPs) in insects. Here we tested an hypothesis that Toll signal downstreamfactors, MyD88 and Pelle, influence eicosanoid signal by activating phospholipase A2 (PLA2) activity in the beet armywormSpodoptera exigua. We knocked-down MyD88 and Pelle expressions using specific dsRNAs and assessed their effect onPLA2 activity in immune-challenged S. exigua larvae. The RNAi-treated larvae failed to induce AMP gene expressionand PLA2 activity under immune challenge. These results shed light on cross-talk between Toll pathway and eicosanoidsignal.

Spodoptera exigua (Hübner) is an important pest with wide host range including cabbage, beans, potato etc. We evaluate the virulence of Beauveria bassiana ARP14, B. bassiana GHA, and Isaria japonica with concentration of 1×108 conidia/mL against larval stage of S. exigua using glass scintillation vial. Lethal median time (LT50) of these three fungi was not found significantly different in both fourth and second instar. Infection rate was higher in fourth instar than second instar. Infection rate was highest by B. bassiana ARP14 (84.0%) followed by B. bassiana GHA (63.6%) and I. japonica (45.8%) in fourth instars of S. exigua. Our results suggest that B. bassiana ARP14 can be a new component of integrated pest management of S. exigua.

Phospholipase A2 (PLA2) hydrolyzes phospholipids at sn-2 position to release free fatty acids. PLA2 consists of a superfamily mainly categorized as secretory PLA2 (sPLA2), cellular Ca 2+ -dependent PLA2 (cPLA2), and cellular Ca 2+ -independent PLA2 (iPLA2). We are the first to report iPLA2 in insect. Here an objective of our study is to purify a recombinant iPLA2 protein (SeiPLA2) of Spodoptera exigua using bacterial expression system. An open reading frame of SeiPLA2 was cloned into an expression vector and then transformed into Escherichia coli BL21. Over-expression with IPTG yielded recombinant SeiPLA2 (rSeiPLA2), which was then purified by an affinity chromatography using Ni-NTA column. The purified rSeiPLA2 gave significant PLA2 activity using a pyrene substrate. Its activity was inhibited by an iPLA2 specific inhibitor (BEL), but not by sPLA2 inhibitor (BPB) or cPLA2 inhibitor (MAFP

RNAi (RNA interference) is a tool for silencing of target genes through sequence-specific manner. Spodoptera exigua belongs to Noctuidae family of Lepidoptera and is serious threat to crops of economic importance. One of S. exigua chymotrypsin gene (SeCHY2) was cloned into the L4440 vector to produce sequence specific dsRNAs (double-stranded RNAs). Recombinant L4440 vectors were transformed into Escherichia coli strain HT115 (DE3). Oral delivery of bacterially expressed dsRNA gave significant larval mortality. Quantitative real-time PCR results showed that expression level of target SeCHY2 gene in the larval gut tissue was significantly down-regulated. Pretreatment with an ultra-sonication and heating to disrupt bacterial cell wall/membrane significantly increased the insecticidal activity of the transformed bacteria

A blast search using SPODOBASE gave 10 chymotrypsin (CHY) candidate genes with complete open reading frames (ORFs) of Spodoptera exigua. Among these candidates, Se-CHY2 was chosen and further analyzed in expression and RNAi. Se-CHY2 was expressed only in larval stage and especially in midgut. Its RNAi was successfully performed by double injection of specific double-stranded RNA (301 bp). qPCR showed a decrease of Se-CHY2 expression level by ~50% at 48 h and ~83% at 72 h. RNAi resulted in significant larval mortality. All treated larvae died at 96 h after dsRNA injection. In contrast, control dsRNA treatment did not give any mortality. Enzyme activity of gut juice was measured with N-succinyl-alanine-alanine-prolin-phenylalanine-p-nitroanilide substrate and significantly decreased in larvae treated with dsRNA. These results indicate that Se-CHY2 is a chymotrypsin that is secreted into midgut lumen and plays a role in nutrient digestion.

Phospholipase A2 (PLA2) catalyzes the committed step for eicosanoid biosynthesis. Two cellular PLA2 genes (iPLA2 and cPLA2) have been identified in Spodoptera exigua. PLA2 activity varied among developmental stages and different tissues. There was a positive correlation between PLA2 activities and their expression levels. Interestingly, there was significant inhibition by sPLA2 inhibitor, suggesting a presence of sPLA2 in S. exigua. An entomopathogenic bacterium, Xenorhabdus hominickii, infection suppressed expression of PLA2 genes and its extract inhibited enzyme activities. Oxindole of a bacterial metabolite of X. hominikii inhibited PLA2 activity and expression.

아이코사노이드는 탄소수 20 개의 다가불포화지방산 산화물로 구성된다. 이들 다가불포화지방산의 생합성 전구물질을 탐지하기 위해 파 밤나방(Spodoptera exigua)의 서로 다른 조직으로부터 지방산을 분리하여 GC/MS로 조성을 분석하였다. 파밤나방 5령 유충에서 소화관, 지방체, 혈구 및 체벽을 분리하고, 각 조직에서 지질을 추출하여 각각 중성지질, 당지질 및 인지질로 분리하였다. 대부분의 조직은 palmitic acid (16:0), stearic acid (18:0), oleic acid (18:1), linoleic acid (18:2) 그리고 linolenic acid (18:3)를 주요 지방산으로 함유하였다. 그러나 이들 지방산의 조성은 조직과 지질 종류에 따라 상이하였다. 지방체와 혈구세포는 이들 주요 지방산 이외에 myristic acid (14:0)와 3 종류의 미동정 지방산들이 추가로 검출되었다. 서로 다른 지질 종류 가운데 인지질은 중성지질이나 당지질에 비해 상대적으로 높은 linolenic acid를 지닌 반면 포화지방산의 함유량은 낮았다. 전체 불포화지방산의 조성도 조직과 지질 종류에 따라 상이하였다. 인지질은 지방체, 혈구 및 소화관에서 높은 불포화지방산 함유 량을 나타냈다. 세포성 인지질분해효소인 calcium-independent phospholipase A2 (iPLA2)는 지방산 조성을 조절하는 데 역할을 담당하였다. 이 유전자의 RNA 간섭은 중성지질과 인지질에서 지방산 조성의 변화를 유발하였다. 본 연구는 아이코사노이드 생합성의 전구물질로 여겨지는 아라키도닉산을 검출하지 못했다. 이는 곤충에 있어서 아이코사노이드는 포유동물과는 다른 새로운 생합성 과정을 통해 형성되는 것으로 추정된다.

Chitin synthase (CHS) is an important enzymatic component, which is required for chitin formation in the cuticles and cuticular linings of other tissues in insects. CHSs have been divided into two classes, class A and B, based on their amino acid sequence similarities and functions. Class A CHS (CHS-A) is specifically expressed in the epidermis and related ectodermal cells such as tracheal cells, while expression of class B CHS (CHS-B) is expressed in gut epithelial cells that produce peritrophic matrices. In this study, we cloned the CHS-A gene from the beet armyworm, Spodoptera exigua. The SeCHS-A mRNA was expressed in all developmental stages and specifically in the epidermis and tracheae tissue by RT-PCR analysis. Expression of SeCHS-A gene was suppressed by feeding double-stranded RNA (dsRNASeCHS-A, 150 ng/larva) in the fifth instar of S. exigua. The suppression of SeCHS-A gene expression significantly induced mortality on pupal stage. Also, larvae fed with dsRNASeCHS-A significantly enhanced pathogenicity of an entomopathogenic fungus, Beauveria bassiana ANU1. These results suggest that the SeCHS-A gene plays an important role in development of S. exigua and dsRNA, which is a specific to SeCHS-A gene, may be applied to effective pest control with B. bassiana.

Microbe have been considered as potential control agents for pest, as alternative to chemical control methods. Among entomopathogens, fungi cause the mortality by penetrating the cuticle of pest and/or by metabolites such as toxin. Not only this direct control effect of fungi, but repellency of fungi also may be used to prevent the pest. Repellence effect of fungi is considered as inhibitory factor to control termite. A study was reported in Japan that termite was able to detect and remove the conidia of fungi on their surface. The termite can escape from fungal infection and protect their colony. There is few study that insect pest such as moth can detect and avoid the virulence fungi. Therefore, we has been conducting the detection and avoidance of beet armyworm to high pathogenic fungi, Paecilomyces fumosoroseus. Adult of the beet armyworm avoided oviposition at Chinese cabbage treated with P. fumosoroseus compare to control. This result may be used to prevent the infestation of moth in crop production.