담배가루이는 전세계의 온대 및 아열대 지방에 넓게 분포하고 있으며, 한국에서는 시설재배작물의 주요 해충 이다. 1998년 충북 친천군 장미재배지에서 처음 확인되었으며, 이후 전국적으로 확산된 것으로 추정된다. 담배가 루이는 고추, 토마토, 오이 등 300종이 넘는 넓은 기주범위를 가지며, 식물체를 흡즙하여 작물의 생산성을 저하시 키고 토마토황화잎말림바이러스(tomato yellow leaf curl virus, TYLCV) 등 100여종이 넘는 바이러스를 매개한다. 담배가루이는 주로 살충제를 이용한 방제가 이루어진다. 하지만 살충제를 이용한 방제법은 살충제 저항성 개체 를 발생시킨다. 살충제 저항성 개체가 발생하게 되면 방제 효율이 급감하여 농가에 추가적인 경제적 피해를 야기 한다. 본 연구는 국내 지역별 담배가루이를 대상으로 약제별 살충제 저항성의 발생 현황을 조사하여, 지역별 효과적인 약제를 탐색하고 향후 방제 전략 수립에 기여하고자 한다. 담배가루이는 전국 15지역(파주, 양평, 화성, 양구, 횡성, 평창, 당진, 천안, 공주, 예천, 구미, 사천, 남원, 나주, 고흥)에서 채집되었다. 살충제는 작용기작 별 사용량이 많은 8종을 선정하였으며, 엽침지법을 사용하여 살충률을 확인하였다. 곤충생장조절제(insect growth regulators, IGR) 피리프록시펜계 약제는 알을 대상으로, 그 외 7개 약제는 2령약충을 대상으로 살충률을 확인하였 다. 모든 지역에서 높은 살충력을 보인 약제는 아버멕틴과 밀베마이신계, 스피노신계, METI살충제, 디아마이드 계 약제이다. 특히 아버멕틴과 밀베마이신계 약제는 살충제 권장사용농도(10ppm)보다 낮은 8.9ppm이하의 LC90 값을 보여 감수성으로 추정되며, 예외적으로 천안 지역의 25.6ppm으로 상대적으로 높은 LC90값을 보였다. 낮은 살충력을 보인 약제는 네오니코티노이드계, 설폭시민계, 피리프록시펜계, 테트론산 및 테트람산 유도체 약제이 다 특히 네오니코티노이드계 약제는 모든 지역에서 살충제 권장사용농도(50ppm)보다 높은 715ppm이상의 LC90 값을 보여, 모든 지역에서 살충제 저항성이 발생한 것으로 추정된다. 실험결과를 통해 살충제 별 살충력의 차이와 지역별 살충제 저항성을 검정하였다. 연구결과를 통해 지역별/약제별 저항성관리 및 대응전략을 수립하여 농업 생산성을 향상시키는 것에 도움이 되고자 한다.

복숭아혹진딧물(Myzus persicae)은 다식성으로 담배, 감자, 고추, 배추, 복숭아 등에 심각한 피해를 입히는 대표적인 농업해충이다. 본 연구에 서는 국내 복숭아혹진딧물 야외개체군의 λ-cyhalothrin, imidacloprid 및 flupyradifurone에 대한 약제 저항성 발달 수준과 점 돌연변이(R81T, L1014F, M918L)의 발생 여부를 확인하였다. 또한, qRT-PCR을 통해 사이토크롬 P450 유전자인 CYP6CY3 발현량을 확인하였다. 그 결과, λ -cyhalothrin은 저항성비(Resistance Ratio, RR)가 12개 모든 지역이 > 200으로 높은 저항성을 보였다. Imidacloprid와 flupyradifurone은 YS, UR, HY, 그리고 WJ 개체군에서 > 200의 저항성비로 높은 저항성을 나타냈다. R81T는 12개 집단 중 약 50%, L1014F는 약 33.3%, M918L은 100%에서 발현하였다. 또한 qRT-PCR을 통해 imidacloprid 저항성 개체에서 subunit CYP6CY3의 발현량이 높게 나타난 것을 확인하였다. 이러 한 결과는 M918L 점 돌연변이는 λ-cyhalothrin 저항성 진단마커로, R81T와 CYP6CY3의 높은 발현은 imidacloprid 저항성 진단마커로 활용이 가능하다는 것을 시사한다.

담배가루이(Bemisia tabaci)는 국내 주요 농업 해충으로서 고추, 오이, 토마토 등의 시설재배 작물에 큰 피해를 입힌다. 담배가루이는 작물을 흡즙하여 식물체의 상태를 악화시키고 TYLCV(tomato yellow leaf curl virus)등 100 여 종의 바이러스를 매개하며, 배설물을 통해 그을음병을 유발한다. 가장 일반적인 담배가루이의 방제방법은 빠르고 높은 효과를 지닌 살충제 살포이다. 하지만, 지속적인 화학적 방제는 해충의 살충제 저항성을 야기한다. 살충제 저항성은 방제 효율 감소와 농가의 경제적 손실을 일으킨다. 본 연구는 국내 지역별 시설 고추 재배지에서 발생한 담배가루이의 약제별 저항성을 조사하여, 지역별로 효과적인 약제 선정 및 대체 약제 추천 등 지속 가능한 해충관리법을 제시하고자 한다. 담배가루이는 전국 13개(파주, 양평, 화성, 횡성, 당진, 천안, 공주, 예천, 구미, 사천, 남원, 나주, 고흥) 지역에서 채집되었다. 작용기작 별 사용량이 많은 8종을 선정하였으며, 엽침지법을 사용 하여 살충률을 확인하였다. 곤충생장조절제(insect growth regulators, IGR)인 피리프록시펜계 약제는 알을 대상 으로, 그 외 7개 약제는 2령약충을 대상으로 살충률을 확인하였다. 높은 살충력을 보인 약제는 아버멕틴, 밀베마 이신계, 스피노신계, METI살충제, 디아마이드계 약제이다. 아버멕틴, 밀베마이신계는 권장사용농도로 처리 시 최소 84.5%, 최대 100% 살충률을 보였다. 스피노신계는 최소 86.3%, 최대 90.6% 살충률을 보였으나 천안, 파주, 사천에서 각각 59.7%, 66.6%, 79%로 다른 지역에 비해 상대적으로 낮은 살충률을 보였다. METI살충제는 최소 81.1%, 최대 98% 살충률을 보였으나 나주, 예천, 천안, 사천이 각각 61.6%, 68.8%, 68.9%, 69.2%의 상대적으로 낮은 살충률을 보였다. 디아마이드계는 최소 83.9%, 최대 91.9% 살충률을 보였으나 천안과 구미가 각각 56.6%, 59.8%로 상대적으로 낮은 살충률을 보였다. 낮은 살충력을 보인 약제는 네오니코티노이드계, 설폭시민계, 테트 론산 및 테트람산 유도체, 피리프록시펜계 약제이다. 네오니코티노이드계는 최소 14.1%, 최대 41.9% 살충률을 보였다. 설폭시민계는 최소 17%, 최대 42.8% 살충률을 보였다. 테트론산 및 테트람산 유도체는 최소 30.5%, 최대 54.9% 살충률을 보였으나 천안이 14.4%로 특히 낮은 살충률을 보였다. 피리프록시펜계는 최소 55.3%, 최대 64.3% 살충률을 보였으나 횡성이 72.2%로 상대적으로 높은 살충률을 보였으며, 파주가 35%로 상대적으로 낮은 살충률을 보였다. 실험 결과를 통해 살충제 별 살충력의 차이와 지역별로 살충률의 차이를 확인하였다. 본 연구결 과를 통해 효과적인 약제 추천과 지역에 따라 다른 방제 전략 제시에 도움이 되고자 한다.

담배가루이(Bemisia tabaci (Gennadius))는 전세계적으로 다양한 작물을 가해하고 있는 흡즙성 해충이다. 국내에서는 1998년에 최초로 유입이 보고되었고 현재 전국에 널리 퍼져 농가에 경제적으로 큰 손실을 주고 있다. 담배가루이의 주 방제방법인 화학적 방제를 위해서는 정확한 약제 저항성 진단을 바탕으로 한 약제 선택이 중요하다. 따라서 본 연구에서는 2020년 6월부터 10월까지 전국 7개도 12지역의 온실에서 담배가루이를 채집하여 잔류접촉법 기반 생물검정 및 분자마커를 통해 약제 저항성을 진단하였다. 그 결과, 작용기작이 다른 5종 약제(dinotefuran, spinosad, emamectin benzoate, chlorfenapyr 및 bifenthrin)에 대해 전국적으로 높은 저항성을 지니고 있으며 유기인계 및 피레스로이드계 약제에 대한 저항성 점돌연변이가 포화상태임을 확인하였다. 이는 약제 저항성 모니터링에 기반한 담배가루이 약제 저항성 관리가 시급하다는 것을 시사한다.

For our survey of insecticidal resistance of Palm thrips (Thrips palmi Karny), we established the discriminating time (DT) and concentration (DC) of nine insecticides, and we conducted a bioassay about seven local populations via leaf-dipping methods. The discriminating times of the recommended concentration (RC) were 24 h at emamectin benzoate EC and spinetoram SC, 48 h at chlorfenapyr EC, 72 h at spinosad SC, cyantraniliprole EC, acetamiprid WP, dinotefuran WG, imidacloprid WP and thiacloprid SC after treatment. The DC estimated the concentration which showed the difference within the mortalities of these local populations. The DCs were emamectin benzoate EC 0.013 mg L-1 (RC, 10.8 mg L-1), spinetoram SC 0.125 mg L-1 (RC, 25.0 mg L-1), chlorfenapyr EC 0.25 mg L-1 (RC, 50.0 mg L-1), spinosad SC 0.083 mg L-1 (RC, 50.0 mg L-1) and cyantraniliprole EC 5.0 mg L-1 (RC, 50.0 mg L-1), and DCs of neonicotinoids were their RCs, that is, acetamiprid WP (RC, 40.0 mg L-1), dinotefuran WG (RC, 20.0 mg L-1), imidacloprid WP (RC, 50.0 mg L-1) and thiacloprid SC (RC, 50.0 mg L-1). From our investigation into the resistance of the local populations with DT and DC application, the neonicotinoid insecticides have shown a high resistant level for all the local populations, and the other insecticides have demonstrated low or non-resistance. In the use of neonicotinoid insecticides to control Palm thrips, one must take caution. As a result, the establishment of DT and DC in the single dose bioassay method was helpful for surveying the insecticide response dynamics and the development of an insecticide resistance management strategy.

Aedes albopictus is an important vector for yellow fever, dengue fever, chikungunya fever and Zika virus. This mosquitohas been exposed to organophosphates and pyrethroids in Republic of Korea for a long time. Using the direct contactmortality bioassay, susceptibility strain and two field populations of Ae. albopictus from Busan, and Damyang. Insecticidalproperty were indicated to RR ratio values (Resistant ratio to strain of susceptibility) of LC50 (Midian Lethal concentration)to organophosphats and pyrethroids. Genomic DNA was used for Allele-specific PCR (AS-PCR) genotyping of the acethylcholineesterase to detect Gly119Ser mutations and kdr gene to detect Phe1534Cys mutations. Detoxification enzyme activitiesof Ae. albopictus from Busan and Damyang were assessed using microplate enzyme activity assays. Activities of fourdetoxification enzymes eg., glutathione S-transferase(GST), Non specific esterases (α-naphthyl acetate and β-naphthyl acetate),and cytochrome C oxidase were determined for each Ae. albopictus strain. This study might suggest that Ae. albopictuscontrol programs should be prepared for the management of organophosphate and pyrethroid insecticide resistance.

The small brown planthopper (SBPH), Laodelphax striatellus Fallen, is an important pest that causes severe yield losses by transmitting plant viruses to rice. For the efficient control of SBPH in Korea, the respective resistance levels in the immigrant and indigenous populations need to be discerned. The resistance levels to 10 insecticides (three carbamates, an organophosphate, four neonicotinoids, and a phenylpyrazole) were evaluated in 21 field populations collected from either SBPH-immigrating or indigenous regions during two different seasons (early spring vs. late summer). Imidacloprid resistance was most widely observed in many regional populations, followed by thiamethoxam resistance. Interestingly, the resistance level to imidacloprid was significantly higher in both immigrant and late-summer-collected populations than in indigenous and early spring-collected populations, respectively [3.3- (p = 0.018) and 2.6-fold (p = 0.026)]. Moreover, the late summer immigrant population exhibited higher imidacloprid resistance (2.4-fold) than the early spring-collected population from the same region, suggesting that the migratory SBPH that immigrated into Korea already exhibited imidacloprid resistance traits and were further selected after inhabitation. All field populations showed little resistance to fipronil (0.1- to 0.7-fold), suggesting that it is the most effective among the tested insecticides to control field populations of SBPH. The coefficient of variation of the resistance ratio (RR) among different regional populations and the correlation coefficient of RR among different insecticides have been suggested as supplementary parameters when determining appropriate insecticides as respective indicators for the dispersion status of resistance among SBPH populations and the possibility of cross resistance among tested insecticides.

Tetranychus urticae was collected from greenhouse roses to monitor the development of acaricide resistance. Dose-mortality lines were estimated on 16 regional populations with 13 acaricides. For each acaricide, LC50s of the populations were plotted to check normality. LC50s of eight acaricides showed normal distribution and five others did not. An index of Ln (recommended dose/LC50 for each acaricide) checked the development of resistance to populations. The index is based upon recommended dose to control a pest stage and empirical LC50 got from serial dilution range for each pesticide. We tried to categorize acaricides by index due levels of effectiveness to mites: tebufenpyrad, fenpyroximate, bifenthrin, and fenbutatin oxide as non-effective acaricides with less than index 1.0, chlorpyrifor+bifenthrin and milbemectin as alert level placed between 1.0 and 2.0, acequincyl as caution between 2.0 and 3.0, and abamectin, cyflumetofen, bifenazate, chlorfenapyr+fulacrypyrim and propargite as effective over 3.0. We also tested the similarity of acaricide actions for choosing effective acaricides against resistant mites and of populations for resistance management. We could make several acaricides groups: group1 including abamectin, cyflumetofen, and bifenazate group 2 propargite and chlorpyrifos+bifenthrin group 3 chlorfenapyr and acequinocy and group 4 mibemectin, fenyroximate, and bifenthrin, by which we can suggest not to use acaricide within the same group to avoid the resistance development. Populations grouping would imply similar practices of acaricide use, so we can manage pesticide usage, effectively. Group A includes Gimhae 2, 3, 4, 5, Jincheon2, and Taean, and Group B includes Goyang, Gangjin1, Paju, Gangin2 and Namwon.

An easy and rapid resistance detection protocol for the Western flower thrips Frankliniella occidentalis was established based on the residual contact vial bioassay (RCV), in which insecticide resistance levels can be estimated at 8 h-post treatment of diagnostic doses. The RDA strain was used as a reference susceptible strain, which has been reared under laboratory conditions over 10 years without exposure to any insecticides. Seven insecticides were tested for the determination of diagnostic dose. Among them, five insecticides (chlorfenapyr, acrinathrin, spinosad, emmamectin benzoate and thiamethoxam, ranged as 0.03 ~ 0.42 μg-1cm2) were applicable to the RCV. However, two insecticides (omethoate and imidacloprid) were not able to be used for the RCV because the treated inner surface of glass vials by these insecticides were too viscous, causing non-specific mortality. The RCV detection kit was employed for the estimation of resistance levels for the five insecticides in five local populations. Almost field-collected populations revealed high levels of resistance to the four insecticides (acrinathrin, spinosad, emmamectin benzoate and thiamethoxam) by showing less than 50% mortality. The baseline resistance detection by RCV method will facilitate the selection of proper insecticides for farmers to manage insecticide resistant-populations of F. occidentalis.

Nilaparvata lugens Stål is one of the important migratory pests of rice paddy fields in Korea. Resistance levels to nine insecticides were monitored in 12 local strains and correlation analysis was conducted to determine cross-resistance relationships among the tested insecticides. The local strains revealed 1.3- to 28.0-, 1.6- to 6.0-, 2.8- to 237.0-, 0.6- to 0.9-, and 0.7- to 1.3-fold resistance to carbamates, organophosphates, neonicotinoids, fipronil and etofenprox, respectively. Organophosphates revealed moderate correlations with benzofuranyl methylcarbamate (r = 0.566 - 0.614, p > 0.01). Three neonicotinoids were not correlated each other, but imidacloprid and clothianidin were moderately correlated with several benzofuranyl methylcarbamate and organophosphate insecticides (r = 0.590 - 0.705, p > 0.05), indicating that unknown common factors (such as detoxification enzymes) might contribute to resistance to both insecticides. Fipronil and etofenprox exhibited low levels of resistance and cross-resistance with other insecticides, suggesting their potential as an effective insecticide for field application. Resistance level monitoring and correlation analysis would be valuable for the selection of appropriate insecticides to control insecticide-resistant N. lugenes, a typical migratory pest in Korea.

Vector mosquitoes are related with various diseases on human, such as malaria (Anopheles sinensis (L.), An. sinensis), Japanese encephalitis (Culex tritaeniorhynchus (L.), Cx. tritaeniorhynchus), yellow fever (Aedes albopictus (L.)), dengue hemorrhagic fever (Ae. albopictus) and filariasis (Ae. albopictus), as well as nuisance insect pests (Culex pipiens pallens Foskal, Cx. p. pallens). Continued and repeated use of conventional insecticides such as organophosphorus and carbamates, pyrethroids has often resulted in the widespread development of resistance and has undesirable effects on non-target organisms and environment. Particularly, widespread insecticide resistance has been a major obstacle in the cost-effective integrated vector management program. In theRepublic of Korea (ROK), studies on mass vector mosquito control using insecticides were mainly carried out until 1970’s such as a fuselage-mounted ULV spray system to C-46 aircraft, effectiveness of ground aerosols ULV premiumsumihtion, a helicopter application of ULV Dibrom and residual toxicity of organophosphates in animal shelters for Japanese Encephalitis (JE) and Plasmodium vivax malaria vector mosquito control. Regional insecticide resistance of vector mosquitoes has been annually evaluated since ,early 1980’s, the first detailed study on susceptibility of seven mosquito species to 13 organophosphorus insecticides revealed the DDT and lindane resistance of Cx. tritaeniorhyunchus. Marked annual and regional variations of insecticide resistance have been observed. In comparative resistance, the resistance ratios of various insecticides were compared among each other using two strains of vector mosquitoes that were collected from same locality. An. sinensis from Koyang-si in 1992 and 1981 demonstrated >100 folds and >3000 folds of higher resistance to DDT and to bio-resmethrin. An. sinensis from Paju-si in 2008 demonstrated 30 folds of higher susceptibility to fenthion than those collected in seven years ago. Cx. tritaeniorhynchus from Gwanju in 2010 and in 1992 showed >100 folds of higher resistance to pyrethroids. In comparative regional resistance monitoring, Cx. p. pallens from six localities in the ROK in 2008 and six geospatially-distant field Cx. tritaeniorhynchus strains in 2011 showed marked regional resistance variations. Field populations of Cx. tritaeniorhynchus from agricultural areas demonstrated extremely higher insecticide resistance to pyrethroids than those from non-agricultural areas. Cx. tritaeniorhynchus populations from rice paddies had been under heavy selection pressure due to pyrethroids, which were used for a long time to control agricultural pests in the localities. These results indicate that careful selection and rotational use of these insecticides may results in continued satisfactory control against field populations of vector mosquitoes

The cotton aphid, Aphis gossypii (Glover) (Hemiptera:Aphididae), is a highly polyphagous pest that directly or indirectly damages cultivated plants. Six field-collected populations of cotton aphid, A. gossypii (BY-A, BY-B, YJ-A, YJ-B, CJ-A, and CJ-B) were tested for susceptibility to 14 different insecticides. Most population exhibited high to very high levels of resistance to neonicotinoid. Among them, a strain showing resistance to imidacloprid were selected and showed 1,543-fold in resistance as compared with susceptible strain. Piperonyl butoxide (PBO), diethyl maleate (DEM), and S,S,S tributyl-phosphorothiolate (DEF) failed to synergize imidacloprid in this resistant population. In addition, the activity of detoxification enzymes (P450, EST, GST) were no differences between susceptibility and imidalcoprid resistance strain. However, by analyzing the nicotinic acetylcholine receptor β1 subunit loop D, R81T point mutation was detected in BY-A, BY-B, YJ-A, and YJ-B strain.

The resistance levels to carbamate (CB) and organophosphate (OP) insecticides were determined by topical application in 14 field strains of Nilaparvata lugens. The resistance levels to CB and OP were 1.3~47.5-fold and 1.4~14.4-fold higher than a susceptible strain, respectively. A quantitative sequencing (QS) protocol was established to determine the allele frequencies of four acetylcholinesterase point mutations putatively associated with CB and OP resistance. The allele frequencies of the four mutations (G119A, F/Y330S, F331H and I332L) in field strains ranged from ca. 0.0~51.7%, 0.0~88.9%, 2.5~47.7%, 5.1~56.0% and 6.7~57.3%, respectively. The F331H and I332L were tightly linked each other, suggesting these mutations may occur simultaneously. In the correlation analysis, G119A was not well correlated with actual resistance levels (r2= < 0.232), whereas F331H and I332L showed a better correlation with the resistance levels of benzofuranyl methylcarbamates (r2= 0.595). This finding indicates that F331H and I332L mutation frequencies may be used as molecular markers for detecting carbamate resistance in N. lugens. A QS protocol detecting the F331H and I332L mutation frequencies would be employed as a supportive tool for rapid monitoring of CB insecticide resistance levels in N. lugens

The green peach aphid (Myzus persicae) is a serious pest of agricultural and horticultural crops all over the world. M. persica has rapidly developed resistance to a wide variety of insecticides, including carbamates. The E4/FE4 carboxylesterase is known to be involved in carbamate resistance. To compare the E4/FE4 carboxylesterase gene copy number, as a genetic resistance marker, between seven field strains, quantitative real-time PCR (qPCR) was performed. In addition, quantitative sequencing (QS) was employed to predict the frequencies of acetylcholinesterase (AChE) mutations (A301S and S431F) that are associated with target site insensitivity. All M. persica strains examined possessed the S431F mutation in the heterozygous state except for a susceptible strain, implying the possibility of AChE duplication. In contrast, no A301S mutation was found. Frequency prediction equation was generated from the plots of signal ratios and amplification critical time, which showed a high correlation (r2>0.996). QS analysis of M. persicae populations revealed that the allele frequency of S431F ranged 4% to 63%. Taken together, the AChE resistance allele frequencies determined by QS and the E4/FE4 gene copy number by qPCR should facilitate the detection and monitoring of carbamate resistance in M. persicae in the field.