To identify whether higher expression of carboxylesterase (CbE) E4 in Myzus persicae is due to gene duplication, gene copy number was determined by quantitative real-time PCR. In addition, to determine the actual protein concentration of CbE E4 and it activity, Western blotting and activity staining were conducted. CbE gene copy number was highly correlated with carbamate resistance ratio (r2=0.934). However, CbE E4 expression level was little correlated with insecticide resistance ratio (r2<0.046) and no apparent correlation was observed among the gene copy number, protein quantity and total activity of CbE E4. Therefore, it was assumed that not only quantitative changing but also qualitative alteration of CbE E4 occurred in M. persicae. To investigate any potential alteration of CbE E4, mutation survey was conducted by sequencing of CbE E4 from various local strains of M. persicae. G137D and W251L mutations have been known as the main mutations associated with structural change leading to resistance. Interestingly, a new G134C mutation, which is in proximity of G137D mutation, was identified in the oxyanion hole of CbE E4. To predict the functional role of this mutation in resistance, 3-dimensional structure modeling was conducted. In summary, CbE E4 appears to be involved in resistance to both pyrethroids and carbamates as a nonspecific hydrolase or sequestration protein in M. persicae.

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 green peach aphid, Myzus persicae (Sulzer), is one of the most serious pests in cabbage cultivation. Field survey was carried out to know the insecticide resistance levels and to develop the applicable insecticide resistant markers in five main cabbage cultivation regions (Pyeong-chang, Hong-cheon, Bong-wha, Mu-ju and Je-ju) during 2009 to 2011. M. persicae can resist a wide range of insecticides in five surveyed local populations. Therefore multi resistant (MR) strain was selected from these five local populations and esterase over-expression, modified AChE (MACE) and mutation(s) in para-type sodium channel were analyzed using native IEF and quantitative sequencing with five local populations. Esterase over-expression and MACE (StoF mutation) were observed in all populations including MR strain. LtoF mutation is well known as a kdr mutation in para-type sodium channel. However, even though LC50 values of MR strain noted over 2,000 times higher than that of susceptible strain against bifenthrin, LtoF mutation was not detected in para type sodium channel and also local populations. We found another mutation (MtoL) in para and that mutation highly correlated between mutation ratio and bioassay data. For preliminary resistance monitoring, we developed quantitative sequencing (QS) to detect the frequencies of point mutation as a population genotyping. These methods can apply to manage M. persicae resistant populations in field.

The green peach aphid, Myzus persicae (Sulzer), is one of the most serious pest in cabbage cultivation. Field survey was carried out to know the insecticide resistance levels in five main cabbage cultivation regions (Pyeong-chang, Hong-cheon, Bong-wha, Mu-ju and Je-ju) in 2009~2011. The green peach aphid can resist a wide range of insecticides in five surveyed local populations. Among the nine tested insecticides, four chemicals (methomyl, bifenthrin, pymetrozine and flonicarmid) showed less than 60% mortality in the recommended concentration in most years of local populations. Multi resistant (MR) strain was selected from these populations and modified AChE (MACE: StoF mutation), MtoL mutation in para-type sodium channel, esterase over-expression were observed in almost of all populations including MR strain. Especially, StoF and MtoL mutations were highly correlated with resistance ratio and population based quantitative sequencing results. Therefore, these results suggested that molecular-biology based resistance monitoring can applied resistance management in M. persicae.

Toxicities of 10 insecticides were examined against late third instars of Culex tritaeniorhynchus using the direct-contact mortality bioassay. Six geospatially distant field mosquitoes were collected from Chuncheon-si (designated CC-CT), Hwaseong (HS-CT), Seosan (SS-CT) Jeonju (JJ-CT), Daegu (DG-CT), and Busan (BS-CT) in the Republic of Korea (ROK). Marked regional variations of insecticide susceptibility were observed. Field populations of SS-CT, JJ-CT and DG-CT from agricultural areas showed higher to extremely higher insecticide susceptibility to pyrethroids than those of CC-CT, HS-CT and BS-CT strains from none agricultural areas. Extremely high to low levels of susceptibility were measured: bifenthrin, susceptible ratio (SR) = 2.7–896.3; β-cyfluthrin, SR = 1.8–633.3; α-cypermethrin, SR = 1.2–1,051.9; deltamethrin, SR = 1.3–711.1; permethrin, SR = 1.5–1,053.4 etofenprox, SR = 2.2–29.3; chlorfenapyr, SR = 5.1–103.6; chlorpyrifos, SR = 2.3–337.0; fenitrothion, SR = 2.0–142.3 and fenthion, SR = 1.4–186.2. Culex tritaeniorhynchus populations from rice paddies had been under heavy selection pressure due to the agricultural insecticides and that’s why the mosquito species demonstrated high resistance 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 mayresult in continued satisfactory control against field populations of Japanese encephalitis vector mosquitoes.

Five field-collected populations (TR-CP, UR-CP, JR-CP, GR-CP, and BR-CP colonies) of northern house mosquito, Culex pipiens pallens, identified by polymer chain reaction (PCR) were tested for susceptibility to 10 different insecticides by direct-contact mortality bioassay in comparison with an insecticide-susceptible KS-CP strain. Marked regional variations of insecticide susceptibility were observed. Extremely high to high levels of resistance were obtained: bifenthrin, resistance ratio (RR) = 1–520; β-cyfluthrin, RR = 16–397; α -cypermethrin, RR = 9–343; deltamethrin, RR = 1–40; etofenprox, RR = 2–42; permethrin, RR = 2–12; chlorpyrifos, RR = 2–675; fenitrothion, RR = 0.5–364; and fenthion, RR = 2–360. All of the colonies were susceptible to one or more of the insecticides examined. These results indicate that careful selection and rotational use of these insecticides should result in continued satisfactory control against field populations of northern house mosquito.

The green peach aphid, Myzus persicae (Sulzer), is one of the most serious pest in cabbage cultivation. Field survey was carried out to know the insecticide resistance levels in five main cabbage cultivation regions (Pyeong-chang, Hong-cheon, Bong-wha, Mu-ju and Je-ju) in 2009. The green peach aphid can resist a wide range of insecticides in five surveyed local populations. Among the nine tested insecticides, four chemicals (methomyl, bifenthrin, pymetrozine and flonicarmid) showed less than 50% mortality in the recommended concentration in all populations. Multi resistant (MR) strain was selected from these populations and esterase over-expression, modified AChE (MACE) and mutation(s) in para-type sodium channel were analysed using native IEF and quantitative sequencing with five local populations. Esterase over-expression and MACE (StoF mutation) were observed in all populations including MR strain. LtoF mutation is well known as a kdr mutation in para-type sodium channel. However, even though LC50 values of MR strain noted over 2,000 times higher than that of susceptible strain against bifenthrin, any mutation was not detected in para type sodium channel and also local populations. These results suggested that unusual case could be existed in pyrethroid resistance mechanism in green peach aphid.

Insecticide resistance development of insect pests is one of the main threatening factors against stable crop production and keeping human life safe from insect pest disease vector. To know properties of insecticides resistance is very important to develop a management program against insecticide resistance pests. Insecticide resistance development is an event of evolutionary process. We have to deal with insecticide resistance of insect pests by interaction between insecticide and insect pests in their chemical, physical and biological environment. We can glimpse at and infer the evolutionary process of insects from investigating and comparing the resistance level of insect pests to insecticides. Huge amount and many kinds of insecticides have been sprayed to control insect pests. Development of insecticide resistance of insect such as housefly and mosquito, known as most common medical insect, is a result of adaptation to environment covered with insecticides. We can easily assume that the amount and kind of insecticides sprayed the evolutionary force to the diversity of insecticide resistance. The resistance diversity will be very good character of good markers for determining geographical isolation.

The stable fly, Stomoxys calcitrans L., is an important pest of livestock. Stable flies are considered as mechanical vectors of veterinary disease. Pyrethroids and organophosphates have been widely used for stable fly control. To establish resistance monitoring molecular tool, we isolated the partial cDNA and genomic fragments of voltage-sensitive sodium channel and acetylcholinesterase genes encompassing the well known conserved sites for resistance-associated mutations. To examine the current status of stable fly resistance to pyrethroids and organophosphates mediated by the nerve insensitivity mechanism in Korean population of S. calcitrans, DNA-based genotyping in conjunction with residual contact vial (RCV) bioassay were conducted with 11 representative regional field populations. No resistance-associated mutations were detected in these S. calcitrans populations, suggesting that these populations are likely still susceptible to both pyrethroids and organophosphates. Establishment of RCV bioassay protocol and availalbility of target site sequence information will greatly facilitate resistance monitoring of S. calcitrans in the field.

An attempt was made to stimulate future research by providing exemplary information, which would integrate published knowledge to solve specific pest problem caused by resistance. This review was directed to find a way for delaying resistance development with consideration of chemical(s) nature, of mixture, rotation, or mosaics, and of insecticide(s) compatible with the biological agents in integrated pest management (IPM). The application frequency, related to the resistance development, was influenced by insecticide activity from potentiation, residual period, and the vulnerability to resistance development of chemical, with secondary pest. Chemical affected feeding, locomotion, flight, mating, and predator avoidance. Insecticides with negative cross-resistance by the difference of target sites and mode of action would be adapted to mixture, rotation and mosaic. Mixtures for delaying resistance depend on each component killing very high percentage of the insects, considering allele dominance, cross-resistance, and immigration and fitness disadvantage. Potential disadvantages associated with mixtures include disruption of biological control, resistance in secondary pests, selecting very resistant population, and extending cross-resistance range. The rotation would use insecticides in high and low doses, or with different metabolic mechanisms. Mosaic apply insecticides to the different sectors of a grid for highly mobile insects, spray unrelated insecticides to sedentary aphids in different areas, or mix plots of insecticide-treated and untreated rows. On the evolution of pest resistance, selectivity and resistance of parasitoids and predator decreased the number of generations in which pesticide treatment is required and they could be complementary to refuges from pesticides To enhance the viability of parasitoids, the terms on the insecticides selectivity and factors affecting to the selectivity in field were examined. For establishment of resistant parasitoid, migration, survivorship, refuge, alternative pesticides were considered. To use parasitoids under the pressure of pesticides, resistant or tolerant parasitoids were tested, collected, and/or selected. A parasitoid parasitized more successfully in the susceptible host than the resistant. Factors affecting to selective toxicity of predator are mixing mineral oil, application method, insecticide contaminated prey, trait of individual insecticide, sub-lethal doses, and the developmental stage of predators. To improve the predator/prey ratio in field, application time, method, and formulation of pesticide, reducing dose rate, using mulches and weeds, multicropping and managing of surroundings are suggested. Plant resistance, predator activity, selective insect growth regulator, and alternative prey positively contributed to the increase of the ratio. Using selective insecticides or insecticide resistant predator controlled its phytophagous prey mites, kept them below an economic level, increased yield, and reduced the spray number and fruits damaged.

The susceptibility of the Blattella germcanica (L.) in the Republic of Korea (ROK) to insecticides was evaluated under laboratory conditions using 12 insecticides currently used by the local public health centers and/or pest control operators in the ROK. The insecticides included seven pyrethroids and five organophosphages. Based on their LD50values, the order of susceptibility of B. germanica adults to the insecticides was chlorpyrifos-methyl, profenofos, and chlorpyrifos with the values of 0.07, 0.29, and 0.88 ㎍/♀, respectively. The least susceptibility was obtained with tetramethrin at LD50 of 7.39 ㎍/♀. In the comparative resistance test, the resistance ratios (RR) of 12 insecticides were compared to each other using field collected B. germanica adults in Seoul between 1993 and 2007. Blattella germanica demonstrated higher RRs to pyrethroids such as λ-cyhalothrin, and low RRs among the organophosphates. Among the pyrethroids, λ-cyhalothrin had the highest RRs of 111 and 129 fold differences at LD50 and LD90 values, respectively. Among the organophosphates, profenofos was observed to have the highest RRs of 4 and 15 fold differences at LD50 and LD90 values, respectively. However, there were no significant differences in susceptibility to tetramethrin, chlorpyrifos, and fenitrothion. Blattella germanica was more susceptible to pyridafenthion showing a 0.7 fold difference in a resistance rate (RRLD50), respectively. Resistance ratio of tetramethrin was low, but susceptibility was also not high.

Six kinds of insecticides, fenpropathrin, dicofol, milbemectin, tebufenpyrad, monocrotophos and propargite, were selected for investigating LC50s to local two spotted spider mite(TSSM) strains. The TSSMs were collected from peach (Cheongdo area) and apple orchards (Chungju, Kunwi and Sobo areas) in 2005, from strawberry plastic house (Nonsan and Damyang areas) in 2006 for experiment and some insecticides selected TSSM strains for more than 20 generations were also used. Each strain was clustered by LC50 value got from six selected insecticides treatment. Every insecticide showed about three-fourfold LC50 difference among TSSMs caught in same area. There are no specific differences in LC50 patterns of TSSM to insecticides within peach orchard and among apple orchards. But TSSMs of apple orchard and peach orchard were clustered into different group each other. Grouping of selected TSSM strains with insecticides in lab artificially showed good relationship between action mechanisms of insecticides. In strawberry strains TSSM showed regional differences between Damyan and Nonsan. The TSSMs of Nonsan showed more resistance to Tebyfenpyrad and Propargite, but TSSM of Damyang was more resistant to fenpropathrin than that of Nonsan. The TSSMs selected with monocrotophos and bifenazate, known as acetylcholinesterase inhibitors, and also pyridaben and chlorfenapyr, known as inhibitors of mitochondrial electron transportation, were grouped close respectively.

에 의한 살충제 저항성 사험결과 지역에 따라 pirimicarb가 49~830배 phosphamidon이 31~536배, demeton S-methyl이 5~204배의 약제 저항성이 확인됨으로서 사과에 조팝나무진딧물이 포장에서 방제결과가 없는 난방제해충이 되었음을 알 수 있다. 이와 같이 약제에 따라, 지역에 따라 저항성차가 크게 나타나는 원인을 공시충을 채집한 과수원의 살충제 산포상황을 조사한 결과 저항성이 가장 높았던 예산과 이천-1의 개체군을 채집한 과수원에서는 살충제를 4회 이상산포하였고, 저항성이 가장 낮았던 개체별 esterase 활성을 활성지수별로 분포비율을 비교한 결과 활성지수가 원주는 낮은 곳에, 이천-1과 예산은 높은 곳에 집중적으로 분포하였으며, 효소의 활성지수로써 저항성도를 계산한 결과 원주는 44.5, 이천-1과 예산은 92.0이었고 기타의 개체군들은 약제산포회수가 적어 이 사이에 모두 분포하였음을 알 수 있어 포장에서 약제산포회수가 많을수록 효소의 활성이 높게 나타났고 저항성비도 높았다. 이들중 광주(조팝나무)를 S크론, 과천(사과)을 R크론으로 하여 AChE의 조해제에 대한 감수성을 조사한 결과 pirimicarb는 299.2배, phosphamidon은 186배나 저하되었다. 이로써 약제저항성이 높았던 크론에서 에스테라제의 활성이 높았고, AChE의 감수성이 저하되는 것으로 보아 조팝나무진딧물의 약제저항성기작에는 적오도 두가지 이상의 요인이 작용하고 있음이 확인되었다.