Two-spotted spider mite (Tetranychus urticae Koch; TSSM) is an agriculturally serious pest tat has acquired strong resistance against acaricide. Alternatively, mite-pathogenic fungi could be used to control the mites. The spider mite has symbiotic microorganisms which could be involved in the physiological and ecological adaption to biotic stress. In this study, mite-pathogenic fungi were used to control female adults, and the change of microbiome in the fungus-infected mites was analyzed. The acaricidal activity of fungal isolates was tested, and Akanthomyces attenuatus JEF-147 with the highest acaricidal activity was determined, followed by semi-field assay. Microbiome in the female adults which was infected by A. attenuatus JEF-147 was analyzed, and composition of microorganism was changed by fungal treatment. In bacteria abundance, the arthropod defense-related Rickettsia increased, but reproduction-associated Wolbachia decreased. The change in major bacterial abundance in the infected mites could be explained by the trade-off between reproduction and immunity. This work describes that a possible trade-off in arthropods against fungal pathogens could be predicted by the microbiome analysis.

The two-spotted spider mite, Tetranychus urticae Koch, is one of the most important agricultural pests. Therefore, we screened fifteen compounds from natural products for their spatial repellent and oviposition deterrent activities against T. urticae in the laboratory by using two-choice and no-choice tests. In the bridge two-choice test, nine compounds showed the spatial repellent effects on T. urticae at 20 mg dose, resulting in reduced numbers of eggs. Among the nine compounds, at 2 mg dose, two compounds were selected as having more spatial repellent activity than the others. The two compounds also showed spatial repellent and oviposition deterrent effects in the two-choice test from hosts. In the no-choice test from a host, the spatial repellent effects of the two compounds to T. urticae were significantly stronger than that of controls. These results suggest that the findings can be used as potential agents for the prevention and population control of T. urticae in the field.

The two-spotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae), is one of the most important pest species, because it devastates many horticultural and ornamental crops and fruit trees. The resistance ratios calculated for the LC50 value in acequinocyl- and pyridaben-resistant strain was 4,237- and 5,555-fold higher than that of the susceptible strain, respectively. The objective of this study was to evaluate the cross-resistance to several acaricides and to identify the mechanisms associated with acequinocyl- and pyridaben-resistant strain of T. urticae.

Tetranychus urticae and Myzus persicae are one of the most serious insect pests in many crops, vegetables, flowers, and fruit trees worldwide. Many insecticides have been developed to control green peach aphid and two spotted spider mite, but resistance to almost all insecticides has reduced their control effect. Particular groups of plant-beneficial microbials are not only root colonizers that provide plant disease suppression, but in addition are able to infect and kill insect larvae. Antimicrobial compounds produced by biocontrol microbes are effective weapons against a vast diversity of organisms such as fungi, nematodes, and viruses. In this study, we evaluated the effectiveness of mixtures plant extracts and improvement of culture process biocontrol microbials on insecticidal activity. Azadirachta indica and Derris elliptica mixed with micorbials, which are nutrient sources of mung bean extract and lecithin, were more effective than other the mixtures. Leaf spraying with the mixture of Pseudomonas fluorescens significantly showed the highest insecticidal power in vivo for 24 hours after treatment. The effect of spraying mixture was more than 50% at 2000 times dilution, and the spraying concentration of 90% or more showed a dilution of up to 500 times. Our results indicated that the nutrient sources of microbe act as a key antimicrobial metabolite in biocontrol of insect pests, and mixing with plant extracts can provide synergistic effects as an optimal usage of the biocontrol agents.

The two-spotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae), is one of the most important pest species devastating many horticultural and ornamental crops and fruit trees. In the present study, a field strain was collected in January 2001 and has been selected for sixteen years with acequinocyl. The resistance ratio calculated from the LC50 of eggs and adults was 191- and 4,237-fold, respectively. The laboratory-selected acequinocyl resistance (LSAR16) strain was screened with 11 acaricides for cross-resistance. The detoxifying enzyme activities and quantitative real-time PCR analysis were performed in the LSAR16 strain. Crossing experiments revealed that LSAR16 strain was inherited maternally, incompletely dominant and monogenic. Most importantly, we identified two new point-mutations at mitochondrial cytb from acequinocyl resistant T. urticae.

The two-spotted spider mite, Tetranychus urticae, is one of the most important pests of crops. However, chemical acaricides are not always effective and their continuous use has resulted in resistance among two-spotted spider mite populations. To reduce or replace use of chemical acaricides, the entomopathogenic fungi having acaricidal activity were investigated. As results, 12 isolates could be selected from 342 isolates of fungi against two-spotted spider mite. To show the multiple roles of entomopathogenic fungi against plant pathogen, the antimicrobial activity of selected 12 isolates were evaluated to plant pathogenic bacterium Ralstonia solanacearum and fungus Botrytis cinerea. Various antimicrobial activities were observed against these plant pathogens. Consequently, these entomopathogenic fungi would be offered an alternative for dual control agents against the two-spotted spider mite and plant diseases.

The two-spotted spider mite Tetranychus urticae is being steadily problematic on strawberry. To effective control of this pest, one combination application using predatory mite Phytoseiulus persimilis plus a pesticide Acequinocyl suggests. Prepared six beds (1.5 × 3 m; 20 strawberry plants) on glass house, 100 T. urticae was inoculated on per 20 plants. Four plots prepared - Acequinocyl treatment, P. persimilis application, Acequinocyl+P. persimilis double application, and the control. The change of T. urticae density was investigated in the seven-day intervals. In an Acequinocyl treated plot, T. urticae density was decreased after Acequinocyl sprayed, but began to increase steadily one month later. In a P. persimilis applied plot, T. urticae density began to decrease at 20 days after P. persimilis inoculation. In an Acequinocyl + P. persimilis double applied plot, T. urticae density was dramatically decreased after application and then no found the T. urticae. To practical use in farmhouse, it will be possible to control by inoculation of P. persimilis only when low density of T. urticae; however, double application with spray pesticide Acequinocyl plus apply the natural enemy P. persimilis will be good when high density of T. urticae.

Expression of hairpin RNA corresponding to the part of COPA transcript was done by agroinfiltration in soybean plants and was confirmed by qRT-PCR. In a pot experiment, T. urticae was infested on agroinfiltrated soybean plants and T. urticae mortality was observed and compared with control plants overtime. Significantly higher mortalities of T. urticae were observed in the COPA-agroinfiltrated soybean plants from post-infestation day 2 (15 ±5%), day 4 (50 ±10 %). At post-infestation day 6, mortality reached to (70 ± 15%). To validate the observed COPA silencing effect in T. urticae fed on the agroinfiltrated soybean plant expressing COPA hairpin RNAs, qRT-PCR analysis was performed. The transcript level of COPA gene was decreased in T. urticae fed on agroinfiltrated soybean plants expressing COPA hairpin RNA from post-infestation day 2. At post-infestation day 2, 4 and 6, COPA transcript levels were reduced by 23.8, 20.7 and 18.8 fold, respectively compared to post-infestation day 1 (control). The results obtained in this study also ruled that the plant mediated production and uptake of silencing (dsRNAs/siRNAs) is an effective way to trigger RNAi in the T. urticae.

The two spotted mite, Tetranychus urticae Koch, is a global pest, and has developed severe resistance to several types of acaricides. Acequinocyl-resistant strain and bifenazate-resistant strain of T. urticae showed higher relative activity level of glutathion-S-transferases (GSTs) than susceptible strain. Studies of resistance in T. urticae have focused on classical detoxifying enzyme system, such as GSTs. We examined the expression of six GSTs genes in T. urticae belong to five different cytosolic classes, including two in delta, one in omega, one in mu, one in zeta, and one mitochondrial GSTs class, kappa. RT-qPCR results showed that, among these six GSTs genes, the relative expression level of delta 1 was significantly higher in bifenazate-resistant strain than in other strains (susceptible, abamectin, acequinocyl, etoxazole and pyridaben strain). Expression level of the delta 2 was also higher in acequinocyl-resistant strain than in other strains. The elevated GSTs gene transcripts following exposure to acequinocyl, bifenazate might be one of the mechanisms involved in detoxification of these acaricide. These results may be caused by different mechanisms in the resistance development of T. urticae to these two different types of acaricides.

The detoxification enzymes activities were investigated to Tetranychus urticae Koch using five acaricide-resistant strains. Activities of detoxification enzymes which are glutathione S-transferase (GST), general esterases (α-naphthyl acetate and β-naphthyl acetate), and cytochrome C oxidase were determined to each resistant strain mite. Acequinocyl-resistant strain and bifenazate-resistant strain of T. urticae were showed 2.1 folds and 1.6 folds higher relative activity (RA) level of GST than susceptible strain. Other three resistant strain mites were not significant different to susceptible strain mite. General esterases and cytochrome C oxidase were not significant to all the strain of T. urticae. Acaricidal activities of acequinocylresistant strain and bifenazate-resistant strain of T. urticae showed cross-resistant both acequinocyl and bifenazate. However, other strain mites were showed susceptible acaricidal activities to two acaricides. In this result suggests that resistance of two acaricides (acequinocyl and bifenazate) might be influenced by glutathione S-tansferase activity.

Emergence of resistant two-spotted spider mite (TSSM) can induce the over usage of standard amount of acaricides and result in various side effects. Rapid resistance monitoring is essential step for the efficient management of resistant populations by enabling the selection of appropriate acaricides. Here, we evaluated the 19 acaricides to determine its suitability for residual contact vial bioassay (RCV) by using PyriF strain as a reference. Twelve acaricides (Amitraz Abamectin, Bifenthrin, Bifenazate, Chlorfenapyr, Cyenopyrafen, Cyflumetofen, Endosulfan, Fenothiocarb, Monocrotophos, Omethoate and Tebufenpyrad) revealed the dose-dependent mortality within 8 h, whereas other remaining acaricides (Dicofol, Etoxazole, Fenbutatin oxide, Fenpyroxymate, Flufenoxuron, Spiromesifen and Pyridaben) did not. This finding suggests that the application of RCV method is limited depending on the mode of action and physicochemical properties of each acaricide. Resistance levels to 12 acaricides were determined for four field populations of TSSM by using RCV diagnostic kit. All TSSM populations showed the highest sensitivity to cyflumetofen, indicating that it would be most effective in controling field populations. RCV diagnostic kit would enable to provide crucial information for choosing the most appropriate acaricides in the field.

For replacement of methyl bromide(MB) in Korea, VapormateTM, a gas formulation of ethyl formate(EF) with carbon dioxide, are firslty introduced in import bananas and VivakilTM, a gas formulation of phosphine(PH3) with carbon dioxide, are ready to commercial use in export cut flowers. Their applications are being extended for other import and export commodities. Ethyl formate have been shown to have effectiveness short exposure time(<4hr) but it has a limited application at low temperature for TSM(Two spotted spider mites) due to some phytotoxic to strawberry. Phosphine gas has demonstrated their efficacy to TSM at low temperature but only meets quarantine requirement when applying longer fumigation time(>24hr) to control CA(cotton aphids). We firstly evaluated two MB alternatives, EF and PH3, have enhansive effect within short exposure time(4hr) at low temperature(5℃) for controlling both TSM and CA without any injury to export strawberries.

Emergence of resistant two-spotted spider mite (TSSM) can induce the over usage of standard amount of acaricides and result in various side effects. Rapid resistance monitoring is essential step for the efficient management of resistant populations by enabling the selection of appropriate acaricides. Here, we evaluated the 10 acaricides to determine its suitability on residual contact vial bioassay (RCV) by using PyriF strain as standard. Five acaricides (Abamectin, monocrotophos, tebufenpyrad, bifenazate and cyflumetofen) revealed the dose-dependent mortality within 8 h, whereas other remaining acaricides (Fenbutatin oxide, fenpyroxymate, flufenoxuron, spiromesifen and etoxazole) did not. This finding suggests that the application of RCV method is limited depending on the mode of action and physicochemical properties of each acaricide. Resistance levels to five acaricides (Abamectin, monocrotophos, tebufenpyrad, bifenazate and cyflumetofen) were determined for four field populations of TSSM by using RCV diagnostic kit. All TSSM populations showed the highest sensitivity to cyflumetofen, indicating that it would be most effective in controling field populations. RCV diagnostic kit would enable to provide crucial information for choosing the most appropriate acaricides in the field.

The two-spotted spider mite Tetranychus urticae is a worldwide crop pest with a high insecticide resistance and an extensive host range. The aim of the present study was to evaluate the effect of PaeciPora®, which was formulated from the aerial conidia of an entomopathogenic fungus Paecilomyces lilacinus strain HY-4, to control T. urticae in cucumber field. In the field study, conidia of P. lilacinus HY-4 and a chemical acaricide azocyclotin were investigated for their control of the adult females of T. urticae. The strain produced a mortality of 56.0% on day 3 and 63.6% on day 7 post-treatment respectively at 1×107 conidia/mL, and no evidence of a mortality benefit was seen in the control group. Additionally, in the pesticide injury test, no agrochemical damage was found in hot pepper, watermelon, Chinese cabbage, oriental melon or strawberry by spraying PaeciPora® on them. The results indicated the possibility of the use of P. lilacinus HY-4 as a microbiological control agent against T. urticae in the Integrated Pest Management program.

The two-spotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae), is one of the most important pest species devastating many horticultural, ornamental crops and fruit trees. Difficulty in managing this mite is largely attributed to its ability to develop resistance to many acaricides. Development of 3,700 folds resistance to etoxazole was found in the population of T. urticae collected from rose greenhouses in Buyeo, Chungnam Province in August 2000. This population has been selected for eleven years with etoxazole (over 500 times), and increased over 5,000,000 fold in resistance as compared with susceptible strain (S). Etoxazole-resistant strain was shown to be maternally inherited. The objective of this study was to determine whether resistance of T. urticae to etoxazole was linked with point mutations in the mitochondrial gene. DNA sequencing of cytochrome c oxidase subunit I (COX1), COX2, COX3, cytochrome b (CYTB), NADH dehydrogenase subunit 1 (ND1), ND2, ND3, ND4, ND5, and ND6 were analyzed by comparing two isogenic etoxazole-susceptible (EtoS) and etoxazole-resistant (EtoR) strains. As a result, all genes revealed no point mutations between the two strains.

Two-spotted spider mite, Tetranychus urticae (Koch) is one of the serious pests in economically important crops such as strawberry and cucumber and so on. Acaricides have been used as the main control agents. This study was conducted to test the synergistic effects of the Beauveria bassiana GHA, which has been registered for whitefly and thrips, and five pesticides (abamectin, acrinathrin, bifenthrin imidacloprid, dinotefuran, and indoxacarb, which are commonly used pesticides in strawberry in Korea), on the two-spotted spider mite T. urticae. Five tested pesticides did not inhibit spore germination and mycelial growth of B. bassiana. Pesticides were applied to potted strawberry plants at four different treatments (recommended concentration, 1/5 recommended concentration, 1/5 recommended concentration + GHA(108 conidia/ml), and only GHA(108 conidia/ml). Mortality in larvae of two-spotted spider mite was 12% in GHA 5 day after treatment. Mortality in abamectin treated T. urticae was 98.5% and 100% at 1/5 recommended concentration and recommended concentration, respectively, 3 days after treatment. Acrinathrin, indoxacarbe, dinotefuran and difenthrin imidacloprid caused 60%, 14%, 16% and 91% mortality at recommended concentration 5 day after treatment. The tested five pesticides and B. bassiana GHA had no synergistic effect.

The two-spotted spider mite, Tetranychus urticae Koch, is an economically important pest of crops of plant grown in the field or greenhouse worldwide. It has recently become a serious problem because of the continuous use of acaricides resulting in resistance among spider mite population. Thus, there is a need to find alternative control measures to suppress spider mite populations. In this study, we report the screening result of pathogenic fungi for the control of spider mite. Initial screenings were performed using 352 isolates of putative pathogenic fungi from Korea soils. As results, 11 strains of acaropathogenic fungi were isolated from 8 cadavers of spider mite supporting fungal conidiation. These isolated were identified as four isolates of Beauveria bassiana (6, 2R-3-3-1, 2R-4-5, 2R-4-7), two isolates of Metarhizium anisopliae (4-2, 2-2), one isolate of Clonostachys rosea 5-2, one isolate of Lecanicillium attenuatum 4-1, one isolate of Pochonia suchlasporia 2R-3-1, one isolate of Aspergillus flavus 7 and one isolate of Isaria lilacinus 2R-4-6 by microscopic examination and genetic sequencing of the ITS region. Based on the screening results, eleven isolates were tested for their virulence against adult spider mites. All fungal isolates were pathogenic to spider mite but mortality varied with isolates. These acaropathogenic fungi may be useful to develop eco-friendly acaricide to control two-spotted spider mite.