Phenotypic- and Genotypic-Resistance Detection for Adaptive Resistance Management in Tetranychus urticae Koch

Rapid resistance detection is necessary for the adaptive management of acaricide-resistant populations of Tetranychus urticae. Detection of phenotypic and genotypic resistance was conducted by employing residual contact vial bioassay (RCV) and quantitative sequencing (QS) methods, respectively. RCV was useful for detecting the acaricide resistance levels of T. urticae, particularly for on-site resistance detection; however, it was only applicable for rapid-acting acaricides (12 out of 19 tested acaricides). QS was effective for determining the frequencies of resistance alleles on a population basis, which corresponded to 12 nonsynonymous point mutations associated with target-site resistance to five types of acaricides [organophosphates (monocrotophos, pirimiphos-methyl, dimethoate and chlorpyrifos), pyrethroids (fenpropathrin and bifenthrin), abamectin, bifenazate and etoxazole]. Most field-collected mites exhibited high levels of multiple resistance, as determined by RCV and QS data, suggesting the seriousness of their current acaricide resistance status in rose cultivation areas in Korea. The correlation analyses revealed moderate to high levels of positive relationships between the resistance allele frequencies and the actual resistance levels in only five of the acaricides evaluated, which limits the general application of allele frequency as a direct indicator for estimating actual resistance levels. Nevertheless, the resistance allele frequency data alone allowed for the evaluation of the genetic resistance potential and background of test mite populations. The combined use of RCV and QS provides basic information on resistance levels, which is essential for choosing appropriate acaricides for the management of resistant T. urticae.     

Resistance to acaricides in Italian strains of <Emphasis Type="Italic">Tetranychus urticae</Emphasis>: toxicological and enzymatic assays

Problems with Tetranychus urticae are frequently reported in protected crops in Italy, particularly in roses where many introduced acaricides show a progressive loss of effectiveness. We have conducted bioassays to assess the response of some Italian strains of T. urticae to a number of acaricides. These include compounds that were widespread and frequently used in the past, but also some recently registered compounds. We investigated two T. urticae strains collected from rose growers where control failures were reported (SAN and PSE), together with a strain collected from unsprayed vegetables (BOSA). Adult females of the rose strains (SAN and PSE) were resistant to tebufenpyrad (Resistant Ratio—RR, RR₅₀ = 48.4 and 163.6) and fenpyroximate (RR₅₀ = 74.1 and 25.9) when compared to the susceptible BOSA strain. Lethal concentrations for these products were higher than the registered field rate. The PSE strain proved to be highly resistant to abamectin (RR₅₀ = 1,294.1). Variation in bifenazate susceptibility was detected amongst strains, but LC₉₀ values of SAN and PSE were still in the range of the registered field rate. In egg bioassays, the SAN and PSE strains exhibited high resistance levels to clofentezine (RR₅₀ = 66,473 and 170,714), hexythiazox (RR₅₀ = 70,244 and 159,493) and flufenoxuron (RR₅₀ = 61.9 and 117.9). But the recently introduced ovi/larvicides etoxazole and spirodiclofen exhibited high activity on all strains. The activity of detoxifying enzymes such as esterases, glutathione-S-transferases (GSTs) and cytochrome P450 monooxygenases (MFOs) was determined in these strains as a preliminary attempt to identify potential resistance mechanisms. Enzymatic assays showed that the rose strains exhibited 2.66 and 1.95-fold increased MFOs activity compared to the susceptible strain. Assays for GSTs revealed that only the SAN strain exhibited a significantly higher activity. In contrast, only the PSE strain showed a significant higher hydrolysis of 1-naphthyl acetate.     

Milbemectin resistance in <Emphasis Type="Italic">Tetranychus urticae</Emphasis> (Acari: Tetranychidae): selection,stability and cross-resistance to abamectin

Studies on artificial laboratory selections with milbemectin, stability of milbemectin resistance and possible cross-resistance with abamectin were carried out with Tetranychus urticae Koch to provide basic information for a milbemectin resistance management program. Selections for resistance and susceptibility to milbemectin were performed in a population of T. urticae, collected from a commercial chrysanthemum field in the State of São Paulo, Brazil. After six selections for resistance and five selections for susceptibility, susceptible (S) and resistant (R) strains of T. urticae to milbemectin were obtained. The resistance ratio (R/S) at the LC₅₀ reached 409-fold value. The stability of milbemectin resistance was also studied under laboratory conditions, using a population with initial frequency of 75% of resistant mites. The frequencies of milbemectin resistance were evaluated monthly for a period of 7 months. In order to observe possible correlation between milbemectin and abamectin resistance, the frequencies of abamectin resistance were also evaluated for that population, during the same period. The frequency of milbemectin resistance decreased from 75 to 14.5%, while the percentage of abamectin resistant mites decreased from 57 to 9.1%, in 7 months. The frequencies of milbemectin and abamectin resistance were also evaluated in 25 field populations of T. urticae, collected from several crops in the State of São Paulo. The frequencies of milbemectin resistance varied from 4.1 to 89.5%, and of abamectin, from 7.0 to 90.5%. A positive and significant correlation was observed between the frequencies of milbemectin and abamectin resistance, indicating positive cross-resistance between these acaricides. The results indicate that abamectin should be avoided for managing milbemectin resistance in T. urticae. This is the first report on milbemectin resistance in T. urticae in Brazil.     

Residual contact vial bioassay for the on-site detection of acaricide resistance in the two-spotted spider mite

To develop a less technique-dependent bioassay technique that can be conveniently used by practitioners or farmers in the field for the monitoring of acaricide resistance of the two-spotted spider mite, Tetranychus urticae Koch, a residual contact vial (RCV) method was established using a 5-ml glass vial coated with acaricides. The RCV bioassay procedures were optimized by using abamectin and tebufenpyrad, two widely used acaricides. The diagnostic concentrations causing 100% mortality within 8 h post-treatment in a susceptible strain of T. urticae was set at 30 and 60 ppm for abamectin and tebufenpyrad, respectively. The vial-coated pesticides were stable at least one year when stored at ? 20 °C as determined by HPLC. There was no significant difference in the bioassay results in repeated RCV bioassay by three different experimenters, indicating its high reproducibility and reliability. RCV-based resistance monitoring of 15 field populations of mites revealed that abamectin resistance begins to spread but tebufenpyrad resistance is already prevalent in Korea. The RCV diagnostic kit, when used by practitioners or farmers on site, should provide crucial information for the selection of most suitable acaricides for different field populations of T. urticae.     

Multiple Resistance and Biochemical Mechanisms of Dicofol Resistance in Tetranychus urticae (Acari: Tetranychidae)

A field colony of Tetranychus urticae (Koch) resistant to dicofol was selected with dicofol successively for 20 generations to produce the DR-20 strain. Resistance and multiple resistance levels of the DR-20 strain to 15 acaricides were determined using a spray bioassay. The DR-20 strain was extremely resistant to dicofol [resistance ratio (RR), 465]. The strain showed extremely strong resistance to acrinathrin (RR, 373) and benzoximate (RR, 197) and strong resistance to bromopropylate (RR, 136), fenbutatin oxide (RR, 65), fenpropathrin (RR, 70), fenpyroximate (RR, 68), and pyridaben (RR, 63). A RR of 11–29 was observed with abamectin, fenazaquin, milbemectin, propagite, and tebufenpyrad. The DR-20 strain exhibited low levels of resistance (RR<3) to azocyclotin and chlorfenapyr. In comparative assays with detoxifying enzymes, the DR-20 strain showed 4.7-fold higher activity in p-nitroanisole-O-demethylation and 1.6-fold higher activities in both α- and β-naphthyl acetate hydrolysis. Synergist experiments with different metabolic inhibitors revealed that piperonyl butoxide, iprobenfos, triphenyl phosphate, and 4, 4-dichloro-α-methyl benzhydrol had little or no synergistic activity in the susceptible and DR-20 strains. These results suggest that employment of certain acaricides with little multiple resistance will be useful for the management of dicofol resistance in the field.     

Adaptation of acaricide stress facilitates Tetranychus urticae expanding against Tetranychus cinnabarinus in China

The two‐spotted spider mite, Tetranychus urticae, and the carmine spider mite, Tetranychus cinnabarinus, are invasive and native species in China, respectively. Compared with T. cinnabarinus, T. urticae has expanded into most parts of China and has become the dominant species of spider mite since 1983, when it was first reported in China. However, the mechanism of the demographic conversion has not been illuminated. In this study, one T. urticae field population and one T. cinnabarinus field population were isolated from the same plant in the same field, and the toxicological characteristics were compared between these two species. Laboratory bioassays demonstrated that T. urticae was more tolerant to commonly used acaricides than T. cinnabarinus. The activities of detoxification enzymes were significantly greater in T. urticae, and the fold changes of enzymes activities in T. urticae were also greater following exposure to acaricides. Furthermore, more metabolism‐related genes were upregulated at a basal level, and more genes were induced in T. urticae following exposure to acaricides. The comparison of proteins and genes between both species led credence to the hypothesis that T. urticae was more resistant to acaricides, which was the reason explaining the expansion of invasive T. urticae against native T. cinnabarinus. Laboratory simulation experiments demonstrated that following the application of acaricides, the composition of a mixed T. urticae/T. cinnabarinus population would change from a T. cinnabarinus‐dominant to a T. urticae‐dominant population. This study not only reveals that T. urticae possesses stronger detoxification capacity than its sibling species T. cinnabarinus, which facilitated its persistent expansion in China, but also points to the need to accurately identify Tetranychus species and to develop species‐specific management strategies for these pests.     

A mutation in the PSST homologue of complex I (NADH:ubiquinone oxidoreductase) from Tetranychus urticae is associated with resistance to METI acaricides

The acaricidal compounds pyridaben, tebufenpyrad and fenpyroximate are frequently used in the control of phytophagous mites such as Tetranychus urticae, and are referred to as Mitochondrial Electron Transport Inhibitors, acting at the quinone binding pocket of complex I (METI-I acaricides). Because of their very frequent use, resistance evolved fast more than 20 years ago, and is currently wide-spread. Increased activity of P450 monooxygenases has been often associated with resistance, but target-site based resistance mechanisms were never reported.Here, we report the discovery of a mutation (H92R) in the PSST homologue of complex I in METI-I resistant T. urticae strains. The position of the mutation was studied using the high-resolution crystal structure of Thermus thermophilus, and was located in a stretch of amino acids previously photo-affinity labeled by fenpyroximate. Selection experiments with a strain segregating for the mutant allele, together with marker-assisted back-crossing of the mutation in a susceptible background, confirmed the involvement of the mutation in METI-I resistance. Additionally, an independent genetic mapping approach; QTL analysis identified the genomic region of pyridaben resistance, which included the PSST gene. Last, we used CRISPR-Cas9 genome editing tools to introduce the mutation in the Drosophila PSST homologue.     

Molecular analysis of resistance to acaricidal spirocyclic tetronic acids in Tetranychus urticae: CYP392E10 metabolizes spirodiclofen,but not its corresponding enol

Spirodiclofen is one of the most recently developed acaricides and belongs to the new family of spirocyclic tetronic acids (ketoenols). This new acaricidal family is an important chemical tool in resistance management strategies providing sustainable control of spider mites such as Tetranychus urticae. Spirodiclofen targets lipid biosynthesis mediated by direct inhibition of acetyl coenzyme A carboxylase (ACCase). In this study, we investigated two genetically distant spider mite strains with high resistance to spirodiclofen. Despite the strong resistance levels to spirodiclofen (up to 680-fold), only limited cross-resistance with other members of this group such as spiromesifen and spirotetramat could be detected. Amplification and sequencing of the ACCase gene from resistant and susceptible strains did not reveal common non-synonymous mutations, and expression levels of ACCase were similar in both resistant and susceptible strains, indicating the absence of target-site resistance. Furthermore, we collected genome-wide expression data of susceptible and resistant T. urticae strains using microarray technology. Analysis of differentially expressed genes revealed a broad response, but within the overlap of two resistant strains, several cytochrome P450s were prominent. Quantitative PCR confirmed the constitutive over-expression of CYP392E7 and CYP392E10 in resistant strains, and CYP392E10 expression was highly induced by spirodiclofen. Furthermore, stage specific expression profiling revealed that expression levels were not significantly different between developing stages, but very low in eggs, matching the age-dependent resistance pattern previously observed. Functional expression of CYP392E7 and CYP392E10 confirmed that CYP392E10 (but not CYP392E7) metabolizes spirodiclofen by hydroxylation as identified by LC–MS/MS, and revealed cooperative substrate binding and a K_m of 43 μM spirodiclofen. CYP392E10 also metabolizes spiromesifen, but not spirotetramat. Surprisingly, no metabolism of the hydrolyzed spirodiclofen-enol metabolite could be detected. These findings are discussed in the light of a likely resistance mechanism.     

Diversity of Wolbachia in Natural Populations of Spider Mites (genus Tetranychus): Evidence for Complex Infection History and Disequilibrium Distribution

Wolbachia are endosymbiotic bacteria that commonly infect arthropods and cause reproductive disorders in host. Within several Tetranychus species, Wolbachia have been detected and shown to affect their reproduction. However, little is known about their transmission and distribution patterns in natural populations of Tetranychus species. Here, we used multilocus sequence typing to confirm Wolbachia infection status and examined the relationship between Wolbachia infection status and host phylogeny, mitochondrial diversity, and geographical range in five Tetranychus species (Tetranychus truncatus, Tetranychus urticae, Tetranychus pueraricola, Tetranychus phaselus, and Tetranychus kanzawai) from 21 populations in China. The prevalence of Wolbachia within the five Tetranychus species ranged from 31.4 to 100 %, and the strains were remarkably diverse. Together, these observations indicate that Wolbachia was introduced to these populations on multiple separate occasions. As in other arthropods, the same Tetranychus species can accommodate very different strains, and identical Wolbachia occasionally infect different species. These observations suggest that Wolbachia are transmitted both vertically and horizontally. Horizontally, transmission is probably mediated by the host plants. The distribution patterns of Wolbachia were quite different among populations of the same species, suggesting that the dynamics of Wolbachia in nature may be affected by ecological and other factors.     

Monitoring of acaricide resistance of Tetranychus urticae (Acari: Tetranychidae) from Korean apple orchards

Tetranychus urticae populations were collected from ten commercial apple orchards and their susceptibilities to 12 acaricides were tested using a leaf disc bioassay. The resistance of each T. urticae population was reported as the LC₅₀ value, the resistance ratio (RR) and the slope of the probit–concentration regression. Cross resistances of T. urticae populations were estimated using the Spearman's rank correlation coefficient. Most local populations showed low resistance levels (RR ≤ 10). Development of resistance to METI and pyrethroid acaricides differed among local populations. The highest RR value (154.6) was found in the Uiseong population to tebufenpyrad. The Geochang population was highly resistant, especially to METI and pyrethroid acaricides. T. urticae populations collected from Suwon, Chungju, Yeongju and Geochang showed moderate resistance (10 < RR ≤ 40) to more than two acaricides. Resistance ratios to abamectin, chlorfenapyr, fenbutatin-oxide and milbemectin were low (RR ≤ 10) in all populations. The LC₅₀ values of abamectin, chlorfenapyr, fenbutatin-oxide and milbemectin ranged from 0.06 to 0.2 mg/l, from 0.67 to 3.38 mg/l, from 10.12 to 40.85 mg/l and from 0.47 to 3.01 mg/l, respectively. We discuss possible cross-resistance to acaricides using Spearman's rank correlation coefficient.     

Impact of living with kin/non-kin on the life history traits of Tetranychus urticae (Acari: Tetranychidae)

In many vertebrates and invertebrates, living in a group may influence the life history traits, physiology and behaviour of its individual members, whereas genetic relatedness affects social interactions among individuals in a group. The two-spotted spider mite Tetranychus urticae is characterised by a communal organization, in which silk production plays a key role. A silken web protects the colony against biotic and abiotic agents such as predators, competitors, humidity, wind, rain and acaricides. To evaluate the potential costs and benefits of being associated with genetically distant vs genetically close individuals in T. urticae, we assessed various fitness indicators (faecal pellet production, fecundity, death rate) in pure and mixed groups of two distinct populations of T. urticae: a red-form population from Tunisia and a green-form population from Belgium. If genetic origin had no influence, the values of fitness indicators in mixed groups composed of green and red individuals, would be intermediate between those of the pure green-form and red-form groups. Our results show that in a mixed group, faecal pellet production and death rate were statistically similar to the values obtained in the pure group of green-form individuals. Therefore, our study suggests that strain recognition ability may occur in T. urticae and that the genetic background of an individual may have a great impact on several of its life history traits.     

A critical review on some closely related species of <Emphasis Type="Italic">Tetranychus</Emphasis> sensu stricto (Acari: Tetranychidae) in the public DNA sequences databases

Taxonomic misidentification of the specimens used to obtain DNA sequences is a growing problem reported for different groups of organisms, which threatens the utility of the deposited sequences in public DNA databases. This paper provides new evidence of misidentifications in molecular DNA public databases in phytophagous mites of the Tetranychidae family belonging to the group Tetranychus (Tetranychus). Several species in this group are of economic and quarantine importance in agriculture and among them Tetranychus urticae, a highly polyphagous mite causing outbreaks in many crops worldwide, is certainly the most studied. We analyzed and evaluated the identity of 105 GenBank accessions of ITS2 rDNA and 138 COI mtDNA sequences which were deposited as T. urticae and those of 14 other taxa morphologically closely related to Tetranychus sensu stricto. In addition, ITS2 and COI sequences of 18 T. urticae samples collected for this study and identified by morphological criteria, were generated and included in the analyzed dataset. Among the deposited sequences in the GenBank database, numerous cases of apparently mistaken identities were identified in the group Tetranychus s. str., especially between T. urticae, T. cinnabarinus, T. kanzawai and T. truncatus. Unreliable sequences (misidentified or dubious) were estimated at nearly 30%. In particular the analysis supports the invalidity of the controversial species status of T. cinnabarinus. More generally, it highlights the need of using combined morphological and molecular approaches to guarantee solid species diagnostics for reliable sequence accessions in public databases.     

Development of acaricide resistance in Pacific spider mite (<Emphasis Type="Italic">Tetranychus pacificus</Emphasis>) from California vineyards

In recent years, grape growers in California reported failures of acaricides against Tetranychus pacificus McGregor. We collected T. pacificus populations from four vineyards and tested them for resistance to bifenazate, propargite and pyridaben. In addition, we sequenced part of the cytochrome b gene of bifenazate-resistant and -susceptible T. pacificus to test for the presence of mutations reported to confer resistance to the congeneric T. urticae. None of the mutations conferring resistance to bifenazate in T. urticae were present in resistant T. pacificus. Resistance levels ranged from full susceptibility to statistically significant 11-fold resistance to pyridaben, sevenfold resistance to bifenazate and fourfold resistance to propargite compared to a susceptible population. Despite the relatively low levels of resistance detected, we estimated that under the conditions of our study the highest field rates of bifenazate and pyridaben application would cause less than 58 and 66% mortality of adult females in the most resistant populations, respectively. In contrast, field rates of propargite application would cause close to 100% mortality in the least susceptible population. These results highlight a potential link between resistance development and reduced field effectiveness for bifenazate and pyridaben. Finally, T. pacificus may be more tolerant to bifenazate and propargite than T. urticae, since the LC₅₀ values for the susceptible population of T. pacificus were several times higher than LC₅₀’s reported for susceptible T. urticae.     

The invasive spider mite Tetranychus evansi (Acari: Tetranychidae) alters community composition and host-plant use of native relatives

The tomato spider mite Tetranychus evansi Baker and Pritchard (Acari: Tetranychidae), is a worldwide pest of solanaceous crops that has recently invaded many parts of the world. In the present study we examined the ecological impact of its arrival in the Mediterranean region. The spider mite and phytoseiid mite assemblages in various crop and non-crop plants in three areas of Valencia (Spain) were studied a few months before and 10 years after the invasion of T. evansi. According to rarefaction analyses, the invasion of T. evansi did not affect neither the total number of species in the mite community examined (spider mite and phytoseiid species) nor the number of species when the two communities were examined separately. However, after the invasion, the absolute and relative abundance of the native Tetranychus species was significantly reduced. Before the invasion, T. urticae and T. turkestani were the most abundant spider mites, accounting for 62.9 and 22.8 % of the specimens. After the invasion, T. evansi became the most abundant species, representing 60 % of the total spider mites recorded, whereas the abundance of T. urticae was significantly reduced (23 %). This reduction took place principally on non-crop plants, where native species were replaced by the invader. Null model analyses provided evidence for competition structuring the spider mite community on non-crop plants after the invasion of T. evansi. Resistance to acaricides, the absence of efficient native natural enemies, manipulation of the plant defenses and the web type produced by T. evansi are discussed as possible causes for the competitive displacement.     

Ambulatory dispersal of the susceptible and propargite-resistant strains of Tetranychus urticae and its influence on pesticide resistance dynamics

The distribution of resistant individuals is determined by the amount of movement between populations. The differential rate of dispersal of a susceptible and a pesticide-resistant strain could influence the resistance dynamics under field conditions. The dispersal rate and dispersal efficiency of the susceptible and propargite-resistant strains of Tetranychus urticae were measured in separate-release and mixed-release experiments. The diffusion coefficient (D) of both strains did not differ significantly (P > 0.344) and an estimate of the asymptotic rate of advance (2 √ rD) (for one generation) was estimated at 0.1047 and 0.0930 cm per degree day for the susceptible and propargite-resistant strains, respectively. The dispersal efficiency of the two strains differed significantly (P < 0.005) as more susceptible mites than propargite-resistant mites crossed into specified zones more quickly after 290 and 366 degree days. Significantly (P < 0.05) higher number of susceptible adults, immatures and eggs were found in the outer most zone of an arena as compared to that of the propargite-resistant mites. The bioassay of the two strains showed a similar pattern of the spread of the adult females across the specified zones in the mixed-release experiment. The relatively lower dispersive tendency of the propargite-resistant T. urticae and the smaller proportion of adult females exhibiting that behaviour increase the chances of developing resistant ‘hotspots’ in field specially after an acaricide application.     

The Complete Mitochondrial Genomes of Six Species of Tetranychus Provide Insights into the Phylogeny and Evolution of Spider Mites

Many spider mites belonging to the genus Tetranychus are of agronomical importance. With limited morphological characters, Tetranychus mites are usually identified by a combination of morphological characteristics and molecular diagnostics. To clarify their molecular evolution and phylogeny, the mitochondrial genomes of the green and red forms of Tetranychus urticae as well as T. kanzawai, T. ludeni, T. malaysiensis, T. phaselus, T. pueraricola were sequenced and compared. The seven mitochondrial genomes are typical circular molecules of about 13,000 bp encoding and they are composed of the complete set of 37 genes that are usually found in metazoans. The order of the mitochondrial (mt) genes is the same as that in the mt genomes of Panonychus citri and P. ulmi, but very different from that in other Acari. The J-strands of the mitochondrial genomes have high (∼84%) A+T contents, negative GC-skews and positive AT-skews. The nucleotide sequence of the cox1 gene, which is commonly used as a taxon barcode and molecular marker, is more highly conserved than the nucleotide sequences of other mitochondrial genes in these seven species. Most tRNA genes in the seven genomes lose the D-arm and/or the T-arm. The functions of these tRNAs need to be evaluated. The mitochondrial genome of T. malaysiensis differs from the other six genomes in having a slightly smaller genome size, a slight difference in codon usage, and a variable loop in place of the T-arm of some tRNAs by a variable loop. A phylogenic analysis shows that T. malaysiensis first split from other Tetranychus species and that the clade of the family Tetranychoidea occupies a basal position in the Trombidiformes. The mt genomes of the green and red forms of T. urticae have limited divergence and short evolutionary distance.     

Fitness costs associated with low-level dimethoate resistance in Phytoseiulus macropilis

Phytoseiulus macropilis Banks (Acari: Phytoseiidae) is an effective predator of tetranychid mites, but there are no data on its response to pesticides. We investigated the resistance of the predatory mite P. macropilis to the acaricides abamectin and dimethoate, and we examined the fitness costs associated with resistance. Two populations were tested: one from conventional cultivation and another from an area not commercially exploited. After the application of acaricides to the predator, we determined the lethal effects of the acaricides, the instantaneous rate of population increase (r _i ), the predation on Tetranychus urticae Koch (Acari: Tetranychidae) and its ability to locate prey in an olfactometer. P. macropilis exhibited resistance to dimethoate only. The low level of resistance (9.4x) of the predator did not affect their ability to locate prey. However, the dimethoate resistant population was not as effective in contatining prey population when in lower density and exhibited a more pronounced decrease of r _i in the presence of this acaricide, due to the reduced oviposition of the predator, a likely consequence of the different genetic background of this population.     

Genetic and biochemical comparisons of organophosphate resistance between strains of spider mites (Tetranychus species: Acari)

The results of genetic and biochemical studies of resistance to parathion in strains of Tetranychus urticae are presented. Included in the study is the well-known Leverkusen-R strain, as well as two resistant strains from New Zealand. For the toxicological part of the genetic investigation, a topical application technique is employed. Both genetically and biochemically, the resistances of the two independent New Zealand strains appear identical to the Leverkusen resistance, viz., due to the same major dominant gene which lowers the inhibition rate of the cholinesterase enzyme. Comments are made upon the universal significance of this basic resistance mechanism.

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Genetische und biochemische vergleiche der organophosphatresistenz von verschiedenen spinnmilben-stämmen (Tetranychus-species: Acari)

Zusammenfassung Die Ergebnisse genetischer und biochemischer Untersuchungen der Resistenz gegenüber Parathion an Stämmen von Tetranychus urticae Koch werden dargestellt. In die Untersuchung waren sowohl der wohl bekannte Leverkusener R-Stamm als auch zwei resistente Stämme von Neuseeland einbezogen. Im toxikologischen Teil der genetischen Untersuchungen wurde eine topische Applikationstechnik angewendet. Genetisch wie biochemisch scheint die Resistenz der zwei voneinander unabhängigen Neuseeländer Stämme mit der Resistenz des Leverkusener Stammes identisch, d.h. von dem gleichen dominanten Hauptgen verursacht zu sein, welches die Hemmungsrate der Cholinesterase-Enzyme senkt. Bemerkungen über die universelle Bedeutung dieses grundlegenden Resistenzmechanismus werden angefügt.

     

Olfactory response of Phytoseiulus persimilis (Acari: Phytoseiidae) to untreated and Beauveria bassiana-treated Tetranychus urticae (Acari: Tetranychidae)

Determination of attraction and avoidance behavior of predators is important in concomitant use of multiple natural enemies to control a pest. The olfactory response of the predatory mite Phytoseiulus persimilis was studied to odors related to Tetranychus urticae adults infected by Beauveria bassiana DEBI008 in 0, 24, 48 and 72 h intervals, both in absence and in presence of plants. In plant-present experiments, P. persimilis attraction was neither towards adults of T. urticae infected by 0.02 % Tween 80 (as control), nor to the ones infected by B. bassiana for 0 or 24 h, whereas significant attraction towards the control was observed when tested against T. urticae infected by B. bassiana for 48 or 72 h. In absence of plants, P. persimilis displayed significant avoidance of T. urticae infected by B. bassiana for 48 or 72 h, when their alternative option was 0.02 % Tween 80-infected T. urticae adults. These results indicate that P. persimilis can recognize the presence of B. bassiana and that the predator avoids the fungus. This suggests that the two natural enemy species can be used together in biological control programmes.     

Variation in Susceptibility to Bacillus thuringiensis Toxins among Unselected Strains of Plutella xylostella

So far, the only insect that has evolved resistance in the field to Bacillus thuringiensis toxins is the diamondback moth (Plutella xylostella). Documentation and analysis of resistant strains rely on comparisons with laboratory strains that have not been exposed to B. thuringiensis toxins. Previously published reports show considerable variation among laboratories in responses of unselected laboratory strains to B. thuringiensis toxins. Because different laboratories have used different unselected strains, such variation could be caused by differences in bioassay methods among laboratories, genetic differences among unselected strains, or both. Here we tested three unselected strains against five B. thuringiensis toxins (Cry1Aa, Cry1Ab, Cry1Ac, Cry1Ca, and Cry1Da) using two bioassay methods. Tests of the LAB-V strain from The Netherlands in different laboratories using different bioassay methods yielded only minor differences in results. In contrast, side-by-side comparisons revealed major genetic differences in susceptibility between strains. Compared with the LAB-V strain, the ROTH strain from England was 17- to 170-fold more susceptible to Cry1Aa and Cry1Ac, respectively, whereas the LAB-PS strain from Hawaii was 8-fold more susceptible to Cry1Ab and 13-fold more susceptible to Cry1Da and did not differ significantly from the LAB-V strain in response to Cry1Aa, Cry1Ac, or Cry1Ca. The relative potencies of toxins were similar among LAB-V, ROTH, and LAB-PS, with Cry1Ab and Cry1Ac being most toxic and Cry1Da being least toxic. Therefore, before choosing a standard reference strain upon which to base comparisons, it is highly advisable to perform an analysis of variation in susceptibility among field and laboratory populations.