Resistance to binapacryl and tetradifon, and the genetic background, in fruit tree red spider mite, Panonychus ulmi, from English apple orchards

Assays on eggs of Panonychus ulmi held on leaf discs of Myrobalan plum were used to characterise strains (1) multi-resistant to organophosphates (OPs) and binapacryl (strain BR), (2) multi-resistant to OPs and tetradifon (strain TR), and (3) susceptible to OPs, binapacryl and tetradifon (strain EM). By crosses of EM females and BR males, and of EM females and TR males, and by repeated backcrossing with EM males under selection pressure of the acaricides concerned, two strains were produced in which the specific genes for resistance to binapacryl and tetradifon were isolated in the genome of the EM strain. Reciprocal crosses of the resulting backcrossed strains with EM, assays on the F₁ eggs and the F₂ haploid eggs, confirmed the presence of single major genes for resistance to binapacryl and tetradifon, and the absence of maternal effects. The expression of gene BR was incompletely dominant in response to binapacryl, whilst gene TR was effectively dominant in response to tetradifon. Eggs of the field-derived strain BR showed c. 50× resistance to binapacryl, 8–19 × to dinobuton, but only c. 2× to dinocap and dinoseb. The backcrossed strain was less resistant (26 ×) to binapacryl implying that resistance in strain BR was augmented by the effects of minor genes. Resistance to tetradifon (150–200 × on eggs) involved cross-resistance to tetrasul (23 ×) and to chlorfensulphide (27×); resistance in the field-derived strain appeared to depend on the major gene alone. Resistance mechanisms were highly specific in their effects, so that there was no cross-resistance to unrelated acaricides, including amitraz, dicofol, cyhexatin, formetanate, chlordimeform, and quinomethionate. Fecundity of strain BR was 44% less than that of EM; laboratory and field evidence suggest that multi-resistant mites tend to be less fit than susceptible mites. The practical implications are discussed.     

Resistance to organophosphates, and the genetic background, in fruit tree red spider mite, Panonychus ulmi, from English apple orchards

Assays on adult females and eggs of Panonychus ulmi , on leaf discs of Myrobalan plum, were used to characterise and establish homogenous organophosphateresistant (OR) and susceptible strains. Reciprocal crosses of these strains, and assays on adult mites and eggs of the parent, F₁ and F₂ generations confirmed that resistance was controlled by a single major gene, the expression of which was dominant in the response to parathion, and incompletely dominant to vamidothion, dimethoate and demeton-S-methyl. Reproductive incompatibility, between strains, as in Tetranychus urticae , was not found.
Bioassays on larvae hatched from winter eggs, collected from several orchards in Kent, Essex and Somerset, were used to evaluate the spectrum of cross-resistance to a range of 20 organophosphates (OPs). Together with assays on adult females, these tests showed 10- to 100-fold resistance to most of the OPs used for several years including those noted above, but less than 10-fold to azinphos compounds and to several recently-introduced OPs, e. g. dialifos, triazophos, dioxathion. The resistance spectrum in strain OR and five field strains was similar, but some had higher levels to parathion and demeton-S-methyl, and others to dioxathion after 3–4 years' exposure. Resistance levels to individual OPs may be enhanced by specific selection. Strains resistant to OPs showed only 2- to 5-fold resistance to two carbamates, and were fully susceptible to dicofol.     

Genetics of resistance to parathion and demeton-S-methyl inTetranychus urticae Koch (Acarina)

Out of a strain, resistant to organophosphorous insecticides, a dominant factor has been isolated by repeated backcrossings with a susceptible strain under selection pressure. This factor, calledOP, gives a large resistance to parathion and a much smaller resistance to demeton-S-methyl. Modifiers toOP increase the demeton-S-methyl resistance to a high level. Evidence is given that the resistance to the mentioned insecticides of the well known Leverkusen resistant strain has a similar inheritance.Abbreviations used OP organophosphorus - R resistant - S susceptible     

Effects of insecticides on life history parameters of the aphid parasitoidAphidius rhopalosiphi [Hym.: Aphidiidae]

The effects of four insecticides (two organophosphates, a pyrethroid and a carbamate), applied at different dosages, on different life history parameters of the aphid parasitoidAphidius rhopalosiphi (De Stefani-Perez) were tested. The methods used simulated field conditions in the laboratory. After application on the mummies, only two treatments (parathion and the high dosage of pirimicarb) caused a significant reduction of adult hatching. Fertility and longevity of the wasps were not affected by any of the tested insecticides. Dried residues of parathion, but not of the other insecticides, killed a considerable number ofA. rhopalosiphi females. The searching behaviour of the wasps was affected by demeton-S-methyl and fenvalerate. A direct application of demeton-S-methyl, parathion, and the high dosage of pirimicarb onA. rhopalosiphi females caused high mortality.     

Preliminary evaluation of <Emphasis Type="Italic">Cladosporium cladosporioides</Emphasis> (Fresen.) de Vries in laboratory conditions,as a potential candidate for biocontrol of <Emphasis Type="Italic">Tetranychus urticae</Emphasis> Koch

A survey for natural entomopathogenic fungi of two spotted spider mite (Tetranychus urticae) adults was made in Erzurum, Turkey, during the period 2006. Tetranychus urticae (65.8%) infected with a strain of the fungus Cladosporium cladosporioides were found. Thirteen isolates of C. cladosporioides were assessed against T. urticae, in a single dose (8 × 10⁶ conidia ml ⁻¹), laboratory bioassay on bean leaflets. The total mortality percentage caused by C. cladosporioides isolates varied from 50.95 to 74.76% and LT₅₀ values ranged from 2.34 to 3.90 days. The results revealed that isolates of C. cladosporioides were effective against two spotted spider mite. This is the first record of natural infection of T. urticae by C. cladosporioides.     

European populations of Diabrotica virgifera virgifera are resistant to aldrin,but not to methyl‐parathion

The western corn rootworm, Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae), is a major pest of cultivated corn in North America and has recently begun to invade Europe. In addition to crop rotation, chemical control is an important option for D. v. virgifera management. However, resistance to chemical insecticides has evolved repeatedly in the USA. In Europe, chemical control strategies have yet to be harmonized and no surveys of insecticide resistance have been carried out. We investigated the resistance to methyl‐parathion and aldrin of samples from nine D. v. virgifera field populations originating from two European outbreaks thought to have originated from two independent introductions from North America. Diagnostic concentration bioassays revealed that all nine D. v. virgifera field populations were resistant to aldrin but susceptible to methyl‐parathion. Aldrin resistance was probably introduced independently, at least twice, from North America into Europe, as there is no evident selection pressure to account for an increase of frequency of aldrin resistance in each of the invasive outbreaks in Europe. Our results suggest that organophosphates, such as methyl‐parathion, may still provide effective control of both larval and adult D. v. virgifera in the European invasive outbreaks studied.     

Insecticidal properties of nonactic acid and homononactic acid,the precursors of macrotetrolide antibiotics

(±)-Nonactic acid (1) and (±)-homononactic acid (2), the non-antibiotic precursors of macrotetrolide antibiotics, showed significant insecticidal effects (comparable with those of the commercial synthetic pesticide Metathion) onLeptinotarsa decemlineata, Epilachna varivestis andEuproctis chrysorrhoea. Anisoplia austriaca, Aphis fabae andCalandra granaria were less sensitive in this respect. A low acaricidal activity againstTetranychus urticae (not reaching that of the commercial miticide Acarition) was also found. Both1 and2 exhibited growth-stimulating properties for plants.     

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.

     

Construction of an engineered strain free of antibiotic resistance gene markers for simultaneous mineralization of methyl parathion and ortho-nitrophenol

Pseudomonas sp. strain NyZ402, a native soil organism that grows on para-nitrophenol (PNP), was genetically engineered for the simultaneous degradation of methyl parathion (MP) and ortho-nitrophenol (ONP) by integrating mph (methyl parathion hydrolase gene) from Pseudomonas sp. strain WBC-3 and onpAB (ONP 2-monooxygenase and ONP o-benzoquinone reductase genes) from Alcaligenes sp. strain NyZ215 into the genome of strain NyZ402. Methyl parathion hydrolase (MPH), ONP 2-monooxygenase (OnpA) and o-benzoquinone reductase (OnpB) were constitutively expressed in the engineered strain NyZ-MO. Strain NyZ-MO was free of exogenous antibiotic resistance gene markers and the introduced genes were genetically stable. Degradation experiments showed that strain NyZ-MO could utilize MP or ONP as the sole carbon and energy source, and mineralize 0.1 mM MP–0.1 mM ONP simultaneously. This method may serve as a useful strategy for the construction of engineered strains in the degradation of multiple environmental pollutants.     

Acaricidal properties of essential oils from agro‐industrial waste products from citric fruit against Tetranychus urticae

Tetranychus urticae is a major agricultural pest with worldwide distribution that has caused considerable damage to vegetable crops in north‐eastern Brazil. The aim of the present study was to investigate the chemical and lethal/sublethal effects of essential oils from the peels of the lime (Citrus aurantiifolia), lemon (C. limon), mandarin orange (C. reticulata) and (C. reticulata × C. sinensis) as well as selected constituents (linalool, α‐terpineol, α‐pinene, β‐pinene, terpinolene and limonene) against T. urticae. The greatest yield was achieved with the mandarin and tangerine peel oils. The chemical analysis (gas chromatography‐mass spectrometry) of the essential oils from the Citrus fruit peels enabled the identification of 127 compounds, revealing a predominance of monoterpenes. Limonene was the major constituent, and α‐pinene, β‐pinene, linalool and α‐terpineol were found in substantial quantities. Regarding the susceptibility of T. urticae, the Citrus oils and selected constituents were more effective by fumigation than residual contact. The C. reticulata oil was the most toxic by fumigation, and the C. limon oil was the most toxic by residual contact. The constituent α‐terpineol exhibited the highest toxicity with both methods. At a sublethal concentration, the oils and selected constituents had significant effects on the fecundity, feeding preference and oviposition of the mite. Citrus oils and their constituents are potentially useful for the future integrated management of T. urticae due to their lethal and sublethal properties. However, further studies are needed to evaluate the action of these essential oils against non‐target organisms and determine the cost–benefit ratio for the formulation of an acaricide harvested from agro‐industrial waste from citric fruit processing activities for use in the integrated control of T. urticae.     

Spider mite resistance to miticides in South Carolina strawberry and implications for improved integrated pest management

Tetranychus urticae Koch (Acari: Tetranychidae), twospotted spider mite, is a major secondary pest of strawberry and can cause significant yield loss. Tetranychus urticae is typically controlled using miticides, which has led to rapid resistance development. In South Carolina (USA), extension agents and growers have reported field failures of miticides (inadequate pest suppression), but resistance has not been quantitatively determined. In 2018, we determined the level of miticide resistance of six T. urticae populations found on strawberry across South Carolina. We examined efficacy of all miticides registered for use on US strawberry by conducting an initial diagnostic bioassay at 20% of the maximum labeled field rate. Any population?×?active ingredient combination resulting in?<?55% mortality was identified as ‘potentially resistant’ and concentration–response bioassays were then conducted to calculate LC₅₀ values for an individual population. These values were compared with those of a known-susceptible laboratory population to calculate resistance ratios (RR). Our results indicate that examined South Carolina populations of T. urticae from strawberry were highly resistant to bifenthrin (RR?=?100–60,000) and there was reduced susceptibility to fenbutatin-oxide (RR?=?25–123). The 'Sardinia' population had decreased abamectin susceptibility (RR?=?25). No resistance to hexythiazox, etoxazole, acequinocyl, bifenazate, fenpyroximate, spiromesifen, or cyflumetofen was found. Based on available data, it appears that miticide resistance is not a likely cause of field failures and issues related to application error and coverage should be investigated. Overall, this work supports the need to reduce the use of broad-spectrum pesticides and older products, in favor of newer miticide chemistries due to resistance issues.

     

Sublethal effects of Thymus vulgaris essential oil on life-table parameters of two-spotted spider mite,Tetranychus urticae Koch (Acari: Tetranychidae)

Sublethal effects of Thymus vulgaris were investigated on life-table parameters of Tetranychus urticae Koch (Acari: Tetranychidae) in the laboratory conditions at 25?±?1?°C, 70?±?10% RH and a 14:10?h (L:D) photoperiod. Bioassay was conducted to assess the effect of different concentrations of T. vulgaris essential oil on two-spotted spider mite. LC₅₀ and LC₂₀ of T. vulgaris on T. urticae were calculated as 18.86 and 6.24?μl/litre air for 24?h after treatment. In the sublethal effects, after 24?h exposure to LC₅₀ concentration, mortality, fecundity and longevity of the females that survived from treatment were determined. Results showed that T. vulgaris caused significant reductions in fecundity and longevity of adults. Life-table assay indicated that the intrinsic rate of increase (r _m), net reproductive rate (R ₀), finite rate of increase (λ), mean generation time (T), significantly reduced in treated females compared to control. These results suggest that T. vulgaris could be incorporated in integrated pest management programmes of T. urticae.     

Lethal activity of beauvericin,a Beauveria bassiana mycotoxin,against the two‐spotted spider mites,Tetranychus urticae Koch

The entomopathogenic fungi Beauveria bassiana (Balsamo) Vuillemin is known to produce a broad range of secondary metabolites. Beauvericin, a cyclic hexadepsipeptide, is the best known mycotoxin produced by B. bassiana; however, reports discussing the insecticidal activity of beauvericin per se are limited. In this study, we assessed the lethal activity of beauvericin against Tetranychus urticae Koch (Acarina: Tetranychidae). In addition, screening for suitable application of the mycotoxin against T. urticae on greenhouse strawberries is discussed. Beauvericin was able to control successfully T. urticae where concentrations of 10, 100 and 1,000 µg/g recorded mortalities of 84%, 100% and 100%, respectively, against motile stages. Furthermore, beauvericin inhibited egg hatching up to 83.3%, 69.3% and 53.3%, respectively, using the same concentrations under laboratory conditions. Under greenhouse conditions, the efficacy recorded was 52.6%, 85.7%, 72.4% and 72.4% at 1, 3, 7 and 10 days post‐inoculation, respectively. Beauvericin was efficacious under greenhouse conditions since the application increased strawberry yields while showing no phytotoxicity and ecotoxicological risk. Resistance to beauvericin was not detected initially at the unselected strain of T. urticae. Yet, the laboratory selection of populations of T. urticae exposed to beauvericin resulted in relatively resistant T. urticae strain that displayed no cross‐resistance to cyflumetofen and bifenazate. The acaricidal activity of beauvericin documented in this study would increase the efficacy of integrated pest management strategies.     

The development of bifenthrin resistance in two-spotted spider mite (Acari: Tetranychidae) from Australian cotton

Bifenthrin (Talstar^®) was registered at the beginning of the 1993/94 season for control of Helicoverpa spp. and Tetranychus urticae in Australian cotton. Resistance was monitored in T. urticae following registration but despite the implementation of resistance management, resistance evolved in the 1996/97 cotton-growing season after only four seasons use. During the following three seasons resistance increased progressively both in level (from 1.2- to 109-fold at LC₅₀) and abundance (from 20 to 90% strains) and was linked to field control failures. The evolution of resistance has now reduced the reliability of bifenthrin for T. urticae control in Australian cotton. The impact of bifenthrin resistance in T. urticae and its continued management in Australian cotton is discussed.     

Interactions of two natural enemies of Tetranychus urticae,the fungal entomopathogen Beauveria bassiana and the predatory mite,Phytoseiulus persimilis

The effect of either untreated or treated adults of the spider mite, Tetranychus urticae Koch (Acari: Tetranychidae) by Beauveria bassiana (Bals.) Vuillemin (Ascomycota: Hypocreales) DEBI008 at 1×10⁶ (conidia/ml) was investigated on developmental stages and life table parameters of Phytoseiulus persimilis Athias-Henriot (Acari: Phytoseiidae) under laboratory conditions. Four time intervals (0, 24, 48 and 72 h post-inoculation of spider mites) were considered for studying the predator characteristics as different treatments. Duration of each life stage, longevity, reproduction rate, intrinsic rate of natural increase (r _m ), net reproductive rate (R ₀), mean generation time (T) and finite rate of increase (λ) of the P. persimilis were calculated on both untreated and B. bassiana treated spider mite adults. Data analysis showed that longevity and fecundity of predatory mites fed on untreated and treated mites (time interval 0) were higher in comparison with other time intervals after inoculation. The entomopathogenic fungus adversely affected longevity and fecundity of the predatory mite. Fertility life table parameters of predatory mites fed on T. urticae treated by B. bassiana at different time intervals showed that T, R ₀, λ and r _m are strongly affected by the fungus presence and these parameters had significant differences among time treatments. The least r _m value was observed in the time interval of 72 h post-inoculation. The fitness of T. urticae was affected by B. bassiana 24, 48 and 72 h post-inoculation of mite adults, and consequently it caused decreased longevity of P. persimilis and accordingly a decrease in the intrinsic rate of natural increase of the predator.     

A region of barley chromosome 6H harbors multiple major genes associated with net type net blotch resistance

Net type net blotch (NTNB), caused by Pyrenophora teres f. teres Drechs., is prevalent in barley growing regions worldwide. A population of 118 doubled haploid (DH) lines developed from a cross between barley cultivars ‘Rika’ and ‘Kombar’ were used to evaluate resistance to NTNB due to their differential reaction to various isolates of P. teres f. teres. Rika was resistant to P. teres f. teres isolate 15A and susceptible to isolate 6A. Conversely, Kombar was resistant to 6A, but susceptible to 15A. A progeny isolate of a 15A × 6A cross identified as 15A × 6A#4 was virulent on both parental lines. The Rika/Kombar (RK) DH population was evaluated for disease reactions to the three isolates. Isolate 15A induced a resistant:susceptible ratio of 78:40 (R:S) whereas isolate 6A induced a resistant:susceptible ratio of 40:78. All but two lines had opposite disease reactions indicating two major resistance genes linked in repulsion. Progeny isolate 15A × 6A#4 showed a resistant:susceptible ratio of 1:117 with the one resistant line also being the single line that was resistant to both 15A and 6A. An RK F₂ population segregated in a 1:3 (R:S) ratio for both 15A and 6A indicating that resistance is recessive. Molecular markers were used to identify a region on chromosome 6H that harbors the two NTNB resistance genes. This work shows that multiple NTNB resistance genes exist at the locus on chromosome 6H, and the recombinant DH line harboring the resistance alleles from both parents will be useful for the development of NTNB-resistant barley germplasm.     

Insecticide resistance in Brazilian Triatoma infestans and Venezuelan Rhodnius prolixus

Resistance levels to insecticides used in control of Chagas Disease vectors were assessed in two species of bugs (Hemiptera: Reduviidae): Triatoma infestans (Klug) from Brazil and Rhodnius prolixus Stål from Venezuela. The resistance ratios, compared to susceptible laboratory strains, were determined by topical application bioassays. The T. infestans PA strain exhibited resistance ratios of 7× to deltamethrin, 3.6× to β‐cyfluthrin and 3.3× to cypermethrin, but was susceptible to β‐cypermethrin and lambda‐cyhalothrin. Rhodnius prolixus CA strain showed resistance to all the pyrethroids evaluated, the resistance ratios ranging between 4.5× to lambda‐cyhalothrin and 12.4× to cypermethrin. Deltamethrin resistance in both strains was decreased by piperonyl butoxide, suggesting oxidative metabolism as cause of resistance.     

Laboratory evaluation of resistance to pesticides in the phytoseiid predator Typhlodromus pyri from English apple orchards

Typhlodromus pyri, reared on plate cultures and fed on pollen of Vicia faba, were bioassayed using a taped-slide technique. Mite stocks from isolated unsprayed orchards were used to establish base-line susceptibility to azinphos-methyl, parathion, carbaryl and permethrin. Stocks from English orchards with a spray history of organophosphates and carbaryl showed resistance of 4 to 6x to azinphos-methyl, c 50 x to parathion, >20x to carbaryl, and no resistance to permethrin. An orchard population derived from T. pyri imported from New Zealand in 1977 responded similarly. In other tests, mites exposed to spray residues on glass and on apple leaves were killed by lower concentrations than in taped-slide tests (glass < apple leaf < taped-slide technique); but for all three assay techniques the resistance factors to azinophos-methyl and carbaryl, comparing two stocks, were similar. All stocks from sprayed orchards were resistant to both azinphos-methyl and carbaryl, suggesting cross-resistance; and resistance to both these pesticides appeared to be stable in the field when selection pressure was relaxed. The results are discussed in relation to earlier work on T. pyri and two other resistant orchard-inhabiting phytoseiid species.     

Acaricidal resistance and genetic affinities of some two‐spotted spider mite populations in New Zealand

Toxicological tests on the present status of resistance to acaricides in six strains of the two‐spotted spider mite (Tetranychus urticae Koch) in New Zealand are reported. Resistance to parathion appears widespread; cross‐resistance to formetanate is developed. Resistance to Plictran has not developed, and there is no indication of high levels of Kelthane resistance. The levels of genetic incompatibility operating between the strains are also investigated. The causes of such incompatibilities and their possible exploitation in integrated control programmes are discussed.     

Field evaluation of Beauveria bassiana efficacy for the control of Tetranychus urticae Koch (Acari: Tetranychidae)

Trials with eggplants were conducted to evaluate field efficacy of Beauveria bassiana (Bb) applications against Tetranychus urticae, during spring of 2006 and 2007. In the 2006‐Trial, a rate of ca. 0.7 × 10¹² conidia/ha was sprayed onto plants at intervals. In the 2007‐Trial, the rates of ca. 0.8 × 10¹² and 1.6 × 10¹² conidia/ha were sprayed at 1‐ or 2‐week intervals. Based on population densities of mites and leaf damage assessed throughout the trial, the Bb sprays with 1‐ or 2‐week interval showed better control than those of 3 or 4 weeks. In both trials there was no statistical difference between 1‐week and 2‐week spray interval for all parameters measured. Furthermore, the higher rate performed significantly better than the lower rate, regardless of the intervals. No interaction was found between the spraying interval and the applied rate. Ultimately, to suppress the population density of T. urticae and prevent the eggplant damage in spring, the higher rate applied at 2‐week interval was effective.