Genetic transfer of a twospotted spider mite (Acari: Tetranychidae) repellent in tomato hybrids

Lycopersicon hirsutum Dunal is very resistant to arthropod herbivory, and research on causes of resistance has often implicated trichomes and their secretions. To better understand relationships among resistance, repellency, and 2,3-dihydrofarnesoic acid, a trichome-borne sesquiterpenoid spider mite repellent, two tomato, Lycopersicon esculentum Miller, varieties were interbred with a highly resistant, spider mite repellent accession (LA1363) of L. hirsutum. Backcross and F2 generations were produced with each tomato variety. Whole leaves of 99 hybrids were bioassayed with twospotted spider mites, Tetranychus urticae Koch, allowing selection of six hybrids (two susceptible and four resistant) for each generation of each family. When these 24 hybrids were characterized for spider mite repellency with thumbtack bioassays, two hybrids had repellent leaflets, demonstrating that repellency was genetically transferred to interspecific tomato hybrids. Leaflet washes containing trichome secretions from each of three hybrids, including the two having repellent leaflets, were repellent in bridge bioassays. For the two hybrids having repellent leaflets and leaflet washes, removal of trichome secretions by dipping leaflets in methanol eliminated leaflet repellency. 2,3-Dihydrofarnesoic acid was present in trichome secretions of the hybrids having leaflet repellency, and it also was present in secretions of other hybrids, indicating that its presence is essential, but not sufficient for leaflet repellency. With regard to resistance, 16 of the hybrids tested had been identified as resistant in a whole leaf bioassay, but only two had repellent leaflets, indicating that other mechanisms of resistance are present in the resistant L. hirsutum parent.     

Approaches for sampling the twospotted spider mite (Acari: Tetranychidae) on clementines in Spain

Tetranychus urticae Koch (Acari: Tetranychidae) is an important pest of clementine mandarins, Citrus reticulata Blanco, in Spain. As a first step toward the development of an integrated crop management program for clementines, dispersion patterns of T. urticae females were determined for different types of leaves and fruit. The study was carried out between 2001 and 2003 in different commercial clementine orchards in the provinces of Castelló and Tarragona (northeastern Spain). We found that symptomatic leaves (those exhibiting typical chlorotic spots) harbored 57.1% of the total mite counts. Furthermore, these leaves were representative of mite dynamics on other leaf types. Therefore, symptomatic leaves were selected as a sampling unit. Dispersion patterns generated by Taylor's power law demonstrated the occurrence of aggregated patterns of spatial distribution (b > 1.21) on both leaves and fruit. Based on these results, the incidence (proportion of infested samples) and mean density relationship were developed. We found that optimal binomial sample sizes for estimating low populations of T. urticae on leaves (up to 0.2 female per leaf) were very large. Therefore, enumerative sampling would be more reliable within this range of T. urticae densities. However, binomial sampling was the only valid method for estimating mite density on fruit.     

Impact of twospotted spider mite (Acari: Tetranychidae) on growth and productivity of glasshouse cucumbers

Experiments were conducted to assess the damage of the glasshouse cucumber by twospotted spider mite, Tetranychus urticae Koch, and to investigate when the economic yield begins to decrease after T. urticae infestations. To assess the damage, dry matter partitioning in the cucumber plant was quantified and plant growth analyses were conducted at four different T. urticae infestation levels. T. urticae infestations decreased leaf productivity by reducing the total number of leaves per plant. Approximately 14% reductions of total leaf areas could result in significant yield loss. The decreased leaf productivity by T. urticae feeding caused biomass reductions and altered the pattern of dry matter partitioning in the plant; damaged plants accumulated more dry matter in the leaf, and partitioning of dry matter to fruits was hindered. The economic yield of cucumber began to significantly decrease as early as 4 wk after heavy mite infestations. This study also showed the seasonal differences in T. urticae-cucumber damage interactions among mite infestation levels.     

Leaf cell and tissue damage of cucumber caused by twospotted spider mite (Acari: Tetranychidae)

To quantify the damage caused by twospotted spider mite, Tetranychus urticae Koch, feeding on cucumber, Cucumis sativus L., leaf-cell and -tissue damage was assessed. On the abaxial leaf surface, adult T. urticae could feed through the spongy parenchyma and part of the palisade parenchyma of the leaf, while immature T. urticae could feed only through the sponge parenchyma. T. urticae punctured individual epidermal cells and consumed the contents of the mesophyll cells. Injured leaves had more empty space in the spongy parenchyma and fewer chloroplasts per cell. Damage also occurred even in the adjacent uninjured parenchyma cells without additional T. urticae feeding injury. Net photosynthetic rate, total chlorophyll content, and greenness of the leaf were significantly affected by feeding as quantified by mite-days. The percent loss of these parameters increased linearly or nonlinearly as mite-days increased, regardless of mite developmental stages. At 1,000 mite-days per 6 cm2, net photosynthetic rate was reduced by approximately 50 and 95%, total chlorophyll content was reduced by approximately 55 and 80%, and greenness was reduced by approximately 50 and 80% by feeding by immature and adult T. urticae, respectively.     

Efficacy and persistence of rosemary oil as an acaricide against twospotted spider mite (Acari: Tetranychidae) on greenhouse tomato

Efficacy of rosemary, Rosmarinus officinalis L., essential oil was assessed against twospotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae), as well as effects on the tomato, Lycopersicum esculatum Mill., host plant and biocontrol agents. Laboratory bioassay results indicated that pure rosemary oil and EcoTrol (a rosemary oil-based pesticide) caused complete mortality of spider mites at concentrations that are not phytotoxic to the host plant. The predatory mite Phytoseiulus persimilis Athias-Henriot is less susceptible to rosemary oil and EcoTrol than twospotted spider mite both in the laboratory and the greenhouse. Rosemary oil repels spider mites and can affect oviposition behavior. Moreover, rosemary oil and rosemary oil-based pesticides are nonpersistent in the environment, and their lethal and sublethal effects fade within 1 or 2 d. EcoTrol is safe to tomato foliage, flowers, and fruit even at double the recommended label rate. A greenhouse trial indicated that a single application of EcoTrol at its recommended label rate could reduce a twospotted spider mite population by 52%. At that rate, EcoTrol did not cause any mortality in P. persimilis nor did it affect their eggs. In general, EcoTrol was found to be a suitable option for small-scale integrated pest management programs for controlling twospotted spider mites on greenhouse tomato plants.     

Characterization of vitellin protein in the twospotted spider mite, Tetranychus urticae (Acari: Tetranychidae)

In mites, vitellogenin synthesis, regulation and uptake by the oocytes as vitellin remain practically unknown. Although a partial sequence of the gene is now available, no previous studies have been conducted that describe the native vitellin protein in mites. The objective of this study was to characterize vitellin in the twospotted spider mite, Tetranychus urticae. The native twospotted spider mite vitellin migrated as a single major band with a molecular weight of 476 ± 14.5 kDa as compared to 590 ± 25.5 kDa for vitellin from the American dog tick, Dermacentor variabilis. However, isoelectric focusing analysis of native spider mite vitellin showed five bands with pI values slightly acidic to neutral (pH 5.8, 6.2, 6.7, 7.0 and 7.2), as is the case for insect and tick vitellins. Reducing conditions (SDS-PAGE) also revealed multiple subunits ranging from 290.9 to 3.6 kDa and was similar to that found in D. variabilis. Spider mite vitellin weakly bound lipids and carbohydrates compared to the tick. Unlike D. variabilis, the spider mite egg yolk protein does not bind heme. The significance of non-heme binding in mites is discussed.     

Fecundity in twospotted spider mite (Acari: Tetranychidae) is increased by direct and systemic exposure to imidacloprid

The effect of imidacloprid on fecundity in twospotted spider mites, Tetranychus urticae Koch, was investigated in laboratory experiments using individual females on bean leaf discs. Mites were directly exposed to spray formulations of imidacloprid or fed on discs cut from a systemically treated bean plant. Imidacloprid-treated T. urticae produced 10-26% more eggs during the first 12 d of adult life and 19-23% more during adulthood compared with a water-only treatment. Increased egg production occurred immediately after exposure and lasted for about 15 d in sprayed mites. In mites exposed to imidacloprid by ingestion, increased egg production was not apparent until after 6 d and lasted until about day 18. Longevity was significantly greater in mites that ingested imidacloprid but not in sprayed mites. The significance and importance of imidacloprid-stimulation of fecundity in T. urticae to pest management in crop systems like hops, which routinely use this insecticide, is discussed.     

Host plant-induced changes in detoxification enzymes and susceptibility to pesticides in the twospotted spider mite (Acari: Tetranychidae)

Adult female twospotted spider mites, Tetranychus urticae Koch, reared on lima bean plants were moved to cucumber, maize, or new lima bean plants (the latter being a control) and evaluated after 24 h or 7 d for changes in susceptibility to three pesticides and in levels of related detoxification enzymes. The largest and most consistent changes were observed in mites feeding on cucumber. Susceptibility of mites on cucumber to the synthetic pyrethroids bifenthrin and lambda-cyhalothrin was greater than that of mites reared on lima bean and maize after only 24 h on the plants, and remained higher after 7 d. Mites on cucumber also were more susceptible to the organophosphate dimethoate than were mites on lima bean, but only after 7 d on the host. Susceptibility was inversely related to activities of both general esterase and glutathione S-transferase (GST) in mites on cucumber; general esterase and GST activities were 60 and 25% lower, respectively, than activities of twospotted spider mite on lima bean after 7 d of feeding. Mites on maize were slightly but significantly more susceptible than those on lima bean to bifenthrin, but not to lambda-cyhalothrin, after 7 d and to dimethoate after 24 h but not after 7 d. General esterase and GST activities in twospotted spider mite fed on maize for 24 h were 20 and 16% higher, respectively, than activities in twospotted spider mite on lima bean, but general esterase activity was 30% lower than lima bean-fed mites and GST was not different after 7 d. Thus, plant-induced changes in general esterase activity, perhaps in combination with GST activity, in twospotted spider mite appear to be inversely related to, and possibly responsible for, changes in susceptibility of twospotted spider mite to several pesticides, particularly the synthetic pyrethroids. General esterases appear to play less of a role in the detoxification of the organophosphate insecticide dimethoate.     

Effects of soil moisture and temperature on reproduction and development of twospotted spider mite (Acari: Tetranychidae) in strawberries

The effects of soil moisture and temperature on the reproduction of twospotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae), were examined in laboratory and field tests in strawberries, Fragaria x ananassa Duchesne, in Florida. Different soil moisture levels (low, moderate, and high) were compared to determine how soil moisture affects the reproduction and development of twospotted spider mite. In addition to soil moisture, different irrigation techniques (drip versus drip/overhead) were compared to determine their effects on twospotted spider mite reproduction as well as the incidence of angular leaf spot, Xanthomonas fragaria Kennedy & King disease. Similar studies were conducted to determine how different temperatures (18, 27, and 35 degrees C) affect the reproduction and development of twospotted spider mites. In the laboratory, low soil moisture as well as temperatures >27 degrees C promoted twospotted spider mite development. A similar trend was observed in a field study with low soil moisture promoting twospotted spider mite reproduction during the early season (11 November--8 December). Irrespective of moisture levels, a significantly higher incidence of X. fragaria was recorded in treatments with drip/overhead irrigation systems compared with drip irrigation. Implications for management of soil moisture levels are discussed with respect to the abundance of twospotted spider mite and X. fragaria in strawberries.     

A comparison of the toxic and sub-lethal effects of fluvalinate and esfenvalerate on the twospotted spider mite (Acari: Tetranychidae)

There was no difference in the direct toxicity of fluvalinate and esfenvalerate to twospotted spider mite (TSSM), Tetranychus urticae Koch. adults. The residual toxicity LC₅₀ of esfenvalerate was lower. Neither pyrethroid was toxic (<10% mortality) to TSSM eggs or adults at their recommended field concentrations. Fluvalinate was twice as toxic (45% mortality) than esfenvalerate to TSSM larvae at 0.01 g.a.i L^-1. The toxicity of the pyrethroids to TSSM protonymphs and deutonymphs was similar (16–28% mortality at 0.1 g a.i. L^-1). Dispersal from the treated surface was the main response to both pyrethroids by TSSM protonymphs, deutonymphs and adults. Maximum run-off by TSSM adults from fluvalinate and esfenvalerate treated surfaces was found with 0.01 and 0.005 g a.i. L^-1 respectively. Spin-down from pyrethroid treated surfaces was positively correlated with concentration. Oviposition was negatively correlated with concentration. Fluvalinate caused greater reductions in oviposition than esfenvalerate. Both pyrethroids reduced TSSM development rate from larval, protonymph and deutonymph stages, but fluvalinate caused larger reductions. Both pyrethroids prevented mating: for ten days oviposition 93% and 98% of offspring were male for esfenvalerate and fluvalinate respectively at 0.1 g a.i. L^-1. These findings are discussed with respect to the incidence of pyrethroid induced mite outbreaks.     

Chlorfenapyr resistance in two-spotted spider mite (Acari: Tetranychidae) from Australian cotton

The responses of Tetranychus urticae Koch from Australian cotton to chlorfenapyr has been monitored since the 1997–1998 growing season. Resistance was first detected in the 2001–2002 season and then increased quickly in both level and proportion of resistant strains detected. In response, the resistance management strategy for chlorfenapyr use in cotton was altered and now recommends a further restriction of use from two to one spray per season. There was no evidence of negative cross-resistance to the pyrethroid bifenthrin, but chlorfenapyr was associated with an undefined negative cross-resistance.     

Functional response of Neoseiulus barkeri Hughes on two-spotted spider mite (Acari: Tetranychidae)

Generalist predatory mites are the common phytoseiid fauna in many agroecosystems, but little attention has been paid to their potential as biological control agents. In this study, we determined the functional responses of adult females of the generalist predator Neoseiulus barkeri Hughes on eggs, larvae, and adults of the two-spotted spider mite, Tetranychus urticae Koch, in the laboratory. Predation experiments were conducted on pepper leaf discs over a 24 h period at 25±1°C, 70–80% RH and 16L:8D photoperiod. Prey densities ranged 5 to 80 eggs, or 5 to 40 larvae, or 1 to 8 female adults of T. urticae per disc. The predation rate of N. barkeri adult females on T. urticae eggs was the same as on its larvae, but the predation rate on adult females was much lower. The role of generalist predatory mites in integrated and biological control of greenhouse pests was discussed.     

Acaricidal activity and sublethal effects of an oxymatrine-based biopesticide on two-spotted spider mite (Acari: Tetranychidae)

Lethal and sublethal effects of the biopesticide Kingbo (oxymatrine 0.2 % + psoralen 0.4 %) on the two-spotted spider mite (Tetranychus urticae Koch) were investigated in laboratory bioassays. The biopesticide was applied to bean leaf discs or primary leaves by using a Potter spray tower. Acute toxicity tests showed no significant ovicidal action: toxic effect (LC₅₀ = 55.49 μl/l) was the result of a residual activity against larvae that hatched from the treated eggs. Preovipositional females and female teleiochrysales showed similar susceptibility (LC₅₀ = 52.68 and 59.03 μl/l, respectively), whereas larvae, protonymphs and female deutonymphs were the most susceptible stages (LC₅₀ = 6.88, 13.03, and 8.80 μl/l, respectively). In a choice test, females preferred the untreated halves of leaves over the halves treated with 2,000, 1,000, and 500 μl/l in the first 24 h, and their oviposition in those treatments was significantly greater on the untreated halves after 24 and 48 h, as well as the summed oviposition over 72 h. Viability and reproduction of survivors, as well as population growth, were strongly affected after the treatments of preovipositional females and female teleiochrysales with 100, 50 and 25 μl/l. On the other hand, sublethal effects on the females that survived treatment at the egg stage or reached adulthood from the eggs laid on the treated surface (treatments with 50 and 25 μl/l) were significantly weaker. Acaricidal and sublethal effects of the biopesticide Kingbo were discussed as a starting point for further research aimed to improve management of T. urticae populations. Regulatory issues and safety concerns regarding further commercialization of this biopesticide are addressed as well.     

Linkage between one of the polygenic hexythiazox resistance genes and an etoxazole resistance gene in the twospotted spider mite (Acari: Tetranychidae)

Genetic linkage between hexythiazox and etoxazole resistance loci was analyzed by crossing experiments. Two strains, one resistant (R) and the other susceptible (S) to both chemicals were established from field-collected Tetranychus urticae Koch (Acari: Tetranychidae) populations that were further selected in the laboratory. To analyze the recombination rate of the loci associated with resistance, we tested the ovicidal effects of a mixed solution of hexythiazox and etoxazole on haploid F2 eggs laid by F1 females from an R female x S male cross. This revealed tight or complete linkage between the hexythiazox and etoxazole resistance loci. We then assessed the number of loci associated with resistance to each acaricide based on mortality in the haploid F3 progeny (eggs) of F2 females from an F1 female (R x S) x S male testcross. The mortality rate indicated that etoxazole resistance was largely controlled by a single major locus, whereas hexythiazox resistance was controlled by more than one locus. Thus, one hexythiazox resistance locus was tightly or completely linked to the etoxazole resistance locus.     

Potentiation of a propargite and fenpyroximate mixture against two-spotted spider mite, Tetranychus urticae (Acari: Tetranychidae)

Using susceptible and resistant reference strains and standard bioassay methods a new miticide product was screened for efficacy and cross-resistance against two-spotted spider mite, Tetranychus urticae Koch. Experimental product NN1 850.0 EW (fenpyroximate and propargite 3:10 ratio) was significantly (P<0.05) more toxic than either compound tested alone (co-toxicity index 1029). Non-overlapping 95% CIs at the LC₅₀ and LC₉₉ level suggest that there is no cross-resistance to NN1 850.0 EW from existing organophosphate and pyrethroid resistance. NN1 850.0 EW is worthy of further development for T. urticae control.     

"Inert" formulation ingredients with activity: toxicity of trisiloxane surfactant solutions to twospotted spider mites (Acari: Tetranychidae)

Organosilicone molecules are important surfactant ingredients used in formulating pesticides. These methylated silicones are considered inert ingredients, but their superior surfactant properties allow them to wet, and either suffocate or disrupt important physiological processes in mites and insects. Aqueous solutions of the tri-siloxane surfactants Silwet L-77, Silwet 408, and Silwet 806 were bioassayed against adult female two-spotted spider mites, Tetranychus urticae Koch, with leaf dip methods to compare their toxicity with organosilicone molecules containing bulkier hydrophobic components. All three tri-siloxanes in aqueous solutions were equivalently toxic (LC50 = 5.5-8.9 ppm), whereas Silwet L-7607 solutions were less toxic (LC50 = 4,800 ppm) and Silwet L-7200 was nontoxic to mites. In another experiment, the toxicity of Silwet L-77 was affected by the wettability of leaf surfaces. The LC50 shifted from 22 to 84 ppm when mites were tested on bean and strawberry leaf disks, respectively. Droplet spreading on paraffin and surface tension were both related to the toxicity of surfactant solutions. Surface tensions of solutions below 23 mN/m caused > 90% mite mortality in leaf dip bioassays. A field test of Conserve SC and its formulation blank, with and without Dyne-Amic adjuvant (a vegetable oil-organosilicone surfactant mixture) revealed that Dyne-Amic had the greatest miticidal contribution, reducing mite populations by 70%, followed by formulation inactive ingredients. Spinosad, the listed active ingredient in Conserve, only contributed miticidal activity when synergized by Dyne-Amic. Researchers should include appropriate surfactant or formulation blank controls when testing insecticides or miticides, especially when using high spray volumes.     

Factors determining harem ownership in a subsocial spider mite (Acari,Tetranychidae)

Several factors which may affect harem ownership by males ofSchizotetranychus miscanthi Saitō (a new subsocial species of theSchizotetranychus celarius complex) were experimentally tested. A marking method was developed and used for studying spider mite behavior. Nest ownership (longer male residence in a nest) had little influence on the consequence of male-male combat. Furthermore, age differences in males never affected the outcome of the combats. Length of male leg I, however, appeared to be an important factor. Factors affecting variation in male leg size were examined. Length of leg I was strongly affected by host plant quality but not by male age.     

Control of phytoseiids in a spider mite mass-rearing system (Acari: Phytoseiidae,Tetranychidae)

Phytoseiid mites which contaminated the spider mite colony and interfered with the mass-rearing of spider mites were controlled by dipping in hot water. Immersion for 60 s in water of 50°C killed all stages ofAmblyseius fallacis (Garman),A. womersleyi Schicha andPhytoseiulus persimilis (Athias-Henriot) (Acari: Phytoseiidae). Only approximately 0.3% of theA. womersleyi eggs hatched, and this seems negligible. The populations ofTetranychus kanzawai Kishida andT. urticae (Koch) (Acari: Tetranychidae) were reduced. However, they recovered well. Although this treatment resulted in the withering of some soybean seedlings, the next trifoliate leaf to be produced was normal. A very satisfactory result was obtained when this technique was applied to the mass-rearing system.