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.     

Temperature-dependent fumigant activity of essential oils against twospotted spider mite (Acari: Tetranychidae)

Fumigant activity of 34 commercial essential oils was assessed on female adults and eggs of twospotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae) at three temperatures (5, 15, and 25 degrees C). Common thyme, cinnamon, and lemongrass oils were equally effective on twospotted spider mite adults showing 85.8-100% mortality at 5 and 10 microl/liter air at 25 degrees C. At a lower temperature of 15 degrees C, lemongrass and peppermint resulted in > or =90% mortality of adults at 10 microl/liter air. Only lemongrass was relatively active at 5 microl/liter air, at 15 degrees C. At 5 degrees C, lemongrass and peppermint caused significantly higher adult mortality than controls but only at 10 microl/liter air. Common thyme oil showed the highest ovicidal activity at 5 microl/liter air at 25 degrees C. Among the main components of common thyme and lemongrass oils, citral was lethal to twospotted spider mite adults at all tested temperatures. Carvacrol, thymol, and citral caused the same inhibitory effects on the hatch of twospotted spider mite eggs at 25 degrees C. However, citral was more active than other compounds to twospotted spider mite eggs at 15 degrees C. Therefore, we conclude that citral has the best potential for development as a fumigant against twospotted spider mite on agricultural products harvested late in the growing season.     

Effects of neonicotinoid insecticides on the bionomics of twospotted spider mite (Acari: Tetranychidae)

Previous reports indicate that applications of imidacloprid, a neonicotinoid insecticide, can lead to population buildups of twospotted spider mite, Tetranychus urticae Koch, in the field. Moreover, laboratory studies showed enhanced fecundity of T. urticae after an imidacloprid treatment. In this study, experiments were conducted in the greenhouse to investigate the potential effects of imidacloprid and several other neonicotinoid insecticides on fecundity, egg viability, preimaginal survivorship, and sex ratio of T. urticae (German strain WI) on French beans, Phaseolus vulgaris L. Four insecticides, i.e., imidacloprid (Confidor 200SL), thiacloprid (Calypso 480 SC), acetamiprid (Mospilan 70 WP), and thiamethoxam (Actara 25 WG), were tested at field-relevant (100, 120, 125, and 95 ppm) and sublethal doses (10, 12, 12.5, and 9.5 ppm), respectively. Both spray and drench applications were tested. At field-relevant doses, fecundity of T. urticae decreased and was lower in the treatments compared with the untreated control, whereas preimaginal survivorship and proportion of female offspring (i.e., sex ratio) were lower compared with the control. At sublethal doses, no significant differences were found among the treatments. Data on egg viability, preimaginal survivorship, and sex ratio at sublethal doses followed the same trends as at field-relevant doses. In an additional experiment, the metabolism of imidacloprid into monohydroxy-imidacloprid, olefine, guanidine, and 6-chloronicotinic acid was compared with the oviposition pattern of T. urticae. These findings are discussed with regard to previous laboratory and field observations of imidacloprid-induced fertility increases in T. urticae.     

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.     

Comparative susceptibility and possible detoxification mechanisms for selected miticides in banks grass mite and two-spotted spider mite (Acari: Tetranychidae)

The susceptibility and possible detoxification mechanisms of the Banks grass mite (BGM), Oligonychus pratensis (Banks), and the two-spotted spider mite (TSM), Tetranychus urticae Koch, to selected miticides were evaluated with and without synergists. BGM was 112-fold more susceptible to the organophosphate dimethoate, and 24-fold more susceptible to both the pyrethroids bifenthrin and -cyhalothrin than TSM. The synergist triphenyl phosphate (TPP) enhanced the toxicities of bifenthrin and -cyhalothrin against BGM by 3.0- and 4.2-fold, respectively, and enhanced the toxicities of bifenthrin, -cyhalothrin, and dimethoate against TSM by 6.2-, 1.9-, and 1.7-fold, respectively. The synergist diethyl maleate (DEM) enhanced the toxicities of bifenthrin and -cyhalothrin against BGM by 2.2- and 2.9- fold, respectively, and enhanced the toxicity of bifenthrin against TSM by 4.1-fold. On the other hand, the synergist piperonyl butoxide (PBO) increased the toxicities of bifenthrin and -cyhalothrin by 6.0- and 2.6-fold, respectively, against BGM, and by 4.5- and 1.9-fold, respectively, against TSM. The significant synergism with these pyrethroids of all three tested synergists (except for DEM with -cyhalothrin against TSM) suggests that esterases, glutathione S-transferases, and cytochrome P450 monooxygenases all play important roles in their detoxification. However, the toxicity of dimethoate was not enhanced by these synergists in either mite species (except for TPP against TSM). Apparently, these metabolic enzymes play less of a role in detoxification of this organophosphate in these mites.     

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.     

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.     

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.     

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.     

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.     

Host plant associations in the spider mite Tetranychus urticae (Acari: Tetranychidae): insights from molecular phylogeography

This article integrates studies on the genetic variation of T. urticae populations and the interspecific variation of several tetranychid species. It aims at obtaining insights into the roles of the historical, geographical and ecological factors in the partitioning of variation of species. Two types of molecular markers were used to determine whether the patterns of genetic variation in mites inhabiting different host plants can shed light on the existence of host plant associations. The ribosomal sequences of the second internal transcribed spacer (ITS2), which evolves through concerted evolution, are good indicators of long-term isolation between populations and reveal exceptional homogeneity in a worldwide sampling of T. urticae. The mitochondrial cytochrome oxidase I (COI) sequences do not disclose old divergences related to host plant in this mite but rather suggest recent geographic colonization patterns of the species. Allozyme variation on a fine scale gives some evidence of host associations in the case of citrus trees. However, if any divergence of mites related to this host plant exists, it probably prevails in local populations only and it should not be old enough to be revealed by a phylogenetic analysis of mitochondrial COI sequences. The phyletic constraint in the evolution of feeding specificity in the family Tetra-nychidae is investigated based on a phylogenetic analysis of mitochondrial sequences. The results provide some support for the hypothesis that an evolutionary trend towards polyphagy has occurred in the family. Overall, it seems that the major characteristic of T. urticae is its high colonization potential. Polyphagy has enhanced its successful spread and may have led to connectivity between populations worldwide. © Rapid Science Ltd. 1998     

Influence of pesticides and neuroactive amines on cAMP levels of two-spotted spider mite (Acari: Tetranychidae)

The interaction of several formamidine pesticides, chlordimeform (CDM), N-demethylchlordimeform (DCDM) and amitraz with octopamine receptor(s) and the resulting enhancement of cyclic-AMP (cAMP) production in vitro were investigated in the two-spotted spider mite, Tetranychus urticae Koch. DCDM and amitraz clearly stimulated the production of cAMP when added to a homogenate of the spider mite. Among various biogenic amines tested, octopamine and synephrine were most active but dopamine (DA) and 5-hydroxytroptamine showed only marginal potency to elevate cAMP production. An additivity study was devised to find whether these formamidines interact with the same target site as octopamine. The results indicate that all these chemicals act on the same receptor which functions to transduce the signal of certain biogenic amines to elevate the intracellular cAMP level. Phentolamine (PH) and propranolol (PR) showed an antagonistic effect against the portion of cAMP production which was elevated by DCDM. Among pesticides tested, deltamethrin, fenvalerate, DDT and benzenehexachloride showed no such effect, whereas dicofol, chlorobenzilate, parathion and aldicarb showed slight stimulatory effects on cAMP production. Both DCDM and octopamine cause an increase in the phosphorylation of proteins that are also phosphorylated by exogenous cAMP-dependent protein kinase. The results of pharmacological characterization studies confirmed the overall theory that the agonistic effects of formamidines are expressed primarily through the octopamine-sensitive adenylate cyclase.     

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.     

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.     

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.     

Yield loss to corn from feeding by the Banks grass mite and two-spotted spider mite (Acari: Tetranychidae)

Our research was designed to determine the effects of a mite complex consisting of the Banks grass mite (BGM),Oligonychus pratensis (Banks), and the two-spotted spider mites (TSM),Tetranychus urticae Koch, on corn yield and plant lodging. For BGM, mite days and damage rating for the whole plant and leaves in the lower third of a corn plant had the best correlation with corn yield. The best correlation with yield for TSM was plant damage ratings. The percentage loss per unit for most independent variables (mite densities, mite days, or percentage of the leaf area damaged on a plant) was very similar for BGM and TSM. Therefore, the same economic threshold can be used for either mite species. When TSM fed on corn in the dent growth stage, yield was not reduced, and their feeding did not influence corn plant lodging.     

Genetic basis of resistances to chlorfenapyr and etoxazole in the two-spotted spider mite (Acari: Tetranychidae)

We studied the genetic basis of resistance to two new acaricides, chlorfenapyr and etoxazole, which have different chemical structures and modes of action in the two-spotted spider mite, Tetranychus urticae Koch. The resistance ratios calculated from the LC50s of resistant and susceptible strains were 483 for chlorfenapyr and >100,000 for etoxazole. Mortality caused by the two acaricides in F1 progeny from reciprocal crosses between the resistant and susceptible strains indicated that the modes of inheritance of resistance to chlorfenapyr and etoxazole were completely dominant and completely recessive, respectively. Mortality in F2 progeny indicated that for both acaricides, the resistance was under monogenic control. Repeated backcross experiments indicated a linkage relationship among the two acaricide resistances and malate dehydrogenase, although phosphoglucoisomerase was not linked with them. The recombination ratio between the resistances was 14.8%. From this result, we suggest that heavy spraying of the two acaricides will lead to apparent cross-resistance as a consequence of crossing over; the two resistance genes are so close to each other that it would be difficult to segregate them once they came together on the same chromosome.