Resistance to the two-spotted spider mite (<Emphasis Type="Italic">Tetranychus urticae</Emphasis>) by acylsucroses of wild tomato (<Emphasis Type="Italic">Solanum pimpinellifolium</Emphasis>) trichomes studied in a recombinant inbred line population

Trichome-based host plant resistance is a complex mechanism that could be used in tomato breeding to control arthropod pests. The aims of this work were to evaluate the plant traits (density of trichomes and acylsucrose production) and the functional relationships of these traits with mortality, repellence, and oviposition of Tetranychus urticae Koch (Acari: Tetranychidae). We used a population of recombinant inbred lines (RILs) derived from the cross between the wild tomato, Solanum pimpinellifolium L. ‘TO-937’, and the cultivated tomato, Solanum lycopersicum L. Multiple regression analyses showed that high acylsucrose content and high type-IV trichome density increased mortality and repellence, and reduced oviposition of T. urticae. Single regression analyses showed that a logistic model best explained the relationship between mortality or repellence and acylsucrose content, whereas a negative-exponential model best described the relationship between oviposition and acylsucrose content. Linear models were the best-fits for the three resistance variables with trichome IV density. Probit analysis was used to estimate acylsucrose effective doses, and revealed that 31 and 10% of the RILs produced acylsucrose above the effective doses for 90% mortality or repellence, respectively. Altogether, these results indicate that S. pimpinellifolium may be a suitable genetic source of resistance to spider mites to be used in cultivated tomato.     

UV-B susceptibility and photoreactivation in embryonic development of the two-spotted spider mite, <Emphasis Type="Italic">Tetranychus urticae</Emphasis>

Developmental errors are often induced in the embryos of many organisms by environmental stress. Ultraviolet-B radiation (UV-B) is one of the most serious environmental stressors in embryonic development. Here, we investigated susceptibility to UV-B (0.5 kJ m^?2) in embryos of the two-spotted spider mite, Tetranychus urticae, to examine the potential use of UV-B in control of this important agricultural pest worldwide. Peak susceptibility to UV-B (0% hatchability) was found in T. urticae eggs 36–48 h after oviposition at 25 °C, which coincides with the stages of morphogenesis forming the germ band and initial limb primordia. However, hatchability recovered to?~?80% when eggs irradiated with UV-B were subsequently exposed to visible radiation (VIS) at 10.2 kJ m^?2, driving photoreactivation (the photoenzymatic repair of DNA damage). The recovery effect decreased to 40–70% hatchability, depending on the embryonic developmental stage, when VIS irradiation was delayed for 4 h after the end of exposure to UV-B. Thus UV-B damage to T. urticae embryos is critical, particularly in the early stages of morphogenesis, and photoreactivation functions to mitigate UV-B damage, even in the susceptible stages, but immediate VIS irradiation is needed after exposure to UV-B. These findings suggest that nighttime irradiation with UV-B can effectively kill T. urticae eggs without subsequent photoreactivation and may be useful in the physical control of this species.     

Antixenosis and antibiosis response of common bean (<Emphasis Type="Italic">Phaseolus vulgaris</Emphasis>) to two-spotted spider mite (<Emphasis Type="Italic">Tetranychus urticae</Emphasis>)

The two-spotted spider mite, Tetranychus uticae Koch (Acari: Tetranychidae), is globally one of the most devastating pests that feed on numerous crops, including common bean (Phaseolus vulgaris L.). This study was aimed to evaluate the effects of genotype and morphological attributes of common bean on T. uticae. Forty common bean accessions were used to investigate antixenosis and antibiosis through assessing mite feeding preference and reproduction under laboratory conditions. Three resistant (i.e., 56, 63, 238) and two susceptible (i.e., 182, 236) accessions, along with cultivars Naz (resistant) and Akhtar (susceptible), were used in a life-table study. Both antixenosis and antibiosis mechanism were observed in all of the accessions, albeit a negative correlation occurred. Significant differences were observed for all traits of T. urticae: developmental time of immature stages, reproduction, adult longevity and life-table parameters. Based on the intrinsic rate of increase, the accessions 56, 63, 182, 238, and cv. Naz impose high antibiotic effects on T. urticae. Although significant variation existed among accessions for morphological factors, only glandular trichomes correlated with mite fecundity and feeding preference.     

Effects of azadirachtin on the two-spotted spider mite, <Emphasis Type="Italic">Tetranychus urticae</Emphasis> (Acari: Tetranychidae)

Effects of sublethal azadirachtin exposure to the biological performance of Tetranychus urticae Koch was studied under laboratory conditions. Bioassay was used to asses the effect of different concentrations of azadirachtin on longevity, fecundity, fertility, and offspring development. Azadirachtin (64 and 128 ppm) affected fecundity and mortality but had no effect on fertility and offspring development. A subsequent life-table study with 80 ppm of azadirachtin found that the compound caused a reduction of 50% in survival to adult stage. The peak of reproduction was reached at 5 days causing a decrease in the mean fecundity to almost eight times than of untreated females. The net reproductive rate (R ₀), the intrinsic rate of increase (r _m), and the finite rate of increase () of treated females were lower. Treatment showed a negative value of r _m, resulting in a declining population. These results suggest that azadirachtin could be incorporated in Integrated Pest Management (IPM) programmes of T. urticae.This revised version was published online in May 2005 with a corrected cover date.     

Vasa expression and germ-cell specification in the spider mite <Emphasis Type="Italic">Tetranychus urticae</Emphasis>

The specification of germ cells is an important process during the development of all animals. Expression of an evolutionarily conserved gene such as vasa can be used as a marker for germ cell fate. We have isolated a vasa-related gene from the two-spotted spider mite (Tetranychus urticae) and used it to examine the segregation of germ cells in this animal. In spider mites, vasa expression first appears in a group of cells that do not join the initial blastoderm surface. Instead, these cells remain in the interior of the blastoderm and then migrate to posterior regions of the embryo, where they form a cluster that appears in regions of the embryo consistent with the gonads. The expression pattern of this spider mite vasa homologue implies a novel process acts to specify germ cells in this species and that the specification of germ cells is an evolutionarily labile process.     

Consumption rate and functional response of the predaceous mite<Emphasis Type="Italic"> Kampimodromus aberrans</Emphasis> to two-spotted spider mite <Emphasis Type="Italic">Tetranychus urticae</Emphasis> in the laboratory

Prey stage preference of female Kampimodromus aberrans (Oudemans) (Phytoseiidae) at constant densities of different stages of Tetranychus urticae Koch (Tetranychidae), functional response types and parameters of the predator females to the varying densities of eggs, larvae, protonymphs and deutonymps of T. urticae were determined in order to establish its potential for the mite biological control. Experiments were conducted at 25 ± 1°C, 65 ± 10% RH and 16:8 (L:D) photoperiod. Our results indicated that the predator consumed significantly more prey larvae than other prey stages. Functional response type of predator was determined by a logistic regression model. The predator exhibited a Type II response on all prey stages. The attack rate (α) and handling time (T _h ) coefficients of a Type II response were estimated by fitting a “random-predator” equation to the data. The lowest estimated value α and the highest value of T _h (including digestion) were obtanined for the predator feeding on deutonmph. The lowest value of T _h were obtained for the predator feeding on prey larvae, but the attack rate value obtained on larva wasn’t different than that obtained on egg and protonymph. According to our results, K. aberrans could be an efficient biological control agent of T. urticae at least at low prey densities. However, further field based studies are needed to draw firm conclusions.     

Chemical defense,mycorrhizal colonization and growth responses in <Emphasis Type="Italic">Plantago lanceolata</Emphasis> L.

Allelochemicals defend plants against herbivore and pathogen attack aboveground and belowground. Whether such plant defenses incur ecological costs by reducing benefits from plant mutualistic symbionts is largely unknown. We explored a potential trade-off between inherent plant chemical defense and belowground mutualism with arbuscular mycorrhizal fungi (AMF) in Plantago lanceolata L., using plant genotypes from lines selected for low and high constitutive levels of the iridoid glycosides (IG) aucubin and catalpol. As selection was based on IG concentrations in leaves, we first examined whether IG concentrations covaried in roots. Root and leaf IG concentrations were strongly positively correlated among genotypes, indicating genetic interdependence of leaf and root defense. We then found that root AMF arbuscule colonization was negatively correlated with root aucubin concentration. This negative correlation was observed both in plants grown with monocultures of Glomus intraradices and in plants colonized from whole-field soil inoculum. Overall, AMF did not affect total biomass of plants; an enhancement of initial shoot biomass was offset by a lower root biomass and reduced regrowth after defoliation. Although the precise effects of AMF on plant biomass varied among genotypes, plants with high IG levels and low AMF arbuscule colonization in roots did not produce less biomass than plants with low IG and high AMF arbuscule colonization. Therefore, although an apparent trade-off was observed between high root chemical defense and AMF arbuscule colonization, this did not negatively affect the growth responses of the plants to AMF. Interestingly, AMF induced an increase in root aucubin concentration in the high root IG genotype of P. lanceolata. We conclude that AMF does not necessarily stimulate plant growth, that direct plant defense by secondary metabolites does not necessarily reduce potential benefits from AMF, and that AMF can enhance concentrations of root chemical defenses, but that these responses are plant genotype-dependent.     

Biological control of twospotted spider mite, <Emphasis Type="Italic">Tetranychus urticae</Emphasis>, with predatory mite, <Emphasis Type="Italic">Neoseiulus californicus</Emphasis>, in strawberries

Greenhouse and field experiments were conducted from 2005 to 2007 to determine the effectiveness of different release times with the predatory mite, Neoseiulus californicus (McGregor), for control of the twospotted spider mite (TSSM), Tetranychus urticae Koch, in strawberries (Fragaria x ananassa Duchesne). The effect of N. californicus releases over time and on development of TSSM populations during a growing season were evaluated. Our hypothesis was that repeated applications of N. californicus, which is currently recommended by biological control companies, might be unnecessary to attain season-long control of TSSM. In greenhouse trials, three treatments consisting of releases of N. californicus at five-day intervals: day 0, day 5, and day 10, and an untreated control were evaluated. The treatment releases significantly reduced TSSM below the control within five days of each release. Neoseiulus californicus significantly reduced TSSM in treatments with high densities (leaflets with ≥ 40 TSSM) below that of treatments with lower densities (leaflets with ≤ 10 TSSM) demonstrating that if released at a predator: prey ratio of 1:10, timing of release does not alter the effectiveness of N. californicus in controlling TSSM. However, we found that if the ratio of predator: prey remains adequate, N. californicus is a more efficient predator at high TSSM densities. Field studies included three treatments consisting of releases of N. californicus at one-month intervals. All treatments significantly reduced TSSM compared with the control plots (no releases). Releases applied early in the season sustained TSSM significantly below those in the control plots for the whole season. Our results indicate that one release of N. californicus is able to sustained control of TSSM in strawberry throughout a growing season if released when TSSM populations are low early in the season in the southeastern United States.     

Effects of anoxia and hypoxia on the two-spotted spider mite, <Emphasis Type="Italic">Tetranychus urticae</Emphasis> (Acari: Tetranychidae)

We investigated the potential use of anoxic (0% O₂) and hypoxic (lower O₂ concentration than in the atmosphere) conditions for controlling the two-spotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae). Adult T. urticae females were exposed to O₂ concentrations of 0, 0.5, 1, 2, or 21% (control) with a constant CO₂ concentration of 0.05% at 1 atm and 25 °C under continuous darkness for 24 h. The survival and fecundity at 8 days after treatment significantly decreased when the O₂ concentration was lower than 0.5% and 1%, respectively; the lethal concentration at 50% survival (LC₅₀) was 0.55%. The miticidal hypoxic condition (0.5% O₂) led to physiological disorders in host plants. The degree of physiological disorders differed among the plant species tested. Although tomato seedlings died after the hypoxia treatment, in kidney bean and cucumber seedlings the primary leaves remained and lateral buds developed instead of the apical buds that ceased. Hypoxia treatment could be useful as a physical measure for controlling spider mites depending on plant species or cultivars.     

Sublethal effects of spirodiclofen on life history and life-table parameters of two-spotted spider mite (<Emphasis Type="Italic">Tetranychus urticae</Emphasis>)

Laboratory bioassays were conducted to evaluate the effects of spirodiclofen on life history and life-table parameters of two-spotted spider mite (Tetranychus urticae Koch) females treated at pre-ovipositional period with a series of acaricide concentrations starting with the concentration discriminative for eggs and immatures i.e. the lowest concentration that causes 100% mortality of those stages. After a 24 h exposure, the proportion of females that survived treatments was 0.71 (6 mg/l), 0.51 (12 mg/l), 0.41 (24 mg/l), 0.30 (48 mg/l) and 0.25 (96 mg/l). At the end of the trial, the survival rate of females treated with the lowest concentration was significantly lower than the survival rate of untreated females but it remained above that of females treated with higher concentrations. Total fecundity/fertility significantly decreased as concentrations of spirodiclofen increased. Viable eggs were laid by females treated with 6, 12 and 24 mg/l, and total fertility was reduced by 42, 84 and 97%, respectively. Compared with control, the gross fecundity/fertility of the treated females was significantly reduced throughout the trial, except in females treated with 6 mg/l. All concentrations caused a significant reduction in the net fecundity/fertility throughout the trial. The females treated with 12 mg/l had significantly reduced net reproductive rate (R ₀ = 6.45), compared to females treated with 6 mg/l (R ₀ = 23.35) and to untreated females (R ₀ = 28.92); there was no significant difference between the last two treatments. The intrinsic rate of increase (r _m ) and finite rate of increase (λ) were significantly reduced in treated females (r _m = 0.141, λ = 1.156 and r _m = 0.214, λ = 1.232; 12 and 6 mg/l, respectively), compared to control (r _m = 0.251, λ = 1.276). The reduction was significantly greater in females treated with the highest concentration. As a result of the lowered r _m , the doubling time in treated females was significantly extended. Sublethal effects of spirodiclofen and its impact on T. urticae management are discussed.     

Preference and performance of the two-spotted spider mite <Emphasis Type="Italic">Tetranychus urticae</Emphasis> (Acari: Tetranychidae) on strawberry cultivars

The first record of human infestation and feeding by the native tick species Ixodes australiensis is reported in Australia based on a specimen collected from an adult male. Human infestation by ticks in Australia is reviewed and a concise list of anthropophagic ticks occurring in Australia is presented.     

Function of the web box as an anti-predator barrier in the spider mite,<Emphasis Type="Italic"> Schizotetranychus recki</Emphasis>

Spider mites inhabiting Sasa bamboo show considerable variation in traits believed to be the result of coevolution between predator and prey. In Schizotetranychus recki Ehara inhabiting the hairy leaves of a dwarf bamboo, Sasa senanensis, all quiescent stages, including eggs, appear within web boxes in the leaf hairs of their host plant, and this habit is thought to be a trait involved in predator avoidance. To test this hypothesis, the survival rates of S. recki eggs inside a web box and those freed of a web box were assessed in relation to six predator species that co-occur with the spider mite in the field. The results clearly show that the webbing behavior (web box) of S. recki has a function in avoiding five predator species. However, one predator species, Agistemus summersi Ehara, preyed more on eggs protected by a web box. This suggests that this predator species has a special trait to overcome the nest barriers.     

Effects of extending the light phase on diapause induction in a Japanese population of the two-spotted spider mite, <Emphasis Type="Italic">Tetranychus urticae</Emphasis>

Artificial lighting is a merit of a ‘plant factory’, which might be utilized to suppress an increase in pest population. We investigated the effects of extending the light phase on diapause induction in the two-spotted spider mite (TSSM), Tetranychus urticae. TSSM were reared at 18°C under light phases ranging from 2 to 64 h combined with a constant dark phase of 16 h in aluminum bottles, with white light emitting diodes attached inside to minimize fluctuations in air temperature between the light and dark phases. Diapause was induced in adult TSSM females when the light phase was 24 h or shorter, and diapause induction was inhibited when the light phase extended over 32 h. The development of deutonymphs was delayed under a diapause-inducing photoperiod. Diapause inducing photoperiods may suppress an increase in the TSSM population, by slowing down development and reproduction.     

<Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> is a susceptible host plant for the holoparasite <Emphasis Type="Italic">Cuscuta </Emphasis>spec

Arabidopsis thaliana and Cuscuta spec. represent a compatible host–parasite combination. Cuscuta produces a haustorium that penetrates the host tissue. In early stages of development the searching hyphae on the tip of the haustorial cone are connected to the host tissue by interspecific plasmodesmata. Ten days after infection, translocation of the fluorescent dyes, Texas Red (TR) and 5,6-carboxyfluorescein (CF), demonstrates the existence of a continuous connection between xylem and phloem of the host and parasite. Cuscuta becomes the dominant sink in this host–parasite system. Transgenic Arabidopsis plants expressing genes encoding the green fluorescent protein (GFP; 27 kDa) or a GFP–ubiquitin fusion (36 kDa), respectively, under the companion cell (CC)-specific AtSUC2 promoter were used to monitor the transfer of these proteins from the host sieve elements to those of Cuscuta. Although GFP is transferred unimpedly to the parasite, the GFP–ubiquitin fusion could not be detected in Cuscuta. A translocation of the GFP–ubiquitin fusion protein was found to be restricted to the phloem of the host, although a functional symplastic pathway exists between the host and parasite, as demonstrated by the transport of CF. These results indicate a peripheral size exclusion limit (SEL) between 27 and 36 kDa for the symplastic connections between host and Cuscuta sieve elements. Forty-six accessions of A. thaliana covering the entire range of its genetic diversity, as well as Arabidopsis halleri, were found to be susceptible towards Cuscuta reflexa.     

A key volatile infochemical that elicits a strong olfactory response of the predatory mite <Emphasis Type="Italic">Neoseiulus californicus</Emphasis>, an important natural enemy of the two-spotted spider mite <Emphasis Type="Italic">Tetranychus urticae</Emphasis>

Herbivore-induced plant volatiles (HIPVs) emitted from lima bean leaves infested with the two-spotted spider mites Tetranychus urticae strongly attract the predatory mites Neoseiulus californicus. Among these HIPVs, methyl salicylate and linalool can attract the predators. Three green-leaf volatiles (GLVs) of (Z)-3-hexen-1-ol, (Z)-3-hexenyl acetate and (E)-2-hexenal, found in the odor blends from T. urticae-infested leaves and physically damaged leaves, can also attract the predators. To search for a strong predator attractant, the olfactory responses of N. californicus to each synthetic compound or their combinations were investigated in a Y-tube olfactometer. When presented a choice between a mixture of the five compounds (i.e. the two HIPVs and the three GLVs) and T. urticae-infested leaves, N. californicus did not discriminate between these odor sources. The same trend was observed when either a mixture of the two HIPVs or methyl salicylate vs. T. urticae-infested leaves were compared. In contrast, the predators preferred T. urticae-infested leaves to linalool, each of the three GLVs, or a mixture of the three GLVs. These results indicated that methyl salicylate is a strong predator attractant, and its potential attractiveness almost equaled that of the blend of HIPVs from T. urticae-infested leaves.     

The predatory mite <Emphasis Type="Italic">Neoseiulus womersleyi</Emphasis> (Acari: Phytoseiidae) follows extracts of trails left by the two-spotted spider mite <Emphasis Type="Italic">Tetranychus urticae</Emphasis> (Acari: Tetranychidae)

As it walks, the two-spotted spider mite Tetranychus urticae Koch (Acari: Tetranychidae) spins a trail of silk threads, that is followed by the predatory mite, Neoseiulus womersleyi Schicha (Acari: Phytoseiidae). Starved adult female N. womersleyi followed T. urticae trails laid down by five T. urticae females but did not follow a trail of one T. urticae female, suggesting that the amount of spun threads and their chemical components should correlate positively with the number of T. urticae individuals. To examine whether chemical components of T. urticae trails are responsible for the predatory mite’s trail following, we collected separate T. urticae threads from the exuviae and eggs, and then washed the threads with methanol to separate chemical components from physical attributes of the threads. Female N. womersleyi did not follow T. urticae trails that had been washed with methanol but contained physical residues, but they did follow the direction to which the methanol extracts of the T. urticae trails was applied. These results suggest that the predatory mite follows chemical, not physical, attributes of T. urticae trails.     

Gene silencing in the spider mite <Emphasis Type="Italic">Tetranychus urticae</Emphasis>: dsRNA and siRNA parental silencing of the <Emphasis Type="Italic">Distal-less</Emphasis> gene

A major prerequisite to understanding the evolution of developmental programs includes an appreciation of gene function in a comparative context. RNA interference (RNAi) represents a powerful method for reverse genetics analysis of gene function. However, RNAi protocols exist for only a handful of arthropod species. To extend functional analysis in basal arthropods, we developed a RNAi protocol for the two-spotted spider mite Tetranychus urticae focusing on Distal-less (Dll), a conserved gene involved in appendage specification in metazoans. First, we describe limb morphogenesis in T. urticae using confocal and scanning electron microscopy. Second, we examine T. urticae Dll (Tu-Dll) mRNA expression patterns and correlate its expression with appendage development. We then show that fluorescently labeled double-stranded RNA (dsRNA) and short interfering RNA (siRNA) molecules injected into the abdomen of adult females are incorporated into the oviposited eggs, suggesting that dsRNA reagents can be systemically distributed in spider mites. Injection of longer dsRNA as well as siRNA induced canonical limb truncation phenotypes as well as the fusion of leg segments. Our data suggest that Dll plays a conserved role in appendage formation in arthropods and that such conserved genes can serve as reliable starting points for the development of functional protocols in nonmodel organisms.     

<Emphasis Type="Italic">Wolbachia</Emphasis>-induced cytoplasmic incompatibility in Japanese populations of <Emphasis Type="Italic">Tetranychus urticae</Emphasis> (Acari: Tetranychidae)

Intracellular bacteria of the genus Wolbachia (alpha Proteobacteria) induce cytoplasmic incompatibility (CI) in many arthropod species, including spider mites, but not all Wolbachia cause CI. In spider mites CI becomes apparent by a reduced egg hatchability and a lower daughter:son ratio: CI in haplodiploid organisms in general was expected to produce all-male offspring or a male-biased sex ratio without any death of eggs. In a previous study of Japanese populations of Tetranychus urticae, two out of three green-form populations tested were infected with non-CI Wolbachia strains, whereas none of six red-form populations harbored Wolbachia. As the survey of Wolbachia infection in T. urticae is still fragmentary in Japan, we checked Wolbachia infection in thirty green-form populations and 29 red-form populations collected from a wide range of Japanese islands. For Wolbachia-infected populations, we tested the effects of Wolbachia on the reproductive traits and determined the phylogenetic relationships of the different strains of Wolbachia. All but one green-form populations were infected with Wolbachia and all strains belonged to the subgroup Ori when the wsp gene was used to determine the phylogenetic relationships of different strains of Wolbachia. Six out of 29 red-form populations harbored Wolbachia and the infected strains belonged to the subgroups Ori and Bugs. Twenty-four of 29 infected green-form populations and five of six infected red-form populations induced CI among the hosts. Thus, CI-Wolbachia strains are widespread in Japan, and no geographical trend was observed in the CI-Wolbachia. Although three red-form populations harbored other intracellular bacteria Cardinium, they did not affect host reproduction.