Electrophoretic detection of enzyme variation among Japanese red-coloured spider mites of the genus Tetranychus (Acari: Tetranychidae)

The genetic variation in phosphoglucoisomerase (PGI) and malate dehydrogenase (MDH) was studied in red-pigmented Japanese spider mites of the genus Tetranychus by means of poly-acrylamide gel electrophoresis. The analysis revealed (1) that Tetranychus kanzawai possesses five and three alleles for PGI and MDH, respectively and that PGI allele frequencies clearly differ between the Hokkaido and the Honshu populations (2) that Tetranychus urticae and Tetranychus pueraricola, two closely related species, have different alleles for PGI and (3) that two populations of Tetranychus piercei, a species which had only been found on the islands of Okinawa, were obtained from Honshu and that the PGI locus is fixed for different alleles in the two populations. The two enzyme systems are apparently useful not only for discrimination of spider mite species but also for the detection of intraspecific variation. © Rapid Science Ltd. 1998     

Sexual selection and mating behavior in spider mites of the genus Tetranychus (Acari: Tetranychidae)

As sexual selection is a coevolutionary process between males and females, various morphological and behavioral traits have evolved in each sex. In the tetranychid mites Tetranychus urticae Koch and T. kanzawai Kishida (Acari: Tetranychidae), males can mate repeatedly, whereas females normally accept only the first copulation for fertilization. Since early times, it had been reported that males engage in precopulatory mate guarding and combat against conspecifics for females to enhance their reproductive success. On the other hand, it was believed that females do not have opportunities to choose their mates. In the last 10 or so years, however, several new findings related to mating behavior were reported. Some of the findings reinforce our established knowledge, whereas some of them explode it. Here, I review the mating behavior of T. urticae and T. kanzawai by incorporating recent findings and then propose a new direction for future research.     

Desiccation tolerance in diapausing spider mites Tetranychus urticae and T. kanzawai (Acari: Tetranychidae)

We investigated the effects of changes in vapor pressure deficit (VPD) on the survival of diapausing (winter form) and non-diapausing (summer form) spider mites Tetranychus urticae Koch and Tetranychus kanzawai Kishida (Acari: Tetranychidae). Adult females of both species were kept without food at VPDs of 0.0, 0.4, 0.7, 1.5, 1.9, or 2.7 kPa for 3, 6, 9, 12, or 15 days at 25 °C. Diapausing females of both species kept at a VPD of ≥0.4 kPa for ≥6 days clearly tolerated desiccation. Under water-saturated conditions (VPD = 0.0 kPa), in which no desiccation occurred, diapausing females showed high starvation tolerance: 90 % survived for up to 15 days. No interspecific differences in tolerance to desiccation or starvation were observed under most conditions. These results indicate that diapause functions increase tolerance to desiccation and starvation. Such multiple tolerances to harsh environments might support winter survival in spider mites.     

Non-destructive direct polymerase chain reaction (direct PCR) greatly facilitates molecular identification of spider mites (Acari: Tetranychidae)

Spider mites (Acari: Tetranychidae) include serious agricultural pests and some species have spread globally as invasive species.. For this reason, rapid and simple identification of spider mite species is necessary for agricultural field and plant quarantine inspection. DNA sequence-based molecular techniques can rapidly identify spider mites. However, extracting DNA from minute invertebrates is difficult, expensive and time consuming. Here, we describe a polymerase chain reaction (PCR) technique in which the whole body of a spider mite adult is non-destructively soaked in PCR solution. The mite is then removed intact and can be used as a voucher specimen, leaving the PCR solution as the template and avoiding the need for a DNA extraction kit. For this study, we used six common spider mite species from four genera [Tetranychus urticae Koch (red form), Tetranychus kanzawai Kishida, Tetranychus parakanzawai Ehara, Oligonychus gotohi Ehara, Eotetranychus smithi Pritchard & Baker and Panonychus citri (McGregor)]. A portion of the mitochondrial cytochrome c oxidase subunit I (COI) gene was amplified with a universal primer pair from all individuals examined and sequenced. This method shortened the time for molecular identification from 9 to 5 h and eliminated the cost of commercial kits for DNA extraction [ca. 600 yen (~5.3 USD) per sample].     

Resurgences of spider mites (Acari: Tetranychidae) induced by synthetic pyrethroids

Causes of spider mite (Acari: Tetranychidae) population resurgences consequent upon exposure to synthetic pyrethroid (SP) treatments are reviewed. Resurgences may be seen as soon as 1 week, or even as late as a whole season, post-treatment. Synthetic pyrethroids vary in their adverse effects on spider mites, and also differ in their ability to invoke resurgences of different spidermite species on diverse plants. These pesticides are lethal as well as repellent to phytoseiids and other predators that prey on spider mites, may inhibit fungi which attack the latter, and affect phytophagous competitors. Spider mites are likewise repelled by SPs, thus becoming more evenlydistributed and less web-restricted, with a resultant increase in fecundity. Spider-mite development is shortened due to SPs and the sex ratio becomes more female-biased; onset of winter diapause also seems to be delayed. Synthetic pyrethroids appear to sensitize to spider-mite infestation plants which have not hitherto been attacked. Some SP effects (whether on spider mites, natural enemies or competitors) appear to be direct, whereas others may be mediated through the host plants. The effect of SPs on the other Acari is variable within the Prostigmata and Astigmata. Most Mesostigmata and Metastigmata (ticks) are very sensitive, whilst the Cryptostigmata (Oribatei) appear to be insensitive. Synthetic pyrethroids-induced resurgences of Homoptera are comparatively reviewed, with the conclusion that some of the phenomena may be similar to those observed in spider mites. Various resurgence models are discussed, as well as the three main causes of variation (SPs, spider-mite species, host plants) in the observed phenomena. The need for more rigorous and carefully controlled experimentation is emphasized.     

Induced feeding preferences in dicofol-resistant two-spotted spider mites (Acari: Tetranychidae)

Preadult rearing conditions affected the behavior of dicofol-resistant two-spotted spider mites (Tetranychus urticae). Resistant spider mites reared on dicofol-treated leaves initiated a significantly greater number of feeding bouts on dicofol-treated leaves than did genetically identical spider mites reared on residue-free leaves. Therefore the prior exposure of resistant spider mites resulted in induced feeding preferences that could exacerbate the potential outcome of the resistance by resulting in greater amounts of feeding by resistant individuals on dicofol-treated areas. Since resistant individuals that had not experienced dicofol in their lifetime did not display this feeding preference, avoidance of this phenomenon of induced feeding preference may be an undescribed value of rotations of pesticides.     

Functional response ofAllothrombium pulvinum deutonymphs (Acari: Trombidiidae) on two-spotted spider mites (Acari: Tetranychidae)

Functional responses of deutonymphs of the predatory mite,Allothrombium pulvinum Ewing, on eggs and adult females of two-spotted spider mite,Tetranychus urticae Koch, were determined in the laboratory. Predation experiments were conducted on lima bean leaf discs over a 24-h period at 25±2°C, 30–50% RH and 24L: 0D photoperiod. Prey densities ranged from 10 to 120T. urticae eggs per disc or 2 to 32 adult females per disc.Allothrombium pulvinum deutonymphs were more effective againstT. urticae eggs than its adult females. The role ofA. pulvinum deutonymphs in integrated and biological pest control is discussed.     

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     

Evaluation of a damage threshold for two-spotted spider mites, Tetranychus urticae Koch (Acari: Tetranychidae), in hop culture

Damage caused by two‐spotted spider mites (Tetranychus urticae) at harvest to yield, quality (measured in percentage α‐acids content) and cone infestation was assessed on hop cvs Hallertauer Magnum, Hallertauer Tradition and Perle. Acaricide‐untreated hop plants with known levels of T. urticae infestation were compared with neighbouring acaricide‐treated plants. Although in 24 of the 36 experimental harvests the untreated hop plants had spider mite infestations of > 100 mites leaf^?1, yields and α‐acids content from the untreated plants were significantly lower than the treated plants in only four instances. However, although mite infestation of cones from untreated hops were significantly higher than acaricide‐treated plants in 27 of the 36 cases, in only one instance did that cause economic loss. Spider mite infestation levels of c. 90 mites leaf^?1 are tolerable at harvest time with little or no risk of causing economic loss to hop growers.     

ITS2 sequences as barcodes for identifying and analyzing spider mites (Acari: Tetranychidae)

The use of DNA barcodes, short DNA sequences from a standardized region of the genome, has recently been proposed as a tool to facilitate species identification and discovery. Here we show that second internal transcribed spacer of nuclear ribosomal DNA (rDNA-ITS2) barcodes effectively discriminate among 16 species of spider mites (Acari: Tetranychidae) from Israel. The barcode sequences of each species were unambiguously distinguishable from all other species and formed distinct, nonoverlapping monophyletic groups in the maximum-parsimony tree. Sequence divergences were generally much greater between species than within them. Using a 0.02 (2%) threshold for species diagnosis in our data set, 14 out of 16 species recognized by morphological criteria would be accurately identified. The only exceptions involved the low divergence, 0.011–0.015 (1.1–1.5%), between Tetranychus urticae and Tetranychus turkestani, where speciation may have occurred only recently. Still, these species had fixed alternative rDNA-ITS2 variants, with five diagnostic nucleotide substitutions. As a result, we tentatively conclude that rDNA-ITS2 sequence barcodes may serve as an effective tool for the identification of spider mite species and can be applicable as a diagnostic tool for quarantine and other pest management activities and decision-making. We predict that our work, together with similar efforts, will provide in the future the platform for a uniform, accurate, practical and easy-to-use method of spider mite species identification.     

DNA preparation method in eggs, immature stages, and diapausing females of Tetranychus spider mites (Acari: Tetranychidae) for diagnostic PCR-RFLP

A DNA preparation method for diapausing females, immature stages, and eggs of spider mites for diagnostic analyses using restriction fragment length polymorphisms after polymerase chain reaction (PCR-RFLP analysis) of the internal transcribed spacer (ITS) region including 5.8 S of the nucleic ribosomal DNA was developed using Tetranychus urticae Koch as a typical example of tetranychid mites. In diapausing females and deutonymphs, the method of crushing an individual mite with a glass rod was eminently successful and a useful method to obtain the DNA template for PCR. For protonymphs, larvae, and eggs older than 48?h after oviposition at 24?°C, the DNA preparation method for a single nematode, crushing a tiny sample with a filter paper chip, was also successful and a useful method to obtain the DNA template from mite individuals (eggs) for PCR. Consequently, we found a single distinctive band of an amplified DNA fragment after electrophoresis in all developmental stages, and digestion of the PCR product by a restriction endonuclease, DraI, resulted in identical banding patterns being exhibited in all developmental stages. Our findings indicate that PCR-RFLP analysis of the ITS region can be used for species identification of diapausing females, immature stages, and eggs of Tetranychus species.     

The locomotor activities on sites covered by silk produced by related and unrelated spider mites in Tetranychus urticae (Acari: Tetranychidae)

The two-spotted spider mite Tetranychus urticae is a silk producer known to live in groups with a common silk web that can cover entire plants and protect mites against predators, rain and wind. Silk also plays an important role during collective migration by aerial dispersal or by walking. In this context, we studied the locomotor activity i.e. time in movement, in resting, and in static exploration (probing behaviour) of an individual confronted to clean places and/or places covered by silk coming from its own population or from another. Two populations were tested, one of the red form (Tunisia) and another of the green form (Belgium). The experimental results showed that the presence or the absence of silk did not influence the relative proportion of each behavioural item either for the red or the green population. Individuals of the green form population spent more time moving and less time resting than individuals of the red form population and this, whatever the substrate (red/green silk, clean). Moreover, the silk from the red form population attracted individuals from both populations, whereas the silk produced by the green form population did not attract any individual either from the red or the green form. This surprising result might have been due to a difference in the quantity and/or quality of silk woven by the two forms. We discuss how the differences observed in the behaviour of these two populations may result from differences in their strategy to rapidly increase the population of the colony and reinforce the silk nest.     

Double-leaf-disk residue assay for assessing the toxicity of repellent acaricides to spider mites (Acari: Tetranychidae)

A closed double-leaf-disk (DLD) assay is presented which prevents escape (runoff) of spider mites and exposes them to treated leaf tissue during the entire test period. The physiological (direct) toxicity of acaricide residue can be estimated with this assay without the interference of behavioral factors. In contrast, runoff is often a major component of the total mortality in open single-leaf-disk (SLD) assays, in addition to direct mortality. Fenpropathrin, a highly repellent pyrethroid acaricide, caused primarily runoff and little direct mortality in SLD assays withPanonychus ulmi (Koch). When runoff and direct mortality were combined, concentration-response lines andlc ₅₀ values were identical to those obtained with the closed DLD assay. Assays with the organotin fenbutatin oxide gave similar results, suggesting that the runoff response should be included when estimating the residual toxicity of these compounds with the standard SLD assay. In fact, excluding runoff mortality from the analysis may underestimate the toxicity of foliar residue. Providing residue-free non-leaf surface inside the DLD cage caused a shift to the right of the concentration-response line, resulting in higherlc ₅₀s. The DLD assay can be employed for mite species which are difficult to confine, and in toxicological studies of repellent or irritating acaricides. This assay may be particularly useful for elucidating the relationship between physiological toxicity and mite behavior in response to acaricide residue.Technical Paper 9167 of the Oregon State University Agricultural Experiment State, Corvallis, Oregon, USA.     

Molecular approach in spider mites (Acari: Tetranychidae): Preliminary data on ribosomal DNA sequences

DNA sequence data were used to examine phylogenetic relationships between six species of economically important Tetranychidae mites:Eotetranychus carpini (Oudemans),E. pruni (Reck),Tetranychus pacificus McGregor,T. mcdanieli McGregor,T. turkestani Ugarov & Nikolski andT. urticae Koch. With primers directed toward conserved elements flanking the target region, the Polymerase Chain Reaction was used to amplify the ITS2 spacer of the ribosomal DNA molecule. The nucleotide sequence of a 300-bp fragment of the ITS2 was determined by direct sequencing and nucleotide divergence used for intra-generic comparison in mites. The resulting phylogenetic tree expressing interspecific relationships in genusTetranychus agrees with morphological data. The study demonstrates the usefulness of the approach in the assessment of the systematics and evolution of the group.     

Life history parameters of three phytophagous spider mites,Tetranychus piercei,T. truncatus and T. bambusae (Acari: Tetranychidae)

Tetranychus piercei McGregor and Tetranychus truncatus Ehara are major pests of many crops in Bangladesh, while Tetranychus bambusae Wang and Ma is found only on plants in the family Bambusaceae. We compared the development, survivorship, and life table parameters of T. piercei and T. truncatus on bean leaves and of T. bambusae on bamboo leaves at 25 °C, 60–70% RH and a photoperiod of 16:8 h (light:dark). The development times of T. piercei and T. truncatus from eggs to adult females were 10.2 and 8.8 days, respectively, when fed on bean leaves, while it took 9.5 days for T. bambusae feeding on Bambusa sp. leaves to develop from eggs to adult females. Mated T. piercei, T. truncatus, and T. bambusae females laid 186.9, 132.5, and 46.3 eggs on average over oviposition periods averaging 22.3, 13.5, and 20.2 days, respectively. The intrinsic rate of increase (r_m) was 0.275 for T. piercei, 0.301 for T. truncatus, and 0.182 for T. bambusae; meanwhile the net reproductive rate (R₀) was 114.5/female for T. piercei, 101.9/female for T. truncatus, and 25.4/female for T. bambusae. The doubling time (t_D) was lowest in T. truncatus (2.3 days) compare to T. piercei (2.5 days) and T. bambusae (3.8 days). The two polyphagous Tetranychus mites collected in Bangladesh were found to share similar life table parameters with Tetranychus mites from various other regions irrespective of food sources. This is the first report of the life table parameters of T. bambusae reared on bamboo leaves.     

Effects of chlorpyrifos and sulfur on spider mites (Acari: Tetranychidae) and their natural enemies

In many agricultural systems spider mites are believed to be induced pests, only reaching damaging densities after pesticides decimate predator populations. Wine grapes typically receive two types of pesticides, insecticides and fungicides. Chemicals in either class could impact spider mite densities both directly through spider mite mortality, and indirectly by negatively affecting natural enemies. The impact of a broad-spectrum insecticide (chlorpyrifos) and an inorganic fungicide (sulfur) on mites and their natural enemies was monitored in replicate open-field experiments conducted in an abandoned vineyard in Washington State. In both experiments, chemicals were applied within a 2 × 2 factorial design, allowing assessment of both main and interactive effects of the two chemicals. Following typical management practices on wine grapes in Washington State, we made a single insecticide application early in the season, but repeatedly applied sulfur throughout the season. In the absence of sulfur, chlorpyrifos application led to higher spider mite densities. The main effect of chlorpyrifos appeared to be indirect, perhaps mediated through mortality of generalist phytoseiid mites; generalists appeared to be unable to recover following even a single insecticide application, while there was no evidence for harmful effects of chlorpyrifos on specialist phytoseiid mites. Sulfur had direct suppressive effects on both pest and predatory mites, although in the second experiment the suppressive effect of sulfur on spider mites was weaker when chlorpyrifos was also applied. These field experiments suggest that a complex mix of direct and indirect effects of the two chemicals impacted spider mite population dynamics in our system.     

Molecular identification and phylogenetic analysis of spider mites (Prostigmata: Tetranychidae) of Turkey

The family Tetranychidae includes many agriculturally important species known as spider mites. Their morphological identification is quite difficult due to the tiny size of their taxonomic characters and the requirement for high-level expertise. This may lead to pest misidentification and thus failure in pest management. DNA-based species identification seems to offer an alternative solution to overcome these issues. In the present study, two common molecular markers—Cytochrome oxidase subunit I (COI) and Internal transcribed spacer 2 (ITS2)—were used to identify 10 spider mite species from Turkey. Furthermore, genetic distances for several of them were assessed. Panonychus ulmi and Bryobia kissophila had the lowest (1.1%) and highest (4.5%) intra-specific genetic distances, respectively. In addition, integrative taxonomy allowed to identify Eotetranychus quercicola in Turkey as a new record. The sequences herein obtained will allow rapid species identification using molecular techniques and will contribute to resolve the phylogenetic history of spider mites.