Biogenic amines in the two-spotted spider mite

Whole body extracts of the two-spotted spider mite (Tetranychus urticae Koch) were analyzed using a 16-channel electrochemical array high performance liquid chromatography (HPLC)-based detection system that allows the simultaneous isolation and identification of a variety of biogenic amines. The spider mite extracts were found to contain the biogenic amines octopamine, dopamine, and 5-hydroxytryptamine (5-HT), as well as several precursors and metabolites including tyrosine, tyramine, tryptophan, and N-acetyl octopamine. Differences in the levels of biogenic amines were observed between eggs and the adult stages and between males and females. This is the first direct determination of biogenic amines in the Tetranychidae and the first demonstration of 5-HT in any mite species. © 1994 Wiley-Liss, Inc.     

Multiple resistance mechanisms to abamectin in the two-spotted spider mite

The two-spotted spider mite, Tetranychus urticae, is one of the most destructive pests of various orchard trees and garden plants. Biochemical mechanisms of abamectin resistance in two T. urticae strains (PTF, 239-fold resistance; AbaR, ca. 4753-fold resistance) were investigated. The involvement of both esterase (Est) and mixed function oxidase (MFO) in abamectin resistance was suggested by synergistic bioassays, in which median lethal time (LT₅₀) values were significantly reduced by pretreatment with triphenyl phosphate (TPP) and piperonyl butoxide, respectively. Detoxification enzyme assays confirmed that Est and MFO were related to abamectin resistance as metabolic factors. Moreover, some Est bands on a native isoelectric focusing gel were specifically inhibited by TPP, implying their association with resistance. Pretreatment with verapamil in synergistic bioassays did not reduce the LT₅₀ to abamectin, suggesting that the ABC transporter is not likely involved in resistance. However, enhanced MFO and Est activities in the AbaR strain were not enough to account for the extremely high level of abamectin resistance, which suggests the involvement of additional resistance mechanisms, such as target site insensitivity.     

Mate choice promotes inbreeding avoidance in the two-spotted spider mite

Since inbreeding in Tetranychus urticae can reduce offspring fitness, sexual selection may favour disassortative mate choice with respect to relatedness of the mating partners. We tested whether T. urticae shows this preference for mating with unrelated partners. We chose an experimental set-up with high potential for female choosiness, since females only mate once and are therefore expected to be the choosier gender. An adult virgin female was placed together with two adult males from the same population. One male was unrelated and the other male was related—a brother with whom she had grown up. Significantly more copulations (64%) took place with the unrelated male. Time to mating did not depend on the female-to-male relatedness. The remaining (non-copulating) male tried to interfere with the ongoing mating in the majority of cases, but this interference did not depend on the female-to-male relatedness. These results imply that T. urticae (a) can recognize kin (via genetic and/or environmental similarity) and (b) has the potential to avoid inbreeding through mate choice.     

Wolbachia-induced 'hybrid breakdown' in the two-spotted spider mite Tetranychus urticae Koch

The most common post-zygotic isolation mechanism between populations of the phytophagous mite Tetranychus urticae is 'hybrid breakdown', i.e. when individuals from two different populations are crossed, F1 hybrid females are produced, but F2 recombinant male offspring suffer increased mortality. Two-spotted spider mites collected from two populations, one on rose and the other on cucumber plants, were infected with Wolbachia bacteria. These bacteria may induce cytoplasmic incompatibility in their hosts: uninfected (U) females become reproductively incompatible with infected (W) males. We report on the effect of Wolbachia infections in intra- and interstrain crosses on (i) F1 mortality and sex ratios (a test for cytoplasmic incompatibility), and (ii) the number of haploid offspring and mortality in clutches of F1 virgins (a test for hybrid breakdown). U x W crosses within the rose strain exhibited partial cvtoplasmic incompatibility. More interestingly, F2 males suffered increased mortality, a result identical to the hybrid breakdown phenomenon. The experiments were repeated using females from the cucumber strain. In interstrain U x W and U x U crosses, hybrid breakdown was much stronger in the former (80 versus 26%). This is the first report of a Wolbachia infection causing a hybrid breakdown phenotype. Our results show that Wolbhachia infections can contribute to reproductive incompatibility between populations of T. urticae.     

Alternative phenotypes of male mating behaviour in the two-spotted spider mite

Severe intraspecific competition for mates selects for aggressive individuals but may also lead to the evolution of alternative phenotypes that do not act aggressively, yet manage to acquire matings. The two-spotted spider mite, Tetranychus urticae, shows male mate-guarding behaviour and male–male combat for available females. This may provide opportunity for weaker males to avoid fighting by adopting alternative mating behaviour such as sneaker or satellite tactics as observed in other animals. We investigated male precopulatory behaviour in the two-spotted spider mite by means of video-techniques and found three types of male mating behaviour: territorial, sneaker and opportunistic. Territorial and sneaker males associate with female teleiochrysales and spend much time guarding them. Territorial males are easily disturbed by rival males and engage themselves in fights with them. However, sneaker males are not at all disturbed by rival males, never engage in fights and, strikingly, never face attack by territorial males. Opportunistic males wander around in search of females that are in the teleiochrysalis stage but very close to or at emergence. To quickly classify any given mate-guarding male as territorial or sneaker we developed a method based on the instantaneous response of males to disturbance by a live male mounted on top of a brush. We tested this method against the response of the same males to natural disturbance by two or three other males. Because this method proved to be successful, we used it to collect territorial and sneaker males, and subjected them to morphological analysis to assess whether the various behavioural phenotypes are associated with different morphological characters. However, we found no statistical differences between territorial and sneaker males, concerning the length of the first legs, the stylets, the pedipalps or the body.     

Settlement decisions by the two-spotted spider mite Tetranychus urticae

In silk-spinning arthropods, silk can be used for web building, protection, and communication. Silk is an informative material about the presence of conspecifics. It can therefore inform on habitat suitability and hence assist in habitat choice. In this context, we investigated the influence of silk on microhabitat choice by the two-spotted spider mite, Tetranychus urticae. Three factors that could potentially influence habitat choice were manipulated: the strain, number, and the stage of mites. Our study showed that these factors all influence the choice of microhabitat. The tendency of whether to settle on a silk-covered area was influenced by the origin of mites (strain effect). Adult females showed a higher tendency to settle on an area covered with the silk laid by numerous congeners (number effect). Moreover, larvae seemed to be more responsive to the presence of silk than adults (stage effect). This suggests that individuals use silk as a social cue in selecting their microhabitat and that the spatial organization and group behaviour seem to be shaped by the individuals’ response to social cues, such as the amount of silk already present.     

UV tolerance in the two-spotted spider mite, Tetranychus urticae

The two-spotted spider mite, Tetranychus urticae was exposed to UV-C (250 nm), UV-B (300 nm), and UV-A (350 nm). In non-diapausing females, the median effective doses for 50% mortality plus escape incidence (ED₅₀) were 21 (UV-C) and 104 kJ m⁻² (UV-B); those for 50% oviposition rate in continuous darkness-treated mites were 6.2 (UV-C) and 41 kJ m⁻² (UV-B). No significant effects of UV-A on mortality and oviposition rate were observed. The ED₅₀ values for UV-B were similar to the natural UV-B observed for 2-5 days in summer when T. urticae inhabits the undersides of leaves. Therefore, T. urticae possibly uses leaves as a filter to avoid the deleterious effects of UV-B. In diapausing females, low mortality was observed even at high doses of UV radiation, but more than half escaped even at low doses. The orange body color of diapausing females results from accumulation of carotenoids, a scavenger for UV-induced reactive oxygen species; this may explain the low mortality of diapausing females. Diapausing females may overcome the deleterious effects of UV-B during winter in the absence of leaves by emigrating to UV-free environments and by accumulating carotenoids.     

Identification of Wolbachia-responsive microRNAs in the two-spotted spider mite,Tetranychus urticae

Background

The two-spotted spider mite, Tetranychus urticae, is infected with Wolbachia, which have the ability to manipulate host reproduction and fitness. MicroRNAs (miRNAs) are small non-coding RNAs that are involved in many biological processes such as development, reproduction and host-pathogen interactions. Although miRNA was observed to involve in Wolbachia-host interactions in the other insect systems, its roles have not been fully deciphered in the two-spotted spider mite.

Results

Small RNA libraries of infected and uninfected T. urticae for both sexes (in total four libraries) were constructed. By integrating the mRNA data originated from the same samples, the target genes of the differentially expressed miRNAs were predicted. Then, GO and pathway analyses were performed for the target genes. Comparison of libraries showed that Wolbachia infection significantly regulated 91 miRNAs in females and 20 miRNAs in males, with an overall suppression of miRNAs in Wolbachia-infected libraries. A comparison of the miRNA and mRNA data predicted that the differentially expressed miRNAs negatively regulated 90 mRNAs in females and 9 mRNAs in males. An analysis of target genes showed that Wolbachia-responsive miRNAs regulated genes with function in sphingolipid metabolism, lysosome function, apoptosis and lipid transporting in both sexes, as well as reproduction in females.

Conclusion

Comparisons of the miRNA and mRNA data can help to identify miRNAs and miRNA target genes involving in Wolbachia-host interactions. The molecular targets identified in this study should be useful in further functional studies.

Electronic supplementary material

The online version of this article (doi:10.1186/1471-2164-15-1122) contains supplementary material, which is available to authorized users.     

Fertilization in the two-spotted spider mite (Tetranychus urticae: Acari)

Different aspects of the fertilization of the two-spotted spider mite (Tetranychus urticae Koch) were studied. It appeared that after the mating act of virgin females, haploid eggs were produced first before the appearance of diploid eggs. From this and other facts, it was concluded that fertilization of the oocytes occurs in a very early stage of egg development. The fertilization is assumed to take place in the ovary and not in the oviduct.By using genetic markers, it could be proved that in most cases the first mating is effective. Later matings usually are ineffective. It is suggested that the sperm supply in the first mating determines the success of later matings.

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Zusammenfassung Die Befruchtung der Bohnenspinnmilbe wurde unter verschiedenen Gesichtspunkten untersucht. Nach der Begattung jungfräulicher Weibchen wurden zunächst haploide Eier abgelegt, bevor diploide Eier produziert wurden. Daraus und aus anderen Grüunden wird geschlossen, daß die Befruchtung der Oocyten in einem sehr frühen Stadium der Entwicklung erfolgt. Die Befruchtung findet vermutlicht im Ovar und nicht im Ovidukt statt.Unter Verwendung genetischer Markierung wurde nachgewiesen, daß in den meisten Fällen die erste Begattung ausschlaggebend ist. Spätere Kopulationen bleiben gewöhnlich wirkungslos. Es wird vermutet, daß die Spermafüllung bei der ersten Begattung den Erfolg späterer Begattungen bestimmt.

     

Chemical investigation of aggregation behaviour in the two-spotted spider mite Tetranychus urticae

The silk produced by the group-living mite Tetranychus urticae provides group protection and is used as an informative material during habitat settlement, egg laying, mating, and dispersal events. In this context, cues contained in the silk and other materials produced by mites [eggs, black faeces (BF) and white faeces + silk (WFS)] were investigated. Chemical compounds were extracted by hexane or methanol, and choice tests were used to determine the individual attractiveness of each extract. For both solvents, individuals did not respond to the extract from eggs and WFS. BF extracts were attractive for both solvents. After separating the BF methanol extract into four different chemical components using thin layer chromatography, no component was determined to be responsible for mite attraction. This work supports the evidence that the faeces of T. urticae do contain substances that promote behavioural changes. Not particular chemical compounds but combinations of them seem to induce the mites’ preference. Moreover, the response of mites to chemicals seemed to be context dependent as mites belonging to populations with different densities differed in their attraction to BF extracts.     

Males mate with females even after sperm depletion in the two-spotted spider mite

Generally, males increase their reproductive success by mating with as many females as possible, whereas females increase their reproductive success by choosing males who provide more direct and indirect benefits. The difference in reproductive strategy between the sexes creates intense competition among males for access to females, therefore males spend much energy and time for competition with rival males for their reproduction. However, if they do not need to engage themselves into male competition and females are in no short supply, how many females can a male mate with and fertilize? We address this question in the two-spotted spider mite, Tetranychus urticae Koch. In this study, we investigated how many females a young, virgin male mated in 3 h, and checked whether the mated females were fertilized. We found that on average males mated with 12–13 females (range: 5–25). As latency to next mating did not change with the number of matings, the males are predicted to engage in even more matings if the mating trial were continued beyond 3 h. Copulation durations decreased with the number of matings and typically after 11 copulations with females any further copulations did not lead to fertilization, suggesting that males continued to mate with females even after sperm depletion. We discuss why spider mite males continue to display mating and copulation behaviour even after their sperm is depleted.

     

Demographic examination of sex ratio in the two-spotted spider mite, Tetranychus urticae

Primary sex ratio of arrhenotokous tetranychid mites depends on the age-specific schedule of ratios of progeny produced by individual females. Since primary sex ratio is conditional upon female age and since the proportion of females of a given age in a population is influenced by the rate of increase, both the agespecific ratio of males to females and the overall ratio of males to females depends on the rate of increase. We given an expression for this dependence and compare it to experimental results for the two-spotted spiter mite. These results imply that there is no standard sex ratio of spider mites because of interpopulation variation in rates of increase.

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Résumé Le taux sexuel des acariens tétranychidés arrhénotoques est défini comme le rapport de la somme des nombres de mâles sur la somme des nombres de femelles pour chaque âge de la distribution en âges. Comme le taux sexuel primaire dépend de l'âge de la femelle et comme la proportion de femelles d'un âge donné dans la population est influencée par le taux d'accroissement, alors le rapport mâle/femelle spécifique d'un âge déterminé et le rapport global mâle/femelle dépendent du taux d'accroissement. Nous avons cherché à exprimer cette dépendance et à la comparer aux résultats expérimentaux pour Tetranychus urticae. Ces résultats signifient qu'il n'y a pas de taux sexuel caractéristique de Tétranyques par suite des variations du taux d'accroissement suivant les populations.

     

Genetics of diapause supression in the two-spotted spider mite,Tetranychus urticae Koch

Starting from the Sambucus strain of the two-spotted spider miteTetranychus urticae Koch, the strains ND₁ and ND₂ were selected for lack of diapause under a short-day regimen (LD 1014, 18°C). The genetic basis of the suppression of diapause in ND₁ and ND₂ was analysed by means of mendelian crosses and backcrosses, using the photoperiodic cycle LD 1014 as diagnostic for the distinction of diapausing and non-diapausing phenotypes. Suppression of diapause is inherited as a recessive trait. From backcross analysis it was concluded that the suppression of diapause in ND₁ and ND₂ is under monogenic control. The major genes for suppression of diapause in ND₁ and ND₂ are alleles at a locus designated asd. A mutation for albinism of the locusa-p is pleiotropic for suppression of diapause. It was shown from crosses between ND₁ and the albino strain, thatd anda-p are distinct loci.     

The fine-scale genetic structure of the two-spotted spider mite in a commercial greenhouse

The fine-scale genetic structure of Tetranychus urticae Koch was studied to estimate local gene flow within a rose tree habitat in a commercial greenhouse using seven microsatellite markers. Two beds of rose trees with different population densities were selected and 18 consecutive quadrats of 1.2 m length were sequentially established in each bed. Heterozygote deficiency was positive within quadrats, which was most likely a result of the Wahlund effect because the mites usually form small breeding colonies. Low population density and frequent inbreeding could also accelerate genetic differentiation among the breeding colonies. A short-range (2.4–3.6 m) positive autocorrelation and clear genetic cline among quadrat populations was detected within a bed. This suggests that gene flow was limited to a short range even if population density was substantially increased. Therefore, large-scale dispersal such as aerial dispersal contributed very little to gene flow in the greenhouse.     

Residual contact vial bioassay for the on-site detection of acaricide resistance in the two-spotted spider mite

To develop a less technique-dependent bioassay technique that can be conveniently used by practitioners or farmers in the field for the monitoring of acaricide resistance of the two-spotted spider mite, Tetranychus urticae Koch, a residual contact vial (RCV) method was established using a 5-ml glass vial coated with acaricides. The RCV bioassay procedures were optimized by using abamectin and tebufenpyrad, two widely used acaricides. The diagnostic concentrations causing 100% mortality within 8 h post-treatment in a susceptible strain of T. urticae was set at 30 and 60 ppm for abamectin and tebufenpyrad, respectively. The vial-coated pesticides were stable at least one year when stored at ? 20 °C as determined by HPLC. There was no significant difference in the bioassay results in repeated RCV bioassay by three different experimenters, indicating its high reproducibility and reliability. RCV-based resistance monitoring of 15 field populations of mites revealed that abamectin resistance begins to spread but tebufenpyrad resistance is already prevalent in Korea. The RCV diagnostic kit, when used by practitioners or farmers on site, should provide crucial information for the selection of most suitable acaricides for different field populations of T. urticae.     

Comparative toxic and sublethal effects of fluvalinate on two-spotted spider mite and European red mite

Four fluvalinate formulations differed in their residual toxicity to female two-spotted spider mite (TSM), Tetranychus urticae adults; the emulsifiable concentrate (EC) was the most toxic. In contrast, there was little difference in toxicity between the formulations with the European red mite (ERM) Panonychus ulmi with the exception of the EC formulation which was the least toxic. Fluvalinate 2F caused minimal (<10%) TSM and ERM egg mortality. Fluvalinate 2F was more toxic and caused greater larval dispersal for the TSM compared to the ERM at the field concentration and below. The toxicity of fluvalinate 2F to TSM and ERM protonymphs, deutonymphs and adults was low, approximately <20% at field concentration. Dispersal was the main response to fluvalinate and this was positively correlated with increasing concentration. The combined mortality and dispersal LC₅₀ was five times lower for ERM protonymphs and adults, but 11 times higher for ERM deutonymphs compared to equivalent TSM life stages. Fluvalinate 2F reduced TSM development from the protonymph and deutonymph stages to a greater extent compared to the ERM. The mortality response to fluvalinate 2F was unaffected by host type (peach or apple) for the TSM whereas ERM mortality was higher on apple compared to peach. TSM dispersal was higher from apple compared to peach whereas ERM dispersal was similar on both host types. Oviposition by both mite species was lower on apple than peach leaves. A 1 h exposure to fluvalinate 2F reduced ERM oviposition for 12 days.