Cold storage of the predatory mite Neoseiulus californicus is improved by pre-storage feeding on the diapausing spider mite Tetranychus urticae

Low air temperature accompanied with high humidity is effective for long-term cold storage of the predatory mite Neoseiulus californicus (McGregor) (Acari: Phytoseiidae). To further improve this storage method, we investigated the effect of pre-storage nutrition on survival during storage and on post-storage quality in terms of survival, oviposition, and progeny viability. The predatory mite was fed from the egg to adult stage on the diapausing two-spotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae), non-diapausing spider mites, or Japanese pear pollen, Pyrus pyrifolia Nakai. Newly emerged N. californicus adult females and males were mated, and then both were stored at 7.5 °C and a vapor pressure deficit of 0.0 kPa for up to 75 days. Survival during storage and post-storage quality was significantly better with the diapausing spider mite diet than with the other diets. No effects on the survival or sex ratio of the progeny of the stored adults were observed, regardless of diet or storage duration. Providing diapausing spider mites as a pre-storage diet therefore significantly improves the long-term storage of N. californicus. We discuss the possibility that ingestion of the cryoprotectants, antioxidants, and energy reserves that are present in rich amounts in diapausing spider mites mitigates chilling injury.     

Behavioural response and winter survival of mated and unmated diapausing females of the Tetranychus atlanticus-urticae complex (Acari, Tetranychidae)

Behavioural responses were studied in mated and unmated females of a Dutch strain of the spider mite Tetranychus urticae which were destined to enter diapause. Observations were confined to the period between acquirement of the winter coloration by the females (a few days after the last moult) and their entering of artificial hibernation sites offered to the mites on the host leaves. Unmated females were found to stay longer on the leaf surface than mated females; they entered the hibernation sites significantly later than mated females. The delay in the search for hibernation sites shown by unmated females may be seen as a behavioural adaptation to enhance the chance of being fertilized before hibernation. Winter survival of mated and unmated diapausing females of the same strain of mites was studied both in the laboratory at a constant temperature of 2±1°C and outdoors under natural climatic conditions in Amsterdam during the winter of 1990–1991. Survival was high under both conditions for mated as well as unmated females; no significant differences in survival were found between both types of female. Observations on post-diapause females of Tetranychus atlanticus (a mite belonging to the T. urticae complex) sampled from strawberry fields near Moscow in spring, showed that at most 10% of the females of this natural spider mite population were unmated. Both mated and unmated females had survived winter temperatures of -28 to -30°C.     

The effects of prestarvation diet on starvation tolerance of the predatory mite Neoseiulus californicus (Acari: Phytoseiidae)

Understanding the factors affecting stress tolerance in phytoseiid mites is critical for their integration into biological control programs. In the present study, the effects of diet (varying in prey species, physiological status and phenotype) are examined on the future starvation tolerance of the predatory mite Neoseiulus californicus McGregor (Acari: Phytoseiidae). The predators are fed from egg to adulthood on diapausing or nondiapausing Tetranychus urticae, Tetranychus kanzawai (wild and albino strains) or the nondiapausing species Panonychus citri (wild and albino strains). Thereafter, 3‐day‐old mated adult females are held without food at 25 ± 1 °C and a relative humidity of 98 ± 2%. The survival of these starved females is observed daily until all females have died. The survival curves and mean survival times of N. californicus are found to vary among prey types and are significantly longer when the predator is fed with diapausing prey. This enhanced survival is consistent with high concentrations of glycogen and triacylglyceride in the body of the predator at the onset of starvation. The predators fed nondiapausing prey have shorter survival times, and the glycogen and triacylglyceride contents in their bodies are low or undetectable. The protein contents of the predator's body are similar after consuming different prey types, except for a high concentration when fed the albino strain of P. citri. Protein content is unlikely to play a direct role in starvation tolerance, although it may affect the response to varying glycogen and triacylglyceride levels. These findings indicate that nutritional value of prey has a strong impact on the starvation tolerance of N. californicus.     

Overwintering survival and postdiapause fecundity in a population of the Kanzawa spider mite <Emphasis Type="Italic">Tetranychus kanzawai</Emphasis> (Acari: Tetranychidae) on <Emphasis Type="Italic">Orixa japonica</Emphasis> (Rutaceae)

In several adult-diapausing insects and mites, long duration of diapause decreases egg production after diapause termination. However, such nonlethal effects are much less studied than overwintering survival. In this study, lethal and nonlethal effects of different periods of diapause were investigated in an adult-diapausing spider mite, Tetranychus kanzawai Kishida (Acari: Tetranychidae), through field experiments. Diapausing females produced in the laboratory were kept in their natural habitat from October or November to the next April, and survival rate and postdiapause fecundity were compared between the months. The survival rate was lower, but not significantly, in the October treatment. Postdiapause fecundity was also not significantly different between the months. These results suggest that the effects of diapause on survival and reproduction are quite small after October, when diapausing mites are increasing in their natural habitat.     

Deciphering the Metabolic Changes Associated with Diapause Syndrome and Cold Acclimation in the Two-Spotted Spider Mite Tetranychus urticae

Diapause is a common feature in several arthropod species that are subject to unfavorable growing seasons. The range of environmental cues that trigger the onset and termination of diapause, in addition to associated hormonal, biochemical, and molecular changes, have been studied extensively in recent years; however, such information is only available for a few insect species. Diapause and cold hardening usually occur together in overwintering arthropods, and can be characterized by recording changes to the wealth of molecules present in the tissue, hemolymph, or whole body of organisms. Recent technological advances, such as high throughput screening and quantification of metabolites via chromatographic analyses, are able to identify such molecules. In the present work, we examined the survival ability of diapausing and non-diapausing females of the two-spotted spider mite, Tetranychus urticae, in the presence (0 or 5°C) or absence of cold acclimation. Furthermore, we examined the metabolic fingerprints of these specimens via gas chromatography-mass spectrophotometry (GC-MS). Partial Least Square Discriminant Analysis (PLS-DA) of metabolites revealed that major metabolic variations were related to diapause, indicating in a clear cut-off between diapausing and non-diapausing females, regardless of acclimation state. Signs of metabolic depression were evident in diapausing females, with most amino acids and TCA cycle intermediates being significantly reduced. Out of the 40 accurately quantified metabolites, seven metabolites remained elevated or were accumulated in diapausing mites, i.e. cadaverine, gluconolactone, glucose, inositol, maltose, mannitol and sorbitol. The capacity to accumulate winter polyols during cold-acclimation was restricted to diapausing females. We conclude that the induction of increased cold hardiness in this species is associated with the diapause syndrome, rather than being a direct effect of low temperature. Our results provide novel information about biochemical events related to the cold hardening process in the two-spotted spider mite.     

Effects of acclimation and diapause on the thermal tolerance of the two-spotted spider mite Tetranychus urticae

The two-spotted spider mite, Tetranychus urticae, is a worldwide pest species that overwinters as diapausing females. Cold hardening is presumed to start during diapause development to ensure the successful overwintering of this species. To address this hypothesis, we compared cold tolerance between non-diapausing and diapausing females. We measured supercooling point (SCP) and survival to acute cold stress by exposing the mites at a range of sub-zero temperatures (from −4 to −28 °C for 2 h). The mean SCPs of non-diapausing and diapausing females were −19.6±0.5 and −24.7±0.3 °C respectively, and freezing killed the mites. Diapausing females were significantly more cold tolerant than non-diapausing ones, with LT₅₀ of −19.7 and −13.3 °C, respectively. Further, we also examined the effects of cold acclimation (10 d at 0 or 5 °C) in non-diapausing and diapausing females. Our findings indicated that diapause decreased SCP significantly, while cold acclimation had no effect on the SCP except for non-diapausing females that were acclimated at 5 °C. Acclimation at 5 °C enhanced survival to acute cold stress in diapausing and non-diapausing females, with LT₅₀ of −22.0 and −17.1 °C, respectively. Altogether, our results indicate that T. urticae is a chill tolerant species, and that diapause and cold acclimation elevate cold hardiness in this species.     

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.     

Effects of mite age, mite density, and host quality on aerial dispersal behavior in the twospotted spider mite

Mite age, population density, and host leaf quality affect various life history traits in spider mites. We investigated the effects of these factors on the aerial dispersal behavior of adult female twospotted spider mites,Tetranychus urticae Koch (Acari: Tetranychidae). The proportion of adult females exhibiting the dispersal behavior dropped significantly with age following adult emergence, particularly in the first 3 days. Sixty to eighty percent of female mites 2-days old or younger displayed the behavior under test conditions, whereas less than 20% of female mites older than 3-days-old showed the behavior. Younger adult females also exhibited shorter latency for the behavior, although this trend was not as clear. Leaf quality experienced during deutonymph development had no effect on the behavior adults subsequently displayed. On the other hand, adult females that fed on poor quality leaves after emergence were twice as likely to display the behavior (90% vs. 45%), and with shorter latency (37 vs. 77 min), than those that fed on high quality leaves. When newly emerged adult females encountered high mite density and dry leaves, the incidence of the behavior increased (69% vs. 47%) and latency decreased (69 vs. 93 min) compared to mites that encountered low density on well watered leaves. Our results suggest that both starvation and desiccation of adult females may enhance their dispersal behavior.     

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.     

Dispersal of diapausing Tetranychus urticae and T. kanzawai

The spider mites Tetranychus urticae Koch and Tetranychus kanzawai Kishida (Acari: Tetranychidae) overwinter mostly as mated adult diapausing females. Their overwintering survival depends in part on their dispersal towards suitable habitats. We investigated the dispersal behaviour of diapausing females of T. urticae and T. kanzawai with respect to factors known to affect the dispersal of non‐diapausing mites: light, population density, gravity, and humidity. In general, diapausing females of T. urticae showed a stronger tendency to disperse than did those of T. kanzawai under all test conditions. High population density promoted the dispersal of diapausing T. urticae, but not of T. kanzawai. Dispersal of diapausing females of both species was not significantly affected by gravity, humidity, or whether feeding damage was caused by conspecifics or heterospecifics. On plants, more T. urticae than T. kanzawai moved downward. We propose that dispersal after the onset of diapause may be an important life‐history strategy in T. urticae, but not in T. kanzawai.     

Relationship between body colour, feeding, and reproductive arrest under short-day development in Tetranychus pueraricola (Acari: Tetranychidae)

In Tetranychus spider mites (Acari: Tetranychidae), diapausing females have a conspicuous orange body colour, which is used as an indicator of diapause induction in many laboratory studies. However, to which extent body colour reflects reproductive activity is scarcely investigated. In this study, we investigated the relationship between body colour, reproductive arrest, and food intake in the inbred strain of T. pueraricola individually reared at 20 °C with a 10:14 h light: dark photoperiod. Our results showed that (1) body colour is a good indicator of reproductive arrest 11 days after adult emergence but does not completely reflect reproductive status at an earlier age; (2) even orange females intermittently feed, and the arrest of feeding comes after the change in body colour; and (3) reproducing females have a higher risk of death than non-reproducing females. These results suggest that measurement of diapause incidence by body colour alone may miss the variation in reproductive status in early adult life.     

Imaginal induction of diapause in several 'adult-female diapausing' spider mites

Abstract. The possibility of imaginal induction of diapause in adult female spider mites is suggested in Stigmaeopsis miscanthi. Here, several spider mites are studied to investigate the generality of this phenomenon. The existence of imaginal induction of reproductive diapause is established in Stigmaeopsis longus, two forms of S. miscanthi and Tetranychus kanzawai adult females. The females that develop under diapause-averting (long days and 18 °C) conditions are induced to diapause under short-daylength conditions at 18 °C after moulting to the adult stage. However, the adult females of Tetranychus urticae never diapause under these conditions.     

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.     

Are winter-active species vulnerable to climate warming? A case study with the wintergreen terrestrial orchid,Tipularia discolor

In the eastern United States, winter temperature has been increasing nearly twice as fast as summer temperature, but studies of warming effects on plants have focused on species that are photosynthetically active in summer. The terrestrial orchid Tipularia discolor is leafless in summer and acquires C primarily in winter. The optimum temperature for photosynthesis in T. discolor is higher than the maximum temperature throughout most of its growing season, and therefore growth can be expected to increase with warming. Contrary to this hypothesis, experimental warming negatively affected reproductive fitness (number of flowering stalks, flowers, fruits) and growth (change in leaf area from 2010 to 2012) in T. discolor. Temperature in June–July was critical for flowering, and mean July temperature greater than 29 °C (i.e., 2.5 °C above ambient) eliminated reproduction. Warming of 1.2 °C delayed flowering by an average of 10 days and fruiting by an average of 5 days. Warming of 4.4 °C reduced relative growth rates by about 60 %, which may have been partially caused by the direct effects of temperature on photosynthesis and respiration. Warming indirectly increased vapor pressure deficit (VPD) by 0.2–0.5 kPa, and leaf-to-air VPD over 1.3 kPa restricted stomatal conductance of T. discolor to 10–40 % of maximum conductance. These results highlight the need to account for changes in VPD when estimating temperature responses of plant species under future warming scenarios. Increasing temperature in the future will likely be an important limiting factor to the distribution of T. discolor, especially along the southern edge of its range.     

A molecular‐genetic understanding of diapause in spider mites: current knowledge and future directions

During unfavourable conditions, many arthropods have the ability to enter into diapause and synchronize their development and reproduction to seasonal patterns. Diapause or winter hibernation in insects and mites is set off by a number of cues, with photoperiod being the most well‐defined and strongest signal. This review focuses on the current knowledge of ‘‐omics’ data and the genetics of diapause in the two‐spotted spider mite Tetranychus urticae, a member of the family Tetranychidae (Arthropoda: Chelicerata: Arachnida: Acari). This species is a serious polyphagous pest and females undergo a reproductive facultative diapause when immature stages are exposed to long nights. Winter hibernation induces different physiological processes characterized by a metabolic suppression, different energy use, increased stress tolerance and the production of cryoprotectants, all initiated by a complex signal transduction pathway. Keto‐carotenoids are known to cause the deeply orange colour typical for diapausing females. Furthermore, research with colour mutants of T. urticae has shown the need for carotenoids with respect to the induction of diapause, even though the molecular‐genetic mechanisms underlying these colour phenotypes are still unknown. In addition, marked latitudinal variation in diapause incidence among populations has been observed in nature, with modes of inheritance ranging from recessive to dominant, as well as monogenic to polygenic. We end by highlighting the emerging opportunities for functional studies that aim to unravel the complex factors underlying diapause in spider mites.     

Effects of air temperature and water vapor pressure deficit on storage of the predatory mite Neoseiulus californicus (Acari: Phytoseiidae)

To determine the optimum air temperature and water vapor pressure deficit (VPD) for the storage of the predatory mite, Neoseiulus californicus, 3-day-old mated females were stored at air temperatures of 0, 5, 10, or 15?°C and VPDs of 0.1, 0.3, or 0.5?kPa for 10, 20, or 30?days. At 10?°C and 0.1?kPa, 83?% of females survived after 30?days of storage; this percentage was the highest among all conditions. VPDs of 0.3 and 0.5?kPa regardless of air temperature, and an air temperature of 0?°C regardless of VPD were detrimental to the survival of the females during storage. Since the highest survival was observed at 10?°C and 0.1?kPa, the effect of the storage duration on the post-storage quality of the stored females and their progeny was investigated at 25?°C to evaluate the effectiveness of the storage condition. The oviposition ability of the stored females, hatchability, and sex ratio of their progeny were not affected even when the storage duration was extended to 30?days. Although a slight decrease in the survival during the immature stages of progeny was observed when the storage duration was ≥20?days, the population growth of N. californicus may not be affected when individuals stored in these conditions are applied to greenhouses and agricultural fields. The results indicate that mated N. californicus females can be stored at 10?°C and 0.1?kPa VPD for at least 30?days.     

Diapause induction and duration in the phytoseiid miteTyphlodromus pyri

A photoperiod of 8L/16D for two weeks was used to distinguish between diapausing and non-diapausingTyphlodromus pyri Scheuten. A diet ofPanonychus ulmi orTetranychus urticae, or pollen ofVicia faba did not influence preovipositional periods of diapausing mites. In mid-September, 88% ofT. pyri collected from an insectary were in diapause. The critical day-length appeared to be between 12.5 and 13.5 h. Diapause duration was greatest in mites collected in September–November, becoming progressively less in mites collected later in the winter. By mid-April, ca. 50% of mites collected from the orchard and insectary oviposited promptly when cultured in the laboratory.Typhlodromus pyri eggs and larvae were present on leaves in early May. At this time, only 4% ofP. ulmi winter eggs had hatched. Diapause terminated most quickly inT. pyri kept in an 18L/6D photoperiod, followed by 24L/0D and 0L/24D. Longest preoviposition periods were recorded for mites kept in 8L/16D photoperiods.     

Food scarcity reduces female longevity of <Emphasis Type="Italic">Neoseiulus californicus</Emphasis> (Acari: Phytoseiidae)

A commercial strain of Neoseiulus californicus (Spical^®) has a conspicuously long postoviposition period in comparison with other strains of N. californicus or other phytoseiid mites. In many insects and mites, life span is shorter for multiple-mated females than for single-mated females, and is shorter under poor prey conditions than under ample prey conditions. We previously showed that the postoviposition period of multiple-mated females was 40% shorter than that of single-mated females, but that it was never shorter than 30 days. Here we focused on the effect of prey abundance on the postoviposition period. We examined three groups of multiple-mated females: mites that were fed ample prey (group I), mites that were subjected to repeated cycles of 2 days of fasting followed by 2 days of ample prey (group II), and mites that were subjected to repeated cycles of 4 days of fasting followed by 2 days of ample prey (group III). The postoviposition periods of groups II and III were 90% shorter than that of group I. Also the total adult longevity was significantly shorter in groups II and III than in group I. Total egg production in group III was about half that in groups I and II, although the oviposition periods in groups II and III were significantly longer than that in group I. These results suggest that the prolonged postoviposition period in single-mated females of the Spical strain may only appear under laboratory conditions, without multiple mating or starvation.     

Species composition and arthropod pest feeding of phytoseiid mites in a Japanese pear greenhouse

A population survey of spider mites and phytoseiid mites was conducted on Japanese pear leaves in a greenhouse. For the survey, the method to estimate phytoseiid mite species composition using quantitative sequencing was modified to be applicable for phytoseiid mite species inhabiting in the greenhouse. Results show the dominant appearance of Neoseiulus californicus (McGregor), Neoseiulus womersleyi (Schicha), and Neoseiulus makuwa (Ehara) from the end of June to late September and their contribution in spider mite control. PCR-based method to detect the ribosomal internal transcribed spacer (ITS) sequences of spider mites from phytoseiid mites was developed. The method shows sensitivity to detect the ITS sequences of Tetranychus urticae Koch from single N. californicus adult at 168 h after ingestion of the spider mite. PCR-based method to detect the cytochrome c oxidase subunit I sequences of several arthropod pests belonging to Hemiptera, Thysanoptera, and Acari from phytoseiid mites was also developed. Results show that phytoseiid mites prey on Eriophyes chibaensis (Kadono) and Aphis gossypii (Glover), in addition to spider mites.     

Spatial distribution of predators and prey affect biological control of twospotted spider mites by Phytoseiulus persimilis in greenhouses

We evaluated the effects of predator release pattern and prey distribution on rate of suppression of the twospotted spider mite, Tetranychus urticae Koch (Acari, Tetranychidae) and visual damage to the ornamental plant, Impatiens wallerana Hook.f., in a greenhouse. Sixteen impatiens plants were arranged in a square and infested with the same total number of spider mites distributed either evenly (equal numbers on all plants) or clumped (divided equally among the 4 central plants), simulating a “hot spot.” The predatory mite, Phytoseiulus persimilis Athias-Henriot, was released at a 1:4 predator:prey ratio based on total spider mites in the experimental unit, but the pattern of release was either even or clumped, which simulated broadcast or point-release strategies, respectively. Nine days after predator release, spider mite populations were reduced in all treatments, but only in the clumped pest-clumped predator treatment were spider mites undetectable. Poorest pest suppression occurred in the clumped spider mite-even predator treatment. Eighteen days after predator release, spider mites were eliminated in all treatments, but a reduction in average plant damage occurred only in treatments in which the predator release pattern matched the spider mite distribution (i.e., even-even or clumped-clumped) with the greatest reduction in the even-even treatment. Results suggest that there is an advantage to releasing predators in “hot spots” provided that the recommended predator:prey ratio is maintained within infested patches. If more uniform predator releases are planned, overall predator numbers need to be kept sufficiently high so that the predator:prey ratio of 1:4 shown to prevent damage on impatiens is achieved in higher-density spider mite patches.