Spider mite herbivory induces an ABA-driven stomatal defense.

Arthropod herbivory poses a serious threat to crop yield, prompting plants to employ intricate defense mechanisms against pest feeding. The generalist pest 2-spotted spider mite (Tetranychus urticae) inflicts rapid damage and remains challenging due to its broad target range. In this study, we explored the Arabidopsis (Arabidopsis thaliana) response to T. urticae infestation, revealing the induction of abscisic acid (ABA), a hormone typically associated with abiotic stress adaptation, and stomatal closure during water stress. Leveraging a Forster resonance energy transfer (FRET)-based ABA biosensor (nlsABACUS2-400n), we observed elevated ABA levels in various leaf cell types postmite feeding. While ABA's role in pest resistance or susceptibility has been debated, an ABA-deficient mutant exhibited increased mite infestation alongside intact canonical biotic stress signaling, indicating an independent function of ABA in mite defense. We established that ABA-triggered stomatal closure effectively hinders mite feeding and minimizes leaf cell damage through genetic and pharmacological interventions targeting ABA levels, ABA signaling, stomatal aperture, and density. This study underscores the critical interplay between biotic and abiotic stresses in plants, highlighting how the vulnerability to mite infestation arising from open stomata, crucial for transpiration and photosynthesis, reinforces the intricate relationship between these stress types.

QTL analysis on a lemon population provides novel insights on the genetic regulation of the tolerance to the two-spotted spider mite attack.

BACKGROUND: Among the Citrus species, lemon (Citrus limon Burm f.) is one of the most affected by the two-spotted spider mite (Tetranychus urticae Koch). Moreover, chemical control is hampered by the mite's ability to develop genetic resistance against acaricides. In this context, the identification of the genetic basis of the host resistance could represent a sustainable strategy for spider mite control. In the present study, a marker-trait association analysis was performed on a lemon population employing an association mapping approach. An inter-specific full-sib population composed of 109 accessions was phenotyped through a detached-leaf assays performed in modified Huffaker cells. Those individuals, complemented with two inter-specific segregating populations, were genotyped using a target-sequencing approach called SPET (Single Primer Enrichment Technology), the resulting SNPs were employed for the generation of an integrated genetic map. RESULTS: The percentage of damaged area in the full-sib population showed a quantitative distribution with values ranging from 0.36 to 9.67%. A total of 47,298 SNPs were selected for an association mapping study and a significant marker linked with resistance to spider mite was detected on linkage group 5. In silico gene annotation of the QTL interval enabled the detection of 13 genes involved in immune response to biotic and abiotic stress. Gene expression analysis showed an over expression of the gene encoding for the ethylene-responsive transcription factor ERF098-like, already characterized in Arabidopsis and in rice for its involvement in defense response. CONCLUSION: The identification of a molecular marker linked to the resistance to spider mite attack can pave the way for the development of marker-assisted breeding plan for the development of novel selection coupling favorable agronomical traits (e.g. fruit quality, yield) with a higher resistance toward the mite.

A conserved protein disulfide isomerase enhances plant resistance against herbivores.

Herbivore-associated molecular patterns (HAMPs) enable plants to recognize herbivores and may help plants adjust their defense responses. Here, we report on herbivore-induced changes in a protein disulfide isomerase (PDI) widely distributed across arthropods. PDI from the spider mite Tetranychus evansi (TePDI), a mesophyll-feeding agricultural pest worldwide, triggered immunity in multiple Solanaceae plants. TePDI-mediated cell death in Nicotiana benthamiana required the plant signaling proteins SGT1 (suppressor of the G2 allele of skp1) and HSP90 (heat shock protein 90), but was suppressed by spider mite effectors Te28 and Te84. Moreover, PDIs from phylogenetically distinct herbivorous and nonherbivorous arthropods triggered plant immunity. Finally, although PDI-induced plant defenses impaired the performance of spider mites on plants, RNAi experiments revealed that PDI genes are essential for the survival of mites and whiteflies. Our findings indicate that plants recognize evolutionarily conserved HAMPs to activate plant defense and resist pest damage, pointing to opportunities for broad-spectrum pest management.

A new spider mite elicitor triggers plant defence and promotes resistance to herbivores.

Herbivore-associated elicitors (HAEs) are active molecules produced by herbivorous insects. Recognition of HAEs by plants induces defence that resist herbivore attacks. We previously demonstrated that the tomato red spider mite Tetranychus evansi triggered defence in Nicotiana benthamiana. However, our knowledge of HAEs from T. evansi remains limited. Here, we characterize a novel HAE, Te16, from T. evansi and dissect its function in mite-plant interactions. We investigate the effects of Te16 on spider mites and plants by heterologous expression, virus-induced gene silencing assay, and RNA interference. Te16 induces cell death, reactive oxygen species (ROS) accumulation, callose deposition, and jasmonate (JA)-related responses in N. benthamiana leaves. Te16-mediated cell death requires a calcium signalling pathway, cytoplasmic localization, the plant co-receptor BAK1, and the signalling components SGT1 and HSP90. The active region of Te16-induced cell death is located at amino acids 114-293. Moreover, silencing Te16 gene in T. evansi reduces spider mite survival and hatchability, but expressing Te16 in N. benthamiana leaves enhances plant resistance to herbivores. Finally, Te16 gene is specific to Tetranychidae species and is highly conserved in activating plant immunity. Our findings reveal a novel salivary protein produced by spider mites that elicits plant defence and resistance to insects, providing valuable clues for pest management.

Limits to the adaptation of herbivorous spider mites to metal accumulation in homogeneous and heterogeneous environments.

Metal accumulation is used by some plants as a defence against herbivores. Yet, herbivores may adapt to these defences, becoming less susceptible. Moreover, ecosystems often contain plants that do and do not accumulate metals, but whether such heterogeneity affects herbivore adaptation remains understudied. Here, we performed experimental evolution to test whether the spider mite Tetranychus evansi adapts to plants with high cadmium concentrations, in homogeneous (plants with cadmium) or heterogeneous (plants with or without cadmium) environments. For that we used tomato plants, which accumulate cadmium, thus affecting the performance of these spider mites. We measured mite fecundity, hatching rate, and the number of adult offspring after 12 and 33 generations and habitat choice after 14 and 51 generations, detecting no trait change, which implies the absence of adaptation. We then tested whether this was due to a lack of genetic variation in the traits measured and, indeed, additive genetic variance was low. Interestingly, despite no signs of adaptation, we observed a decrease in fecundity and number of adult offspring produced on cadmium-free plants, in the populations evolving in environments with cadmium. Therefore, evolving in environments with cadmium reduces the growth rate of spider mite populations on non-accumulating plants. Possibly, other traits contributed to population persistence on plants with cadmium. This calls for more studies addressing herbivore adaptation to plant metal accumulation.

Spider mite infestation triggers coordinated hormonal trade-offs enabling plant survival with a fitness cost.

Tetranychus urticae is an important pest that causes severe damage to a wide variety of plants and crops, leading to a substantial productivity loss. Previous research has been focused on plant defence response to T. urticae to improve plant resistance. However, plant growth, development and reproduction throughout the infestation process have not been previously studied. Through physiological, biochemical, transcriptomic and hormonomic evaluation, we uncover the molecular mechanisms directing the defence-growth trade-off established in Arabidopsis upon T. urticae infestation. Upon mite attack, plants suffer an adaptation process characterized by a temporal separation between the defence and growth responses. Jasmonic and salicylic acids regulate the main defence responses in combination with auxin and abscisic acid. However, while the reduction of both auxin signalling and gibberellin, cytokinin and brassinosteroid biosynthesis lead to initial growth arrest, increasing levels of growth hormones at later stages enables growth restart. These alterations lead to a plant developmental delay that impacts both seed production and longevity. We demonstrate that coordinated trade-offs determine plant adaptation and survival, revealing mite infestation has a long-lasting effect negatively impacting seed viability. This study provides additional tools to design pest management strategies that improve resistance without penalty in plant fitness.

Evaluation of the temperature adaptation of three rubber tree pest mites based on their two-sex life table.

To determine the optimal temperature range for the development and reproduction of three spider mites (Eotetranychus sexmaculatus, Eotetranychus orientalis, and Oligonychus biharensis), this study investigated their developmental period, survival rate, lifespan, and reproduction under five temperatures, 21, 24, 27, 30, and 33°C, to predict and control in the field. With the gathered data, a two-sex life table was constructed for each of them. The results revealed that as the temperature increased, both O. biharensis and E. orientalis displayed a gradual reduction in their generation period. Furthermore, an inverse relationship was observed between lifespan and temperature for all three spider mite species. When examining the survival rates at varying temperatures, E. sexmaculatus exhibited the highest rate (98%) at 33°C, while E. orientalis and O. biharensis demonstrated their highest survival rates at 24°C, reaching 90% and 100% respectively. Regarding reproduction, O. biharensis displayed the highest oviposition rates at 30°C with an average of 17.45 eggs per individual. Conversely, E. sexmaculatus and E. orientalis exhibited the highest oviposition rates at 33°C, averaging at 15.22 and 21.38 eggs per individual respectively. Significantly higher intrinsic growth rates were observed for O. biharensis and E. orientalis at 33°C, with rates of 0.22 and 0.26 respectively. In contrast, E. sexmaculatus demonstrated the highest intrinsic growth rate at 27°C. The temperature of 27°C was more suitable for the growth of the E. sexmaculatus, while 33°C was the optimal temperature for the E. orientalis and O. biharensis. The current findings provide valuable guidance for the control and prevention of these three spider mites.

Microbiome changes in Akanthomyces attenuatus JEF-147-infected two-spotted spider mites.

The two-spotted spider mite (Tetranychus urticae Koch) is an agriculturally serious polyphagous pest that has acquired strong resistance against acaricides because of its short life cycle and continuous exposure to acaricides. As an alternative, mite-pathogenic fungi with different modes of action could be used to control the mites. The spider mite has symbiotic microorganisms that could be involved in the physiological and ecological adaptations to biotic stresses. In this study, mite-pathogenic fungi were used to control female adults, and the microbiomes changes in the fungus-infected mites were analyzed. The acaricidal activity of 77 fungal isolates was tested, and Akanthomyces attenuatus JEF-147 exhibited the highest acaricidal activity. Subsequently a dose-response assay and morphological characterization was undertaken For microbiome analysis in female adults infected with A. attenuatus JEF-147, 16S rDNA and ITS1 were sequenced using Illumina Miseq. Infected mite showed a higher Shannon index in bacterial diversity but lower index in fungal diversity. In beta diversity using principal component analysis, JEF-147-treated mites were significantly different from non-treated controls in both bacteria and fungi. Particularly in bacterial abundance, arthropod defense-related Rickettsia increased, but arthropod reproduction-associated Wolbachia decreased. The change in major bacterial abundance in the infected mites could be explained by a trade-off between reproduction and immunity against the early stage of fungal attack. In fungal abundance, Akanthomyces showed up as expected. Foremost, this work reports microbiome changes in a fungus-infected mite and suggests a possible trade-off in mites against fungal pathogens. Future studies will focus on gene-based investigations related to this topic.

Reproduction in the spider mite Oligonychus yothersi (Acari: Tetranychidae).

In this work we reported injuries caused by the spider mite Oligonychus yothersi on Vitis vinifera leaves and we also investigate the sex ratio of this species under laboratory conditions. To access the aspects regarding reproduction, females were placed individually in arenas made of V. vinifera leaves to oviposit and all progeny were mounted on slides when they reached adulthood to confirm the offspring sex. Our study showed that O. yothersi reproduces by thelytokous /arrhenotokous parthenogenesis, generating low number of males. Additionally, we found vine plants with leaf browning, particularly on the adaxial surface, where mites were usually found.

Machine learning-based hyperspectral wavelength selection and classification of spider mite-infested cucumber leaves.

Two-spotted spider mite (Tetranychus urticae) is an important greenhouse pest. In cucumbers, heavy infestations lead to the complete loss of leaf assimilation surface, resulting in plant death. Symptoms caused by spider mite feeding alter the light reflection of leaves and could therefore be optically detected. Machine learning methods have already been employed to analyze spectral information in order to differentiate between healthy and spider mite-infested leaves of crops such as tomatoes or cotton. In this study, machine learning methods were applied to cucumbers. Hyperspectral data of leaves were recorded under controlled conditions. Effective wavelengths were identified using three feature selection methods. Subsequently, three supervised machine learning algorithms were used to classify healthy and spider mite-infested leaves. All combinations of feature selection and classification methods yielded accuracy of over 80%, even when using ten or five wavelengths. These results suggest that machine learning methods are a powerful tool for image-based detection of spider mites in cucumbers. In addition, due to the limited number of wavelengths, there is also substantial potential for practical application.

Plant, pest and predator interplay: tomato trichomes effects on Tetranychus urticae (Koch) and the predatory mite Typhlodromus (Anthoseius) recki Wainstein.

Trichomes are well-known efficient plant defense mechanisms to limit arthropod herbivory, especially in Solanaceae. The present study aims to evaluate the impact of trichome types on the development, survival and dispersal of Tetranychus urticae, and the phytoseiid predatory mite Typhlodromus (Anthoseius) recki. Six Solanum lycopersicum cultivars and two wild Solanum species, S. cheesmaniae and S. peruvianum, presenting contrasting densities and types of trichomes, were considered. Cultivars and species were characterized by counting each trichome type on leaves, petioles and stems. Mites stuck on petiole and stem and alive mites on the leaflet used for mite release and in the whole plant were counted three weeks after T. urticae plant infestation. Tetranychus urticae settlement and dispersal were differently affected by trichomes. Trichome types V and VI did not affect settlement and dispersal, whereas trichome types I and IV on the petiole had the highest impacton mites. Trichomes on leaves slightly affected mite establishment, there appears to be a repellent effect of trichome types I and IV. The low densities of both T. urticae and its predator detected for the cv. Lancaster could not be clearly associated to the trichome types here considered. The predator did not seem to be affected by plant characteristics, but rather by T. urticae numbers on the plant. The trichome traits unfavorable to T. urticae, did not affect the predator which showed high efficiency to control this pest on all the plant genotypes considered, but at a favorable predator:prey ratio (1:1). Altogether, these results are encouraging for the use of T. (A.) recki as a biological control agent of T. urticae regardless of the trichome structure of the tomato cultivars, but other conditions should be tested to conclude on practical implementations.

Influence of abiotic factors on diapause termination and temperature requirements for postdiapause development in the European red mite, Panonychus ulmi (Acari: Tetranychidae).

The European red mite Panonychus ulmi (Koch) is widely distributed and it can severely affect pome fruit crops, particularly apple. Pest outbreaks are related to an overuse of non-selective pesticide treatments that lead to the development of resistance and the absence of natural enemies in the orchard. A key aspect to optimize the use of pesticide treatments in the context of IPM is to increase the knowledge on the biology and ecology of the pest to better predict population dynamics and outbreaks. For the European red mite, knowledge on the conditions that lead to diapause breaking by winter eggs is essential to model population dynamics. To increase this knowledge, winter eggs were collected during field surveys in northen Spain during three years and egg hatching was monitored under controlled temperature and photoperiod conditions in the laboratory. The "number of days exposed to cold temperatures" was the most significant factor that positively affected hatching of overwintering eggs. The time required for 50% of the egg population to hatch (T50%) was also negatively modulated by the duration of exposure to cold temperature. The temperature threshold for postdiapause eggs development collected from the field was estimated between 5 and 6 ºC in 2005 and 2007, respectively. Moreover, the degree-days required for post diapause development were estimated between 263.2 and 270.3, depending on the year of collection. Collectively, we provide additional information on the diapause termination and postdiapause development of the European red mite that may effectively contribute to optimize pest population models.

An exploratory analysis of the structure of tetranychid and phytoseiid assemblages in walnut orchards in California.

Spider mites were considered secondary pests of walnut production in California, under the control of phytoseiid predators. Due to increased importance as walnut pests in recent decades there is renewed interest in the structure and function of the associated phytoseiid assemblage. In this study we report the results from a 3 year survey of the tetranychid and phytoseiid assemblages in walnut orchards in the Central Valley of California. The survey was conducted to determine the range and dominance of web-spinning Tetranychus species present, to investigate the species richness and dominance of the phytoseiid species present, and to explore whether patterns of variation in the relative abundance of phytoseiid species could be explained by one or more explanatory variables. Tetranychus urticae was the dominant spider mite in all growing regions and years with T. pacificus and T. turkestani also present in orchards in the southern San Joaquin Valley. Phytoseiid species richness declined with latitude among the three walnut growing regions and of the 13 species found Amblyseius similoides, Euseius stipulatus, Galendromus occidentalis and Typhlodromus caudiglans were the most abundant and widespread species present. Mean proportional abundance significantly increased from early (mid May-July) to late (August-mid October) season and from southern to northern growing regions for Type II and IV predators, but significantly decreased from early to late season and from southern to northern growing regions for Type III predators. The mean proportional abundance of Type II predators, particularly G. occidentalis, significantly increased and that of Type III predators significantly decreased with mean Tetranychus density in individual orchards. The current survey provides a more in-depth analysis of mite assemblages in walnut orchards in California and can be used to better inform adaptive management strategies for integrated mite management in the future.

Prey stage preference of Amblyseius paraaerialis (Acari: Phytoseiidae) on varied life stages of the spider mites Tetranychus urticae, Tetranychus macfarlanei and Oligonychus biharensis (Acari: Tetranychidae) and exploring the mass rearing possibilities of this predatory mite on alternative diets.

Spider mites Tetranychus urticae, Tetranychus macfarlanei and Oligonychus biharensis are considered to be highly polyphagous in nature and causes severe damage to a wide range of plants around the world. Amblyseius paraaerialis is an efficient phytoseiid predator of spider mites with a potential to survive on both natural and alternative diets. Evaluation of predatory potential and prey stage preference provides valuable information on the efficacy of the predatory species in controlling mite population. Feeding experiments were conducted on mulberry leaf discs under the laboratory conditions of 30 ± 2 °C and 70 ± 5% relative humidity (RH). After 24 h of feeding experiment, the adult female predator exhibited a significant preference in feeding towards the eggs of T. macfarlanei (42.6%) and the larval stages of T. urticae (46%) and O. biharensis (25.3%). The mass rearing possibilities of A. paraaerialis was tested by tracking and comparing the developmental duration of individual life stages on varied food sources like, honey, castor (Ricinus communis) pollen, honey-pollen mixture and mixed life stages of T. urticae. The predator was failed to complete its development on honey and pollen when supplied separately. However it was successfully developed on honey-pollen mixture and mixed life stages of T. urticae. The developmental studies unravelled a shortest developmental duration and an extended adult longevity and lifespan of A. paraaerialis when reared on the alternative diet, thus opened up the mass rearing possibility of the predatory species under laboratory conditions.

Leaf trichome-mediated predator effects on the distribution of herbivorous mites within a kidney bean plant.

Some predators prefer to settle on leaf patches with microstructures (e.g., trichomes and domatia), leaving traces on the patches. Herbivorous arthropods, in turn, select leaf patches in response to these traces left by predators. It remains unclear whether traces of predators on leaf patches affect the distribution of herbivorous prey within plants through plant microstructure. Therefore, we examined the distribution of herbivorous mite (Tetranychus urticae) and predatory mite (Phytoseiulus persimilis) by investigating their oviposition pattern. We used a kidney bean plant (Phaseolus vulgaris) with two expanded primary leaves and the first trifoliate leaf, focusing on leaf trichomes as the microstructure. The density of trichomes was higher on the first trifoliate leaf than on the primary leaves and on the abaxial surface of the leaves than on the adaxial surface. Adult female P. persimilis laid more eggs on the first trifoliate leaf to the primary leaves. Although adult female T. urticae preferred to oviposit on the abaxial surface of primary leaves, previous exposure of plants to predators diminished this preference. The altered egg distribution would be a response to the traces of P. persimilis rather than eggs of P. persimilis. Our findings indicate that T. urticae reproduces on leaf patches with traces of predators without altering their oviposition preference. Given that the presence of predator traces is known to reduce the reproduction of T. urticae, it may have a substantial effect on the population of T. urticae in the next generations on kidney bean plants.

ß-Cyanoalanine synthase protects mites against Arabidopsis defenses.

Glucosinolates are antiherbivory chemical defense compounds in Arabidopsis (Arabidopsis thaliana). Specialist herbivores that feed on brassicaceous plants have evolved various mechanisms aimed at preventing the formation of toxic isothiocyanates. In contrast, generalist herbivores typically detoxify isothiocyanates through glutathione conjugation upon exposure. Here, we examined the response of an extreme generalist herbivore, the two-spotted spider mite Tetranychus urticae (Koch), to indole glucosinolates. Tetranychus urticae is a composite generalist whose individual populations have a restricted host range but have an ability to rapidly adapt to initially unfavorable plant hosts. Through comparative transcriptomic analysis of mite populations that have differential susceptibilities to Arabidopsis defenses, we identified ß-cyanoalanine synthase of T. urticae (TuCAS), which encodes an enzyme with dual cysteine and ß-cyanoalanine synthase activities. We combined Arabidopsis genetics, chemical complementation and mite reverse genetics to show that TuCAS is required for mite adaptation to Arabidopsis through its ß-cyanoalanine synthase activity. Consistent with the ß-cyanoalanine synthase role in detoxification of hydrogen cyanide (HCN), we discovered that upon mite herbivory, Arabidopsis plants release HCN. We further demonstrated that indole glucosinolates are sufficient for cyanide formation. Overall, our study uncovered Arabidopsis defenses that rely on indole glucosinolate-dependent cyanide for protection against mite herbivory. In response, Arabidopsis-adapted mites utilize the ß-cyanoalanine synthase activity of TuCAS to counter cyanide toxicity, highlighting the mite's ability to activate resistant traits that enable this extreme polyphagous herbivore to exploit cyanogenic host plants.

The Microbiota of a Mite Prey-Predator System on Different Host Plants Are Characterized by Dysbiosis and Potential Functional Redundancy.

Microbiota has diverse roles in the life cycles of their hosts, affecting their growth, development, behavior, and reproduction. Changes in physiological conditions of the host can also impact the assemblage of host-associated microorganisms. However, little is known of the effects of host plant-prey-predatory mite interactions on mite microbiota. We compared the microbial communities of eggs and adult females of the two-spotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae), and of adult females of the predatory mite Neoseiulus californicus (McGregor) (Acari: Phytoseiidae) on four different host plants (cotton, maize, pinto bean, and tomato) by metabarcoding sequencing of the V3-V4 region of the 16S ribosomal RNA gene (16S rRNA), using the Illumina MiSeq platform. Only the egg microbiota of T. urticae was affected by the host plant. The microbiota of the predatory mite N. californicus was very different from that of its prey, and the predator microbiota was unaffected by the different host plant-prey systems tested. Only the microbiota of the eggs of T. urticae carried Serratia as a high fidelity-biomarker, but their low abundance in T. urticae adult females suggests that the association between Serratia and T. urticae is accidental. Biomarker bacteria were also detected in the microbiota of adult females of T. urticae and N. californicus, with different biomarkers in each host plant species. The microbiota associated with eggs and adult females of T. urticae and adult females of N. californicus differed in their functional potential contributions to the host mite.

Rapid cold hardening response of the phytoseiid mite Neoseiulus striatus: increased cold tolerance but not reduced predation.

The predatory mite Neoseiulus striatus (Wu) (Acari: Phytoseiidae), which has been found on maize plants in Inner Mongolia, is regarded as a promising biological control agent of small sucking pests, especially Tetranychid mites. Temperature is an important abiotic factor, and a sudden drop in temperature may affect its performance when released in areas with big circadian temperature differences. Rapid cold hardening is a type of phenotypic plasticity that allows ectotherms to quickly enhance their cold tolerance in response to a brief chilling exposure. However, it is not clear whether N. striatus possesses such plasticity. To understand how this species of phytoseiid mite copes with short-term low-temperature stress, its rapid cold hardening response was studied in the laboratory by first exploring its critical temperature. Then, the effects of exposure to a series of temperatures (0, 5, 10, 15, and 20 °C) for different durations on the survival of N. striatus were investigated to determine the optimal temperature and duration for cold hardening. Moreover, the effect of cold hardening on the consumption of Tetranychus urticae by N. striatus was also investigated, as was the response of immature stages. The critical temperature for N. striatus was - 15 °C, at which its survival dropped below 10%. An exposure to 5 °C for 2 h was optimal for rapid cold hardening, efficiently increasing survival in N. striatus. Rapid cold hardening was found in all developmental stages of N. striatus, and there was no significant effect of exposure on the consumption of spider mites. Our study shows that exposure to low temperatures during a limited period can enhance the cold hardiness but not reduce predation by N. striatus, which will be advantageous to its field applications during a period of large diurnal temperature fluctuations.

Expression and functional analysis of Niemann-Pick C2 gene in Phytoseiulus persimilis.

As a new protein class of semiochemical binding and transporting, Niemann-Pick proteins type C2 (NPC2) in arthropods have received more attentions in recent decade. However, the gene function has not been studied in phytoseiid mites with biocontrol potential. In the current study, we cloned a NPC2 gene PpNPC2a from the transcriptome of Phytoseiulus persimilis Athias-Henriot. By encoding 181 amino acids with a conserved ML domain, PpNPC2a was found a homolog of NPC2-1 in Galendromus occidentalis Nesbitt. We then measured the spatio-temporal expression of PpNPC2a in P. persimilis, and found the highest expression in female adults compared to other stages. Due to the tiny body size of predatory mites, we only examined tissue expressions in two sections: the anterior part (gnathosoma and the first pair of legs) and the posterior part (idiosoma without the first pair of legs). Higher transcription of PpNPC2a was found in the posterior part. To investigate the potential function of PpNPC2a in P. persimilis, we interfered gene expression in female adults by feeding dsRNA, which resulted in a decrease of relative expression by 59.1 and 78.2% after 24 and 72 h, respectively. Compared with the control, dsNPC2a-treated P. persimilis were insensitive to the scent of leaves or plants infested by spider mites, suggesting a role of PpNPC2a in response to plant volatiles. However, the dsNPC2a-interfered mites could still respond to four representative compounds of herbivore-induced plant volatiles, including 4,8,12-trimethyl-1,3,7,11-tridecatetraene (TMTT), methyl salicylate (MeSA), ß-caryophyllene and linalool. In short, our results indicated PpNPC2a may be involved in the chemosensory process of P. persimilis in response to whole-plant volatiles.

Sperm-depleted males of the two-spotted spider mite can replenish sperm in a few hours.

In many animals, males increase their reproductive success by mating with as many females as possible. The number of females a male can fertilize is often limited by male competition for access to females, sperm competition, and the cost of sperm production. Especially, recent studies have shown that sperm production is more costly than previously expected. In the two-spotted spider mite, Tetranychus urticae Koch, the number of females a male can inseminate is limited mainly by male competition for access to females. However, in the absence of rivals, males mate with so many females that they can become sperm-depleted. Mating without sperm transfer does not produce any offspring, although it takes time and energy. Therefore, a question arises as to why males continue to mate even after sperm depletion. In this study, we hypothesized that males continue to mate because sperm is replenished after a short period. To test the hypothesis, we investigated how long it takes for sperm replenishment after sperm depletion. We found that in 3 h, sperm can be replenished enough to inseminate a few females. As 3 h is sufficiently short not to lose the next mating opportunity, the results support the hypothesis. However, copulation duration in the sperm-replenished males was significantly longer than in the sperm-depleted males but shorter than in males before sperm depletion. To explain the differences, further research would be necessary. In addition, anatomical physiology study in males is also required to confirm that sperm is indeed depleted and replenished.