Protocols for the delivery of small molecules to the two-spotted spider mite, Tetranychus urticae

Suzuki, Takeshi; España, María Urizarna; Nunes, Maria Andréia; Zhurov, Vladimir; Dermauw, Wannes; Osakabe, Masahiro; Leeuwen, Thomas Van; Grbić, Miodrag; Grbić, Vojislava

doi:10.1371/journal.pone.0180658

Cited by 32 publications

(38 citation statements)

References 40 publications

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“…[23]. After soaking, mites were transferred onto bean leaf discs (10-mm diameter; 5 larvae/disc or 1 adult/disc) placed on water-soaked cotton in the cup with a polyethylene lid with 4 venting holes each covered with a gas-permeable filter (0.45 micron pore size; Milliseal; EMD Millipore, Billerica, MA), and incubated at L:D 16:8, 26°C and 50% RH.…”

Section: Methodsmentioning

confidence: 99%

RNAi-based reverse genetics in the chelicerate model Tetranychus urticae: A comparative analysis of five methods for gene silencing

et al. 2017

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RNA interference (RNAi) can be used for the protection against agricultural pests through the silencing of genes required for pest fitness. To assess the potential of RNAi approaches in the two-spotted spider mite, Tetranychus urticae, we compared 5 methods for the delivery of double-stranded RNA (dsRNA). These methods include mite feeding on either (i) leaves floating on a dsRNA solution, (ii) dsRNA-expressing plants, (iii) artificial diet supplemented with dsRNA, or (iv) dsRNA-coated leaves, and (v) mite soaking in a dsRNA solution. In all cases, the gene targeted for method validation was the Vacuolar-type H+-ATPase (TuVATPase), encoding a constitutively expressed ATP-driven proton pump located in the membrane. Down-regulation of TuVATPase increased mortality and/or reduced fecundity in all methods, but with variable efficiency. The most efficient methods for dsRNA delivery were direct soaking of mites in the dsRNA solution and mite feeding on dsRNA-coated leaves that mimics dsRNA application as a sprayable pesticide. Both resulted in a dark-body phenotype not observed in mites treated with a control dsRNA. Although with lower efficiency, dsRNA designed for TuVATPase silencing and expressed in transgenic Arabidopsis plants impacted the fitness of mites feeding on these plants. RNAi may thus be a valuable strategy to control spider mite populations, either as a sprayable pesticide or through transgenic crops. This comparative methodological study focusing on the induction of RNAi-based gene silencing in T. urticae paves the way for reverse genetics approaches in this model chelicerate system and prepares large-scale systematic RNAi screens as a first step towards the development of specific RNA-based pesticides. Such alternative molecules may help control spider mites that cause significant damages to crops and ornamental plant species, as well as other chelicerates detrimental to agriculture and health.

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Section: Methodsmentioning

confidence: 99%

RNAi-based reverse genetics in the chelicerate model Tetranychus urticae: A comparative analysis of five methods for gene silencing

et al. 2017

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“…To test the filtration properties of the ventriculus-posterior midgut connection, polystyrene FMs of 50 nm (Polysciences) and a fluorescent tracer dye (Alexa Fluor 555, Thermo Fisher Scientific) were delivered using the method described above. In addition, fluorescein-labeled dextran of various sizes (500, 40, and 4 kDa, MilliporeSigma) were delivered to adult female mites as mixtures with 6% blue dye (erioglaucine; McCormick, Sparks Glencoe, MD) using the leaf coating method (Suzuki et al, 2017a ). To ensure the intactness of fluorescein-labeled dextran, the fluorescein-12-UTP (Roche) was delivered as a control.…”

Section: Methodsmentioning

confidence: 99%

The Digestive System of the Two-Spotted Spider Mite, Tetranychus urticae Koch, in the Context of the Mite-Plant Interaction

et al. 2018

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The two-spotted spider mite (TSSM), Tetranychus urticae Koch (Acari: Tetranychidae), is one of the most polyphagous herbivores, feeding on more than 1,100 plant species. Its wide host range suggests that TSSM has an extraordinary ability to modulate its digestive and xenobiotic physiology. The analysis of the TSSM genome revealed the expansion of gene families that encode proteins involved in digestion and detoxification, many of which were associated with mite responses to host shifts. The majority of plant defense compounds that directly impact mite fitness are ingested. They interface mite compounds aimed at counteracting their effect in the gut. Despite several detailed ultrastructural studies, our knowledge of the TSSM digestive tract that is needed to support the functional analysis of digestive and detoxification physiology is lacking. Here, using a variety of histological and microscopy techniques, and a diversity of tracer dyes, we describe the organization and properties of the TSSM alimentary system. We define the cellular nature of floating vesicles in the midgut lumen that are proposed to be the site of intracellular digestion of plant macromolecules. In addition, by following the TSSM's ability to intake compounds of defined sizes, we determine a cut off size for the ingestible particles. Moreover, we demonstrate the existence of a distinct filtering function between midgut compartments which enables separation of molecules by size. Furthermore, we broadly define the spatial distribution of the expression domains of genes involved in digestion and detoxification. Finally, we discuss the relative simplicity of the spider mite digestive system in the context of mite's digestive and xenobiotic physiology, and consequences it has on the effectiveness of plant defenses.

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“…Working knowledge of the spider mite biology together with the development of the wide range of genomic tools and protocols established for T. urticae [23][24][25][26][27] make this species a model within the phytophagous chelicerate to address different aspects of the plant-spider mite interactions.…”

Section: Introductionmentioning

confidence: 99%

Plant Defenses Against Tetranychus urticae: Mind the Gaps

Santamaría

Arnáiz

Rosa‐Diaz

et al. 2020

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The molecular interactions between a pest and its host plant are the consequence of an evolutionary arms race based on the perception of the phytophagous arthropod by the plant and the different strategies adopted by the pest to overcome plant triggered defenses. The complexity and the different levels of these interactions make it difficult to get a wide knowledge of the whole process. Extensive research in model species is an accurate way to progressively move forward in this direction. The two-spotted spider mite, Tetranychus urticae Koch has become a model species for phytophagous mites due to the development of a great number of genetic tools and a high-quality genome sequence. This review is an update of the current state of the art in the molecular interactions between the generalist pest T. urticae and its host plants. The knowledge of the physical and chemical constitutive defenses of the plant and the mechanisms involved in the induction of plant defenses are summarized. The molecular events produced from plant perception to the synthesis of defense compounds are detailed, with a special focus on the key steps that are little or totally uncovered by previous research.

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Protocols for the delivery of small molecules to the two-spotted spider mite, Tetranychus urticae

Cited by 32 publications

References 40 publications

RNAi-based reverse genetics in the chelicerate model Tetranychus urticae: A comparative analysis of five methods for gene silencing

RNAi-based reverse genetics in the chelicerate model Tetranychus urticae: A comparative analysis of five methods for gene silencing

The Digestive System of the Two-Spotted Spider Mite, Tetranychus urticae Koch, in the Context of the Mite-Plant Interaction

Plant Defenses Against Tetranychus urticae: Mind the Gaps

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