Plant-Herbivore Interactions: A Case of an Extreme Generalist, the Two-Spotted Spider Mite Tetranychus urticae.

Plant-herbivore interactions evolved over long periods of time, resulting in an elaborate arms race between interacting species. While specialist herbivores evolved specific strategies to cope with the defenses of a limited number of hosts, our understanding of how generalist herbivores deal with the defenses of a plethora of diverse host plants is largely unknown. Understanding the interaction between a plant host and a generalist herbivore requires an understanding of the plant's mechanisms aimed at defending itself and the herbivore's mechanisms intended to counteract diverse defenses. In this review, we use the two-spotted spider mite (TSSM), Tetranychus urticae (Koch) as an example of a generalist herbivore, as this chelicerate pest has a staggering number of plant hosts. We first establish that the ability of TSSM to adapt to marginal hosts underlies its polyphagy and agricultural pest status. We then highlight our understanding of direct plant defenses against spider mite herbivory and review recent advances in uncovering mechanisms of spider mite adaptations to them. Finally, we discuss the adaptation process itself, as it allows TSSM to overcome initially effective plant defenses. A high-quality genome sequence and developing genetic tools, coupled with an ease of mite experimental selection to new hosts, make TSSM an outstanding system to study the evolution of host range, mechanisms of pest xenobiotic resistance and plant-herbivore interactions. In addition, knowledge of plant defense mechanisms that affect mite fitness are of practical importance, as it can lead to development of new control strategies against this important agricultural pest. In parallel, understanding mechanisms of mite counter adaptations to these defenses is required to maintain the efficacy of these control strategies in agricultural practices.

Highly virulent Beauveria bassiana strains against the two-spotted spider mite, Tetranychus urticae, show no pathogenicity against five phytoseiid mite species.

Entomopathogenic fungi and predatory mites can independently contribute to suppressing the two-spotted spider mite, Tetranychus urticae Koch. It is important to assess the risk of possible fungal infections in predators when a combination of them are being considered as a tandem control strategy for suppressing T. urticae. The first part of this study tested 12 Beauveria bassiana isolates for virulence in T. urticae. Strains SCWJ-2, SDDZ-9, LNSZ-26, GZGY-1-3 and WLMQ-32 were found to be the most potent, causing 37.6-49.5% adult corrected mortality at a concentration of 1 × 107 m/L conidia 4 days post-treatment. The second part evaluated the pathogenicity of these five strains in five species of predatory phytoseiid mites. The bioassay results indicated that all adult predatory mite mortalities ranged from 7.5 to 9.1% 4 days post-treatment. No viable fungal hyphae were found on predator cadavers. Observations with scanning electron microscopy revealed that conidia were attached to the cuticle of predatory mites within 2-12 h after spraying with strain LNSZ-26, and had germinated within 24-36 h. After 48 h, conidia had gradually been shed from the mites, after none of the conidia had penetrated the cuticular surfaces. In contrast, the germinated conidia successfully penetrated the cuticle of T. urticae, and within 60 h the fungus colonized the mite's body. Our study demonstrated that although several B. bassiana strains displayed a high virulence in T. urticae there was no evident pathogenicity to phytoseiid mites. These findings support the potential use of entomopathogenic fungus in combination with predatory mites in T. urticae control programs.

Mites occurrence on Pachira aquatica Aubl. including aspects of external mouthpart morphology of Brachytydeus formosa (Acari: Tydeidae).

Pachira aquatica Aubl. is commonly used as an ornamental plant in urban areas of Brazil. The objective of the study was to investigate the occurrence of mites on P. aquatica, with emphasis on Brachytydeus formosa (Cooreman), and to describe aspects the external features of its mouthpart. The study was conducted in 2012 in Jaboticabal, State of São Paulo, Brazil. Ten trees of P. aquatica were selected for the experiment. Approximately 130 leaflets were collected from each tree, which were located in different quadrants (north, south, east, and west) and strata (upper, middle, and lower). The leaflets were placed in paper bags and transported to the laboratory. The mites were prepared on optical microscope slides. A total of eleven species of mites were found, belonging to eight different families. The species and genera of the organisms included B. formosa, Eutetranychus banksi (McGregor), Agistemus sp., Tyrophagus putrescentiae (Schrank, 1781), Brevipalpus phoenicis (Geijskes), Brevipalpus sp., Cheletogenes sp., Iphiseiodes zuluagai Denmark & Muma, Euseius sp., Neoseiulus sp., and only one specimen from the Bdellidae family. The predominant species was B. formosa, with 8,142 mites equally distributed among the four quadrants and mostly in the middle and upper strata of the plant. B. formosa mites from leaflets of P. aquatica were separated for the study of the external mouthpart morphology by scanning electron microscopy (SEM).

Mites occurrence on Pachira aquatica Aubl. including aspects of external mouthpart morphology of Brachytydeus formosa (Acari: Tydeidae) / Ocorrência de ácaros em Paquira aquatica Aubl. incluindo aspectos morfológicos do aparato bucal de Brachytydeus formosa (Acari: Tydeidae)

Abstract Pachira aquatica Aubl. is commonly used as an ornamental plant in urban areas of Brazil. The objective of the study was to investigate the occurrence of mites on P. aquatica, with emphasis on Brachytydeus formosa (Cooreman), and to describe aspects the external features of its mouthpart. The study was conducted in 2012 in Jaboticabal, State of São Paulo, Brazil. Ten trees of P. aquatica were selected for the experiment. Approximately 130 leaflets were collected from each tree, which were located in different quadrants (north, south, east, and west) and strata (upper, middle, and lower). The leaflets were placed in paper bags and transported to the laboratory. The mites were prepared on optical microscope slides. A total of eleven species of mites were found, belonging to eight different families. The species and genera of the organisms included B. formosa, Eutetranychus banksi (McGregor), Agistemus sp., Tyrophagus putrescentiae (Schrank, 1781), Brevipalpus phoenicis (Geijskes), Brevipalpus sp., Cheletogenes sp., Iphiseiodes zuluagai Denmark & Muma, Euseius sp., Neoseiulus sp., and only one specimen from the Bdellidae family. The predominant species was B. formosa, with 8,142 mites equally distributed among the four quadrants and mostly in the middle and upper strata of the plant. B. formosa mites from leaflets of P. aquatica were separated for the study of the external mouthpart morphology by scanning electron microscopy (SEM).

Resumo Pachira aquatica Aubl. é frequentemente utilizada como planta ornamental em áreas urbanas no Brasil. O objetivo do trabalho foi conhecer os ácaros associados a P. aquatica com ênfase em Brachytydeus formosa (Cooreman), bem como descrever alguns aspectos morfológicos do seu aparato bucal. O estudo foi realizado em 2012, em Jaboticabal, estado de São Paulo, Brasil. Foram selecionadas dez árvores de P. aquatica para realização do experimento. Cerca de 130 folíolos foram coletados de cada árvore localizados em diferentes quadrantes (norte, sul, leste e oeste) e estratos (superior, médio e inferior). Os folíolos foram colocados em sacos de papel e transportados para o laboratório. Os ácaros foram preparados em lâminas de microscopia óptica. No total foram encontradas onze espécies de ácaros, pertencentes a oito famílias. As espécies encontradas foram B. formosa, Eutetranychus banksi (McGregor), Agistemus sp., Tyrophagus putrescentiae (Schrank, 1781), Brevipalpus phoenicis (Geijskes), Brevipalpus sp., Cheletogenes sp., Iphiseiodes zuluagai Denmark & Muma, Euseius sp., Neoseiulus sp., e apenas um exemplar da família Bdellidae. A espécie predominante foi B. formosa, com 8.142 ácaros igualmente distribuídos nos quatro quadrantes e principalmente nos estratos médio e superior da planta. Ácaros de B. formosa dos folíolos de P. aquatica foram separados para o estudo da morfologia externa do aparato bucal utilizando-se microscopia eletrônica de varredura (SEM).

Assessment of the contact toxicity of methyl palmitate on Tetranychus viennensis (Acari: Tetranychidae).

Previous work demonstrated that methyl palmitate possesses acaricidal activity against Tetranychus viennensis Boisduval (Acari: Tetranychidae) via an unknown mechanism. Here, the symptoms of methyl palmitate toxicity to T. viennensi were studied to determine the acaricidal mechanism of action of this fatty acid methyl ester. Methyl palmitate caused concentration-dependent mortality of T. viennensis, with a moderate concentration (5 mg/ml) eliciting excitement and premature oviposition without spinning shortly after exposure. Tremors of the appendages were subsequently observed, followed by quiescence after approximately 5 h. Mites developed dorsal fluid exosmosis at 15-20 h posttreatment with reduced egg production, followed shortly thereafter by death. Some typical neurotoxic symptoms such as excitement and convulsions were observed in methyl palmitate-exposed mites, suggesting that methyl palmitate may be a neurotoxin. Compared with other neurotoxic acaricides, methyl palmitate poisoning is a slow process in mites. Transmission electron microscopy revealed serious ultrastructural damage in response to 5 mg/ml methyl palmitate exposure. Autolysis of membranous structures was also observed, especially in the mitochondria, suggesting a novel mode of action for methyl palmitate-induced toxicity.

Demographic traits of Tetranychus urticae (Acari: Tetranychidae) on leaf discs and whole leaves.

Life tables of twospotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae), were studied on leaf discs and whole leaves of the common bean plant, Phaseolus vulgaris L. Data were analyzed based on the age-stage, two-sex life table theory. Durations of deutonymph stage, total preadult stage, and preoviposition period of mites reared on whole leaves (1.22, 7.6, and 0.29 d, respectively) were significantly shorter than those reared on leaf discs (1.73, 8.2, and 0.89 d, respectively). The lifetime fecundities were 22.81 and 12.05 offspring on whole leaves and leaf discs, respectively. Although hooked trichomes on the lower surface of the bean leaf contribute to adult mortality, spider mites successfully survive and reproduce on bean plants. The intrinsic rate of increase on whole leaves (0.235 d(-1)) was higher than that on leaf discs (0.159 d(-1). Higher intrinsic rate and fecundity found in mites reared on whole leaves may be due to the higher nutritional quality of whole leaves over leaf discs. We suggest that whole leaves should be used in future studies to better simulate realistic life history characteristics. The advantages of using age-stage, two-sex life tables over female age-specific life tables are discussed.

Gene silencing in the spider mite Tetranychus urticae: dsRNA and siRNA parental silencing of the Distal-less gene.

A major prerequisite to understanding the evolution of developmental programs includes an appreciation of gene function in a comparative context. RNA interference (RNAi) represents a powerful method for reverse genetics analysis of gene function. However, RNAi protocols exist for only a handful of arthropod species. To extend functional analysis in basal arthropods, we developed a RNAi protocol for the two-spotted spider mite Tetranychus urticae focusing on Distal-less (Dll), a conserved gene involved in appendage specification in metazoans. First, we describe limb morphogenesis in T. urticae using confocal and scanning electron microscopy. Second, we examine T. urticae Dll (Tu-Dll) mRNA expression patterns and correlate its expression with appendage development. We then show that fluorescently labeled double-stranded RNA (dsRNA) and short interfering RNA (siRNA) molecules injected into the abdomen of adult females are incorporated into the oviposited eggs, suggesting that dsRNA reagents can be systemically distributed in spider mites. Injection of longer dsRNA as well as siRNA induced canonical limb truncation phenotypes as well as the fusion of leg segments. Our data suggest that Dll plays a conserved role in appendage formation in arthropods and that such conserved genes can serve as reliable starting points for the development of functional protocols in nonmodel organisms.