Luc Tirry

The economic importance of acaricides in the control of phytophagous mites and an update on recent acaricide mode of action research

Acaricides are one of the cornerstones of an efficient control program for phytophagous mites. An analysis of the global acaricide market reveals that spider mites such as Tetranychus urticae, Panonychus citri and Panonychus ulmi are by far the most economically important species, representing more than 80% of the market. Other relevant mite groups are false spider mites (mainly Brevipalpus), rust and gall mites and tarsonemid mites. Acaricides are most frequently used in vegetables and fruits (74% of the market), including grape vines and citrus. However, their use is increasing in major crops where spider mites are becoming more important, such as soybean, cotton and corn. As revealed by a detailed case study of the Japanese market, major shifts in acaricide use are partially driven by resistance development and the commercial availability of compounds with novel mode of action. The importance of the latter cannot be underestimated, although some compounds are successfully used for more than 30 years. A review of recent developments in mode of action research is presented, as such knowledge is important for devising resistance management programs. This includes spirocyclic keto-enols as inhibitors of acetyl-CoA carboxylase, the carbazate bifenazate as a mitochondrial complex III inhibitor, a novel class of complex II inhibitors, and the mite growth inhibitors hexythiazox, clofentezine and etoxazole that interact with chitin synthase I.

The salivary protein repertoire of the polyphagous spider mite Tetranychus urticae: a quest for effectors

The two-spotted spider mite Tetranychus urticae is an extremely polyphagous crop pest. Alongside an unparalleled detoxification potential for plant secondary metabolites, it has recently been shown that spider mites can attenuate or even suppress plant defenses. Salivary constituents, notably effectors, have been proposed to play an important role in manipulating plant defenses and might determine the outcome of plant-mite interactions. Here, the proteomic composition of saliva from T. urticae lines adapted to various host plants—bean, maize, soy, and tomato—was analyzed using a custom-developed feeding assay coupled with nano-LC tandem mass spectrometry. About 90 putative T. urticae salivary proteins were identified. Many are of unknown function, and in numerous cases belonging to multimembered gene families. RNAseq expression analysis revealed that many genes coding for these salivary proteins were highly expressed in the proterosoma, the mite body region that includes the salivary glands. A subset of genes encoding putative salivary proteins was selected for whole-mount in situ hybridization, and were found to be expressed in the anterior and dorsal podocephalic glands. Strikingly, host plant dependent expression was evident for putative salivary proteins, and was further studied in detail by micro-array based genome-wide expression profiling. This meta-analysis revealed for the first time the salivary protein repertoire of a phytophagous chelicerate. The availability of this salivary proteome will assist in unraveling the molecular interface between phytophagous mites and their host plants, and may ultimately facilitate the development of mite-resistant crops. Furthermore, the technique used in this study is a time- and resource-efficient method to examine the salivary protein composition of other small arthropods for which saliva or salivary glands cannot be isolated easily.

Transcriptome profiling of a spirodiclofen susceptible and resistant strain of the European red mite Panonychus ulmi using strand-specific RNA-seq

BackgroundThe European red mite, Panonychus ulmi, is among the most important mite pests in fruit orchards, where it is controlled primarily by acaricide application. However, the species rapidly develops pesticide resistance, and the elucidation of resistance mechanisms for P. ulmi has not kept pace with insects or with the closely related spider mite Tetranychus urticae. The main reason for this lack of knowledge has been the absence of genomic resources needed to investigate the molecular biology of resistance mechanisms.ResultsHere, we provide a comprehensive strand-specific RNA-seq based transcriptome resource for P. ulmi derived from strains susceptible and resistant to the widely used acaricide spirodiclofen. From a de novo assembly of the P. ulmi transcriptome, we manually annotated detoxification enzyme families, target-sites of commonly used acaricides, and horizontally transferred genes implicated in plant-mite interactions and pesticide resistance. In a comparative analysis that incorporated sequences available for Panonychus citri, T. urticae, and insects, we identified radiations for detoxification gene families following the divergence of Panonychus and Tetranychus genera. Finally, we used the replicated RNA-seq data from the spirodiclofen susceptible and resistant strains to describe gene expression changes associated with resistance. A cytochrome P450 monooxygenase, as well as multiple carboxylcholinesterases, were differentially expressed between the susceptible and resistant strains, and provide a molecular entry point for understanding resistance to spirodiclofen, widely used to control P. ulmi populations.ConclusionsThe new genomic resources and data that we present in this study for P. ulmi will substantially facilitate molecular studies of underlying mechanisms involved in acaricide resistance.

Diurnal Temperature Variations Affect Development of a Herbivorous Arthropod Pest and its Predators

The impact of daily temperature variations on arthropod life history remains woefully understudied compared to the large body of research that has been carried out on the effects of constant temperatures. However, diurnal varying temperature regimes more commonly represent the environment in which most organisms thrive. Such varying temperature regimes have been demonstrated to substantially affect development and reproduction of ectothermic organisms, generally in accordance with Jensen’s inequality. In the present study we evaluated the impact of temperature alternations at 4 amplitudes (DTR0, +5, +10 and +15°C) on the developmental rate of the predatory mites Phytoseiulus persimilis Athias-Henriot and Neoseiulus californicus McGregor (Acari: Phytoseiidae) and their natural prey, the two-spotted spider mite Tetranychus urticae Koch (Acari: Tetranychidae). We have modelled their developmental rates as a function of temperature using both linear and nonlinear models. Diurnally alternating temperatures resulted in a faster development in the lower temperature range as compared to their corresponding mean constant temperatures, whereas the opposite was observed in the higher temperature range. Our results indicate that Jensen’s inequality does not suffice to fully explain the differences in developmental rates at constant and alternating temperatures, suggesting additional physiological responses play a role. It is concluded that diurnal temperature range should not be ignored and should be incorporated in predictive models on the phenology of arthropod pests and their natural enemies and their performance in biological control programmes.

Supplemental food for Amblyseius swirskii in the control of thrips: feeding friend or foe?

BACKGROUND In integrated pest management systems in greenhouse crops, the predatory mite Amblyseius swirskii is becoming increasingly important as a biological control agent of various pests, especially thrips and whiteflies. An emerging strategy to promote the predators establishment and retention in the crop consists in providing food supplements. However, when faced with omnivorous pests, such as the western flower thrips, Frankliniella occidentalis, food supplements need to be applied with extreme care, in order not to boost population growth of the pest. This laboratory study was conducted to evaluate the impact of commercial products of Typha angustifolia pollen and decapsulated brine shrimp cysts (Artemia sp.) on populations of both pest and predator and on predator-prey interactions. RESULTS Pollen was highly supportive for both F. occidentalis and A. swirskii, whereas Artemia cysts supported thrips populations to a lesser extent than those of the predator. Furthermore, a less pronounced reduction in thrips consumption by A. swirskii was observed in the presence of Artemia cysts as compared with T. angustifolia pollen. CONCLUSION Artemia might be a valuable alternative to pollen for supporting populations of A. swirskii in order to improve thrips management, as they are less beneficial for the pest but do support population growth of A. swirskii.

Disruption of a horizontally transferred phytoene desaturase abolishes carotenoid accumulation and diapause in Tetranychus urticae

Significance In animals, carotenoid pigments fulfill a number of essential roles in vision, protection from stresses, and signaling. Although carotenoids are synthesized primarily by plants and some bacteria and fungi, carotenoid biosynthetic genes have been acquired by horizontal gene transfer in some insects and mites. In the two-spotted spider mite, Tetranychus urticae, as well as in a related species, we show that a horizontally transferred carotenoid biosynthetic gene is required for normal pigmentation. Spider mites can therefore synthesize their own carotenoids. Further, in a T. urticae strain, the gene is required for the induction of diapause, a key for overwintering of this widespread and important agricultural pest. Carotenoids underlie many of the vibrant yellow, orange, and red colors in animals, and are involved in processes ranging from vision to protection from stresses. Most animals acquire carotenoids from their diets because de novo synthesis of carotenoids is primarily limited to plants and some bacteria and fungi. Recently, sequencing projects in aphids and adelgids, spider mites, and gall midges identified genes with homology to fungal sequences encoding de novo carotenoid biosynthetic proteins like phytoene desaturase. The finding of horizontal gene transfers of carotenoid biosynthetic genes to three arthropod lineages was unprecedented; however, the relevance of the transfers for the arthropods that acquired them has remained largely speculative, which is especially true for spider mites that feed on plant cell contents, a known source of carotenoids. Pigmentation in spider mites results solely from carotenoids. Using a combination of genetic approaches, we show that mutations in a single horizontally transferred phytoene desaturase result in complete albinism in the two-spotted spider mite, Tetranychus urticae, as well as in the citrus red mite, Panonychus citri. Further, we show that phytoene desaturase activity is essential for photoperiodic induction of diapause in an overwintering strain of T. urticae, consistent with a role for this enzyme in provisioning provitamin A carotenoids required for light perception. Carotenoid biosynthetic genes of fungal origin have therefore enabled some mites to forgo dietary carotenoids, with endogenous synthesis underlying their intense pigmentation and ability to enter diapause, a key to the global distribution of major spider mite pests of agriculture.

Molecular analysis of cyenopyrafen resistance in the two-spotted spider mite Tetranychus urticae

BACKGROUND Cyenopyrafen is a recently developed acaricide with a new mode of action as a complex II inhibitor. However, it was recently shown that cross-resistance to cyenopyrafen can occur in resistant field strains of Tetranychus urticae, which might be linked to the previous use of classical METI acaricides. Here, we selected for cyenopyrafen resistance and studied the molecular mechanisms that underlie resistance. RESULTS Selection for cyenopyrafen resistance confers cross-resistance to the complex II inhibitor cyflumetofen, but also to pyridaben, a frequently used complex I inhibitor. Cyenopyrafen resistance is highly synergised by piperonyl butoxide, and a 15-fold higher P450 activity was detected in the resistant strain. Target-site resistance was not detected. Genome-wide gene expression data, followed by a meta-analysis of previously obtained gene expression data, revealed the overexpression specifically of CYP392A11 and CYP392A12. CONCLUSIONS Cyenopyrafen resistance is strongly linked to the overexpression of two P450s, which probably explains the observed cross-resistance. This information is highly valuable, as the novel complex II inhibitors cyenopyrafen and cyflumetofen are in the process of worldwide registration. The role of both CYP392A11 and CYP392A12 should be further supported by functional expression, but they are very promising candidates as molecular diagnostic markers for monitoring cyenopyrafen susceptibility in the field.

High population densities of Macrolophus pygmaeus on tomato plants can cause economic fruit damage: interaction with Pepino mosaic virus?

BACKGROUND The zoophytophagous predator Macrolophus pygmaeus Rambur (Hemiptera: Miridae) is a successful biocontrol agent against several pest species in protected tomato crops. This predator is considered to be harmless for the crop. However, in recent years, Heteroptera feeding punctures on tomato fruit in Belgian and Dutch greenhouses have been misinterpreted as Pepino mosaic virus (PepMV) symptoms. In this study, three hypotheses were tested: (1) M. pygmaeus causes fruit damage that increases with population density and surpasses economic thresholds; (2) the presence of prey or alternative prey reduces the damage; (3) an infection of the tomato plants by PepMV triggers or aggravates M. pygmaeus fruit damage. RESULTS At increasing M. pygmaeus densities, the severity of fruit damage increased from a few dimples towards yellowish discoloration and deformed fruits. A correlation with an infection with PepMV was found. The severity of the symptoms was independent of the presence of prey. A minimum economic density threshold was estimated at 0.32 M. pygmaeus per leaf. CONCLUSION M. pygmaeus can cause economic damage to tomato fruits at densities common in practice. An infection of the plants with PepMV enhances fruit symptoms significantly. Interacting plant defence responses are most likely the key to explaining this, although confirmation is required.

Establishment of Amblyseius swirskii in greenhouse crops using food supplements

Abstract Food supplementation is an emerging strategy to improve the establishment and resilience of generalist predators in greenhouse crops. This study was conducted to assess the pre-establishment of the generalist predatory mite Amblyseius swirskii Athias-Henriot (Phytoseiidae) on two ornamental greenhouse crops, chrysanthemum and ivy. In the first greenhouse trial, we compared two commercial products consisting of Typha angustifolia pollen and decapsulated brine shrimp cysts (Artemia spp.), respectively, with a powdered artificial diet (AD) as food supplements for the predator on chrysanthemum plants at a constant temperature regime of 25°C at a 16:8 h (L:D) photoperiod. A population of A. swirskii established well when T. angustifolia pollen or the artificial diet were applied, but did not when the Artemia product was used as a supplement. A second trial on ivy plants was conducted at two temperature regimes (a constant 25°C and an alternating temperature regime of 30°C/15°C, both at a 16:8 h (L:D) photoperiod), comparing T. angustifolia pollen, the artificial diet and a non-commercial strain of Artemia franciscana cysts as food supplements. At the constant temperature regime, all three food sources allowed the establishment of A. swirskii. At the alternating temperature regime, however, T. angustifolia pollen and the AD but not A. franciscana cysts allowed a population of the predator to establish. Failure of establishment on the latter food supplement was attributed to dehydration effects under low humidity conditions in the greenhouse associated with the higher daytime temperatures at the alternating temperature regime. In conclusion, this study demonstrated the potential of T. angustifolia pollen and an artificial diet as a food supplement to support a population of A. swirskii in different ornamental crops.

Efficacy of entomopathogenic nematodes against larvae of Tuta absoluta in the laboratory.

BACKGROUND Previous studies have indicated the control potential of entomopathogenic nematodes (EPNs) against Tuta absoluta. Here, the potential of Steinernema feltiae, S. carpocapsae and Heterorhabditis bacteriophora is studied when applied against larvae of T. absoluta inside leaf mines in tomato leaf discs by means of an automated spray boom. RESULTS The studied EPN species were effective against all four larval instars of T. absoluta but caused higher mortality in the later instars (e.g. fourth instar: 77.1-97.4% mortality) than in the first instars (36.8-60.0% mortality). Overall, S. feltiae and S. carpocapsae yielded better results than H. bacteriophora. Steinernema carpocapsae and H. bacteriophora performed better at 25 °C (causing 55.3 and 97.4% mortality respectively) than at 18 °C (causing 12.5 and 34.2% mortality respectively), whereas S. feltiae caused 100% mortality at both temperatures. Under optimal spraying conditions and with the use of Addit and Silwet L-77 adjuvants, a reduced dosage of 6.8 infective juveniles (IJs) cm(-2) yielded equally good control as a recommended dosage of 27.3 IJs cm(-2) . CONCLUSION Under laboratory conditions, S. feltiae and S. carpocapsae showed good potential against the larvae of T. absoluta inside tomato leaf mines. Results need to be confirmed in greenhouse experiments.