The Effect of Patch Size and Persistence of Host Plants on the Development of Acaricide Resistance in the Two-spotted Spider Mite Tetranychus Urticae (Acari: Tetranychidae)

Spatial and temporal characteristics of host plants can influence the population biology of the herbivores feeding on them. In this study, I examined the effect of variation in host plant characteristics on the development of acaricide resistance in the two-spotted spider mite Tetranychus urticae, a widely distributed agricultural pest. This investigation examined the geographic variation in the degree of resistance to two new types of acaricide, pyridaben and fenpyroximate. From mortality tests at field-level concentrations of the acaricides, many populations collected from fruit trees and roses had a high frequency of resistant individuals for acaricides while almost all populations collected from herbaceous crops had low frequencies of resistant individuals. These results, combined with those from a previous allozyme study, indicate that patch size and persistence of host plants regulate the population structure of the mites including gene flow between populations and, by extension, the development of acaricide resistance.     

Demographic examination of sex ratio in the two-spotted spider mite, Tetranychus urticae

Primary sex ratio of arrhenotokous tetranychid mites depends on the age-specific schedule of ratios of progeny produced by individual females. Since primary sex ratio is conditional upon female age and since the proportion of females of a given age in a population is influenced by the rate of increase, both the agespecific ratio of males to females and the overall ratio of males to females depends on the rate of increase. We given an expression for this dependence and compare it to experimental results for the two-spotted spiter mite. These results imply that there is no standard sex ratio of spider mites because of interpopulation variation in rates of increase.

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Résumé Le taux sexuel des acariens tétranychidés arrhénotoques est défini comme le rapport de la somme des nombres de mâles sur la somme des nombres de femelles pour chaque âge de la distribution en âges. Comme le taux sexuel primaire dépend de l'âge de la femelle et comme la proportion de femelles d'un âge donné dans la population est influencée par le taux d'accroissement, alors le rapport mâle/femelle spécifique d'un âge déterminé et le rapport global mâle/femelle dépendent du taux d'accroissement. Nous avons cherché à exprimer cette dépendance et à la comparer aux résultats expérimentaux pour Tetranychus urticae. Ces résultats signifient qu'il n'y a pas de taux sexuel caractéristique de Tétranyques par suite des variations du taux d'accroissement suivant les populations.

     

Role of the entomogenous fungus, Neozygites floridana, in population declines of the twospotted spider mite, Tetranychus urticae, on field corn

In field plots in which populations of predaceous species were suppressed using carbaryl, populations of the twospotted spider mite, Tetranychus urticae Koch, on field corn declined in association with aerial dispersal of the mites or with epizootics of the entomogenous fungus, Neozygites floridana Weiser & Muma. The relative importance of these two factors varied from year to year depending on environmental conditions. When moist weather conditions induced epizootics of N. floridana before corn plants became heavily infested with mites, the pathogen appeared to be the major factor causing mite population declines. When dry weather conditions allowed mite populations to expand unchecked until corn plants became entirely infested, mite aerial dispersal was the major factor associated with mite population declines. In every field in which there was an epizootic of N. floridana, routine applications of the fungicide maneb delayed epizootics and reduced or delayed the impact of the pathogen on mite populations. The total number of Neozygites-infected mites produced in maneb-treated plots throughout the test period was significantly greater than that in untreated plots when epizootics were initiated during periods of mite population growth. Fewer total infected mites were produced in maneb-treated plots compared to untreated plots when epizootics were initiated during periods of mite population decline.

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Résumé Dans des parcelles de maïs dont les populations d'espèces prédatrices avaient été éliminées par du carbaryl, des populations de T. urticae ont régressé en fonction de la dispersion aérienne des acariens ou d'épizooties provoquées par N. floridana. L'importance relative de ces deux causes varie d'année suivant les conditions écologiques. Le pathogène paraissait la principale cause de déclin des populations quand le temps humide a induit les épizooties par N. floridana avant que le maïs n'ait été largement contaminé par les acariens. Quand le temps sec a permis aux populations d'acariens de se développer sans frein jusqu'à contamination totale du maïs, la dispersion aérienne des acariens était la principale cause de déclin des populations d'acariens. Dans tous les champs où il y avait eu épizootie de N. floridana, des traitements de routine avec le fongicide meneb ont retardé les épizooties et réduit ou retardé l'impact du pathogène sur les populations d'acariens. Pendant toute la période d'étude, le nombre total d'acariens infestés par Neozygites était significativement plus élevé dans les parcelles traitées au maneb que dans les parcelles non traitées, lorsque les épizooties étaient provoquées pendant la période de croissance des populations d'acariens. Quand les épizooties ont été déclenchées pendant la période de déclin des populations d'acariens, un nombre plus faible d'acariens infestés était observé dans les parcelles traitées au maneb que dans les parcelles sans traitement.

     

Identification of Wolbachia-responsive microRNAs in the two-spotted spider mite,Tetranychus urticae

Background

The two-spotted spider mite, Tetranychus urticae, is infected with Wolbachia, which have the ability to manipulate host reproduction and fitness. MicroRNAs (miRNAs) are small non-coding RNAs that are involved in many biological processes such as development, reproduction and host-pathogen interactions. Although miRNA was observed to involve in Wolbachia-host interactions in the other insect systems, its roles have not been fully deciphered in the two-spotted spider mite.

Results

Small RNA libraries of infected and uninfected T. urticae for both sexes (in total four libraries) were constructed. By integrating the mRNA data originated from the same samples, the target genes of the differentially expressed miRNAs were predicted. Then, GO and pathway analyses were performed for the target genes. Comparison of libraries showed that Wolbachia infection significantly regulated 91 miRNAs in females and 20 miRNAs in males, with an overall suppression of miRNAs in Wolbachia-infected libraries. A comparison of the miRNA and mRNA data predicted that the differentially expressed miRNAs negatively regulated 90 mRNAs in females and 9 mRNAs in males. An analysis of target genes showed that Wolbachia-responsive miRNAs regulated genes with function in sphingolipid metabolism, lysosome function, apoptosis and lipid transporting in both sexes, as well as reproduction in females.

Conclusion

Comparisons of the miRNA and mRNA data can help to identify miRNAs and miRNA target genes involving in Wolbachia-host interactions. The molecular targets identified in this study should be useful in further functional studies.

Electronic supplementary material

The online version of this article (doi:10.1186/1471-2164-15-1122) contains supplementary material, which is available to authorized users.     

Effects of mating status and age on dispersal behavior in the twospotted spider mite, Tetranychus urticae in response to fenvalerate-treated leaf surfaces

Effects of variation in age and mating status on dispersal of the twospotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae), from fenvalerate-treated leaf surfaces were investigated. Unmated and mated female mites, 1 to 6 days in age, were scored for walkoff and spindown dispersal response on bean leaves treated with a sub-lethal, residual dose of fenvalerate. Variation in mite age did not have a pronounced effect on dispersal response. Significant differences in dispersal response due to mating type were observed, yet the mean differences in dispersal response between unmated and mated females was generally small. Our data suggest that the current practice of using mated female mites of unknown age when testing for among-population differences in dispersal response, in most cases, provides an adequate estimate of population response when differences among treatments in dispersal response are large. However, when testing for within-population differences in dispersal response, small treatment effects may be more easily measured using a single age class of mites, because this could decrease within-treatment variance.     

Intraspecific variation in a generalist herbivore accounts for differential induction and impact of host plant defences

Plants and herbivores are thought to be engaged in a coevolutionary arms race: rising frequencies of plants with anti-herbivore defences exert pressure on herbivores to resist or circumvent these defences and vice versa. Owing to its frequency-dependent character, the arms race hypothesis predicts that herbivores exhibit genetic variation for traits that determine how they deal with the defences of a given host plant phenotype. Here, we show the existence of distinct variation within a single herbivore species, the spider mite Tetranychus urticae, in traits that lead to resistance or susceptibility to jasmonate (JA)-dependent defences of a host plant but also in traits responsible for induction or repression of JA defences. We characterized three distinct lines of T. urticae that differentially induced JA-related defence genes and metabolites while feeding on tomato plants (Solanum lycopersicum). These lines were also differently affected by induced JA defences. The first line, which induced JA-dependent tomato defences, was susceptible to those defences; the second line also induced JA defences but was resistant to them; and the third, although susceptible to JA defences, repressed induction. We hypothesize that such intraspecific variation is common among herbivores living in environments with a diversity of plants that impose diverse selection pressure.     

Comparison of methods for estimating the prevalence of Neozygites floridana in Tetranychus urticae populations infesting strawberries

Methods for measuring prevalence of Neozygites floridana in a Tetranychus urticae population collected from strawberries were developed and compared. T. urticae were extracted from leaves using a soapy water solution (0.5 ml washing detergent : 8 L water) and then placed into 80% alcohol for use in Methods 1 and 2. Method 1: N. floridana-sporulating T. urticae cadavers were observed and quantified under a compound microscope (40-80x). Method 2: Adult females were mounted in lactophenol cotton blue and observed for the presence or absence of N. floridana hyphal bodies under a microscope (200-400x). Method 3: Live T. urticae females were incubated at 25 degrees C and 75% RH and observed for mortality and N. floridana infection under a compound microscope (6.4-40x). Method 1 was the most time-efficient method and it also allows processing of samples as time permits. Method 2 quantified significantly higher fungal prevalence than Methods 1 and 3, but Method 2 is not considered to be reliable because hyphal bodies are difficult to detect. No significant differences were found between Methods 1 and 3.     

Sublethal effects of esfenvalerate residues on pyrethroid resistant Typhlodromus pyri (Acari: Phytoseiidae) and its prey Panonychus ulmi and Tetranychus urticae (Acari: Tetranychidae)

The effect of residues of esfenvalerate on oviposition of the resistant strain of the predatory mite Typhlodromus pyri and its main prey, European red mite Panonychus ulmi and two-spotted spider mite Tetranychus urticae, were investigated. T. pyri showed a significant linear reduction in oviposition after 24h in the presence of increasing levels of esfenvalerate residue applied at the field rate. Furthermore, when given a choice, T. pyri preferred to lay eggs on residue-free surfaces. Of the two prey species, only P. ulmi showed significant avoidance of increasing levels of residues of the field rate concentration of esfenvalerate, as measured by runoff mortality, however both P. ulmi and T. urticae, when given a choice, showed a preference for esfenvalerate-free surfaces. As with the predatory mite T. pyri, both prey species showed a significant linear reduction of oviposition with increasing esfenvalerate residues and a preference to lay eggs on esfenvalerate-free surfaces. Esfenvalerate residues as high as 15X field rate were not repellent to pyrethroid-resistant T. pyri. The possible effects of these sublethal effects on predator-prey dynamics and implications for integrated mite control programmes in apple orchards are discussed.     

Evaluation of Vigna angularis (Willd.) Ohwi and Ohashi accessions for resistance to Tetranychus urticae Koch (Acari:Tetranychidae)

Under laboratory conditions 88 azuki bean and three rice bean accessions were evaluated with a leaf-disc technique to select for those which expressed an antibiotic response to the TSSM ovipositional rate. Nine of the most promising accessions determined in the screening and Erimo, a commercial cultivar used as a control, were re-evaluated to determine the ovipositional rate and fecundity. Statistically significant differences between accessions were found: some of the accessions exhibited a reduced TSSM ovipositional rate and fecundity when compared with the susceptible control. The accessions ADZU279 and ADZU514 from Japan and ADZU577 from Ivory Coast demonstrated strong inhibition to TSSM fecundity. These three accessions may possess inherent potential in a breeding programme designed to enhance resistance to T. urticae.     

Life table characteristics of Macrolophus caliginosus preying upon Tetranychus urticae

The life table characteristics of the polyphagous mirid Macrolophus caliginosus Wagner (Heteroptera: Miridae) preying on various stages of Tetranychus urticae Koch (Acari: Tetranychidae) with tomato as host plant were described at 22 °C. The following average parameters were obtained: Female longevity: 28.7 days; fecundity: 0.7 eggs/female/day; egg mortality: 2.6%; pre-oviposition period: 5.5 days; oviposition period: 18.1 days; post-oviposition period: 3.2 days; juvenile development time: 26.8 days; juvenile mortality: 34.9%; and sex ratio (/(+): 0.46. Life table parameters were estimated as net reproduction rate (R ₀): 6.15; intrinsic rate of increase (r _m): 0.031 day^–1; finite rate of increase (): 1.032; mean generation time (T _c): 58.17 days; and doubling time (T ₂) 22.2 days. The parameters obtained were in accordance with those reported for M. caliginosus fed on another mite species (T. turkestani Ugarov & Nikolski (Acari: Tetranychidae)). However, compared to the performance of M. caliginosus fed on common glasshouse insect pests, a diet consisting of only mites appeared to be inferior. However, being a voracious predator, M. caliginosus may be a valuable addition to existing methods of mite control.     

Comparative analysis of proteome changes induced by the two spotted spider mite Tetranychus urticae and methyl jasmonate in citrus leaves

Citrus plants are currently facing biotic and abiotic stresses. Therefore, the characterization of molecular traits involved in the response mechanisms to stress could facilitate selection of resistant varieties. Although large cDNA microarray profiling has been generated in citrus tissues, the available protein expression data are scarce. In this study, to identify differentially expressed proteins in Citrus clementina leaves after infestation by the two-spotted spider mite Tetranychus urticae, a proteome comparison was undertaken using two-dimensional gel electrophoresis. The citrus leaf proteome profile was also compared with that of leaves treated over 0-72 h with methyl jasmonate, a compound playing a key role in the defense mechanisms of plants to insect/arthropod attack. Significant variations were observed for 110 protein spots after spider mite infestation and 67 protein spots after MeJA treatments. Of these, 50 proteins were successfully identified by liquid chromatography-mass spectrometry-tandem mass spectrometry. The majority constituted photosynthesis- and metabolism-related proteins. Five were oxidative stress associated enzymes, including phospholipid glutathione peroxidase, a salt stressed associated protein, ascorbate peroxidase and Mn-superoxide dismutase. Seven were defense-related proteins, such as the pathogenesis-related acidic chitinase, the protease inhibitor miraculin-like protein, and a lectin-like protein. This is the first report of differentially regulated proteins after T. urticae attack and exogenous MeJA application in citrus leaves.     

Interactions in a tritrophic acarine predator-prey metapopulation system III: effects of Tetranychus urticae (Acari: Tetranychidae) on host plant condition

Spider mites are serious pests on many economically important plant species, because they may reduce plant productivity and, at high mite densities, overexploit and even kill the host plants. We have conducted a series of greenhouse experiments to quantify the effects of two-spotted spider mites (Tetranychus urticae) on host plants (Phaseolusvulgaris). The average amount of chlorophyll per cm² leaf area was used as a measure of plant condition. It was shown that chlorophyll concentration decreases with plant age and intensity of spider mite feeding. Damage caused by spider mites was assessed visually, using the Leaf Damage Index (LDI) defined by, and a mathematical relationship between the visual measurements and the amount of chlorophyll/cm² was fitted to data. The relationship may serve as a short-cut to estimate overall plant injury, expressed as the relative loss of chlorophyll/cm² leaf area caused by spider mites (D). D takes values between 0 (no injury) and 1 (all leaves dead). A highly significant positive relationship between the instantaneous spider mite density and D was found, even though D is expected to reflect the cumulated density of mites (mite-days). A model of plant growth incorporating information about plant age and D predicts that plant area has a maximum when plant age is about 60 days, and that plant area decreases exponentially with an increase in D. This revised version was published online in July 2006 with corrections to the Cover Date.     

Soil Sampling and Isolation of Entomopathogenic Nematodes (Steinernematidae,Heterorhabditidae)

Entomopathogenic nematodes (a.k.a. EPN) represent a group of soil-inhabiting nematodes that parasitize a wide range of insects. These nematodes belong to two families: Steinernematidae and Heterorhabditidae. Until now, more than 70 species have been described in the Steinernematidae and there are about 20 species in the Heterorhabditidae. The nematodes have a mutualistic partnership with Enterobacteriaceae bacteria and together they act as a potent insecticidal complex that kills a wide range of insect species.Herein, we focus on the most common techniques considered for collecting EPN from soil. The second part of this presentation focuses on the insect-baiting technique, a widely used approach for the isolation of EPN from soil samples, and the modified White trap technique which is used for the recovery of these nematodes from infected insects. These methods and techniques are key steps for the successful establishment of EPN cultures in the laboratory and also form the basis for other bioassays that consider these nematodes as model organisms for research in other biological disciplines. The techniques shown in this presentation correspond to those performed and/or designed by members of S. P. Stock laboratory as well as those described by various authors.     

In Vitro Reconstitution of Light-harvesting Complexes of Plants and Green Algae

In plants and green algae, light is captured by the light-harvesting complexes (LHCs), a family of integral membrane proteins that coordinate chlorophylls and carotenoids. In vivo, these proteins are folded with pigments to form complexes which are inserted in the thylakoid membrane of the chloroplast. The high similarity in the chemical and physical properties of the members of the family, together with the fact that they can easily lose pigments during isolation, makes their purification in a native state challenging. An alternative approach to obtain homogeneous preparations of LHCs was developed by Plumley and Schmidt in 1987¹, who showed that it was possible to reconstitute these complexes in vitro starting from purified pigments and unfolded apoproteins, resulting in complexes with properties very similar to that of native complexes. This opened the way to the use of bacterial expressed recombinant proteins for in vitro reconstitution. The reconstitution method is powerful for various reasons: (1) pure preparations of individual complexes can be obtained, (2) pigment composition can be controlled to assess their contribution to structure and function, (3) recombinant proteins can be mutated to study the functional role of the individual residues (e.g., pigment binding sites) or protein domain (e.g., protein-protein interaction, folding). This method has been optimized in several laboratories and applied to most of the light-harvesting complexes. The protocol described here details the method of reconstituting light-harvesting complexes in vitro currently used in our laboratory,and examples describing applications of the method are provided.     

Preliminary evaluation of <Emphasis Type="Italic">Cladosporium cladosporioides</Emphasis> (Fresen.) de Vries in laboratory conditions,as a potential candidate for biocontrol of <Emphasis Type="Italic">Tetranychus urticae</Emphasis> Koch

A survey for natural entomopathogenic fungi of two spotted spider mite (Tetranychus urticae) adults was made in Erzurum, Turkey, during the period 2006. Tetranychus urticae (65.8%) infected with a strain of the fungus Cladosporium cladosporioides were found. Thirteen isolates of C. cladosporioides were assessed against T. urticae, in a single dose (8 × 10⁶ conidia ml ⁻¹), laboratory bioassay on bean leaflets. The total mortality percentage caused by C. cladosporioides isolates varied from 50.95 to 74.76% and LT₅₀ values ranged from 2.34 to 3.90 days. The results revealed that isolates of C. cladosporioides were effective against two spotted spider mite. This is the first record of natural infection of T. urticae by C. cladosporioides.     

Accumulation and turnover of 2-tridecanone in Tetranychus urticae and its consequences for resistance of wild and cultivated tomatoes

In this study we assessed the dynamic changes of 2-tridecanone in a herbivorous mite (Tetranychus urticae) on tomato (Lycopersicon esculentum, cv. Moneymaker), a plant with methyl ketones in the tetracellular tips of the glandular trichomes (Type VI). We showed that spider mites accumulate 2-tridecanone when foraging on cultivated tomato. Thus, the rate of mite–trichome contact multiplied by the amount of toxin per trichome tip exceeded the relative rate of toxin turnover multiplied by the amount of toxin per mite. The relative rate of toxin turnover was estimated to be 1.1 per day on cucumber, a plant without this toxin. The amount per trichome tip varied from 0.33 ng for middle-leaf trichomes to 1.26 ng for main-stem trichomes. Hence, to achieve a static level of 2-tridecanone equal to 8–17 ng per mite – representing the level we found in mites on middle leaves – the rate of mite–trichome contact should be 26–57 per day. Because methyl ketone apparently accumulates in the spider mites on tomato, the rate of mite–trichome contact is probably higher than that. We expect the accumulation of ketones to occur especially on the stems of cultivated tomato, since this is the area most densely occupied with glandular hairs and because here the hairs have higher levels of the methyl ketones.Using dose–response relationships assessed earlier (Chatzivasileiadis and Sabelis, 1997, 1998), we estimated that the number of mite–trichome contacts causing 50% mortality per day is equal to 88 on a tomato stem, whereas it equals 70 for another strain of spider mites collected from cucumber. On wild tomato, L. hirsutum f. glabratum (PI 134417), just one to two contacts would suffice to cause 50% mortality per day. We suggest that methyl ketones from glandular hairs on tomato are an important mortality factor for spider mites on wild tomato and probably also on cultivated tomato.