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.

Drivers for mutation in amino acid sequences of two mitochondrial proteins (Cytb and COI) in Phytoseiidae mites (Acari: Mesostigmata).

Mutations in amino acid sequences can affect protein function. Such aspects have been poorly studied for arthropods. As recent studies have shown mutations in cytochrome b (Cytb) associated with geographic locations in several Phytoseiidae species, the present study aims at investigating (i) the mutation pattern in additional species for the Cytb fragment, (ii) the mutation pattern for another mitochondrial amino acid sequence, cytochrome c oxidase subunit 1 (COI), and (iii) factors affecting the mutations observed (taxonomy, plant support, climatic variables, wild vs. commercialised species). Mutations in amino acid sequences were assessed in seven Phytoseiidae species, with populations collected in contrasted environments. The DNA sequences were mainly obtained from published studies and some were newly obtained. Mutations were observed within and between the populations considered for both fragments, with higher mutation rates in Cytb than in COI sequences, confirming the robustness of this former fragment. Plant support and taxonomic position were not related to mutation patterns. A lower number of mutations was observed in commercialised populations than in wild ones. As preliminary tendencies, mutations in Cytb and COI sequences seem associated to temperature and moisture. Such a preliminary approach, attempting to relate mutation to functional adaptations, clearly opens new research tracks for better assessment of the drivers of mite adaptation, in a context of climate change.

Molecular variation of the cytochrome b DNA and protein sequences in Phytoseiulus macropilis and P. persimilis (Acari: Phytoseiidae) reflect population differentiation.

Several phytoseiid mite species are important natural enemies used in biological control strategies. In the present study, Cytb mtDNA sequences of various populations of two species, Phytoseiulus macropolis and P. persimilis, were compared to determine whether the specimens collected in Brazil could belong to P. persimilis as this latter species is reported in South America but not in Brazil. The Cytb marker was used because of its high evolution rate, assumed to capture intraspecific variation. No overlap between intra- and interspecific distances was observed but the distances were quite low for interspecific variation. This can be due to the particular biology of Phytoseiulus species and this shows the difficulty to apply a universal threshold in genetic distances to conclude about the existence of one or several species. Cytb mtDNA sequences were also considered to assess intraspecific variation. The DNA sequences of P. persimilis populations were very similar, probably because they all originated from the West Palearctic region or because of a prevalence of commercialized specimens in natura. For P. macropilis, higher genetic distances were observed and differentiation was noted according to geographic location and, to a smaller extent, pyrethroid resistance. To determine how DNA variation might impact the protein function (CytB fragment considered), the amino acid compositions of the populations studied were compared. No diagnostic mutation was observed between pyrethroid resistant and susceptible populations, whereas four mutations were identified between populations of P. macropilis separated by 1300 km (different climatic conditions). The impact of such mutations is discussed but knowledge is scarce, which makes it difficult to root testable hypotheses. The protein analysis clearly opens new perspectives in Phytoseiidae studies.

Molecular characterization of phytoseiid mites in Turkey based on the internal transcribed spacer (ITS) region, with a new record for the country.

The family Phytoseiidae contains many predatory mite species and some are used in biological control programs worldwide. The identification of phytoseiid mites is based on tiny morphological structures and sometimes species diagnosis is not easy especially for non-taxonomists. DNA-based approaches may offer a fast and accurate diagnosis to overcome these difficulties, nevertheless more DNA sequences are needed to determine intra- and inter-specific variations and to provide accurate decision rules based on genetic distances between the taxa considered. In this study, we provide the molecular characterization of seven phytoseiid species based on the internal transcribed spacer (ITS) region. Several populations of these species collected in Turkey were considered. A phylogenetic tree was also constructed. Finally, we record the presence of Neoseiulus reductus (Wainstein) in Turkey.

Mites of the genus Typhlodromus (Acari: Phytoseiidae) from Southern France: combined morphological and molecular approaches for species identification.

Mites of the family Phytoseiidae are important predators for biological control applications. They occur naturally in ecosystems but their overall distribution is not completely known. This study presents results of surveys carried out in the south of France. It proposes the use of a combination of morphological and molecular approaches for species diagnosis. Eighteen species of the genus Typhlodromus are reported from southern France, of which nine belong to Typhlodromus (Anthoseius) and nine to Typhlodromus (Typhlodromus). Eight of these species are new to the French fauna. The mitochondrial DNA CytB gene from 85 specimens (18 species) and the 12S rRNA gene from 30 specimens (9 species) were partially sequenced and analysed. Based on molecular and morphological comparisons, the synonymy between Typhlodromus (Anthoseius) ilicis and T. (A.) creticus is discussed. High genetic distances between specimens morphologically assigned to T. (A.) rhenanoides suggests the existence of cryptic species. The reliability of integrative approaches for species identification is discussed.

Molecular and morphological characterization of the predatory mite Amblyseius largoensis (Acari: Phytoseiidae): surprising similarity between an Asian and American populations.

The accurate characterization of biological control agents is a key step in control programs. Recently, Amblyseius largoensis from Thailand were introduced in Brazil to evaluate their efficiency for the control of the red palm mite, Raoiella indica. The aim of this study was to confirm their identification and to characterize the population from Thailand, comparing it to populations of the Americas and Indian Ocean islands. In addition, a population of A. largoensis from New Caledonia, Oceania, of which DNA sequences were available, was included in phylogenetic analyses. Morphometric data obtained for the population of A. largoensis from Thailand were compared to those of populations from Reunion Island and the Americas through univariate and multivariate analyses. Two DNA fragments were amplified and sequenced: the nuclear ribosomal region ITSS and the mitochondrial 12S rRNA. Haplotypes (12S rRNA) and genotypes (ITSS) were identified and phylogenetic analyses using both fragments were conducted separately and combined using maximum likelihood and the Bayesian information criterion. The integrative approach reveals morphometric and molecular variabilities among populations of A. largoensis and shows that the population identified as A. largoensis collected in Thailand, as well as that from New Caledonia, are conspecific to the populations of the Americas and Indian Ocean islands. Populations from the Americas and Asia are more related to each other than with that from the Indian Ocean islands. Hypotheses to explain this clustering are proposed. Data on the molecular intraspecific variability of this predatory mite from remote areas will be helpful for the development of molecular diagnosis.

Which molecular markers for assessing which taxonomic level? The case study of the mite family Phytoseiidae (Acari: Mesostigmata).

The use of molecular markers for resolving systematics issues has improved our knowledge of life history. However, for the taxa studied herein-the predatory mite family Phytoseiidae-molecular phylogeny is impeded by a lack of suitable markers for deeper taxonomic levels. This study aims (i) to establish DNA amplification protocols for molecular markers known to resolve supraspecific nodes in other taxa, (ii) to determine their individual performance in assessing the clustering of species, genera, tribes and subfamilies, and (iii) to characterize the additional information provided when markers are concatenated. A new phylogenetic index is proposed based on ecological concepts, considering trees as a community of nodes. New and efficient protocols for DNA amplification of six molecular markers are provided. The concatenated tree globally provides more robust and reliable information, especially for deeper nodes. However, for assessing species identification and within-genera phylogenies, the combined use of six markers does not seem necessary, underlining the need to resize experiments depending on their taxonomic objectives. Finally, this study lays the methodological foundations with which to test the present Phytoseiidae classification as the first phylogeny obtained shows incongruence with the present morphological classification.

Phytoseiidae (Acari: Mesostigmata) of Morocco: new records, descriptions of five new species, re-descriptions of two species, and key for identification.

The family Phytoseiidae includes more than 2,300 species distributed all over the world. However despite the huge numbers of faunistic surveys carried out for more than 60 years, the fauna of some countries and particular ecosystems remain little explored. This paper reports results of surveys carried out in various regions of Morocco (from Sahara to Atlantic and Mediteranean coasts) in 2002 and 2003. A total of 43 species was found. Among them 19 are new for the Moroccan fauna and five are new to science. This paper provides the descriptions of these five new species, Neoseiulus thymeleae, Transeius audeae, Typhlodromus (Typhlodromus) ballotae, T. (T.) leclanti, T. (T.) mazarii, and re-descriptions of two species (Typhlodromus (T.) setubali and Typhlodromus (Anthoseius) clairathiasae. A key to females of the 52 species now known from Morocco is given.

Is Neoseiulus wearnei the Neoseiulus californicus of Australia?

Species of the family Phytoseiidae are known as predatory mites, some of them being used in crops to control mite pests, all around the world. Neoseiulus (=Cydnodromus) californicus is among the most commonly used Phytoseiidae species in biological control programs, especially in vineyards, orchards and vegetable fields. This species is distributed world-wide but has never been reported from Australia. On the other hand, specimens morphologically close to N. californicus have been assigned to a species called Neoseiulus wearnei, only reported from Australia. Investigations based on morphological and molecular comparisons were carried out to investigate whether these two taxa are conspecific. Morphological analyses showed no significant difference between specimens identified as N. wearnei and N. californicus. Similarly, genetic distances between these taxa were null, showing that all these specimens belong to the same species. Although it is not yet possible to conclude that all the specimens identified as N. wearnei are N. californicus, we can conclude that N. californicus is present in Australia. The information about the biology of N. californicus can thus now be applied to the Australian population of this species for biological control purposes.

Phytoseiidae in European grape (Vitis vinifera L.): bio-ecological aspects and keys to species (Acari: Mesostigmata).

The family Phytoseiidae includes many species of predators involved in the control of mite pests of crops all over the world. In European vineyards, these natural enemies play a key role in plant protection as their presence usually makes the use of acaricides unnecessary. Each species has its specific biological features. It is thus of great interest to identify the species reported on grape, Vitis vinifera. The present paper, based on a world database of the family Phytoseiidae and on an analysis of more than 250 publications, presents the 54 species of Phytoseiidae belonging to 15 genera reported on V. vinifera in Europe, and identification keys to species. Online versions of the key (dichotomous and polytomous) with illustrations are available at http://wwwl.montpellier.inra.fr/CBGP/phytoseiidae/sitewebvineyards2/index.htm. An analysis of the biogeographic distribution of these species and their prey has also been carried out. Most species reported on V. vinifera in Europe are rare; only five species are frequently observed: Kampimodromus aberrans, Typhlodromus (Typhlodromus) pyri, Typhlodromus (Typhlodromus) exhilaratus, Euseius finlandicus and Phytoseius finitimus. The 12 countries where Phytoseiidae have been reported from grapes have been unevenly surveyed, the most well-known faunas being from Italy, Greece and France. These species are reported to prey upon the main species of mite pests of grapes.

Statistical approaches to assess intraspecific variations of morphological continuous characters: the case study of the family Phytoseiidae (Acari: Mesostigmata).

Statistical methods for delimiting species with morphological data remain woefully understudied. The present study presents statistical approaches to study the intraspecific variations of continuous characters used in species diagnosis. For this, the data set comprised 14 species of the mite family Phytoseiidae (Acari: Mesostigmata) and 23 continuous characters (seta lengths). The statistical approaches aim to determine (i) how intraspecific variations differ depending on the characters and species considered, and (ii) how to choose the number of specimens to be considered for characterizing intraspecific boundaries. The means of the related confident limits at 95% (rCL95 ) of the 14 species and the 23 setae are around 20%, suggesting that 95% of the specimens of a same species are included in the interval µ ± 0.2µ. No relationship was found between seta variations and the number of specimens considered for each species, nor for seta variations and their position on the mite body. However, a correlation between seta lengths and rCL95 means was observed. An abacus was proposed to determine the coefficient that must be applied to mean seta length to calculate the lower and upper limits of the interval that will include 95% of the specimens of a same species. The higher the precision around the mean, the higher the number of specimens considered. Furthermore, this number varies considerably depending on the characters and species considered for high precisions. However, for a priori error around the mean of 15%, the number of specimens needed to characterize the intraspecific variation is quite homogeneous and ten specimens would be required. I discuss the utility of such results to resolve some persistent problems in the alpha-taxonomy of the family Phytoseiidae but also of other taxa. © The Willi Hennig Society 2012.

Agroforestry management and phytoseiid communities in vineyards in the South of France.

This study deals with the long-term effect of agroforestry management (trees within vine crops) on communities of phytoseiid mites. Several plots were considered: vineyards co-planted with Sorbus domestica or Pinus pinea, monocultures of vines and monocultures of S. domestica or P. pinea. All vine plots included two vine cultivars, Syrah and Grenache. Phytoseiid mites have been surveyed in these plots during several years within the previous 10 years. In 2010, samplings were again carried out in these same plots, from May to September, twice a month. Significantly higher densities of Phytoseiidae were observed on the cultivar Syrah (0.85 phytoseiids per leaf) than on Grenache (0.26 phytoseiids per leaf). Furthermore, significantly higher phytoseiid mite densities were observed in the monocultural grapevine plot than in the two co-planted ones. The main species found was Typhlodromus (Typhlodromus) exhilaratus in all vine plots considered. However, Kampimodromus aberrans was observed in the grapevine plots co-planted with the two trees, but never in the monocultural vine plot. Surprisingly, this phytoseiid species was not found on the co-planted trees, nor in the neighbouring uncultivated vegetation. Several hypotheses are discussed to explain such an unexpected distribution. Furthermore, contrary to what has been observed previously, agroforestry management did not seem to favour phytoseiid mite development, especially on the Grenache cultivar. Again, some hypotheses are developed to explain such observations and density modifications.

Morphological and molecular diagnostics of Phytoseiulus persimilis and Phytoseiulus macropilis (Acari: Phytoseiidae).

This study focuses on the diagnostics of two natural enemy species, belonging to the genus Phytoseiulus in the family Phytoseiidae (sub-family Amblyseiinae): P. macropilis and P. persimilis. These two species are of primary importance in biological control all over the world. However, they are morphologically very similar and specific diagnostics is difficult. This study utilizes mitochondrial molecular markers (12S rRNA and Cytb mtDNA) to differentiate these two species. Morphological analyses showed significant differences between P. persimilis and P. macropilis for 17 morphological characters of the 32 considered. However, despite these significant differences, the ranges of all characters overlap. Only the serration of the macroseta on the basitarsus (StIV) allows the differentiation between P. persimilis and P. macropilis. Despite these small morphological differences, molecular results, for both mitochondrial DNA fragments considered (rRNA and Cytb mtDNA), showed a clear delineation between the specimens of P. macropilis and P. persimilis. This study emphasizes (i) that only one morphological character (serration of the seta StIV) clearly separates these two species, and (ii) the usefulness of an automatical molecular and simple diagnostic tool for accurate differentiation of the two species and ensure the morphological diagnostics. Further studies are proposed, including more DNA sequences especially for P. macropilis.

Influence of ground cover management on diversity and density of phytoseiid mites (Acari: Phytoseiidae) in Guadeloupean citrus orchards.

The abundance and diversity of phytoseiid mites in the vegetal ground cover of a citrus orchard were surveyed by monthly samplings from October 2008 to July 2009. Six different vegetal cover management methods were studied: herbicide application without mowing (Gly), mowing without herbicide application (PV), mowing with herbicide application (AV), late mowing without herbicide application (LMV), cover crop (Neonotonia wightii, Fabaceae) without herbicide application (PNeo) and cover crop with herbicide application (ANeo). Eleven species were present in the ground cover, with Phytoseius rex and Proprioseiopsis mexicanus as major species. Species richness and densities (1.5 ± 4.5) in the Gly treatment were very low, except for one sample 4 months after herbicide treatment. The AV and PV treatments showed poor diversity and very low mite densities (1.2 ± 2.6 and 1.4 ± 1.5, respectively). The LMV treatment showed the highest diversity and high density of phytoseiid mites (6.9 ± 8.8). The ANeo and PNeo treatments also showed generally high diversity, but with variations in time and exhibited the highest density of phytoseiid mites (13.5 ± 12.7 and 13.4 ± 9.1, respectively). Neonotonia wightii as the cover crop seems to act as a reservoir of phytoseiid mites, sustaining abundant and diverse populations all year round. Some naturally occurring plant species such as Achyranthes aspera, Amaranthus dubius and Eleutheranthera ruderalis could also constitute favourable host plants for Phytoseiidae. Results are discussed in relation to the potential of collected phytoseiid species as candidates for biological control of phytophagous mites on Guadeloupean citrus orchards.

Assessment of pesticide susceptibility for Typhlodromus exhilaratus and Typhlodromus phialatus strains (Acari: Phytoseiidae) from vineyards in the south of France.

Two species of Phytoseiidae predominated in and around vineyards in an experimental site in the south of France: Typhlodromus exhilaratus is the dominant species on grapes, whereas T. phialatus occurs in the uncultivated areas surrounding these crops. To help understand their distribution, this study aims to determine their susceptibility to a fungicide (dimethomorph + mancozeb) and an insecticide (chlorpyriphos-ethyl), which have been widely used at the experimental site for at least four years. The hypothesis tested is that the effects of these two pesticides differ between T. exhilaratus and T. phialatus. Two populations of T. exhilaratus, one collected on a vine crop where the two pesticides were applied and the other in an unsprayed orchard of Sorbus domestica, and one population of T. phialatus collected on Viburnum tinus from uncultivated areas adjacent to the vine crop, were tested under laboratory conditions. The fungicide seems to affect the females of T. phialatus more than those of T. exhilaratus from vine. However, at the recommended field concentration the female mortality rates of the three strains were low and female fecundities were not significantly different. Conversely, mortality rates were very high for the three strains at the recommended field concentration of chlorpyrifos-ethyl. The strain of T. exhilaratus from vine seems to be more tolerant and 20% of females survived the recommended field rate. This suggests better survival after insecticide application than females of T. phialatus (100% mortality at a lower concentration than the recommended rate). As extrapolation of laboratory results to the field is often irrelevant, these differences could be just one of several factors affecting the spatial segregation of the two species in the agrosystem considered. Possible explanations other than pesticides for the poor settlement of T. phialatus in vine plots are discussed.

Life tables of the predatory mite Phytoseiulus longipes feeding on Tetranychus evansi at four temperatures (Acari: Phytoseiidae, Tetranychidae).

The tomato red spider mite, Tetranychus evansi, is reported as a severe pest of tomato and other solanaceous crops from Africa, from Atlantic and Mediterranean Islands, and more recently from the south of Europe (Portugal, Spain and France). A population of the predaceous mite Phytoseiulus longipes has been recently found in Brazil in association with T. evansi. The objective of this paper was to assess the development and reproduction abilities of this strain on T. evansi under laboratory conditions at four temperatures: 15, 20, 25 and 30 degrees C. The duration of the immature phase ranged from 3.1 to 15.4 days, at 30 and 15 degrees C, respectively. Global immature lower thermal threshold was 12.0 degrees C. Immature survival was high at all temperatures tested (minimum of 88% at 30 degrees C). The intrinsic rate of increase (r (m)) of P. longipes ranged from 0.091 to 0.416 female/female/day, at 15 and 30 degrees C, respectively. P. longipes would be able to develop at a wide range of temperatures feeding on T. evansi and has the potential to control T. evansi populations.

Cannibalism and intraguild predation in Typhlodromus exhilaratus and T. phialatus (Acari: Phytoseiidae) under laboratory conditions.

Two species of Phytoseiidae are found in the same agroecosystem: Typhlodromus exhilaratus prevails in vine plots, while T. phialatus prevails in uncultivated surrounding areas. The objective of the present paper was to investigate whether the poor settlement of T. phialatus in vine plots can be explained by intraguild predation of these two species and/or cannibalism. Predatory abilities of the females on larvae and protonymphs were studied under laboratory conditions. A first experiment was conducted with only conspecific or heterospecific phytoseiid prey, in a second experiment Tetranychus urticae eggs were added to the phytoseiid prey. Oviposition, prey consumption, and escape rates of females were recorded. Oviposition and intraguild predation rates were higher for T. exhilaratus than for T. phialatus. Typhlodromus exhilaratus consumed fewer conspecifics than heterospecific phytoseiids, and oviposited when feeding on both diets. Typhlodromus phialatus consumed equal amounts of con- and heterospecifics. Although these two generalist predators belong to the type III defined by McMurtry and Croft (Annual Review of Entomology 42:291-321, 1997), our results suggest that they have different predation behaviour. However, because these results were obtained in experiments where no choice was given between the two phytoseiid species, they are difficult to link to previous studies conducted on the intraguild predation of the Phytoseiidae. The greater voracity and prolificacy of T. exhilaratus could partially explain the poor settlement of T. phialatus in vineyards and the predominance of T. exhilaratus. However, a full understanding of this phenomenon will require the study of other factors, such as susceptibility to pesticides and micro-climatic conditions, as well as the ability to cope with different food sources and host plants.

Immigration of phytoseiid mites from surrounding uncultivated areas into a newly planted vineyard.

This study reports (1) a faunistic survey of phytoseiid mites observed inside a vine plot and in neighbouring vegetation (other vine plots and uncultivated areas) and (2) dispersal of phytoseiid mites into the plot studied. These data aim to raise some hypotheses concerning natural colonisation of a vineyard by predatory mites. The study was carried out over 3 years (1999, 2000 and 2001) in an experimental plot planted with two cultivars (Grenache and Syrah) and with Sorbus domestica in a framework of agroforestry investigations. Phytoseiid mites were collected in both cultivated and uncultivated areas surrounding the experimental plot, and their dispersal into the plot studied using "aerial" traps. Densities remained quite low compared to previous studies. The main species encountered in the uncultivated areas and in the traps was Typhlodromus phialatus. Despite the low numbers of phytoseiid mites trapped, densities of phytoseiid mites into the vine field increased during 3 years. Typhlodromus phialatus, the species mainly found in the neighbouring uncultivated areas, was rarely found in vineyards. Another morphologically close species was predominant on vines: Typhlodromus exhilaratus. However, Kampimodromus aberrans the main phytoseiid mite species in vineyards of Southern France was not found in the present survey. Hypotheses for this colonisation process are discussed.

Effects of agroforestry on phytoseiid mite communities (Acari: Phytoseiidae) in vineyards in the South of France.

The abundance and diversity of phytoseiid mites were surveyed from April to September 2003 to 2005 in vineyards (Grenache and Syrah cultivars) co-planted with rows of Sorbus domestica or Pinus pinea and in monoculture plots of grapes in the South of France. Densities of phytoseiid mites were different on the two tree species, with P. pinea a more suitable host than S. domestica. Typhlodromus (Typhlodromus) exhilaratus was the dominant species occurring on grapes and on co-planted rows of S. domestica and P. pinea, whereas T. (T.) phialatus was the most abundant species in monoculture plots of both S. domestica and P. pinea. Factors determining the dominance of T. (T.) phialatus over T. (T.) exhilaratus in monoculture trees are discussed. In this study, agroforestry management did not affect phytoseiid diversity in vineyards, but did affect phytoseiid density, especially in 2005. The results obtained in 2003 and 2004 are not easy to discuss in this regard because of the low densities of mites observed during these 2 years (very dry climatic conditions and pesticide applications).