Project description:DNA barcoding technology using short DNA sequences has emerged as an efficient and reliable tool for identifying, confirming, and resolving closely related taxa. This study used ITS2-rDNA and mtCOI DNA sequences to confirm the identity of eight Oligonychus species, representing 68 spider mite samples, collected mainly from Saudi Arabia (SA) and some from Mexico, Pakistan, USA, and Yemen. The intraspecific nucleotide divergences of the studied Oligonychus species ranged from 0% to 1.2% for ITS2 and 0% to 2.9% for COI. However, the interspecific nucleotide divergences were distinctly higher than the intraspecific ones and ranged from 3.7% to 51.1% for ITS2 and 3.2% to 18.1% for COI. Furthermore, molecular data correctly confirmed the species identity of 42 Oligonychus samples lacking males, including a previously claimed sample of O. pratensis from SA. High genetic variations were detected in two Oligonychus species: O. afrasiaticus (McGregor) (nine ITS2 and three COI haplotypes) and O. tylus Baker and Pritchard (four ITS2 and two COI haplotypes). In addition, ITS2- and COI-based phylogenetic trees confirmed the subdivision of the genus Oligonychus. In conclusion, integrative taxonomic approaches are vital to resolve the closely related Oligonychus species, identify the samples lacking male specimens, and assess phylogenetic relationships within and among species.
Project description:The past 15 years have witnessed a number of molecular studies that aimed to resolve issues of species delineation and phylogeny of mites in the family Tetranychidae. The central part of the mitochondrial COI region has frequently been used for investigating intra- and interspecific variation. All these studies combined yield an extensive database of sequence information of the family Tetranychidae. We assembled this information in a single alignment and performed an overall phylogenetic analysis. The resulting phylogeny shows that important patterns have been overlooked in previous studies, whereas others disappear. It also reveals that mistakes were made in submitting the data to GenBank, which further disturbed interpretation of the data. Our total analysis clearly shows three clades that most likely correspond to the species T. urticae, T. kanzawai and T. truncatus. Intraspecific variation is very high, possibly due to selective sweeps caused by reproductive parasites. We found no evidence for host plant associations and phylogeographic patterns in T. urticae are absent. Finally we evaluate the application of DNA barcoding.
Project description:Phylogenetic trees of spider mites were previously obtained using 18S and 28S rRNA genes. Because some of the bootstrap values were relatively low, these trees were unable to completely resolve the phylogeny. Here, we obtained RNA-Seq data for the 72 known species (73 strains) of spider mites to analyze the phylogeny of the sub-family Tetranychinae. The data were de novo assembled into a total alignment length of 790,047 bases corresponding to 264,133 amino acid residues in 652 genes. The sequence dataset was 200 times larger than the data used in the previous study. The new trees were much more robust and more clearly defined the clades of the tribes and the genera of the sub-family Tetranychinae. The tribe Tetranychini was polyphyletic because a monophyletic clade of Eurytetranychini was placed inside it. The six genera from which two or more species were sampled appeared to be monophyletic, but four genera (Schizotetranychus, Eotetranychus, Oligonychus and Tetranychus) appeared to be polyphyletic. These results strongly support the previous molecular inference of the polyphyletic tribes and genera, although the molecular phylogeny of the sub-family Tetranychinae does not fully agree with the current morphology-based taxonomy. The taxonomy of the sub-family Tetranychinae should be revised according to the molecular relationships revealed by this study.
Project description:Tetranychus cinnabarinus (Acari: Tetranychidae) is a worldwide polyphagous agricultural pest that has the title of resistance champion among arthropods. We reported previously the identification of the acaricidal compound β-sitosterol from Mentha piperita and Inula japonica. However, the acaricidal mechanism of β-sitosterol is unclear. Due to the limited genetic research carried out, we de novo assembled the transcriptome of T. cinnabarinus using Illumina sequencing and conducted a differential expression analysis of control and β-sitosterol-treated mites. In total, we obtained >5.4 G high-quality bases for each sample with unprecedented sequencing depth and assembled them into 22,941 unigenes. We identified 617 xenobiotic metabolism-related genes involved in detoxification, binding, and transporting of xenobiotics. A highly expanded xenobiotic metabolic system was found in mites. T. cinnabarinus detoxification genes-including carboxyl/cholinesterase and ABC transporter class C-were upregulated after β-sitosterol treatment. Defense-related proteins, such as Toll-like receptor, legumain, and serine proteases, were also activated. Furthermore, other important genes-such as the chloride channel protein, cytochrome b, carboxypeptidase, peritrophic membrane chitin binding protein, and calphostin-may also play important roles in mites' response to β-sitosterol. Our results demonstrate that high-throughput-omics tool facilitates identification of xenobiotic metabolism-related genes and illustration of the acaricidal mechanisms of β-sitosterol.
Project description:Spider mites of the genus Tetranychus are difficult to identify due to their limited diagnostic characters. Many of them are morphologically similar and males are needed for species-level identification. Tetranychus urticae is a common interception and non-regulated pest at New Zealand's borders, however, most of the intercepted specimens are females and the identification was left at Tetranychus sp. Consequently, the shipments need to be fumigated. DNA sequencing and PCR-restriction fragment length polymorphism (PCR-RFLP) protocols could be used to facilitate the accurate identification. However, in the context of border security practiced in New Zealand, insect identifications are required to be provided within four hours of receiving the samples; thus, those molecular methods are not sufficient to meet this requirement. Therefore, a real-time PCR TaqMan assay was developed for identification of T. urticae by amplification of a 142 bp Internal Transcribed Spacer (ITS) 1 sequence. The developed assay is rapid, detects all life stages of T. urticae within three hours, and does not react with closely related species. Plasmid DNA containing ITS1 sequence of T. uritcae was serially diluted and used as standards in the real-time PCR assay. The quantification cycle (Cq) value of the assay depicted a strong linear relationship with T. urticae DNA content, with a regression coefficient of 0.99 and efficiency of 98%. The detection limit was estimated to be ten copies of the T. urticae target region. The assay was validated against a range of T. urticae specimens from various countries and hosts in a blind panel test. Therefore the application of the assay at New Zealand will reduce the unnecessary fumigation and be beneficial to both the importers and exporters. It is expected that the implementation of this real-time PCR assay would have wide applications in diagnostic and research agencies worldwide.
Project description:Phytophagous mites and endophytic fungi may interact when sharing a host plant, potentially influencing one another's growth or population dynamics; however, interactions between them are poorly known and remain largely unexplored. In this study, quantitative associations between three species of phytophagous mites and the endophytic fungus Epichloë bromicola Leuchtm. & Schardl (Clavicipitaceae, Ascomycotina) on quackgrass, Elymus repens (L.) Gould are reported. The mites' abundance was assessed on field-collected grass shoots that were either exhibiting choke disease symptoms or without the fungus. Overall, the abundance of Tetranychus urticae and Aculodes mckenziei was significantly lower on quackgrass plants infected by E. bromicola compared to plants without the fungus. Conversely, populations of Abacarus hystrix were significantly larger on plants colonised by the fungus than on uninfected plants. Thus, the presence of this endophytic fungus may have divergent effects on different phytophagous mite species although the basis of these effects is not yet known.
Project description:Amblyseius swirskii Athias-Henriot (Acari: Phytoseiidae) is a predatory mite, effective at controlling whiteflies and thrips in protected crops. However, on tomato its efficacy as a biocontrol agent is hindered, most probably by the plant trichomes and their exudates. Our aim was to characterize the response of A. swirskii to the tomato trichome exudates and identify three major detoxification gene sets in this species: cytochromes P450 (CYPs), glutathione S-transferases (GSTs) and carboxyl/cholinesterases (CCEs). Mites were exposed separately to tomato and pepper, a favourable host plant for A. swirskii, after which their transcriptional responses were analysed and compared. The de novo transcriptome assembly resulted in 71,336 unigenes with 66.1% of them annotated. Thirty-nine A. swirskii genes were differentially expressed after transfer on tomato leaves when compared to pepper leaves; some of the expressed genes were associated with the metabolism of tomato exudates. Our results illustrate that the detoxification gene sets CYPs, GSTs and CCEs are abundant in A. swirskii, but do not play a significant role when in contact with the tomato exudates.
Project description:After a comprehensive taxonomic assessment of descriptions/ illustrations of all known (118) species of the spider mite genus Schizotetranychus Trägårdh (Acari: Prostigmata: Tetranychidae), five species groups are proposed based on the number of tactile setae on tibia II in female, i.e., vermiculatus with four setae (four spp.), schizopus with five setae (52 spp.), spireafolia with six setae (10 spp.), asparagi with seven setae (20 spp.), and bambusae with eight setae on tibia II (22 spp.). The species group schizopus is further divided into three species subgroups based on tactile setae on tibia I: schizopus with eight/ nine setae (21 spp.), andropogoni with seven setae (26 spp.), and taquarae with six tactile setae excluding the solenidion on tibia I (five spp.). Eight Schizotetranychus species were not assigned to any species group because of brief descriptions and/ or illustration and without information on the number of tactile setae on tibiae I and II. Moreover, two Schizotetranychus species, S.gausus Baker & Pritchard and S.luculentus Tseng that have six setae/ structures including a spinneret and a solenidion on the palp tarsus, are provisionally transferred to the genus Stigmaeopsis Banks. Finally, keys to species groups and subgroups of the world species of Schizotetranychus are provided.
Project description:A comprehensive taxonomic assessment of the most agriculturally important and highly diverse spider mite genus, Oligonychus Berlese (Acari: Tetranychidae) was performed. The sub-generic division, species groups, doubtful species, species complexes and the interpretation of a key generic character are discussed. Based on the orientation of the male aedeagus, only two subgenera, namely Oligonychus Berlese (aedeagus downturned) and Reckiella Tuttle & Baker (aedeagus upturned), are valid in the genus Oligonychus. The subgenera Homonychus Wainstein, Metatetranychoides Wainstein, and Wainsteiniella Tuttle & Baker are considered to be synonyms of the subgenus Oligonychus, whereas the subgenus Pritchardinychus Wainstein is proposed as a synonym of the subgenus Reckiella. Moreover, based on female morphological characters, four species groups (coffeae, exsiccator, iseilemae, and peruvianus) and 11 species subgroups (aceris, biharensis, coffeae, comptus, exsiccator, gossypii, iseilemae, peruvianus, pritchardi, smithi, and subnudus) are suggested in the subgenera Oligonychus and Reckiella. Fourteen Oligonychus species are proposed as species inquirendae, and potential cryptic species complexes in the genus Oligonychus are briefly highlighted. It is agreed that the clunal seta h1 is always absent, while the para-anal setae h2 and h3 are always present in the genus Oligonychus. A key to subgenera, species groups, and species subgroups of the genus Oligonychus is provided.
Project description:Oligonychus Berlese (Acari, Prostigmata, Tetranychidae) is an agriculturally important and the largest genus of spider mites, comprised of 211 species (including new species), two subgenera, four species groups, and 11 species subgroups. The present study comprehensively addressed the morphotaxonomic-based identification of world species of the subgenus Reckiella Tuttle and Baker. Five diagnostic keys were developed for identifying Oligonychus (Reckiella) species belonging to five species subgroups: iseilemae, pritchardi, biharensis, gossypii, and exsiccator. Taxonomic notes are provided on intraspecific variations and some closely related Oligonychus (Reckiella) species representing six species complexes, viz. the afrasiaticus complex, the litchii complex, the pratensis complex, the plegas complex, the sacchari complex, and the tylus complex. One new spider mite species, Oligonychus bahaensis sp. nov., is described and illustrated from Grasses (Poaceae) under the subgenus Reckiella.