Reproduction in the spider mite Oligonychus yothersi (Acari: Tetranychidae).

In this work we reported injuries caused by the spider mite Oligonychus yothersi on Vitis vinifera leaves and we also investigate the sex ratio of this species under laboratory conditions. To access the aspects regarding reproduction, females were placed individually in arenas made of V. vinifera leaves to oviposit and all progeny were mounted on slides when they reached adulthood to confirm the offspring sex. Our study showed that O. yothersi reproduces by thelytokous /arrhenotokous parthenogenesis, generating low number of males. Additionally, we found vine plants with leaf browning, particularly on the adaxial surface, where mites were usually found.

Machine learning-based hyperspectral wavelength selection and classification of spider mite-infested cucumber leaves.

Two-spotted spider mite (Tetranychus urticae) is an important greenhouse pest. In cucumbers, heavy infestations lead to the complete loss of leaf assimilation surface, resulting in plant death. Symptoms caused by spider mite feeding alter the light reflection of leaves and could therefore be optically detected. Machine learning methods have already been employed to analyze spectral information in order to differentiate between healthy and spider mite-infested leaves of crops such as tomatoes or cotton. In this study, machine learning methods were applied to cucumbers. Hyperspectral data of leaves were recorded under controlled conditions. Effective wavelengths were identified using three feature selection methods. Subsequently, three supervised machine learning algorithms were used to classify healthy and spider mite-infested leaves. All combinations of feature selection and classification methods yielded accuracy of over 80%, even when using ten or five wavelengths. These results suggest that machine learning methods are a powerful tool for image-based detection of spider mites in cucumbers. In addition, due to the limited number of wavelengths, there is also substantial potential for practical application.

Phylogenetic signals in pest abundance and distribution range of spider mites.

BACKGROUND: Attributes of pest species like host range are frequently reported as being evolutionarily constrained and showing phylogenetic signal. Because these attributes in turn could influence the abundance and impact of species, phylogenetic information could be useful in predicting the likely status of pests. In this study, we used regional (China) and global datasets to investigate phylogenetic patterns in occurrence patterns and host ranges of spider mites, which constitute a pest group of many cropping systems worldwide. RESULTS: We found significant phylogenetic signal in relative abundance and distribution range both at the regional and global scales. Relative abundance and range size of spider mites were positively correlated with host range, although these correlations became weaker after controlling for phylogeny. CONCLUSIONS: The results suggest that pest impacts are evolutionarily constrained. Information that is easily obtainable - including the number of known hosts and phylogenetic position of the mites - could therefore be useful in predicting future pest risk of species.

Geography and major host evolutionary transitions shape the resource use of plant parasites.

The evolution of resource use in herbivores has been conceptualized as an analog of the theory of island biogeography, assuming that plant species are islands separated by phylogenetic distances. Despite its usefulness, this analogy has paradoxically led to neglecting real biogeographical processes in the study of macroevolutionary patterns of herbivore-plant interactions. Here we show that host use is mostly determined by the geographical cooccurrence of hosts and parasites in spider mites (Tetranychidae), a globally distributed group of plant parasites. Strikingly, geography accounts for most of the phylogenetic signal in host use by these parasites. Beyond geography, only evolutionary transitions among major plant lineages (i.e., gymnosperms, commelinids, and eudicots) shape resource use patterns in these herbivores. Still, even these barriers have been repeatedly overcome in evolutionary time, resulting in phylogenetically diverse parasite communities feeding on similar hosts. Therefore, our results imply that patterns of apparent evolutionary conservatism may largely be a byproduct of the geographic cooccurrence of hosts and parasites.

High genetic diversity in a 'recent outbreak' spider mite, Tetranychus pueraricola, in mainland China.

Tetranychus pueraricola is a newly reported spider mite that occurs frequently in mainland China. It is possible that this species was introduced from elsewhere and became a serious pest recently. However, the correct identification of red-pigmented spider mites has repeatedly proven problematic. There is also the possibility that T. pueraricola in China was long misidentified as its sibling species, Tetranychus urticae (red form). To test which of these two scenarios is the more likely, individuals from 14 populations of T. pueraricola and five populations of T. urticae (red form) in China were sampled and genotyped using mitochondrial COI and microsatellite loci. Unlike a recent invasive species, the genetic diversity of T. pueraricola was very high with high mitochondrial genetic diversity (16 haplotypes), high effective alleles (Ne = 2.038 ± 0.081) and expected heterozygosity (He = 0.395 ± 0.016). Surprisingly, we found that all T. urticae (red form) populations shared only one mitochondrial haplotype and showed quite low genetic diversity (Ne = 1.443 ± 0.055; He = 0.234 ± 0.025) which was even lower than that of the green form of T. urticae in mainland China from a previous study. We did not detect significant signals of recent bottlenecks for most populations from both species. These results suggest T. pueraricola is unlikely to be a recent invasive pest but a species that has existed in China for a long time. It is probable that T. pueraricola in China has long been misidentified as T. urticae (red form).

Identification of spider-mite species and their endosymbionts using multiplex PCR.

Spider mites of the genus Tetranychidae are severe crop pests. In the Mediterranean a few species coexist, but they are difficult to identify based on morphological characters. Additionally, spider mites often harbour several species of endosymbiotic bacteria, which may affect the biology of their hosts. Here, we propose novel, cost-effective, multiplex diagnostic methods allowing a quick identification of spider-mite species as well as of the endosymbionts they carry. First, we developed, and successfully multiplexed in a single PCR, primers to identify Tetranychus urticae, T. evansi and T. ludeni, some of the most common tetranychids found in southwest Europe. Moreover, we demonstrated that this method allows detecting multiple species in a single pool, even at low frequencies (up to 1/100), and can be used on entire mites without DNA extraction. Second, we developed another set of primers to detect spider-mite endosymbionts, namely Wolbachia, Cardinium and Rickettsia in a multiplex PCR, along with a generalist spider-mite primer to control for potential failure of DNA amplification in each PCR. Overall, our method represents a simple, cost-effective and reliable method to identify spider-mite species and their symbionts in natural field populations, as well as to detect contaminations in laboratory rearings. This method may easily be extended to other species.

Genotype-specific interactions between parasitic arthropods.

Despite the ubiquity of coinfection, we know little of the effects of intra-specific genetic variability on coinfection by distinct parasite species. Here we test the hypothesis that parasite multiplication depends on the combination of parasite genotypes that coinfect the host (that is Genotype.parasite × Genotype.parasite interaction). To that aim, we infected tomato leaves with the ecto-parasitic mites Tetranychus urticae and Tetranychus evansi. We tested all possible combinations between four T. urticae and two T. evansi populations sampled on different hosts or localities. There was no universal (that is genotype-independent) effect of coinfection on mite multiplication; in many cases the two species had no effect on each other. However, several combinations of T. evansi and T. urticae populations led to elevated T. evansi numbers. Similarly, T. urticae reproduction largely depended on the interaction between T. urticae and T. evansi populations. This evidence for genotype-by-genotype interaction between coinfecting parasites indicates that the effect of coinfection on parasite epidemiology and evolution may vary in space according to the genetic composition of local parasite populations; it further suggests the possibility of coevolution between parasites species that share the same hosts.

Expression characteristics of two novel cytochrome P450 genes involved in fenpropathrin resistance in Tetranychus cinnabarinus (Boisduval).

The carmine spider mite, Tetranychus cinnabarinus, which is also considered as the red form of Tetranychus urticae, is one of the most serious mite pests on crops. It is capable of rapidly developing resistance to acaricides, and has caused difficulty in controlling. However, the resistance mechanism of this mite remains unclear at molecular level. As a member of main detoxification enzymes, cytochrome P450 monooxygenases (CYPs or P450s) play important roles in the development of acaricide resistance in arthropods. In this study, two novel P450 genes (CYP389B1 and CYP392A26) were identified and characterized from T. cinnabarinus. The opening reading frames (ORFs) of CYP389B1 and CYP392A26 contained 1545 and 1488 nucleotides, which encode 514 and 495 amino acids, respectively. Phylogenetic analysis showed that CYP389B1 and CYP392A26 were most closely related to CYP389B1 and CYP392A4 from T. urticae, respectively. When treated with piperonyl butoxide (PBO), an inhibitor of P450s, the resistance ratio of fenpropathrin-resistant (FeR) strain decreased from 101- to 75-fold, which suggested a correlation between P450 and fenpropathrin-resistance in T. cinnabarinus. Furthermore, constitutive over-expressions of CYP389B1 and CYP392A26 were detected in FeR strain. Meanwhile, the expressions of CYP389B1 and CYP392A26 were inducible in FeR strain after treatment in 6, 12 and 24 h with LC30 of fenpropathrin; especially, the expression of CYP392A26 increased to a markedly high level (20.88-fold higher than in the control) after treatment in 6 h. However, there was no significant difference between treatment and control in susceptible strain. Furthermore, stage specific expression profiles of these two genes did not show significant difference among developing stages, except for eggs, in which the mRNA levels were quite low. The results indicate that CYP389B1 and CYP392A26 were involved in the fenpropathrin-resistance in T. cinnabarinus, and the expression of CYP392A26 was more sensitive to fenpropathrin stress. These findings provide clues for further elucidating the function and regulation mechanism of these two cytochrome P450 genes in T. cinnabarinus.

Molecular characterization of vitellogenin and its receptor genes from citrus red mite, Panonychus citri (McGregor).

The production and uptake of yolk protein play an important role in the reproduction of all oviparous organisms. Vitellogenin (Vg) is the precursor of vitellin (Vn), which is the major egg storage protein, and vitellogenin receptor (VgR) is a necessary protein for the uptake of Vg into developing oocytes. In this paper, we characterize the full-length Vg and VgR, PcVg1 and PcVgR, respectively, of the citrus red mite Panonychus citri (McGregor). The PcVg1 cDNA is 5748 nucleotides (nt) with a 5553-nt open reading frame (ORF) coding for 1851 amino acids (aa), and the PcVgR is 6090 nt, containing an intact ORF of 5673 nt coding an expected protein of 1891 aa. The PcVg1 aa sequence shows a typical GLCG domain and several K/RXXR cleavage sites, and PcVgR comprises two ligand-binding domains, two epidermal growth factor (EGF)-like regions containing YWTD motifs, a transmembrane domain, and a cytoplasmic domain. An analysis of the aa sequences and phylogenetics implied that both genes were genetically distinct from those of ticks and insects. The transcriptional profiles determined by real-time quantitative PCR in different developmental stages showed that both genes present the same expressional tendencies in eggs, larvae, nymphs, and adults. This suggested that the biosynthesis and uptake of PcVg occurs coordinately. The strong reproductive capacity of P. citri has been hypothesized as an important factor in its resistance; consequently, understanding the molecular mechanisms regulating Vg and VgR are fundamental for mite control.

Testing for reproductive interference in the population dynamics of two congeneric species of herbivorous mites.

When phylogenetically close, two competing species may reproductively interfere, and thereby affect their population dynamics. We tested for reproductive interference (RI) between two congeneric haplo-diploid spider mites, Tetranychus evansi and Tetranychus urticae, by investigating their interspecific mating and their population dynamics when they competed on the same plants. They are both pests of tomato, but differ in the host plant defences that they suppress or induce. To reduce the effect of plant-mediated interaction, we used a mutant tomato plant lacking jasmonate-mediated anti-herbivore defences in the competition experiment. In addition, to manipulate the effect of RI, we introduced founder females already mated with conspecific males in mild RI treatments or founder, virgin females in strong RI treatments (in either case together with heterospecific and conspecific males). As females show first-male sperm precedence, RI should occur especially in the founder generation under strong RI treatments. We found that T. urticae outcompeted T. evansi in mild, but not in strong RI treatments. Thus, T. evansi interfered reproductively with T. urticae. This result was supported by crossing experiments showing frequent interspecific copulations, strong postmating reproductive isolation and a preference of T. evansi males to mate with T. urticae (instead of conspecific) females, whereas T. urticae males preferred conspecific females. We conclude that interspecific mating comes at a cost due to asymmetric mate preferences of males. Because RI by T. evansi can improve its competitiveness to T. urticae, we propose that RI partly explains why T. evansi became invasive in Europe where T. urticae is endemic.

Population survey of phytoseiid mites and spider mites on peach leaves and wild plants in Japanese peach orchard.

A population survey of phytoseiid mites and spider mites was conducted on peach leaves and wild plants in Japanese peach orchards having different pesticide practices. The phytoseiid mite species composition on peach leaves and wild plants, as estimated using quantitative sequencing, changed during the survey period. Moreover, it varied among study sites. The phytoseiid mite species compositions were similar between peach leaves and some wild plants, such as Veronica persica, Paederia foetida, Persicaria longiseta, and Oxalis corniculata with larger quantities of phytoseiid mites, especially after mid-summer. A PCR-based method to detect the ribosomal ITS sequences of Tetranychus kanzawai and Panonychus mori from phytoseiid mites was developed. Results showed that Euseius sojaensis (specialized pollen feeder/generalist predator) uses both spider mites as prey in the field.

New species and new records of Tetranychidae (Acarina, Prostigmata) from Thailand.

Sampling efforts conducted in several provinces from Thailand disclosed three new species of tetranychid mites. Two of them belong to the genus Tetranychus, namely Tetranychus occultaspina sp. nov. and Tetranychus truncatissimus sp. nov. and the third species belongs to the genus Schizotetranychus, Schizotetranychus krungthepensis sp. nov. They were collected on Ipomoea aquatica, Bambusa multiplex and Saccharum officinarum, respectively. New records and new hosts are also mentioned. 

Diversity of Wolbachia in natural populations of spider mites (genus Tetranychus): evidence for complex infection history and disequilibrium distribution.

Wolbachia are endosymbiotic bacteria that commonly infect arthropods and cause reproductive disorders in host. Within several Tetranychus species, Wolbachia have been detected and shown to affect their reproduction. However, little is known about their transmission and distribution patterns in natural populations of Tetranychus species. Here, we used multilocus sequence typing to confirm Wolbachia infection status and examined the relationship between Wolbachia infection status and host phylogeny, mitochondrial diversity, and geographical range in five Tetranychus species (Tetranychus truncatus, Tetranychus urticae, Tetranychus pueraricola, Tetranychus phaselus, and Tetranychus kanzawai) from 21 populations in China. The prevalence of Wolbachia within the five Tetranychus species ranged from 31.4 to 100 %, and the strains were remarkably diverse. Together, these observations indicate that Wolbachia was introduced to these populations on multiple separate occasions. As in other arthropods, the same Tetranychus species can accommodate very different strains, and identical Wolbachia occasionally infect different species. These observations suggest that Wolbachia are transmitted both vertically and horizontally. Horizontally, transmission is probably mediated by the host plants. The distribution patterns of Wolbachia were quite different among populations of the same species, suggesting that the dynamics of Wolbachia in nature may be affected by ecological and other factors.

DNA-based identification of spider mites: molecular evidence for cryptic species of the genus Tetranychus (Acari: Tetranychidae).

Spider mites are difficult to identify because they are very small and have a limited number of diagnostic characters. Most species of the spider mite genus Tetranychus in Japan are morphologically similar, differing only in the diameter of the aedeagal knob in males. Because this genus contains many important pests, the unambiguous identification of species is crucial for effective pest management and quarantine procedures. DNA-based methods could complement the morphological methods. We examined whether Tetranychus species in Japan could be identified by DNA sequences using the internal transcribed spacer region of nuclear ribosomal DNA and the cytochrome c oxidase subunit I gene of mitochondrial DNA. We determined sequences of the 13 known Tetranychus species in Japan. We could identify 10 of the 13 species in the internal transcribed spacer tree. In the cytochrome c oxidase subunit I tree, we could identify all 13 known Tetranychus species in Japan. Although Tetranychus kanzawai Kishida and T. parakanzawai Ehara were identified by DNA sequences, they were clearly separated into two monophyletic clades each, indicating that a cryptic species existed in each species.

PCR-RFLP analysis for identification of Tetranychus spider mite species (Acari: Tetranychidae).

A polymerase chain reaction (PCR)-restriction fragment length polymorphism (PCR-restriction fragment-length polymorphism)-based method for species identification was applied to 14 Tetranychus spider mite species, which were dominant species intercepted at Japanese import plant quarantine. We sequenced the internal transcribed spacer (ITS) region of the nuclear ribosomal DNA (rDNA), which included the partial ends of the 18S and 28S ribosomal RNA genes, 5.8S ribosomal RNA gene, and two internal transcribed spacers (ITS1 and ITS2) for 15 populations of the 14 species. We analyzed the recognition sites of four restriction endonucleases, which had been proposed for discrimination of Japanese Tetranychus species, and constructed a scheme for Tetranychus species identification by PCR-restriction fragment-length polymorphism. We then applied the scheme to 245 individuals from 199 populations, most of them were from foreign countries. As a result, all 14 species were correctly identified using PCR-restriction fragment-length polymorphism. This demonstrates the usefulness of the PCR-restriction fragment-length polymorphism method for the worldwide identification of Tetranychus species.

A new species of the genus Sonotetranychus (Acari: Tetranychidae) from New Zealand.

The genus Sonotetranychus consisted of seven species prior to this study. Sonotetranychus menziesii sp. nov., collected on Nothofagus menziesii and N. fusca from New Zealand, is described based on females, protonymphs, deutonymphs and a male. An updated key to the species of Sonotetranychus is provided.

Ontogenetic development and redescription of Tribolonychus collyerae (Acari: Tetranychidae).

Tribolonychus collyerae Zhang & Martin is redescribed based on the larva, protonymph, deutonymph and adult male and female specimens from New Zealand. The ontogenetic development of chaetotaxy in Tribolonychus and the number and position of solenidia on tibia I are discussed. Patterns of setal addition on leg III and IV from larva to adult in Trilobonychus are different from the basic pattern of the Tetranychinae and allied genus Neonidulus.

Cysteine peptidases and their inhibitors in Tetranychus urticae: a comparative genomic approach.

BACKGROUND: Cysteine peptidases in the two-spotted spider mite Tetranychus urticae are involved in essential physiological processes, including proteolytic digestion. Cystatins and thyropins are inhibitors of cysteine peptidases that modulate their activity, although their function in this species has yet to be investigated. Comparative genomic analyses are powerful tools to obtain advanced knowledge into the presence and evolution of both, peptidases and their inhibitors, and could aid to elucidate issues concerning the function of these proteins. RESULTS: We have performed a genomic comparative analysis of cysteine peptidases and their inhibitors in T. urticae and representative species of different arthropod taxonomic groups. The results indicate: i) clade-specific proliferations are common to C1A papain-like peptidases and for the I25B cystatin family of inhibitors, whereas the C1A inhibitors thyropins are evolutionarily more conserved among arthropod clades; ii) an unprecedented extensive expansion for C13 legumain-like peptidases is found in T. urticae; iii) a sequence-structure analysis of the spider mite cystatins suggests that diversification may be related to an expansion of their inhibitory range; and iv) an in silico transcriptomic analysis shows that most cathepsin B and L cysteine peptidases, legumains and several members of the cystatin family are expressed at a higher rate in T. urticae feeding stages than in embryos. CONCLUSION: Comparative genomics has provided valuable insights on the spider mite cysteine peptidases and their inhibitors. Mite-specific proliferations of C1A and C13 peptidase and I25 cystatin families and their over-expression in feeding stages of mites fit with a putative role in mite's feeding and could have a key role in its broad host feeding range.

Diversity and recombination in Wolbachia and Cardinium from Bryobia spider mites.

BACKGROUND: Wolbachia and Cardinium are endosymbiotic bacteria infecting many arthropods and manipulating host reproduction. Although these bacteria are maternally transmitted, incongruencies between phylogenies of host and parasite suggest an additional role for occasional horizontal transmission. Consistent with this view is the strong evidence for recombination in Wolbachia, although it is less clear to what extent recombination drives diversification within single host species and genera. Furthermore, little is known concerning the population structures of other insect endosymbionts which co-infect with Wolbachia, such as Cardinium. Here, we explore Wolbachia and Cardinium strain diversity within nine spider mite species (Tetranychidae) from 38 populations, and quantify the contribution of recombination compared to point mutation in generating Wolbachia diversity. RESULTS: We found a high level of genetic diversity for Wolbachia, with 36 unique strains detected (64 investigated mite individuals). Sequence data from four Wolbachia genes suggest that new alleles are 7.5 to 11 times more likely to be generated by recombination than point mutation. Consistent with previous reports on more diverse host samples, our data did not reveal evidence for co-evolution of Wolbachia with its host. Cardinium was less frequently found in the mites, but also showed a high level of diversity, with eight unique strains detected in 15 individuals on the basis of only two genes. A lack of congruence among host and Cardinium phylogenies was observed. CONCLUSIONS: We found a high rate of recombination for Wolbachia strains obtained from host species of the spider mite family Tetranychidae, comparable to rates found for horizontally transmitted bacteria. This suggests frequent horizontal transmission of Wolbachia and/or frequent horizontal transfer of single genes. Our findings strengthens earlier reports of recombination for Wolbachia, and shows that high recombination rates are also present on strains from a restrictive host range. Cardinium was found co-infecting several spider mite species, and phylogenetic comparisons suggest also horizontal transmission of Cardinium among hosts.

Molecular-based identification and phylogeny of Oligonychus species (Acari: Tetranychidae).

The genus Oligonychus has been morphologically divided into two groups based on the direction of curvature of the aedeagus and includes some morphologically similar species that are difficult to distinguish. To develop DNA-based methods for identifying Oligonychus species and to determine the phylogenetic relationships among them, we examined the cytochrome c oxidase subunit I gene of mitochondrial DNA and the internal transcribed spacer and 28S regions of nuclear ribosomal RNA gene for 17 species. Based on the genetic distances (p-distances) of the three DNA regions, the range of intraspecific divergence was found to be below (and not overlap) the range of interspecific divergence, which allowed the 17 species to be discriminated correctly, consistent with their classification based on morphology. Phylogenetic trees constructed by neighbor-joining and Bayesian methods clearly showed two clades, consisting of species whose aedeagi curve ventrally and dorsally, respectively. Three Oligonychus species inhabiting gramineous plants formed clearly defined subclades.