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Article

Molecular-Based Taxonomic Inferences of Some Spider Mite Species of the Genus Oligonychus Berlese (Acari, Prostigmata, Tetranychidae)

by
Hafiz Muhammad Saqib Mushtaq
1,
Amgad A. Saleh
2,
Muhammad Kamran
1 and
Fahad Jaber Alatawi
1,*
1
Acarology Research Laboratory, Department of Plant Protection, College of Food and Agriculture Sciences, King Saud University, P.O. Box 2460, Riyadh 11451, Saudi Arabia
2
Plant Pathology Laboratory, Department of Plant Protection, College of Food and Agriculture Sciences, King Saud University, P.O. Box 2460, Riyadh 11451, Saudi Arabia
*
Author to whom correspondence should be addressed.
Insects 2023, 14(2), 192; https://doi.org/10.3390/insects14020192
Submission received: 11 January 2023 / Revised: 11 February 2023 / Accepted: 12 February 2023 / Published: 15 February 2023
(This article belongs to the Special Issue Mites: Systematics, Ecology, and Evolution)
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Abstract

:

Simple Summary

Spider mites belonging to the genus Oligonychus Berlese are serious pests of various fruits, ornamentals, agronomic crops, and trees. To effectively manage these pests, their accurate identification is crucial. Morphological-based identification of Oligonychus species is very challenging. The present study used molecular data to identify/confirm the species identity of some Oligonychus species, including various samples lacking male specimens. Moreover, phylogenetic analyses validated the morphological-based subdivision of the genus Oligonychus. The integrative taxonomic approaches are vital for accurately identifying closely related Oligonychus species.

Abstract

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.

1. Introduction

In the genus Oligonychus Berlese (Acari, Prostigmata, Tetranychidae), morphological-based differentiation among species always depends on the characterization of the key trait of male aedeagus [1,2,3]; however, in the subdivision of Oligonychus into subgenera and groups/subgroups, the characterization of both male and female is compulsory [3]. Indeed, exact species identification in Oligonychus is usually a challenge because of the minute differences in aedeagus; limited diagnostic traits in females; and the presence of various species complexes, e.g., pratensis complex and punicae complex [1,2,3,4,5,6,7]. Moreover, the key diagnostic trait of the aedeagus becomes unreliable if males are not mounted accurately in their lateral positions, or are either described briefly without illustrations or with insufficient morphological details [8,9]. Furthermore, the identity of some Oligonychus species remains questionable when these species were described based solely on females, and males were absent in the original and subsequent descriptions [10,11]. Therefore, these questionable Oligonychus species are considered species inquirendae [3].
Although morphological-based species identification is the most popular method used by biologists, especially taxonomists, it needs to be fortified by other methods, e.g., DNA-based ones, to be able to delimit species boundaries of closely related species. DNA-based methods include DNA sequences, e.g., nuclear and mitochondrial genes. The mitochondrial cytochrome c oxidase subunit I (COI) gene has been used extensively in identifying organisms, including mites [7,12,13,14,15]. The internal transcribed spacers (ITS1 and ITS2) regions of nuclear ribosomal DNA have also been applied accurately for the identification or confirmation of closely related species in the family Tetranychidae Donnadieu [7,13,14,16,17]. The combined use of morphological- and molecular-based methods for species delineation would greatly help resolve problems of synonymy and the misidentification of closely related tetranychid species [18,19,20]. Recently, an integrative taxonomic study has successfully resolved the long-standing issue of the punicae species complex in the genus Oligonychus [7].
Globally, 212 Oligonychus species have been reported so far [3,21,22]. In Saudi Arabia (SA), previous morphotaxonomic studies have revealed the presence of seven Oligonychus species, viz. O. afrasiaticus (McGregor), O. coniferarum (McGregor), O. dactyloni (Smiley and Baker), O. punicae (Hirst), O. pratensis (Banks), O. tylus (Baker and Pritchard), and O. washingtoniae (Mushtaq, Kamran, and Alatawi) [21,23]. However, the presence of the banks grass mite O. pratensis in SA could be doubtful due to the involvement of the pratensis species complex [1,21].
The present study aimed to apply the DNA-based methods to (1) confirm the morphologically identified Oligonychus species previously reported in SA, (2) identify mite samples that lack males, (3) investigate the intra- and inter-specific genetic variations of some widely distributed Oligonychus species, and (4) validate the subdivision of the genus Oligonychus into two subgenera.

2. Materials and Methods

2.1. Collection, Preservation, and Processing of Spider Mite Samples

In total, 68 spider mite samples were collected from wild and cultivated vegetation in different seasons and localities from Mexico, Pakistan, SA, USA, and Yemen (Table S1; Figure 1). Out of 68 samples, 61 were collected from 10 Saudi provinces: Asir, Baha, Eastern Province, Jouf, Jizan, Makkah, Madinah, Qassim, Riyadh, and Tabuk (Table S1; Figure 2). The Saudi mite samples were collected between 2018 and 2021, except for one sample (sample voucher number/SVN: 39; Table S1), which was collected in 2013 from SA, and was claimed as O. pratensis [23]. The remaining 7 Oligonychus samples were received from 4 other countries: 1 from Mexico (SVN: 53), 1 from USA (SVN: 40), 2 from Yemen (SVN: 49 and 50), and 3 from Pakistan (SVN: 51, 66, and 67) (Table S1; Figure 1). Almost 2/3 of the mite samples had only female specimens (Table S1). Each sample’s collection details were recorded, e.g., sample voucher number (SVN), collection date, locality, host plant, GPS coordinates, collector name, etc. (Table S1). The voucher specimens of male/female representing each collected sample were deposited at the King Saud University Museum of Arthropods (Acarology section), Department of Plant Protection, College of Food and Agriculture Sciences, King Saud University (KSU), Riyadh, SA.

2.2. Mites Identification Based on Morphological Characters

Mite samples of O. coniferarum, O. dactyloni, O. punicae, O. pratensis, O. tylus, and O. washingtoniae were collected from SA and morphologically identified by Alatawi and Kamran [23]; Mushtaq, Kamran, and Alatawi [21]; and Mushtaq, Kamran, Saleh, and Alatawi [7]. The samples received from Mexico (SVN: 53, O. perseae Tuttle et al.), USA (SVN: 40, O. pratensis), and Yemen (SVN: 49 and 50, O. afrasiaticus) were initially identified/labelled and sent to us by our colleagues [24,25] at our request. Mite samples having male and female individuals were morphologically re-examined at the acarology laboratory (KSU) using the phase contrast microscope (BX51, Olympus, Tokyo, Japan) and identified till the species level following the taxonomic literature of the genus Oligonychus [3,26,27]. However, those 42 samples containing only female specimens were only identified up to the level of the genus Oligonychus, following Bolland, Gutierrez, and Flechtmann [26].

2.3. Molecular Analysis

2.3.1. DNA Extraction and Amplification of ITS2/COI Regions

DNA extraction, from mite samples preserved in 99% ethanol, was carried out during the period from 2020 to 2021. The DNeasy mini kit (Qiagen®, Hilden, Germany) was used for DNA extraction from single adult mite females.
The concentration of genomic DNA solutions was assessed by the NanoDropTM One spectrophotometer (Thermofisher Scientific, Waltham, MA, USA). The extracted DNA samples were immediately stored at −20 °C after labelling with the appropriate field information.
The ITS2-rDNA region was amplified from mite samples using PCR primers, ITS2-forward (5′-GTCACATCTGTCTGAGAGTTGAGA-3′) and ITS2-reverse (5′-GTARCCTCACCTRMTCTGAGATC-3′) [16]. In addition, COI-forward primer (5′-TGATTTTTTGGTCACCCAGAAG-3′) and COI-reverse primer (5′-TACAGCTC CTATAGATA AAAC-3′) were also used to amplify the mtCOI region [12] (Table S1). The PCR reaction was performed with a total volume of 30 μL reaction containing 0.4 μL of each 10 μM primer, 15 μL 2× master mix (Molequle-On®, Auckland, New Zealand), ca 20 ng DNA, and appropriate volume of nuclease free water (Promega®, Madison, WI, USA). The PCR cycle conditions were as follows: (i) an initial denaturation cycle at 94 °C for 5 min; followed by (ii) 35 cycles of a denaturation step at 94 °C for 60 s, an annealing step for 90 s at 52 °C for ITS2, and 53 °C for COI and an extension step for 60 s at 72 °C; and (iii) a final extension step for 10 min at 72 °C. The obtained PCR products were run on 1.2% agarose gels in 1× TAE buffer. Gels were stained with acridine orange, observed, and picturized using the gel documentation BioDoc Analyze system (Uvitec, Cambridge, UK).

2.3.2. DNA Sequencing and Analysis

The ITS2 and COI PCR products were purified and sequenced using the same primers at the Macrogen sequencing facility (Macrogen Inc., Seoul, Republic of Korea). The obtained sequences were cleaned and analyzed using BioEdit software [28]. The cleaned sequences were searched using BLASTn against the NCBI GenBank database. The homologous (sequence similarity within and among species were ˃98% and ˃85%, respectively) and closely related ITS2/COI sequences of all available Oligonychus species were retrieved from GenBank based on BLASTn results. The Oligonychus sequences retrieved from GenBank were aligned with their counterpart sequences obtained during the present study using the CLUSTALW multiple alignment tool in BioEdit. The retrieved GenBank ITS2/COI sequences were under the accession numbers: AB683673, OP363224, AB683666, AB683653, AB683662, AB683669, MW491838, AB683658, MN190324, KC009700, AB683660, AB683656, AB683677, LC341206, OP214345, AB683675, AB683655, MZ435900, MZ425483, X80866, AB683665, DQ656485, AB683681, AB683659, KU323485, X80865, GU329963, AB683664, and KC352302. In addition, some ITS2/COI sequences of unidentified Oligonychus species deposited from Australia (MF462136), India (MG429138, MG677944, MK386956, and OL830813), and USA (KP180428) were also retrieved from GenBank. All ITS2/COI sequences of Oligonychus species obtained during the present study were deposited in the NCBI-GenBank database (Table S1).

2.3.3. Phylogenetic and Genetic Distances Analyses

Phylogenetic investigations were performed to assess the evolutionary relationships within and among various Oligonychus species using MEGA-X [29]. Phylogenetic trees were made using the neighbor-joining (NJ) method by the Tamura–Nei model [30]. The tree branch robustness was tested using the bootstrap analysis with 1000 replications [31]. Moreover, the pairwise p-distances (interspecific and intraspecific genetic divergence) were also calculated using MEGA-X.

3. Results

Based on the morphological examination of mite samples having male and female individuals, collected from different hosts and localities from Mexico, Pakistan, SA, USA, and Yemen, seven Oligonychus species viz. O. afrasiaticus, O. coniferarum, O. dactyloni, O. pratensis, O. perseae, O. tylus, and O. washingtoniae were recognized (Figure 1; Table S1). However, the samples containing only female specimens (Table S1) were recognized till the level of the genus Oligonychus.
At the molecular level, out of the 68 samples, 65 ITS2 amplicons were successfully amplified and sequenced. The length of the cleaned ITS2 sequences without ITS2 primers ranged from 405 to 539 bp (Table S1). However, the length of the 16 selected COI sequences obtained during this study was 410 bp (Table S1). Based on the ITS2 and COI data, the taxonomic identity of all 68 spider mite samples was successfully confirmed, which comprised eight Oligonychus species, namely O. afrasiaticus, O. coniferarum, O. dactyloni, O. punicae, O. perseae, O. pratensis, O. tylus, and O. washingtoniae (Figure 3, Figure 4, Figure 5, Figure 6 and Figure 7; Tables S1–S6). The actual taxonomic identity of the previously misidentified O. pratensis sample (SVN: 39; Table S1) that lacked male specimens was recognized as O. afrasiaticus (Figure 3; Table S2).
All the ITS2 and/or COI sequences of O. coniferarum, O. dactyloni, O. punicae, and O. washingtoniae, representing different samples from SA were assigned to a single haplotype (Tables S2 and S3; Figure 3 and Figure 4). Furthermore, the 29 ITS2 sequences of O. afrasiaticus separated into nine haplotypes: H1 (Israel and SA), H2 (SA and Yemen), H3 (SA), H4 (SA), H5 (SA), H6 (SA), H7 (Pakistan), H8 (SA), and H9 (SA) (Figure 5; Table S4). The nine ITS2 sequences of O. tylus were separated into four haplotypes: H1 (India), H2 (Pakistan and SA), H3 (SA), and H4 (SA) (Figure 5; Table S4). Likewise, the COI sequences of O. afrasiaticus and O. tylus were further separated into three and two haplotypes, respectively (Figure 6; Table S5). The three COI haplotypes of O. afrasiaticus were H1 (Israel), H2 (SA), and H3 (Yemen) (Figure 6; Table S5). Whereas the two COI haplotypes of O. tylus were H1 (India and SA) and H2 (SA) (Figure 6; Table S5).
The estimated pairwise p-distances for the ITS2 sequences showed that interspecific genetic divergence in various tested Oligonychus species ranged from 0.037 to 0.511 (3.7% to 51.1%; Tables S2 and S6), and intraspecific divergence ranged from 0.000 to 0.012 (0% to 1.2%; Table S4). The pairwise p-distances of COI sequences among various Oligonychus species showed interspecific divergence ranging from 0.032 to 0.181 (3.2% to 18.1%; Tables S3 and S7); whereas, the intraspecific divergence within same species ranged from 0.000 to 0.029 (0% to 2.9%; Table S5). Exceptionally, three COI sequences retrieved from the GenBank (AB683664 from Japan; GU329963 from China, and X80865 from France), representing three populations of O. ununguis, showed high intraspecific genetic divergence (6.5% to 11%; Table S7), suggesting O. ununguis is a species complex.
According to the ITS2-based NJ phylogenetic tree, the five species O. afrasiaticus, O. dactyloni, O. pratensis, O. tylus, and O. washingtoniae having upturned male aedeagus and belonging to the subgenus Reckiella, clustered as a monophyletic clade with 100% bootstrap value (Figure 3). However, O. coniferarum and O. punicae having downturned male aedeagus and belonging to the subgenus Oligonychus, formed a separate monophyletic clade with 88% bootstrap value (Figure 3). Similarly, in the COI-based NJ tree, the clade of the subgenus Reckiella and the subgenus Oligonychus grouped separately with monophyletic clades (Figure 4).
Moreover, the Oligonychus species of the subgenus Reckiella and species of the subgenus Oligonychus clustered separately into two monophyletic clades with 100% and 93% bootstrap values, respectively (Figure 7). In addition, the subgenus Oligonychus clade (supported with 93% bootstrap value) is further divided into two sub-clades, the b-i sub-clade, representing the peruvianus species group (female with either eight or nine tactile setae on tibia I), and the b-ii sub-clade representing the coffeae species group (female with either five, six, or seven tactile setae on tibia I) (Figure 7). Similarly, Oligonychus species of the subgenus Reckiella (Figure 8, clade a), and species of the subgenus Oligonychus (Figure 8, clade b) grouped separately into two monophyletic clades.

4. Discussion

In the present study, the ITS2 and COI molecular data successfully confirmed the species identity of eight Oligonychus species, namely, O. afrasiaticus, O. coniferarum, O. dactyloni, O. punicae, O. pratensis, O. perseae, O. tylus, and O. washingtoniae, representing 68 different mite samples collected from various hosts and localities in five countries. The genetic divergence is usually higher interspecifically than intraspecifically [7,14,16]. The ranges of interspecific genetic divergences obtained from ITS2 (3.7% to 51.1%), and COI (3.2% to 18.1%) supported the distinction of the eight Oligonychus species. However, the intraspecific genetic divergences ranged between 0% and 1.2% for ITS2 and 0% and 2.9% for COI. The obtained interspecific and intraspecific genetic divergences are aligned with the previous findings on tetranychid species [7,14,16,18,20,32,33], as well as other mites [34,35]. In various tetranychid species, including three Oligonychus species of O. afrasiaticus, O. perseae, and O. mangiferus (=O. punicae), the ITS2-based interspecific nucleotide divergence ranged from 4.4% to 54.8% [16]. Moreover, the interspecific divergence for ITS2 detected among four closely related Oligonychus species belonging to the subgenus Oligonychus ranged from 11.5% to 18.8% [7]. Similarly, the COI-based interspecific nucleotide divergences ranging from 5.4% to 18.3%, were detected among various Oligonychus species [7,14].
The absence of male specimens can lead to the creation of doubtful Oligonychus species, especially in the absence of integrative taxonomic approaches [3]. For example, O. changi (Tseng), O. jiangxiensis (Ma and Yuan), O. longus (Chaudhri, Akbar, and Rasool), and O. pongami (Sivakumar and Kunchithapatham) are suggested as species inquirendae, which were described without males [3,11,36,37,38]. In the present study, the molecular data revealed the correct species identity of 42 mite samples that lacked male specimens. Based on the low intraspecific nucleotide divergence (0% to 1.2%, ITS2; 0% to 2.9%, COI) and phylogenetic analyses, 16 samples of O. afrasiaticus, 14 samples of O. coniferarum, 1 sample of O. dactyloni, 2 samples of O. punicae, 4 samples of O. tylus, and 5 samples of O. washingtoniae were successfully recognized till the level of species. Additionally, the Oligonychus Saudi population, previously claimed O. pratensis [23], was correctly recognized as O. afrasiaticus. The absence of males in the Oligonychus population collected from grasses in the Baha region of SA led to the misidentification as O. pratensis [23]. Indeed, while conducting this study, we have not found any population of O. pratensis from SA. When we compared all the Saudi samples of Oligonychus belonging to the subgenus Reckiella with the Californian sample of O. pratensis collected from the USA (the country of its type locality), high genetic divergence either based on ITS2 (>11%) or COI (>8%), was shown, which is in accordance with separate species in previous molecular studies on tetranychid mites [7,14,16].
The wide distribution and polyphagous behavior of a spider mite species may affect its genetic structure [17,39]. The current study also revealed high genetic variations (number of haplotypes) in two widely distributed oligophagous species O. afrasiaticus and O. tylus collected/analyzed from India, Israel, Pakistan, SA, and Yemen. Nine ITS2 and three COI haplotypes have been identified in the date palm mite O. afrasiaticus. Out of the nine ITS2 haplotypes, representing seven hosts and four countries, six (H3, H4, H5, H6, H8, and H9) were only recovered from SA, one from both SA and Israel (H1), one from SA and Yemen (H2), and one (H7) from Pakistan. The geographic isolation may explain the distinction between the Middle Eastern and Pakistani ITS2 haplotypes of O. afrasiaticus [40]. Geographic isolation was a key factor in generating genetic variations among various populations of the citrus red mite Panonychus citri (McGregor) [41]. Similarly, O. tylus showed four ITS2 haplotypes representing nine localities and two COI haplotypes representing four localities, recovered from seven hosts and two countries. In addition to geographic distribution, host plants can increase genetic variations, as exhibited by the polyphagous spider mite species Eutetranychus orientalis (Klein) [17]. The oligophagous O. coniferarum inhabiting different species of conifers and the monophagous O. washingtoniae inhabiting W. filifera, did not show any genetic variations among their populations collected from different localities of SA.
The phylogenetic trees constructed using ITS2 sequences of 15 Oligonychus species and COI sequences of 30 Oligonychus species confirmed the morphological-based subdivision of the genus Oligonychus into two subgenera and four species groups [3]. The phylogenetic trees showed the monophyly nature of the two subgenera of Oligonychus: Reckiella and Oligonychus. These results are in agreement with previous findings either based on DNA-Sequence data of ITS2, COI, and 28S regions [14,16,32,42], or RNA-Sequence data [43], which separated Oligonychus species into two clades: a group of species with upturned (the subgenus Reckiella) and others with the downturned direction of male aedeagus (the subgenus Oligonychus). Further, in ITS2/COI-based phylogenetic trees, species belonging to the coffeae species group were clustered distantly from O. perseae of the peruvianus species group. It confirmed the morphological-based subdivision of the subgenus Oligonychus into two species groups based on a female character, i.e., the number of tactile setae on tibia I [3].
In addition, the molecular analysis of COI sequences of O. ununguis available on GenBank showed that the three cryptic species are hidden under the ununguis complex. The Japanese (Accession no: AB683664), Chinese (Accession no: GU329963), and French (Accession no: X80865) sequences of O. ununguis showed high genetic divergences (6.5% to 11%) from one another. Such high nucleotide divergences fall within the range of interspecific genetic divergence, as observed in the present (3.2% to 18.1%) and previous studies (5.4% to 18.3%) on Oligonychus species [7,14]. Similarly, the two GenBank ITS2 sequences of O. ununguis from China and Korea were previously highlighted as doubtful [7].
Some ITS2 (Accession no: MG429138, MG677944, OL830813, MK386956, India) and COI (KP180428, USA; MF462136, Australia) sequences available on GenBank identified to the genus Oligonychus level. Molecular data could not assign them to any of the known Oligonychus species. The high genetic divergences of ITS2 (4.4% to 10.4%) and COI (12.6%) clearly indicate their taxonomic identity as separate Oligonychus species. However, it is strongly recommended that the specimens/vouchers of these sequences need to be morphologically and molecularly re-characterized to know the actual global fauna of the genus Oligonychus.

5. Conclusions

In conclusion, the molecular data of ITS2 and COI successfully confirmed the taxonomic identity of 68 spider mite samples collected either with or without male specimens, representing eight Oligonychus species from Mexico, Pakistan, SA, USA, and Yemen. The phylogenetic analyses of various Oligonychus species also validated the subdivision of the genus Oligonychus into subgenera and species groups. The present study highlights the importance of integrated taxonomic approaches, e.g., the combination of morphological and molecular data, to confirm the identity of closely related Oligonychus species, especially in the absence of key morphological characters or individuals. Additionally, molecular data showed that the O. ununguis sequences available on the NCBI-GenBank database might be taxonomically misidentified and need further work. Overall, a taxonomic revision is compulsory to resolve the global issue of species complexes by adopting integrative taxonomic approaches.

Supplementary Materials

The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/insects14020192/s1, Table S1: Geographical distribution, host plant, collection details, and ITS2/COI fragment size of all spider mite samples of seven Oligonychus species viz. O. afrasiaticus, O. coniferarum, O. dactyloni, O. punicae, O. pratensis, O. tylus, and O. washingtoniae, collected from Pakistan, Saudi Arabia, USA, & Yamen, and analyzed in the present study. Table S2: Genetic divergence (pairwise p-distance) based on 33 ITS2 partial sequences, either obtained in the present study or retrieved from GenBank, among seven Oligonychus species (Acari: Prostigmata: Tetranychidae), collected/analyzed from Saudi Arabia and the USA. Table S3: Genetic divergence (pairwise p-distance) based on nine COI partial sequences, either obtained in the present study or retrieved from GenBank, among six Oligonychus species (Acari: Prostigmata: Tetranychidae), collected/analyzed from Saudi Arabia and the USA. Table S4: Genetic divergence (pairwise p-distance) based on 38 ITS2 partial sequences, either obtained in the present study or retrieved from GenBank, showing variations among various populations of Oligonychus afrasiaticus and O. tylus (Acari: Prostigmata: Tetranychidae), collected/analyzed from Israel, India, Pakistan, Saudi Arabia, and Yemen. Table S5: Genetic divergence (pairwise p-distance) based on 12 COI partial sequences, either obtained in the present study or retrieved from GenBank, showing variations among various populations of Oligonychus afrasiaticus and O. tylus (Acari: Prostigmata: Tetranychidae), collected/analyzed from Israel, India, Saudi Arabia, and Yemen. Table S6: Genetic divergence (pairwise p-distance) based on ITS2 sequences among 15 Oligonychus species (Acari: Prostigmata: Tetranychidae), either obtained in the present study or retrieved from GenBank, analyzed from India, Mexico, Saudi Arabia, and the USA. Table S7: Genetic divergence (pairwise p-distance) based on COI sequences among 32 Oligonychus species (Acari: Prostigmata: Tetranychidae), either obtained in the present study or retrieved from GenBank. References [7,23] are cited in the Supplementary Materials.

Author Contributions

Conceptualization, H.M.S.M., F.J.A., A.A.S. and M.K.; methodology, H.M.S.M., A.A.S. and M.K.; software, H.M.S.M. and A.A.S.; validation, F.J.A., A.A.S. and M.K.; formal analysis, H.M.S.M., A.A.S. and M.K.; investigation, H.M.S.M., F.J.A. and A.A.S.; data curation, H.M.S.M., A.A.S. and M.K.; writing—original draft preparation, H.M.S.M.; writing—review and editing, F.J.A., A.A.S. and M.K.; supervision, F.J.A.; project administration, F.J.A.; funding acquisition, F.J.A. All authors have read and agreed to the published version of the manuscript.

Funding

This research was funded by the Deputyship for Research & Innovation, Ministry of Education in Saudi Arabia for funding this research work through the project number IFKSURG-2-1157.

Data Availability Statement

All necessary data is provided in this paper.

Acknowledgments

The authors extend their appreciation to the Deputyship for Research & Innovation, Ministry of Education in Saudi Arabia for funding this research work through the project no. (IFKSURG-2-1157). Special thanks are also due to Teresa Santillan-Galicia (Colegio de Postgraduados, Montecillo, Mexico), Fatemeh Ganjisaffar (University of California, Riverside, CA, USA), Jawwad Hassan Mirza, Jamal Saeed Basahih, Eid Muhammad Khan, Hafiz Muhammad Sajid Ali, Muhammad Waleed, Nasreldeen Elgoni, and Naeem Abbas (King Saud University, Riyadh, Saudi Arabia) who all collected specimens from different countries. With their help, this study was finalized.

Conflicts of Interest

The authors declare no conflict of interest.

References

  1. Pritchard, A.E.; Baker, E.W. A Revision of the Spider Mite Family Tetranychidae; Pacific Coast Entomological Society: San Francisco, CA, USA, 1955; p. 472. [Google Scholar]
  2. Meyer, M.K.P.S. African Tetranychidae (Acari: Prostigmata)—With Reference to the World Genera, Entomology Memoir; Department of Agriculture and Water Supply: Pretoria, South Africa, 1987; pp. 1–175.
  3. Mushtaq, H.M.S.; Alatawi, F.J.; Kamran, M.; Flechtmann, C.H.W. The genus Oligonychus Berlese (Acari, Prostigmata, Tetranychidae): Taxonomic assessment and a key to subgenera, species groups, and subgroups. ZooKeys 2021, 1079, 89. [Google Scholar] [CrossRef] [PubMed]
  4. Meyer, M.K.P.S. A Revision of the Tetranychidae of Africa (Acari) with a Key to the Genera of the World, Entomology Memoir; Department of Agricultural Technical Services: Pretoria, South Africa, 1974; pp. 1–291.
  5. Jeppson, L.R.; Keifer, H.H.; Baker, E.W. Mites Injurious to Economic Plants; University of California Press: Berkeley, CA, USA, 1975; pp. 1–614. [Google Scholar]
  6. Li, J.; Yi, T.-C.; Guo, J.-J.; Jin, D.-C. Ontogenetic development and redescription of Oligonychus pratensis (Banks, 1912) (Acari: Tetranychidae). Zootaxa 2018, 4486, 349–375. [Google Scholar] [CrossRef] [PubMed]
  7. Mushtaq, H.M.S.; Kamran, M.; Saleh, A.A.; Alatawi, F.J. Evidence for Reconsidering the Taxonomic Status of Closely Related Oligonychus Species in punicae Complex (Acari: Prostigmata: Tetranychidae). Insects 2023, 14, 3. [Google Scholar] [CrossRef] [PubMed]
  8. Rahman, K.A.; Sapra, A.N. Mites of the family Tetranychidae from Lyallpur with descriptions of four new species. Indian Acad. Sci. Proc. Sect. B 1940, 11, 177–196. [Google Scholar] [CrossRef]
  9. Mitrofanov, V.I.; Zapletina, A. A new species of the genus Oligonychus from Azerbaijan. Dokl. Akad. Nauk. Azerbaijan SSR 1973, 29, 50–52. [Google Scholar]
  10. Meyer, M.K.P.; Ryke, P.A.J. A revision of the spider mites (Acarina: Tetranychidae) of South Africa with descriptions of a new genus and new species. J. Entomol. Soc. S. Afr. 1959, 22, 330–366. [Google Scholar]
  11. Tseng, Y.H. Mites associated with the conifers of Taiwan, (1) Tetranychoidea, with a checklist of the world. J. Entomol. Natl. Chung Hsien Univ. 1980, 15, 145–168. [Google Scholar]
  12. Navajas, M.; Gutierrez, J.; Lagnel, J.; Boursot, P. Mitochondrial cytochrome oxidase I in tetranychid mites: A comparison between molecular phylogeny and changes of morphological and life history traits. Bull. Entomol. Res. 1996, 86, 407–417. [Google Scholar] [CrossRef] [Green Version]
  13. Matsuda, T.; Fukumoto, C.; Hinomoto, N.; Gotoh, T. DNA-based identification of spider mites: Molecular evidence for cryptic species of the genus Tetranychus (Acari: Tetranychidae). J. Econ. Entomol. 2013, 106, 463–472. [Google Scholar] [CrossRef] [Green Version]
  14. Matsuda, T.; Hinomoto, N.; Singh, R.N.; Gotoh, T. Molecular-based identification and phylogeny of Oligonychus species (Acari: Tetranychidae). J. Econ. Entomol. 2012, 105, 1043–1050. [Google Scholar] [CrossRef] [Green Version]
  15. Arabuli, T.; Gotoh, T. A new species of spider mite, Oligonychus neocastaneae sp. nov. (Acari: Tetranychidae), from Japan. Zootaxa 2018, 4378, 563–572. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  16. Ben-David, T.; Melamed, S.; Gerson, U.; Morin, S. ITS2 sequences as barcodes for identifying and analyzing spider mites (Acari: Tetranychidae). Exp. Appl. Acarol. 2007, 41, 169–181. [Google Scholar] [CrossRef] [PubMed]
  17. Mirza, J.H.; Kamran, M.; Saleh, A.A.; Alatawi, F.J. Molecular and phenotypic variations in Eutetranychus orientalis (Klein) populations from Saudi Arabia. PLoS ONE 2020, 15, e0233389. [Google Scholar] [CrossRef] [PubMed]
  18. Navajas, M.; Gutierrez, J.; Williams, M.; Gotoh, T. Synonymy between two spider mite species, Tetranychus kanzawai and T. hydrangeae (Acari: Tetranychidae), shown by ribosomal ITS2 sequences and cross-breeding experiments. Bull. Entomol. Res. 2001, 91, 117–123. [Google Scholar] [PubMed]
  19. Gotoh, T.; Araki, R.; Boubou, A.; Migeon, A.; Ferragut, F.; Navajas, M. Evidence of co-specificity between Tetranychus evansi and Tetranychus takafujii (Acari: Prostigmata, Tetranychidae): Comments on taxonomic and agricultural aspects. Int. J. Acarol. 2009, 35, 485–501. [Google Scholar] [CrossRef]
  20. Zeity, M.; Srinivasa, N.; Gowda, C.C. Are Tetranychus macfarlanei Baker & Pritchard and Tetranychus malaysiensis Ehara (Acari: Tetranychidae) one species? Morphological and molecular evidences for synonymy between these two spider mite species and a note on invasiveness of T. macfarlanei on okra and eggplant in India. Syst. Appl. Acarol. 2017, 22, 467–476. [Google Scholar] [CrossRef]
  21. Mushtaq, H.M.S.; Kamran, M.; Alatawi, F.J. New species, new records, and re-descriptions of two species of the genus Oligonychus Berlese (Acari: Prostigmata: Tetranychidae) from Saudi Arabia. Syst. Appl. Acarol. 2022, 27, 2568–2582. [Google Scholar] [CrossRef]
  22. Migeon, A.; Dorkeld, F. Spider Mites Web: A Comprehensive Database for the Tetranychidae. Available online: http://www1.montpellier.inra.fr/CBGP/spmweb (accessed on 5 January 2023).
  23. Alatawi, F.J.; Kamran, M. Spider mites (Acari: Tetranychidae) of Saudi Arabia: Two new species, new records and a key to all known species. J. Nat. Hist. 2018, 52, 429–455. [Google Scholar] [CrossRef]
  24. Guzman-Valencia, S.; Santillán-Galicia, M.T.; Guzmán-Franco, A.W.; González-Hernández, H.; Carrillo-Benítez, M.G.; Suárez-Espinoza, J. Contrasting effects of geographical separation on the genetic population structure of sympatric species of mites in avocado orchards. Bull. Entomol. Res. 2014, 104, 610–621. [Google Scholar] [CrossRef]
  25. Ganjisaffar, F.; Perring, T.M. Prey stage preference and functional response of the predatory mite Galendromus flumenis to Oligonychus pratensis. Biol. Control 2015, 82, 40–45. [Google Scholar] [CrossRef]
  26. Bolland, H.R.; Gutierrez, J.; Flechtmann, C.H.W. World Catalogue of the Spider Mite Family (Acari: Tetranychidae); Brill Academic Publishers: Leiden, The Netherlands, 1998; p. 392. [Google Scholar]
  27. Mushtaq, H.M.S.; Alatawi, F.J.; Kamran, M. Keys to World Species of the Genus Oligonychus Berlese (Acari, Prostigmata, Tetranychidae): Taxonomic Notes and New Oligonychus Species, King Saud University: Riyadh, Saudi Arabia, 2023; in prepration.
  28. Hall, T.A. BioEdit: A user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Proc. Nucleic Acids Symp. Ser. 1999, 41, 95–98. [Google Scholar]
  29. Kumar, S.; Stecher, G.; Li, M.; Knyaz, C.; Tamura, K. MEGA X: Molecular evolutionary genetics analysis across computing platforms. Mol. Biol. Evol. 2018, 35, 1547–1549. [Google Scholar] [CrossRef] [PubMed]
  30. Tamura, K.; Nei, M. Estimation of the number of nucleotide substitutions in the control region of mitochondrial DNA in humans and chimpanzees. Mol. Biol. Evol. 1993, 10, 512–526. [Google Scholar]
  31. Felsenstein, J. Confidence limits on phylogenies: An approach using the bootstrap. Evolution 1985, 39, 783–791. [Google Scholar] [CrossRef] [PubMed]
  32. Navajas, M.; Gutierrez, J.; Bonato, O.; Bolland, H.R.; Mapangou-Divassa, S. Intraspecific diversity of the cassava green mite Mononychellus progresivus (Acari: Tetranychidae) using comparisons of mitochondrial and nuclear ribosomal DNA sequences and cross-breeding. Exp. Appl. Acarol. 1994, 18, 351–360. [Google Scholar] [CrossRef] [PubMed]
  33. Khaing, T.M.; Shim, J.K.; Lee, K.Y. Molecular identification of four Panonychus species (Acari: Tetranychidae) in Korea, including new records of P. caglei and P. mori. Entomol. Res. 2015, 45, 345–353. [Google Scholar] [CrossRef]
  34. Yang, B.; Cai, J.; Cheng, X. Identification of astigmatid mites using ITS2 and COI regions. Parasitol. Res. 2011, 108, 497–503. [Google Scholar] [CrossRef]
  35. Navia, D.; Mendonca, R.S.; Ferragut, F.; Miranda, L.C.; Trincado, R.C.; Michaux, J.; Navajas, M. Cryptic diversity in Brevipalpus mites (Tenuipalpidae). Zool. Scr. 2013, 42, 406–426. [Google Scholar] [CrossRef]
  36. Ma, E.P.; Yuan, Y.L. New species and new records of tetranychid mites from China I. (Acarina: Tetranychidae). Acta Zootaxonomica Sin. 1980, 5, 42–45. [Google Scholar]
  37. Sivakumar, K.; Kunchithapatham, R. Phytophagous mite diversity of Tetranychus spp. and Oligonychus spp. (Acari: Tetranychidae) found on different host plants in Coimbatore district and surrounding regions. Trends Biosci. 2014, 7, 4113–4117. [Google Scholar]
  38. Chaudhri, W.M.; Akbar, S.; Rasool, A. Taxonomic Studies of the Mites Belonging to the Families Tenuipalpidae, Tetranychidae, Tuckerellidae, Caligonellidae, Stigmaeidae and Phytoseiidae, PL-480 Project on Mites; University of Agriculture: Lyallpur, Pakistan, 1974; p. 250. [Google Scholar]
  39. Xie, L.; Hong, X.-y.; Xue, X.-f. Population genetic structure of the two spotted spider mite (Acari: Tetranychidae) from China. Ann. Entomol. Soc. Am. 2006, 99, 959–965. [Google Scholar] [CrossRef]
  40. Ge, M.-K.; Sun, E.-T.; Jia, C.-N.; Kong, D.-D.; Jiang, Y.-X. Genetic diversity and differentiation of Lepidoglyphus destructor (Acari: Glycyphagidae) inferred from inter-simple sequence repeat (ISSR) fingerprinting. Syst. Appl. Acarol. 2014, 19, 491–498. [Google Scholar] [CrossRef]
  41. Yuan, M.-L.; Wei, D.-D.; Zhang, K.; Gao, Y.-Z.; Liu, Y.-H.; Wang, B.-J.; Wang, J.-J. Genetic Diversity and Population Structure of Panonychus citri (Acari: Tetranychidae), in China Based on Mitochondrial COI Gene Sequences. Mol. Entomol. 2010, 103, 2204–2213. [Google Scholar] [CrossRef] [PubMed]
  42. Ros, V.I.D.; Breeuwer, J.A.J. Spider mite (Acari: Tetranychidae) mitochondrial COI phylogeny reviewed: Host plant relationships, phylogeography, reproductive parasites and barcoding. Exp. Appl. Acarol. 2007, 42, 239–262. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  43. Matsuda, T.; Kozaki, T.; Ishii, K.; Gotoh, T. Phylogeny of the spider mite sub-family Tetranychinae (Acari: Tetranychidae) inferred from RNA-Seq data. PLoS ONE 2018, 13, e0203136. [Google Scholar] [CrossRef] [PubMed]
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Figure 1. Geographical distribution of the (A) 66 spider mite samples of Oligonychus afrasiaticus, O. coniferarum, O. dactyloni, O. punicae, O. tylus, and O. washingtoniae along with the (B) two samples of O. pratensis and O. perseae, collected from various hosts and localities in Mexico, Pakistan, Saudi Arabia, United States, and Yemen in the present study.
Figure 1. Geographical distribution of the (A) 66 spider mite samples of Oligonychus afrasiaticus, O. coniferarum, O. dactyloni, O. punicae, O. tylus, and O. washingtoniae along with the (B) two samples of O. pratensis and O. perseae, collected from various hosts and localities in Mexico, Pakistan, Saudi Arabia, United States, and Yemen in the present study.
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Figure 2. Geographical distribution of 61 spider mite samples of Oligonychus afrasiaticus, O. coniferarum, O. dactyloni, O. punicae, O. tylus, and O. washingtoniae, reported from various hosts and localities of 10 provinces in Saudi Arabia.
Figure 2. Geographical distribution of 61 spider mite samples of Oligonychus afrasiaticus, O. coniferarum, O. dactyloni, O. punicae, O. tylus, and O. washingtoniae, reported from various hosts and localities of 10 provinces in Saudi Arabia.
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Figure 3. NJ phylogenetic tree based on 33 ITS2 sequences, representing different samples of seven Oligonychus species. Eutetranychus orientalis* was retrieved from GenBank and used as an outgroup taxon. A total of 30 sequences were obtained in the present study from different hosts and regions in Saudi Arabia and one from USA (** including 19 samples that have not been identified morphologically till species due to the absence of males, and *** one sample that was previously claimed as O. pratensis from Saudi Arabia) [23]. One sequence of O. punicae was retrieved from GenBank, previously reported from Saudi Arabia [7]. Numbers on tree branches are bootstrap values obtained from 1000 replicates.
Figure 3. NJ phylogenetic tree based on 33 ITS2 sequences, representing different samples of seven Oligonychus species. Eutetranychus orientalis* was retrieved from GenBank and used as an outgroup taxon. A total of 30 sequences were obtained in the present study from different hosts and regions in Saudi Arabia and one from USA (** including 19 samples that have not been identified morphologically till species due to the absence of males, and *** one sample that was previously claimed as O. pratensis from Saudi Arabia) [23]. One sequence of O. punicae was retrieved from GenBank, previously reported from Saudi Arabia [7]. Numbers on tree branches are bootstrap values obtained from 1000 replicates.
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Figure 4. NJ phylogenetic tree based on 10 COI sequences, representing different samples of six Oligonychus species. Eutetranychus orientalis* was retrieved from GenBank and used as an outgroup taxon. A total of eight sequences were obtained in the present study from different hosts and regions in Saudi Arabia, and one from USA (** including samples that have not been identified morphologically till species due to the absence of males). One sequence of O. punicae was retrieved from GenBank, previously reported from Saudi Arabia [7]. Numbers on tree branches are bootstrap values obtained from 1000 replicates.
Figure 4. NJ phylogenetic tree based on 10 COI sequences, representing different samples of six Oligonychus species. Eutetranychus orientalis* was retrieved from GenBank and used as an outgroup taxon. A total of eight sequences were obtained in the present study from different hosts and regions in Saudi Arabia, and one from USA (** including samples that have not been identified morphologically till species due to the absence of males). One sequence of O. punicae was retrieved from GenBank, previously reported from Saudi Arabia [7]. Numbers on tree branches are bootstrap values obtained from 1000 replicates.
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Figure 5. NJ tree based on 39 ITS2 sequences, representing different populations of the two closely related Oligonychus species (O. afrasiaticus and O. tylus) and one distantly related species, O. punicae (* was retrieved from GenBank and used as an outgroup taxon). A total of 31 sequences were obtained in the present study from different hosts and regions in Saudi Arabia, three from Pakistan, and two from Yemen. In total, 19 samples ** have not been morphologically identified till species level due to the absence of males, and *** one sample previously claimed as O. pratensis from Saudi Arabia [23]. The sequences of O. afrasiaticus from Israel and O. tylus from India were retrieved from GenBank. Numbers on tree branches are bootstrap values obtained from 1000 replicates.
Figure 5. NJ tree based on 39 ITS2 sequences, representing different populations of the two closely related Oligonychus species (O. afrasiaticus and O. tylus) and one distantly related species, O. punicae (* was retrieved from GenBank and used as an outgroup taxon). A total of 31 sequences were obtained in the present study from different hosts and regions in Saudi Arabia, three from Pakistan, and two from Yemen. In total, 19 samples ** have not been morphologically identified till species level due to the absence of males, and *** one sample previously claimed as O. pratensis from Saudi Arabia [23]. The sequences of O. afrasiaticus from Israel and O. tylus from India were retrieved from GenBank. Numbers on tree branches are bootstrap values obtained from 1000 replicates.
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Figure 6. NJ phylogenetic tree based on 13 COI sequences, representing different populations of two closely related Oligonychus species (O. afrasiaticus and O. tylus) and one distantly related species, O. punicae (* was retrieved from GenBank and used as an outgroup taxon). A total of eight sequences were obtained in the present study from different hosts and regions in Saudi Arabia, and two from Yemen (** including samples that have not been identified morphologically till species due to the absence of males). The sequences of O. afrasiaticus from Israel and O. tylus from India were retrieved from GenBank. Numbers on tree branches are bootstrap values obtained from 1000 replicates.
Figure 6. NJ phylogenetic tree based on 13 COI sequences, representing different populations of two closely related Oligonychus species (O. afrasiaticus and O. tylus) and one distantly related species, O. punicae (* was retrieved from GenBank and used as an outgroup taxon). A total of eight sequences were obtained in the present study from different hosts and regions in Saudi Arabia, and two from Yemen (** including samples that have not been identified morphologically till species due to the absence of males). The sequences of O. afrasiaticus from Israel and O. tylus from India were retrieved from GenBank. Numbers on tree branches are bootstrap values obtained from 1000 replicates.
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Figure 7. NJ phylogenetic tree based on 16 ITS2 sequences, representing 15 Oligonychus species. Eutetranychus orientalis* was used as an outgroup taxon. A total of 11 Oligonychus species belong to the subgenus (a) Reckiella, whereas four Oligonychus species (** including a cryptic Oligonychus sp., previously claimed as O. punicae from Mexico) [7] belong to the species groups (b-i) peruvianus and (b-ii) coffeae of the subgenus Oligonychus [3]. Numbers on tree branches are bootstrap values obtained from 1000 replicates.
Figure 7. NJ phylogenetic tree based on 16 ITS2 sequences, representing 15 Oligonychus species. Eutetranychus orientalis* was used as an outgroup taxon. A total of 11 Oligonychus species belong to the subgenus (a) Reckiella, whereas four Oligonychus species (** including a cryptic Oligonychus sp., previously claimed as O. punicae from Mexico) [7] belong to the species groups (b-i) peruvianus and (b-ii) coffeae of the subgenus Oligonychus [3]. Numbers on tree branches are bootstrap values obtained from 1000 replicates.
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Figure 8. NJ phylogenetic tree based on 33 COI sequences, representing 32 Oligonychus species. Eutetranychus orientalis* was used as an outgroup taxon. A total of 12 Oligonychus species belong to the subgenus (a) Reckiella, whereas 18 Oligonychus species (** including a cryptic Oligonychus sp., previously claimed as O. punicae from Mexico) [7] belong to the species groups (b-i) coffeae and (b-ii) peruvianus of the subgenus Oligonychus [3]. Numbers on tree branches are bootstrap values obtained from 1000 replicates.
Figure 8. NJ phylogenetic tree based on 33 COI sequences, representing 32 Oligonychus species. Eutetranychus orientalis* was used as an outgroup taxon. A total of 12 Oligonychus species belong to the subgenus (a) Reckiella, whereas 18 Oligonychus species (** including a cryptic Oligonychus sp., previously claimed as O. punicae from Mexico) [7] belong to the species groups (b-i) coffeae and (b-ii) peruvianus of the subgenus Oligonychus [3]. Numbers on tree branches are bootstrap values obtained from 1000 replicates.
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Mushtaq, H.M.S.; Saleh, A.A.; Kamran, M.; Alatawi, F.J. Molecular-Based Taxonomic Inferences of Some Spider Mite Species of the Genus Oligonychus Berlese (Acari, Prostigmata, Tetranychidae). Insects 2023, 14, 192. https://doi.org/10.3390/insects14020192

AMA Style

Mushtaq HMS, Saleh AA, Kamran M, Alatawi FJ. Molecular-Based Taxonomic Inferences of Some Spider Mite Species of the Genus Oligonychus Berlese (Acari, Prostigmata, Tetranychidae). Insects. 2023; 14(2):192. https://doi.org/10.3390/insects14020192

Chicago/Turabian Style

Mushtaq, Hafiz Muhammad Saqib, Amgad A. Saleh, Muhammad Kamran, and Fahad Jaber Alatawi. 2023. "Molecular-Based Taxonomic Inferences of Some Spider Mite Species of the Genus Oligonychus Berlese (Acari, Prostigmata, Tetranychidae)" Insects 14, no. 2: 192. https://doi.org/10.3390/insects14020192

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