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First DNA phylogeny of Euscorpius Thorell, 1876 (Scorpiones: Euscorpiidae) and its bearing on taxonomy and biogeography of this genus
... As expected, rRNA genes showed less diversity than PCGs. Traditionally, scorpion phylogenetics has relied on the slower evolving 16S rRNA gene and the faster changing COI gene [25][26][27]. While COI is a popular DNA barcoding tool and is often used for intraspecific scorpion studies, our results reveal that other mitochondrial genes are even more variable. ...
While originally intending to explore the venom gland microbiome of the desert hairy scorpion Hadrurus arizonensis Ewing, 1928, nanopore sequencing serendipitously recovered complete mitochondrial genomes for this iconic arachnid. Phylogenetic analysis of these high-quality genomes places Hadrurus as sister to Uroctonus, in agreement with some phylogenomic hypotheses. Additionally, we reveal significant genetic variation among individuals from the same population, highlighting the potential of mitogenomics for population genetics and phylogeography. This study showcases the effectiveness and affordability of nanopore sequencing for research with non-model organisms, opening new avenues for investigating arachnid biodiversity, evolution, and biogeography.
... Scorpion species were traditionally delimited based on diagnostic combinations of morphological characters. However, an increasing number of integrative systematics studies revealed that many widespread species, previously based solely on morphological characters, are actually complexes of more range-restricted species that may be distinguished from one another by a combination of morphological and genetic differences (Gantenbein et al. 1999(Gantenbein et al. , 2000(Gantenbein et al. , 2001Largiadèr 2002, 2003;Gantenbein and Keightley 2004;Parmakelis et al. 2006;Luna-Ramírez et al. 2017;. Similar patterns were revealed in the few studies that applied an integrative approach to the systematics of Iranian scorpions (e.g., Mirshamsi et al. 2010Mirshamsi et al. , 2013Azghadi et al. 2014; Barahoei et al. 2022). ...
Morphological and genetic variation among populations of the buthid scorpion, Hottentotta saulcyi (Simon, 1880), occurring in western and southwestern Iran was explored using morphometric variables, one nuclear marker (28S rDNA) and three mitochondrial markers (12S rDNA, 16S rDNA, and Cytochrome c Oxidase Subunit I). Genetic and morphometric statistical analyses revealed extensive cryptic diversity. Phylogenetic analysis with Bayesian Inference and Maximum Likelihood uncovered two divergent clades, one of which is described as a new species, Hottentotta hatamtiorum sp. nov., from Ilam and Khuzestan Provinces, southwestern Iran. The description of the new species raises the total count of Hottentotta Birula, 1908 species to 61, twelve of which are endemic or subendemic to the Iranian Plateau.
... DNA was extracted from tissue obtained from the pedipalps of each sampled specimen using Qiagen Kit (GENTRA® -Generation ® Capture Column™ Kit) according to the manufacturer's recommendations. Fragments of mtDNA were amplified using primers 1 6 S F ( 5 ' -C G AT T T G A A C T C A G AT C A -3 ' designed by Gantenbein et al. 1999) and LR-J-1 2 8 8 7 ( 5 ' -G TG C A A A G G TA G C ATA ATC A -3 ' ; Simon et al. 1994) for the 16S rRNA gene. For COI amplification the following primers were used: C1-J-2183 (5'-CAACATTTATTTTGATTTTTTGG-3'; S i m o n e t a l . 1 9 9 4 ) a n d C O I K G -R 2 ( 5 ' -GATATTAATCCTAAAAAATGTTGAGG-3'; Tanaka ...
Scorpion envenoming is considered a public health problem in Brazil. A recent study described a variation in the clinical outcome of envenoming by Tityus obscurus in two populations separated by 850 km in the northeastern Amazon region. Our aim was to evaluate whether such clinical and toxinological variations are associated with underlying differences in genetic diversity between these two T. obscurus populations. We obtained DNA from five individuals of each population, in the municipalities of Belém and Santarém, located east and west of the state of Pará, Brazil, respectively. Gene regions encoding mitochondrial DNA (mtDNA) markers cytochrome oxidase subunit I (COI) and ribosomal 16S RNA (16S) were amplified and sequenced. Phylogenetic analyses were performed using maximum likelihood (ML) and Bayesian inferences (BA) for both molecular data (COI and 16S). The sampled T. obscurus populations corresponded to two distinct mtDNA lineages (genetic distance COIK2 P = 0.08 to 0.13; 16SK2 P = 0.10 to 0.11) with no shared mutations between groups and well supported by ML and BA inferences. Based on the divergence values found between eastern and western populations (COI, 0.07 to 0.12; 16S, 0.10), our study confirms the genetic heterogeneity of T. obscurus populations within the state of Pará, which correlates with observed venom and clinical differences, and reinforces the need for mapping the distribution of haplotypes throughout the geographic range of T. obscurus, to aid in future epidemiological, toxinological, and evolutionary studies.
KEYWORDS:
mitochondrial DNA; phylogenetic analyses; cytochrome oxidase subunit I; 16S RNA
... nov., using Qiagen DNA extraction kits following the manufacturer's instructions. The amplified 16S rRNA gene products were purified and sequenced using invertebrate universal primers, as determined, and sequenced on an ABI 3500 automated sequencer (Applied Biosystems Inc., USA) and following Gantenbein et al. (1999). The sequences were edited using BioEdit v. 7.2.5 Hall (1999). ...
A new species, Leiurus hadb Al-Qahtni, Al-Salem, Alqahtani & Badry, sp. nov. , is described and illustrated from the Majami al-Hadb Protected Area in the Riyadh Province of Saudi Arabia. The new species is compared with species of Leiurus distributed in Saudi Arabia, especially L. arabicus Lowe, Yağmur & Kovařík, 2014. The integrated results indicate that the population found in Majami al-Hadb represents a distinct species, which is described herein. Moreover, the molecular analysis is conducted on the mitochondrial gene 16S rRNA to compare L. hadb sp. nov. with samples of L. arabicus and L. haenggii from Saudi Arabia. The analysis revealed a genetic divergence ranging from 6.0 to 12%. The combination of molecular evidence and morphological characteristics provides adequate support for recognizing the Majami al-Hadb population as a distinct species. Additionally, an identification key for the genus Leiurus found in Saudi Arabia is also provided.
... Many scorpion taxa are characterized by high levels of morphological uniformity and conservatism, which hampers straightforward species delimitation [23]. More insights were gained later from studies of mitochondrial and nuclear gene variation [24]. Recent research has employed the molecular phylogeny to interpret the evolutionary relationships of numerous populations of scorpions [25][26][27][28];. ...
Background
The genus Nebo has been identified as a medically important scorpion species distributed across Arabia and the Middle East. However, its taxonomic status remains unclear.
Aim
The molecular phylogeny of two Nebo species from Saudi Arabia and comparative sequences from Palestine is presented based on the mitochondrial cytochrome oxidase subunit I (COI) gene.
Methodology
Scorpion specimens were collected from two different localities, mainly the Southern part of Saudi Arabia. Then, DNA was extracted, amplified using invertebrate universal primers, and sequenced to identify the COI gene. The obtained sequences were analyzed, and phylogenetic trees based on maximum parsimony, neighbor-joining, and Bayesian inference were constructed.
Results
The inferred phylogeny indicates the monophyletic status of the family Diplocentridae and its subfamily Nebinae and Diplocentrinae. Also, the phylogenetic analyses support the existence of interspecific and intraspecific variations among/ within Nebo hierichonticus and Nebo yemenensis which may indicate distinct species.
Conclusion
Further morphological studies with additional specimens from the Arabian Peninsula may reveal possible undiscovered and cryptic species.
... The amplified 16S rRNA gene products were checked and purified (Qiagen) according to the manufacturer's instructions. A fragment of the 16S rRNA gene was amplified via a standard polymerase chain reaction (PCR) using the invertebrate universal primers, as determined and sequenced on an ABI 3500 automated sequencer (Applied Biosystems Inc., USA) and following Gantenbein et al. (1999). The chromatograms and sequences were examined and edited using BioEdit v. 7.2.5 (Hall 1999). ...
We describe and illustrate a new scorpion species, Androctonus tihamicus sp. nov., from the Mecca Province of southwestern Saudi Arabia. The new species is compared to the genus Androctonus Ehrenberg, 1828, which is distributed throughout the Middle East, and especially to A. australis (Linnaeus, 1758). We provide the molecular phylogeny for this species.
... The targeted contiguous nuclear region included the 3 ′ end of the small ribosomal subunit (SSU; 18S), the complete internal transcribed spacer 1 (ITS1), and the 5 ′ end of the 5.8S gene (henceforth called ITS1). In the amplification of the 16S rDNA, either the scorpion-specific primers reported by Gantenbein et al. (1999) or the universal primers (16S_BR_LR_12887/16S_AR_LR_13398) of Simon et al. (1994) were used. For the amplification of the COX1 gene fragment, the universal primers C1-N-2191 and C1-J-1718 (Simon et al., 1994) were used. ...
Iuridae is a family of scorpions that exhibits a highly complex biogeographic and taxonomic history. Iuridae taxa are mainly found in Turkey and Greece, whereas a single species is found in northern Iraq. Several taxonomic revisions have been conducted on this family that initially comprised two genera. The latest taxonomic review, based on morphological and anatomical features, raised the number of Iuridae genera to four, and the number of species to 14.
Sequence data from three molecular markers (COX1, 16S rDNA, ITS1) originating from numerous Iuridae taxa were analyzed within a phylogenetic framework. Divergence time-estimate analyses, species delimitation approaches and estimation of ancestral areas were implemented in order to: 1) reconstruct the phylogenetic relationships of the Iuridae taxa, 2) evaluate the morphological classifications, and 3) obtain insights into the biogeographic history of the family in the East Mediterranean.
The multi-locus phylogeny clearly confirms an ancient division into two clades, Calchinae and Iurinae. Ancient patterns of isolation and dispersal are revealed. Both subfamilies are largely confined to the Anatolian peninsula and its few coastal islands; only the most derived genus Iurus has dispersed westward to Crete and Peloponnese. Based on our findings, three new genera of Iurinae (Metaiurus, Anatoliurus, and Letoiurus) are established. The genus Neocalchas emerges as one of the most ancient scorpion clades, with divergence time about 27 mya.
... A 645-bp fragment of the COI was amplified with universal primers LCO 1490 (5 0 -GGT CAA CAA ATC ATA AAG ATA TTG G-3 0 ) HCO 2198 (5 0 -TAA ACT TCA GGG TGA CCA AAA AAT CA-3 0 ) (Folmer et al. 1994). For amplification of the 354 bp 16S rDNA region, we used the primers LR-J-12887 (5 0 -CGATTTGAACTCAGATCA-3 0 ) (Simon et al. 1994) and (5 0 -GTGCAAAGGTAGCATAATCA-3 0 ) (Gantenbein et al. 1999). ...
With more than 33,000 sting cases and 47 deaths recorded between 2000 and 2016, Panama is the country with the highest incidence of envenomations by scorpions in Central America. Species in the genus Centruroides are responsible for most scorpion sting reports, however, identification at the species level is complicated because the genus has considerable intraspecific morphological variation. To date, no molecular data have been reported from Panama that would help to estimate their genetic diversity and validate morphometric identification methods. We provide here the first genetic diversity data of the two endemic species (C. granosus and C. panamensis) and other two species reported in Panama (C. bicolor and C. limbatus). A total of 41 specimens were sequenced for COI and 16S rDNA mitochondrial genes. The phylogenetic concatenated analysis separates the Panamanian samples into four well-supported clades represented by C. bicolor, C. granosus and (C. panamensis + C. limbatus). The two endemic species are not the closest relatives in the tree. Low diversity in combination with its very narrow distribution suggest that C. panamensis is susceptible to environmental degradation. A single specimen of Coiba island is intermediate in the tree structure between C. bicolor and C. panamensis and may represent an early stage of speciation. The haplotype network is also consistent with the phylogenetic trees.
... Genomic DNA was isolated from leg muscle tissue using the Tissue Genomic DNA kit (Geneaid) following the manufacturer's protocol. Partial fragments of two mitochondrial genes, a barcode locus cytochrome c oxidase subunit I (COI; c. 660 bp) and the ribosomal 16S rRNA gene (16S; c. 370 bp), were amplified by PCR with the primer pairs LCO1490 and HCO2198 (Folmer et al., 1994) for COI, and scorpion-specific primers reported by Gantenbein et al., (1999) for 16S. The PCR protocol, identical for both the gene loci, consisted of initial denaturation step at 95 °C for 5 min, followed by 36 cycles of denaturation at 95 °C for 30 s, annealing at 45 °C for 1 min, extension at 72 °C for 1 min and terminated by final extension at 72 °C for 10 min. ...
To determine the mechanisms of karyotype differentiation in scorpions of the genus Gint, we employed an integrative approach, combining cytogenetic data and sequence-based phylogeny. We cytogenetically examined six species with emphasis on multivalent meiotic configurations, 18S rDNA and (TTAGG)n distribution and compared chromosomal data with genetic divergence based on analysis of 16S rRNA and COI gene markers. Our results show that Gint species exhibit substantial karyotype diversity (2n = 18–45) and a high incidence of chromosome heterozygosity. Meiotic chromosome chains formed by up to six elements were found in 85% of analysed individuals, causing intraspecific chromosome variation in three species. Fluorescence in situ hybridization revealed that the 18S rDNA distribution pattern differed in Gint species, including at the intrapopulation level, but the chromosomal localization of (TTAGG)n motif was stable across species. Conspicuous interspecific differences in chromosome counts broadly corresponded with genetic divergence among Gint species. Our findings indicate that Gint karyotypes have undergone dynamic reorganization through independent fusions, fissions and reciprocal translocations. Owing to present chromosomal polymorphism, such structural changes shaping the genome architecture appear to be still ongoing in the populations of some Gint species.
Populations of Monacha atacis from southern Occitania in France and of M. samsunensis from northern Anatolia in Turkey
(Atakum/Samsun and Kastamonu) were investigated by an integrative approach based on morphological (shell and
genitalia) and molecular (mitochondrial and nuclear gene sequences) features. Morphological examination revealed a
complex pattern of variation within and between geographically separated populations, while molecular analysis showed
strong similarity between the two species, confirming earlier suggestions that the species are conspecific. Pfeiffer’s name
Helix samsunensis introduced in 1868 has priority over the name M. atacis given by Gittenberger & de Winter in 1985.
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