Interspecific Genetic Variation Research Articles

Morpho-molecular characterization and phylogenetic relationships of Encotyllabe percussa n. sp. (Monogenea: Capsalidae) from the spangled emperor Lethrinus nebulosus (Teleostei, Lethrinidae).

Encotyllabe percussa n. sp. is proposed based on morphology and DNA sequences analysis of ribosomal (18S, 28S) and mitochondrial (COI) gene fragments. Encotyllabe percussa n. sp. was found infecting the spangled emperor Lethrinus nebulosus (n = 75) with higher prevalence from Dibba, Musandam (Gulf of Oman) than in Dhofar Salalah (Indian Ocean), Oman (p = 0.03). The general morphology of E. percussa n. sp. resembles E. caballeroi, E. chironemi and E. spari, which exhibit pre-equatorial testes. However, E. percussa n. sp. shows unique morphological characteristics distinguishing from congeneric species: the large hamuli bear notch allocated externally between the first half proximal of the root, and the small hamuli exhibit semicircular shape with undivided roots. Phylogenetic relationships within the Encotyllabe genus remain unresolved. However, the tree topology with the 28S showed overall consistency with the principal component analysis arrangement (PCA) derived from the morphological analysis. Which showed that the large and small hamuli, marginal hooks, ovary, testes (length and width) and peduncle are currently the most important morphological traits within the genus. Cytochrome c oxidase subunit I (COI) gene fragment showed high interspecific genetic divergence adding unambiguous resolution to discriminate/designate species identity. Interrelations within the genus support the identity of Encotyllabe percussa as a new species. This is the first species characterized with three gene fragments, the second congeneric species described in L. nebulosus and the first recorded in Oman.

A chromosome-scale genome assembly of Timorese crabgrass (Digitaria radicosa): a useful genomic resource for the Poaceae.

Timorese crabgrass (Digitaria radicosa) is a grass species commonly found in Southeast Asia and Oceania. Digitaria species have high intraspecific and interspecific genetic and phenotypic diversity, suggesting their potential usefulness as a genetic resource. However, as the only high-quality reference genome available is for a tetraploid Digitaria species, a reference genome of the diploid species D. radicosa would be a useful resource for genomic studies of Digitaria and Poaceae plants. Here, we present a chromosome-level genome assembly of D. radicosa and describe its genetic characteristics; we also illustrate its usefulness as a genomic resource for Poaceae. We constructed a 441.6 Mb draft assembly consisting of 61 contigs with an N50 contig length of 41.5 Mb, using PacBio HiFi long reads. We predicted 26,577 protein-coding genes, reaching a BUSCO score of 96.5%. To demonstrate the usefulness of the D. radicosa reference genome, we investigated the evolution of Digitaria species and the genetic diversity of Japanese Digitaria plants based on our new reference genome. We also defined the syntenic blocks between D. radicosa and 2 Poaceae crops, fonio and rice, and the diverse distribution of representative resistance genes in D. radicosa. The D. radicosa reference genome presented here should help elucidate the genetic relatedness of Digitaria species and the genetic diversity of Digitaria plants. In addition, the D. radicosa genome will be an important genomic resource for Poaceae genomics and crop breeding.

Comparative Plastomics of Plantains (Plantago, Plantaginaceae) as a Tool for the Development of Species-Specific DNA Barcodes.

Plantago (plantains, Plantaginaceae) is a cosmopolitan genus including over 250 species used as functional foods, forage, and traditional medicine. Among them, Plantago lanceolata is commonly used as an ingredient of herbal products, but the close similarity to other Plantago species can cause misidentifications with potentially serious consequences for product safety/quality. To test the possibility of developing species-specific barcoding markers, we de novo assembled plastome sequences of individuals of Plantago argentea, Plantago atrata, P. lanceolata, and Plantago maritima. These genomes were characterized in comparison with both previously sequenced conspecific accessions and other publicly available plastomes, thus providing an assessment of both intraspecific and interspecific genetic variation in Plantago plastomes. Additionally, molecular evolutionary analyses indicated that eleven protein-coding genes involved in different plastid functions in Plantago plastomes underwent positive selection, suggesting they might have contributed to enhancing species' adaptation during the evolutionary history of Plantago. While the most variable mutational hotspots in Plantago plastomes were not suitable for the development of species-specific molecular markers, species-specific polymorphisms could discriminate P. lanceolata from its closest relatives. Taken together, these results highlight the potential of plastome sequencing for the development of molecular markers to improve the identification of species with relevance in herbal products.

Morphological description and genetic analysis of a new black fly species (Diptera: Simuliidae) in the subgenus Asiosimulium from central Thailand

BackgroundBlack flies are among the most medically and veterinary important insects, as adult females of certain species are the sole vector of Onchocerca volvulus. Here, a new black fly species belonging to the subgenus Asiosimulium Takaoka & Choochote, 2005, is described and formally named as Simulium (Asiosimulium) kittipati sp. nov.MethodsPupae and larvae of black flies were collected from available substrates in the stream from central Thailand. Pupae were individually separated in plastic tubes and maintained until adult flies emerged. The emerged adult flies associated with their pupal exuviae and cocoon as well as mature larvae preserved in 85% ethanol were used to describe the new species based on an integrated approach of morphological examination and molecular analysis of the COI gene.ResultsThe new species is characterized in the female by the medium-long sensory vesicle with a medium-sized opening apically, scutum with three faint longitudinal vittae, and the ellipsoidal spermatheca; in the male by the number of upper-eye (large) facets in 20 vertical columns and 21 horizontal rows, hind basitarsus slender, nearly parallel-sided, and median sclerite much wider and upturned apically; in the pupa by the head and thoracic integument densely covered with tiny tubercles, and the pupal gill of arborescent type with 28–30 filaments; and in the larva by the postgenal cleft deep, nearly reaching the posterior margin of the hypostoma, and dark pigmented sheath of the subesophageal ganglion. The DNA barcode successfully differentiated the new species from its congeners with an interspecific genetic divergence of 1.74–18.72%, confirming the morphological identification that the species is a new member of the subgenus Asiosimulium. Phylogenetic analyses also indicated that the new species is genetically closely related to Simulium phurueaense Tangkawanit, Wongpakam & Pramual, 2018, further supporting its morphological classification.ConclusionsThis is the ninth species assigned to the subgenus Asiosimulium within the genus Simulium Latreille, 1802. Taxonomic notes and identification keys are given to distinguish this new species from the eight known species members in its same subgenus. Additionally, a distribution map of all species members in this subgenus occurring in Thailand and other countries is provided.Graphical abstract

Redescription and Molecular Characterization of the External Attaching Fish Parasitic Cymothoid, Nerocila phaiopleura Bleeker, 1857 (Crustacea: Isopoda) off the Southwest Coast of India.

The identification of the external attaching fish parasitic cymothoid, Nerocila phaiopleura Bleeker 1857, is still based on the brief description of Australian specimens provided by Bruce (1987). The present study aimed to provide a redescription and molecular characterisation of Indian specimens of N. phaiopleura. Morphological identification was carried out based on microscopic examinations and taxonomic drawings. mitochondrial DNA cox1 was selected as the target gene for sequencing and molecular identification. Nucleotide genetic divergence (p-distance) and base-pair differences among the different species were determined using MEGA11. Nerocila phaiopleura can be well separated from its congeners by the following combination of characteristics: Body about 2.4 times as long as wide, cephalon broadly rounded anteriorly; coxae posteriorly directed, acute and extending beyond their corresponding pereonite; pereonite 7 posterior angle produced, extending to the pleonite 1; pleonites 1 and 2 ventrolateral process posteriorly directed; uropod exopod straight and elongate, 1.7-2.0 times longer than endopod; uropod endopod lateral margin not serrate, no notch on medial margin; pereopods with short ischium; pleotelson triangular. The p-distance among N. phaiopleura and other available Nerocila spp. ranged from 21 to 19%. This study represents the first detailed taxonomic redescription of Indian specimens of N. phaiopleura. Key taxonomic features of the life stages and molecular data are provided here to identify the species properly. Interspecific genetic divergence between N. phaiopleura and other Nerocila spp. is assessed for the first time. Studies in cymothoid life histories, genetics, and morphology are necessary to understand one of the least understood parasite families.

Population genetics of two sibling species of fiddler crabs (Decapoda, Ocypodidae): Brazilian homogeneity and West Atlantic heterogeneity

Abstract Fiddler crabs are a widespread group of marine invertebrates. Their wide distribution is due to the great capacity of their larvae to disperse among estuarine and shelf waters. Therefore, it is expected that all populations along their distribution show molecular similarities due to extensive gene flow. Thus, we assessed the intra- and interspecific genetic divergence of two sibling species of fiddler crabs, Minuca panema and Minuca mordax, based on COI mtDNA sequences. The molecular data resulted in a highly mixed haplotype network for M. panema, while M. mordax sequences were clustered in two groups, one with Brazilian and other with Costa Rican haplotypes. The highest variance in M. panema was found within populations, while in M. mordax it was between groups and populations. The analyses of nucleotide divergence showed small genetic variance within populations of both species and between Brazilian populations, and higher variance between Brazilian and Costa Rican populations of M. mordax. The highest genetic divergence was found between the two species. Thereby, our results show an intense gene flow along the Brazilian coast, while there is a clear divergence among Brazil and Costa Rica, which may be a result of the great distance between these populations.

A First Approach for the In Vitro Cultivation, Storage, and DNA Barcoding of the Endangered Endemic Species Euonymus koopmannii.

Euonymus koopmannii is a rare and protected species in Kazakhstan, valued for its ecological role in soil stabilization and its ornamental properties. This study presents the first use of micropropagation and phylogenetic analysis for the endemic plant E. koopmannii. Seedlings of E. koopmannii proved to be more effective than internodes as primary explants for plant micropropagation of in vitro culture, with a multiplication coefficient of 28.5 from seedlings and 6.1 from internodes. On MSR I medium supplemented with 0.5 mg/L IBA and 0.05 mg/L IAA, a higher success rate of 67% was achieved for root formation of test tube-grown E. koopmannii plants. Using mannitol as an osmotic agent at a concentration of 8 mg/L prolonged the storage time of E. koopmannii under slow growth conditions when compared to CCC and abscisic acid. Phylogenetic relationships and species identification were analyzed using four DNA-barcoding markers, comparing E. koopmannii with species from NCBI. All candidate barcoding markers showed sufficient levels of interspecific genetic variation among Euonymus species. In addition, ITS region and rbcL gene sequences effectively distinguished E. koopmannii from other species. These results provide fundamental information that will be valuable for future biotechnological and molecular studies.

A taxonomic review of the order Mantodea in Korea based on morphology and DNA barcodes.

A taxonomic study of Korean Mantodea using morphological and molecular characters (COI) is presented. Eight species [Amantisnawai (Shiraki, 1908), Acromantisjaponica Westwood, 1889, Mantisreligiosasinica Bazyluk, 1960, Statiliamaculata (Thunberg, 1784), Tenoderaangustipennis Saussure, 1869, T.sinensis Saussure, 1871, Hierodulachinensis Werner, 1929, H.patellifera (Audinet-Serville, 1838)] belonging to six genera in three families are recognized. Interspecific genetic divergence of COI using uncorrected p-distance ranged from 6.7% to 22.4%, while intraspecific divergence ranged from 0% to 2.2% among eight Korean Mantodea species. All eight species were each strongly supported as a single lineage using COI on both neighbor-joining and parsimony trees. An illustrated key, redescriptions, habitus photographs, and illustrations of diagnostic characters of the species of Korean Mantodea are provided to facilitate identification.

DNA Barcoding of Economically Important Fruit Flies (Diptera: Tephritidae) from the Lower Gangetic Plains of Eastern India

The Lower Gangetic Plains are India's central horticultural crop production regions. Fruit flies (Diptera: Tephritidae) are a significant insect pest that significantly hinders the effective crop production of various commercially valuable fruit and vegetable crops. Therefore, rapid and precise species identification techniques are essential to controlling, managing, and quarantining these pests. Furthermore, reliably identifying fruit flies is exceedingly challenging due to their striking resemblances in morphological characters. In this work, we investigated the effectiveness of mitochondrial cytochrome c oxidase I (COI) gene sequences for identifying seven fruit fly species, involving only the genus Bactrocera from the lower Gangetic Plains in eastern India. The sequences of the studied species showed 99%–100% similarity with sequences from within and outside the country. COI barcodes were able to distinguish between species, with K2P intraspecific genetic divergences ranging from 0-1% and K2P interspecific genetic divergences ranging from 3.65-28.46%, thus maintaining a proper barcode gap. Phylogenetic analyses were performed, and the results showed moderate to high supported monophyly among species. Therefore, COI barcodes have proven a highly successful alternative for quickly identifying fruit flies. This is also the first account of molecular identification of the previously reported tephritid fruit flies infesting fruits and vegetables in eastern India's agriculturally important lower Gangetic Plains.

DNA barcoding for identification of forensically important synanthropic flesh flies (Diptera: Sarcophagidae) from Eastern India

ABSTRACT Sarcophagidae is a significant Diptera family because several of its species are among the first to visit a corpse, and it helps in minPMI estimation. Sarcophagidae species identification for a layman is difficult due to identical morphological appearance and if the fly is a female. Also, fly specimens get damaged due to improper storage of forensic specimens collected from crime scenes. Thus, in this study, we aimed to check the efficiency of cytochrome oxidase 1 (COI) for identifying eleven sarcophagid flies species. Flesh flies were collected from various biogeographic regions of West Bengal. COI barcodes successfully differentiated between species with a very significant K2P interspecific genetic divergence, which ranged between 3.12% and 20.11%, and intraspecific genetic divergences ranged from 0% to 1%. The subgenera Uroxanthisca and Sinonipponia are used in the phylogenetic analysis for the first time in the oriental context. Phylogenetic analysis was performed using a Neighbour Joining (NJ) tree constructed using Kimura two-parameter (K2P) in MEGA X software. Thus, COI barcodes proved to be quite effective as an alternate method of identifying flesh fly species, and future studies should include additional markers for further phylogenetic investigations of the Sarcophagidae family.

Evolution of a novel adrenal cell type that promotes parental care.

Cell types with specialized functions fundamentally regulate animal behaviour, and yet the genetic mechanisms that underlie the emergence of novel cell types and their consequences for behaviour are not well understood1. Here we show that the monogamous oldfield mouse (Peromyscus polionotus) has recently evolved a novel cell type in the adrenal gland that expresses the enzyme AKR1C18, which converts progesterone into 20α-hydroxyprogesterone. We then demonstrate that 20α-hydroxyprogesterone is more abundant in oldfield mice, where it induces monogamous-typical parental behaviours, than in the closely related promiscuous deer mice (Peromyscus maniculatus). Using quantitative trait locus mapping in a cross between these species, we ultimately find interspecific genetic variation that drives expression of the nuclear protein GADD45A and the glycoprotein tenascin N, which contribute to the emergence and function of this cell type in oldfield mice. Our results provide an example by which the recent evolution of a new cell type in a gland outside the brain contributes to the evolution of social behaviour.

Geometric morphometrics to differentiate species and explore seasonal variation in three Mansonia species (Diptera: Culicidae) in central Thailand and their association with meteorological factors.

Mansonia mosquito species are recognised as a significant vector of human pathogens, primarily transmitting the filarial nematode, Brugia malayi. In central Thailand, the three most prevalent Mansonia species are Mansonia annulifera, Mansonia indiana and Mansonia uniformis. This study explored the influence of seasonal changes on the phenotypic variation of these Mansonia species in central Thailand using the geometric morphometrics (GM). To ensure accurate species identification, we integrated GM techniques with DNA barcoding, examining distinctions in both phenotype and genotype among the species. The intraspecific genetic divergence ranged from 0.00% to 1.69%, whereas the interspecific genetic divergence ranged from 10.52% to 16.36%. The clear distinction between intra- and interspecific distances demonstrated the presence of a barcoding gap, confirming the successful differentiation of the three Mansonia mosquito species through DNA barcoding. Similarly, the interspecies GM assessment for classifying Mansonia species demonstrated a high degree of accuracy, with an overall performance of 98.12%. Exploring seasonal variation in the three Mansonia species revealed wing variations across different seasons, and pronounced variations appearing in the cool season. Regarding their association with meteorological factors, Ma. annulifera and Ma. uniformis showed significant positive correlations with temperature (p < 0.05), and Ma. uniformis also displayed a significant negative correlation with atmospheric pressure (p < 0.05). The insights from this study will deepen our understanding of the adaptive patterns of Mansonia mosquitoes in Thailand's central region, paving the way for enhanced disease surveillance related to these vectors.

DNA Barcoding Supports "Color-Pattern''-Based Species of Stictochironomus from China (Diptera: Chironomidae).

The genus Stictochironomus (Diptera: Chironomidae) has an almost worldwide distribution, with more than 30 species. However, species delimitation and identification based on the markings on the wings and legs are controversial and uncertain. In this study, we focused on color patterns to review the adults of the genus from China, and two new species (S. trifuscipes sp. nov. and S. quadrimaculatus sp. nov.) are described and figured. DNA barcodes can accurately separate the two new species with specific color patterns. However, heterospecific individuals form a monophyletic cluster in the phylogeny tree. For example, S. maculipennis (Meigen) and S. pictulus (Meigen), which have a lower interspecific genetic divergence, form a single clade. Sequences with the same species name but with high intraspecific distance form more than one phylogenetic clade, such as S. sticticus (Fabricius) of three clades, S. pictulus of four clades, S. akizukii (Tokunaga) and S. juncaii Qi, Shi, and Wang of two clades, might have potential cryptic species diversity. Species delimitation analysis using ASAP, PTP, and GMYC clearly delineated them as separate species. Consequently, color patterns are a good diagnostic characteristic for species delimitation in Stictochironomus. The distance-based analysis shows that a threshold of 4.5-7.7% is appropriate for species delimitation in Stictochironomus. Additionally, an updated key including color pattern variation for male adults of known Stictochironomus species from China is provided.

The value of hybrid genomes: Building two highly contiguous reference genome assemblies to advance Canis genomic studies.

Previous studies of canid population and evolutionary genetics have relied on high-quality domestic dog reference genomes that have been produced primarily for biomedical and trait mapping studies in dog breeds. However, the absence of highly contiguous genomes from other Canis species like the gray wolf and coyote, that represent additional distinct demographic histories, may bias inferences regarding interspecific genetic diversity and phylogenetic relationships. Here, we present single haplotype de novo genome assemblies for the gray wolf and coyote, generated by applying the trio-binning approach to long sequence reads generated from the genome of a female first-generation hybrid produced from a gray wolf and coyote mating. The assemblies were highly contiguous, with contig N50 sizes of 44.6 and 42.0 Mb for the wolf and coyote, respectively. Genome scaffolding and alignments between the two Canis assemblies and published dog reference genomes showed near complete collinearity, with one exception: a coyote-specific chromosome fission of chromosome 13 and fusion of the proximal portion of that chromosome with chromosome 8, retaining the Canis-typical haploid chromosome number of 2n = 78. We evaluated mapping quality for previous RADseq data from 334 canids and found nearly identical mapping quality and patterns among canid species and regional populations regardless of the genome used for alignment (dog, coyote, or gray wolf). These novel wolf and coyote genome reference assemblies will be important resources for proper and accurate inference of Canis demography, taxonomic evaluation, and conservation genetics.

DNA barcoding of Notopterygii Rhizoma et Radix (Qiang-huo) and identification of adulteration in its medicinal services

Safety concerns, stemming from the presence of complex and unpredictable adulterants, permeate the entire industrial chain of traditional Chinese medicines (TCMs). The Notopterygii Rhizoma et Radix (NReR) from the Apiaceae family, commonly known as “Qiang-huo”, is a widely used herbal medicine. The recent surge in its demand has given rise to a proliferation of counterfeit and substituted products in the market. Traditional identification presents inherent limitations, while DNA mini-barcoding, reliant on sequencing a short-standardized region, has received considerable attention as a new potential means to identify processed medicinal materials. In this study, we constructed a comprehensive Internal Transcribed Spacer 2 (ITS2) matrix encompassing genuine NReR and their commonly found adulterants for the first time. Leveraging this matrix, we conducted a thorough assessment of the genetic profiles and sources of NReR available in the Chinese herbal medicine market. Following established DNA barcoding protocols, the intra-specific genetic divergences within NReR species were found to be lower than the inter-specific genetic divergences from other species. Among the 120 samples that were successfully amplified, ITS2 exhibits an outstanding species-level identification efficiency of 100% when evaluated using both the BLASTN and neighbor-joining (NJ) tree methods. We concluded that ITS2 is a mini-barcode that has shown its potential and may become a universal mini-barcode for the quality control of “Qiang-huo”, thereby ensuring the safety of clinical medication.

A Community-Based Framework Integrates Interspecific Interactions into Forest Genetic Conservation

Forest genetic conservation is typically species-specific and does not integrate interspecific interaction and community structure. It mainly focuses on the theories of population and quantitative genetics. This approach depicts the intraspecific patterns of population genetic structure derived from genetic markers and the genetic differentiation of adaptive quantitative traits in provenance trials. However, it neglects possible interspecific interaction in natural forests and overlooks natural hybridization or subspeciation. We propose that the genetic diversity of a given species in a forest community is shaped by both intraspecific population and interspecific community evolutionary processes, and expand the traditional forest genetic conservation concept under the community ecology framework. We show that a community-specific phylogeny derived from molecular markers would allow us to explore the genetic mechanisms of a tree species interacting with other resident species. It would also facilitate the exploration of a species’ ecological role in forest community assembly and the taxonomic relationship of the species with other species specific to its resident forest community. Phylogenetic β-diversity would assess the similarities and differences of a tree species across communities regarding ecological function, the strength of selection pressure, and the nature and extent of its interaction with other species. Our forest genetic conservation proposal that integrates intraspecific population and interspecific community genetic variations is suitable for conserving a taxonomic species complex and maintaining its evolutionary potential in natural forests. This provides complementary information to conventional population and quantitative genetics-based conservation strategies.

Integrated Identification and Genetic Diversity of Potentially Invasive Clearwing Moths (Lepidoptera: Cossoidea: Sesiidae) in Korea.

The populations of clearwing moth borers in Korea have recently caused extensive and severe damage to pin oaks (Quercus palustris Munchh.). We conducted field monitoring and molecular analyses to identify them in an integrated manner. Morphological examination and molecular analyses of the COI gene, based on intra- and interspecific genetic divergences (GDs), revealed that the borers were identified as two invasive species, Sphecodoptera sheni and Paranthrenella pinoakula sp. nov. The maximum intraspecific GD was found to be 1.9%, whereas the minimum interspecific GD was confirmed as 8.1%, indicating a distinct barcoding gap. Both the MJ network and NJ tree also showed that 18 haplotypes (Hs) were detected from the 52 COI sequences. The borers revealed a total of 17 Hs: (i) H1-H7 were detected in all seven regions with S. sheni; (ii) Wonju and Goyang populations of S. sheni revealed more than three Hs; (iii) H7 was closely connected with H8 of the Chinese population of S. sheni; (iv) H9-H10 were detected in other samples from the Wonju population with P. pinoakula sp. n. and were closely located with congeneric species. A maximum likelihood tree also revealed that P. pinoacula sp. n. nested within the congeneric species, genetically separating from S. sheni.

Morphological differences yet genetic similarity among seasonal cohorts of Japanese anchovy during the early life stages in the Kii Channel

Abstract Species in the genus Engraulis show extensive intraspecific as well as interspecific morphological and genetic diversity. Since morphological differences do not necessarily correspond to genetic differences, it is necessary to clarify the relationship between morphological differences and genetic differences for a better understanding of the population structure. Fish morphology at a given standard length differs between cohorts of Japanese anchovy Engraulis japonicus during the early life stages in the Kii Channel, but it is unknown whether the differences are caused by genetic differences or not. The Kii Channel includes the boundary between the Pacific (southern side of the Kii Channel) and the Seto Inland Sea stocks (northern side), but stock separation is based primarily on demographic characteristics. In the present study, genetic analyses were conducted to examine the genetic differences among samples (month and area) based on mitochondrial DNA cytochrome b region (Cyt b), control region (CR) and microsatellite markers. AMOVA revealed that the percentage of genetic variation among samples was low at 0.11% for Cyt b, 0.30% for CR and 0.00% for microsatellite, and no significant genetic variation was observed among samples. Although two clades were identified in the unrooted neighbour-joining tree for Cyt b and CR, both Cyt b and CR sequences were similar between months and between areas. Accordingly, the morphological differences among cohorts can be attributed to phenotypic plasticity. Additionally, there were no genetic differences between samples from the southern side and the northern side of the Kii Channel, suggesting strong genetic connectivity in these areas.

DNA barcoding and phylogenetic relationships of ten butterfly caterpillars

Cytochrome C oxidase subunit I (COI), known as DNA barcodes, can be employed for the identification of an unknown specimen at the species level. The present study was aimed at performing COI gene-based identification of butterflies using butterfly caterpillars. Consecutively, ten caterpillars of ten different butterfly species from the families Papilionidae, Nymphalidae, Pieridae, Lycaenidae, and Hesperiidae were taken from respective host plants and used to generate COI gene sequences that were about 650 bp long. After BLAST analysis, the sequenced gene revealed 96–100% similarity to the same species from different regions. Then, sequences were submitted to NCBI's GenBank and obtained ten accession numbers. In order to elucidate genetic diversity and evolutionary relationships among ten species of butterfly caterpillars, pairwise distance analysis and the construction of a phylogenetic tree were performed using MEGA10 and BioEdit software. In the analysis, the interspecific genetic divergence among the caterpillars of butterflies was between 0.101-0.164%. A phylogenetic tree was constructed with the assistance of the Neighbor-Joining (NJ) algorithm, which identified two major clades, A and B, as well as indicated that butterflies caterpillars shared a common ancestor. All the species are included in these two clades, except Catopsilia pomona. This species is under the family Pieridae, and the phylogenetic position of Pieridae compared to other butterfly families is ambiguous, necessitating additional research to resolve this issue. The study demonstrated the general applicability of DNA barcoding for rapidly and accurately distinguishing butterfly species, even when using larval stages, as in the present study. Furthermore, it may disclose a higher taxonomic hierarchy of butterfly families. Asian Australas. J. Biosci. Biotechnol. 2023, 8(3), 49-55

Phylogeny, biogeography, and integrative taxonomic revision of the Afro-Arabian rodent genus Ochromyscus (Muridae: Murinae: Praomyini)

Abstract The analyses of Plio-Pleistocene speciation processes in the Horn of Africa are relevant for understanding the evolution of biodiversity patterns of this understudied part of the world. Here we analyse comprehensive genomic and morphological data of the recently delimited murid genus Ochromyscus, one of the few with Afro-Arabian distribution. Using an integrative taxonomic approach, we delimit three species in the genus: two in eastern Africa (O. brockmani and O. niveiventris) and one in southern Arabia (O. yemeni), and detail their distribution, genetic structure, and evolutionary history. Despite their morphological similarity, the three species split before the Pleistocene, and their interspecific genetic divergence level is comparable to that between sister genera of murine rodents. The split between two taxa living on opposite sides of the Red Sea (O. brockmani vs. O. yemeni) is younger than the separation of their ancestor and O. niveiventris living in eastern Africa. The colonization of Arabia can be explained either by the presence of a relatively recent continental bridge or by the past occurrence of the genus along the eastern coast of the Red Sea and subsequent spreading through the Sinai Peninsula.