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Invasive alien plants represent one of the five major threats to biodiversity and the disruption of ecosystems. They are introduced through various routes, starting with commercial trade. Preventing their introduction is essential to avoid the spread of new invasive plants. In this paper, we propose a new early warning DNA barcoding tool for invasive plant detection. Eight invasive alien species of European Union concern (i.e., Ludwigia grandiflora, Elodea nuttallii, Myriophyllum aquaticum, Pontederia crassipes, Ailanthus altissima, Heracleum mantegazzianum, Impatiens glandulifera, Pueraria montana) were selected and analysed. A unique DNA marker for each species was identified and amplified using species-specific primers capable of identifying the presence of alien species. To verify whether the approach could detect the presence of alien plants in urban areas from lawn clippings, mixes with typical urban spontaneous plants and invasive species were tested. In all mixes, only the invasive species was identified. This rapid detection capability will enable environmental operators to intervene promptly to contain the spread of invasive plants before they can cause significant damage to the local ecosystem. This tool could have a significant impact on the protection of local biodiversity and the integrity of urban habitats. [ABSTRACT FROM AUTHOR]

A partial mitochondrial DNA Cytochrome Oxidase subunit I (mtCOI) gene haplotype variant of the coconut rhinoceros beetle (CRB) Oryctes rhinoceros, classed as 'CRB-G (clade I)', has been the focus of much research since 2007, with reports of invasions into new Pacific Island locations (e.g., Guam, Hawaii, Solomons Islands). For numerous invasive species, inference of invasion biology via whole genome is superior to assessments via the partial mtCOI gene. Here, we explore CRB draft mitochondrial genomes (mitogenomes) from historical and recent collections, with assessment focused on individuals associated within the CRB-G (clade I) classification. We found that all Guam CRB individuals possessed the same mitogenome across all 13 protein-coding genes and differed from individuals collected elsewhere, including 'non-Guam' individuals designated as CRB-G (clade I) by partial mtCOI assessment. Two alternative ATP6 and COIII partial gene primer sets were developed to enable distinction between CRB individuals from Guam that classed within the CRB-G (clade I) haplotype grouping and CRB-G (Clade I) individuals collected elsewhere. Phylogenetic analyses based on concatenated ATP6–COIII genes showed that only Guam CRB-G (clade I) individuals clustered together, and therefore Guam was not the source of the CRB that invaded the other locations in the Pacific assessed in this study. The use of the mtCOI and/or mtCOIII genes for initial molecular diagnosis of CRB remained crucial, and assessment of more native CRB populations will further advance our ability to identify the provenance of CRB invasions being reported within the Pacific and elsewhere. [ABSTRACT FROM AUTHOR]

The Asian hornet, Vespa velutina, is an invasive species that has not only expanded its range in Asia but has also invaded European countries, and it incurs significant costs on local apiculture. This phylogeographic study aims to trace the evolutionary trajectory of V. velutina and its close relatives; it aims to identify features that characterize an invasive species. The last successful invasion of Vespa velutina into France occurred in late May, 2002, and into South Korea in early October, 2002, which were estimated by fitting a logistic equation to the number of observations over time. The instantaneous rate of increase is 1.3667 for V. velutina in France and 0.2812 in South Korea, which are consistent with the interpretation of little competition in France and strong competition from local hornet species in South Korea. The invasive potential of two sister lineages can be compared by their distribution area when proper statistical adjustments are made to account for differences in sample size. V. velutina has a greater invasive potential than its sister lineage. The ancestor of V. velutina split into two lineages, one found in Indonesia/Malaysia and the other colonizing the Asian continent. The second lineage split into a sedentary clade inhabiting Pakistan and India and an invasive lineage colonizing much of Southeast Asia. This latter lineage gave rise to the subspecies V. v. nigrithorax, which invaded France, South Korea, and Japan. My software PGT version 1.5, which generates geophylogenies and computes geographic areas for individual taxa, is useful for understanding biogeography in general and invasive species in particular. I discussed the conceptual formulation of an index of invasiveness for a comparison between sister lineages. [ABSTRACT FROM AUTHOR]

Edible mushroom products, encompassing both cultivated and wild varieties, are highly favored by consumers due to their rich nutritional profiles, including significant levels of proteins and amino acids. These mushrooms have extensive applications across the food, pharmaceutical, and cosmetic industries, making the edible mushroom industry a vital component of global poverty alleviation efforts. Taking China as an example, the country produces over 45 million tons of edible mushrooms annually, accounting for 94.01% of the world's total production, thereby establishing itself as the leading global producer of edible mushrooms. However, alongside the rapid expansion of this industry, concerns have emerged regarding counterfeit products and incidents of poisoning resulting from the consumption of toxic wild mushrooms. As follows, to advance the development and integrity of the mushroom production and processing industry: (1) This study presents the situation of counterfeit edible mushrooms and elucidates the factors contributing to the production of fraudulent products from both subjective and non-subjective perspectives. (2) We provide a detailed introduction to 22 varieties of freshly cultivated edible mushrooms and commonly encountered wild edible mushrooms in the Chinese consumer market, proposing the application of DNA barcoding, environmental DNA analysis, and other technologies for the future authentication of counterfeit mushroom products. (3) Concurrently, we present an overview of mushroom poisoning incidents in China from 2010 to 2023, emphasizing the challenges in mitigating the risks associated with wild mushroom consumption and preventing food poisoning, thereby necessitating heightened consumer caution. (4) Finally, we offer four recommendations aimed at ensuring the healthy, stable, and sustainable growth of the edible mushroom industry. [ABSTRACT FROM AUTHOR]

The genus Cernotina Ross, 1938 is represented in the southeastern United States by three nominal species: Cernotina calcea Ross, 1938, Cernotina spicata Ross, 1938, and Cernotina truncona Ross, 1947. Of all Cernotina species, only the larva of C. spicata has been described to date. The goal of this paper is to describe, illustrate, and diagnose the larva of C. truncona using ecologically associated specimens. In addition, we used publicly available mitochondrial DNA barcoding data to evaluate the genetic relationships of these species. The larvae of Cernotina truncona can be distinguished from those of C. spicata by differences in setal placement and number on the meso- and metanota, mandibular morphology, head width, and distal setation of the tarsi. The ultrastructure of the anal claw is figured, highlighting the novel finding of small spines on the concave margin of a larva of Cernotina. With this new description, just the second described larva from this genus, only C. calcea remains unknown in the southeastern United States. The information provided herein enables the in-depth study of the ecology and life history of this diminutive caddisfly. [ABSTRACT FROM AUTHOR]

Simple Summary: Correct taxonomic identification is essential for conducting successful biological research, especially with regard to economically important insects, such as jumping plant lice or psyllids. In the present study, we identify and diagnose the morphologically characterised aphalarid species from Bulgaria using two molecular markers, cytochrome c oxidase I and cytochrome b. A total of 80 sequences of 25 Aphalaridae species were obtained and included in the BOLD and GenBank databases. This should enable even non-experts to identify these species quickly and accurately. The results of the current study show that two barcode genes are sufficient to distinguish most aphalarid species. Psyllids (Hemiptera: Psylloidea) are plant sap-sucking insects whose identification is often difficult for non-experts. Despite the rapid development of DNA barcoding techniques and their widespread use, only a limited number of sequences of psyllids are available in the public databases, and those that are available are often misidentified. Here, we provide 80 sequences of two mitochondrial genes, cytochrome c oxidase I (COI) and cytochrome b (Cytb), for 25 species of Aphalaridae, mainly from Bulgaria. The DNA barcodes for 15 of these species are published for the first time. In cases where standard primers failed to amplify the target gene fragment, we designed new primers that can be used in future studies. The distance-based thresholds for the analysed species were between 0.0015 and 0.3415 for COI and 0.0771 and 0.4721 for Cytb, indicating that the Cytb gene has a higher interspecific divergence, compared to COI, and therefore allows for more accurate species identification. The species delimitation based on DNA barcodes is largely consistent with the differences resulting from morphological and host plant data, demonstrating that the use of DNA barcodes is suitable for successful identification of most aphalarid species studied. The phylogenetic reconstruction based on maximum likelihood and Bayesian inference analyses, while showing similar results at high taxonomic levels to previously published phylogenies, provides additional information on the placement of aphalarids at the species level. The following five species represent new records for Bulgaria: Agonoscena targionii, Aphalara affinis, Colposcenia aliena, Co. bidentata, and Craspedolepta malachitica. Craspedolepta conspersa is reported for the first time from the Czech Republic, while Agonoscena cisti is reported for the first time from Albania. [ABSTRACT FROM AUTHOR]

Narragansett Bay is an estuarine system in the western North Atlantic Ocean that harbors a diverse marine flora, providing structure, habitat, and food for native biodiversity. This area has been the center of numerous environmental, biological, ecological, and oceanographic studies; however, marine macroalgae have not been extensively examined using modern molecular methods. Here, we document the biodiversity of the red algal order Ceramiales based on DNA sequence comparisons of the 3′ end of the RuBisCo large subunit (rbcL-3P) and the universal plastid amplicon (UPA). Thirty-seven distinct species of this order were identified and validated with molecular data, including five new species reports and at least one new report of an introduced species, Antithamnionella spirographidis, in the vicinity of Narraganset Bay. Novel sequence data were generated for numerous species, and it was discovered that the UPA marker, which has been less frequently used in red algal floristics, revealed an identical inventory of ceramialean algae as the rbcL-3P marker. Thus, the shorter length of the UPA marker holds promise for DNA metabarcoding studies that seek to elucidate biodiversity across algal phyla. [ABSTRACT FROM AUTHOR]

Non-indigenous species (NIS) introduction notoriously threatens the Mediterranean Sea. In addition, Mediterranean coastal lagoons play a crucial role as nurseries for marine species, which new NIS arrivals can threaten. Therefore, monitoring and early warning of NIS presence is essential in preserving biodiversity. An innovative technique for rapid and accurate species identification and biodiversity screening is the application of environmental DNA (eDNA) metabarcoding. In this research, different Penaeidae (Arthropoda, Crustacea, Decapoda) NIS specimens were collected from a Mediterranean coastal lagoon after an early warning about a potentially invasive NIS arising from next-generation sequencing data. DNA barcoding of the DNA extracted from tissue samples and amplified with specifically designed primer pairs led to the recognition of Penaeus aztecus in this NATURA 2000 protected ecosystem for the first time. DNA barcoding from DNA isolated from the water where the living specimens were stored further validated the possibility of identifying P. aztecus starting from eDNA. This approach demonstrated the validity of environmental DNA analysis in the early screening of potentially invasive NIS presence in Mediterranean protected areas and ecosystems. This work describes an applicative example of the efficacy in improving the biomonitoring of lagoon ecosystems using molecular tools and it represents a guideline for the validation of eDNA metabarcoding data for the presence of potentially invasive species. [ABSTRACT FROM AUTHOR]

Pyropia yezoensis is the most widely cultivated and economically important alga. Affected by climate change, the cultivation of P. yezoensis has gradually migrated to the northern coast of China, increasing from 6.8% in 2019 to 19.5% in 2023. To date, the genetic impact of northern migration on cultivation resources has not been assessed and analyzed extensively. Here, DNA barcoding (rbcL and cox1) and the presence/absence of intronic-ORFs in mitochondrial regions (rnl and cox1) were applied to investigate genetic diversity in 44 P. yezoensis specimens from 17 aquaculture farms in China, with comparisons to Korean and Japanese cultivated resources. The lower intraspecific variation was 0.31% for the cox1 gene and 0.14% for the rbcL gene, with three haplotypes, indicating that intensive selection and breeding during cultivation had narrowed the germplasm genetic variation. The intron structure of mitochondrial regions showed that the cultivated resources had 17 phenotypes, and the northern specimens shared 35.3% of genotypes with the southern specimens, indicating that the cultivated P. yezoensis is expanding its cultivation ranges through north migration. Even with lower genetic diversity, the northern area of cultivation had already developed 17.6% site-specific specimens. The genetic diversity of cultivated P. yezoensis from the Northwest Pacific is also discussed. Our work provides a preliminary framework for P. yezoensis breeding and cultivation under climate change. [ABSTRACT FROM AUTHOR]

The genus Amomum includes over 111 species, 6 of which are widely utilized as medicinal plants and have already undergone taxonomic revision. Due to their morphological similarities, the presence of counterfeit and substandard products remains a challenge. Accurate plant identification is, therefore, essential to address these issues. This study utilized 11 newly sequenced samples and extensive NCBI data to perform molecular identification of the six medicinal "Doukou" species. The plastomes of these species exhibited a typical quadripartite structure with a conserved gene content. However, independent variation shifts of the SC/IR boundaries existed between and within species. The comprehensive set of genetic sequences, including ITS, ITS1, ITS2, complete plastomes, matK, rbcL, psbA-trnH, and ycf1, showed varying discrimination of the six "Doukou" species based on both distance and phylogenetic tree methods. Among these, the ITS, ITS1, and complete plastome sequences demonstrated the highest identification success rate (3/6), followed by ycf1 (2/6), and then ITS2, matK, and psbA-trnH (1/6). In contrast, rbcL failed to identify any species. This research established a basis for a reliable molecular identification method for medicinal "Doukou" plants to protect wild plant resources, promote the sustainable use of medicinal plants, and restrict the exploitation of these resources. [ABSTRACT FROM AUTHOR]

Snow Lotus Seed (SLS), esteemed for its nutritional and market value, faces challenges of authentication due to the absence of appropriate testing standards and methods. This results in frequent adulteration of SLS sourced from Gleditsia sinensis (G. sinensis) with other plant seeds endosperm. Traditional chloroplast DNA barcoding methods are inadequate for species identification due to the absence of chloroplasts in G. sinensis seeds endosperm. In this study, the homology of 11 ITS genes among 6 common Gleditsia species was analyzed. Universal primers suitable for these species were designed and screened. A DNA barcoding method for distinguishing SLS species was developed using Sanger sequencing technology, leveraging existing GenBank and Barcode of Life Data System (BOLD) databases. Optimized sample pretreatment facilitated effective DNA extraction from phytopolysaccharide-rich SLS. Through testing of commercial SLS products, the species origin has been successfully identified. Additionally, a novel instance of food fraud was uncovered, where the Caesalpinia spinosa endosperm was used to counterfeit SLS for the first time. The study established that the developed DNA barcoding method is effective for authenticating SLS species. It is of great significance for combating food fraud related to SLS, ensuring food safety, and promoting the healthy development of the SLS industry. [ABSTRACT FROM AUTHOR]

Rice cultivation is one of the most important agro-economic activities in many countries, and the correct seed storage between production cycles is essential for crop success. In Guinea-Bissau, mangrove swamp rice (MSR) is a highly productive rice cropping system, thus providing surplus for sale. Depending on storage conditions, rice grains may present moisture, insects, or the development of fungi that might affect crop productivity. Considering different rice varieties and storage conditions, samples were collected from 30 farmers within 13 villages across the main MSR-producing regions. Stored rice was used to isolate and identify seed-borne fungi through DNA barcoding, to test aflatoxin content, and to evaluate seed germination rates. Polyethylene bags (the container), raised wooden platforms, and storage rooms (the structures) are the most used facilities. Hermetic containers were recorded mainly in Oio. A total fungal richness of 18 genera was found, and 16 different species were identified. The most represented fungal genera are Aspergillus, Curvularia, and Bipolaris. Despite the presence of aflatoxin-producing fungal samples, they did not present concerning levels for human consumption, and the germination rate was not affected regardless of storage structures. These results provide a baseline on fungi occurrence in stored MSR seeds from traditional facilities in Guinea-Bissau. [ABSTRACT FROM AUTHOR]

Rhubarb is a significant medicinal herb in China. Its adulteration or fabrication is common in the market. Consequently, it is necessary to establish a comprehensive identification method to accurately identify genuine rhubarb and its adulterants. In this study, the sequences of chloroplast genes rps3-rpl22 and rpl16 from three genuine rhubarbs (Rheum tanguticum, Rh. palmatum and Rh. officinale) and their adulterants (Rumex japonicus and Rumex spp.) were amplified, sequenced and subjected to genetic analyses. The genetic distances for rps3-rpl22 and rpl16 between genuine rhubarbs and their adulterants showed that there was an evident barcoding gap, which allowed the adulterants to be distinguished from the genuine rhubarbs, as demonstrated by a neighbor joining tree. Additionally, Rh. officinale could be distinguished from the other two genuine rhubarbs. The anthraquinone, sennoside, polysaccharide and protein contents were analyzed in seven rhubarbs using high-performance liquid chromatography and ultraviolet spectrophotometry. Cluster and principal component analyses results showed that Rh. tanguticum and Rh. palmatum could be effectively distinguished. The study suggests that DNA barcoding based on rps3-rpl22 and rpl16 sequences coupled with multiple-indicator quantification can be successfully applied to identify rhubarb species and distinguish among the three genuine rhubarbs, and this can provide a scientific foundation for rhubarb quality assurance. [ABSTRACT FROM AUTHOR]

Simple Summary: Due to the instability of interspecific morphological characteristics and the significant convergence phenomenon, it is difficult to identify most species of Neotriplax based on morphology. Through integrated research on the reddish-brown species of Neotriplax, which feature many phenotypic similarities, and by comprehensively utilizing morphology, molecular phylogeny, and geometric morphometrics, the relationship among all these species was reconstructed. The taxonomic status within the genus was partly clarified, with three new species being identified: N. qinghaiensis sp. nov., N. maoershanensis sp. nov., and N. guangxiensis sp. nov. To date, five species of reddish-brown Neotriplax have been described, but their highly similar body color and other phenotypic traits make accurate taxonomy challenging. To clarify species-level taxonomy and validate potential new species, the cytochrome oxidase subunit I (COI) was used for phylogenetic analysis and the geometric morphometrics of elytron, pronotum, and hind wing were employed to distinguish all reddish-brown Neotriplax species. Phylogenetic results using maximum likelihood and Bayesian analyses of COI sequences aligned well with the current taxonomy of the Neotriplax species group. Significant K2P divergences, with no overlap between intra- and interspecific genetic distances, were obtained in Neotriplax species. The automatic barcode gap discovery (ABGD), assemble species by automatic partitioning (ASAP), and generalized mixed Yule coalescent (GMYC) approaches concurred, dividing the similar species into eight molecular operational taxonomic units (MOTUs). Geometric morphometric analysis using pronotum, elytron, hind wing shape and wing vein patterns also validated the classification of all eight species. By integrating these analytical approaches with morphological evidence, we successfully delineated the reddish-brown species of Neotriplax into eight species with three new species: N. qinghaiensis sp. nov., N. maoershanensis sp. nov., and N. guangxiensis sp. nov. Furthermore, we documented the first record of N. lewisii in China. This study underscores the utility of an integrative taxonomy approach in species delimitation within Neotriplax and serves as a reference for the taxonomic revision of other morphologically challenging beetles through integrative taxonomy. [ABSTRACT FROM AUTHOR]

The complete mitochondrial genomes of two Iphiclides species, namely I. podalirius and I. podalirinus, were sequenced, assembled, and reported in this article. Both genomes comprise 37 genes, with 13 protein-coding genes, 22 transfer RNA (tRNA) genes, and two ribosomal RNA (rRNA) genes. The gene orders and alignments agree with the reported mitogenomes of Leptocircini butterflies, while the start codon for the COX1 gene in I. podalirinus is CGA instead of the commonly seen ATN type. Codon preference shows that methionine and tryptophan are the poorest, while arginine, leucine, and serine are the richest. Phylogenetic analysis using Bayesian Inference shows both Iphiclides species are sister to the genus Lamproptera and are basal to all remaining Leptocircini species. The Kimura 2-parameter (K2P) distances of I. podalirinus from I. podalirius exceed 5%, demonstrating its solid species status. The K2P distance between the North African feisthamelii and podalirius exceeds 2%, indicating the reasonable elevation of I. feisthamelii to the full specific level as its type locality is Algeria. Future research is required to tackle the relationship between the Iberian feisthamelii and podalirius using more evidence. [ABSTRACT FROM AUTHOR]

Simple Summary: In Kazakhstan, tulips are not just beautiful flowers but also vital to the environment, with 35 species, 18 of which are protected. Recent research near Kazakhstan's borders uncovered new tulip varieties, highlighting the need for a species inventory. In this study, we identified eight tulip species using both physical traits and advanced DNA testing. By analyzing genetic markers, we discovered that certain DNA regions are particularly useful for identifying tulip species. However, when it comes to closely related tulips, combining data from multiple DNA markers is crucial for accurate classification. This study also unveiled possible natural hybrids, indicating complex interactions between different tulip species. This research sheds light on the diversity and evolutionary relationships among tulips, providing valuable insights for conservation efforts and understanding plant genetics. Further investigations into tulip populations are necessary to confirm these findings and better protect these precious flowers. In Kazakhstan, the genus Tulipa is represented by 35 species, 18 of which are listed in the Red Data Book of Kazakhstan and protected by the state. Recent studies of tulip specimens from regions bordering Kazakhstan emphasize the significance of species inventory and report the discovery of several hybrids. In this study, eight tulip species were identified based on morphological characteristics and using DNA barcoding methods. Molecular genetic markers, including nrDNA (ITS) and cpDNA markers (rbcL, matK), of the studied species were sequenced and analyzed using the Bayesian inference and maximum likelihood phylogenetic analysis methods. Our work demonstrates that DNA barcodes based on the ITS, rbcL, and matK marker regions have successful practical applicability, with ITS being the most informative at the intragenic level. However, for distinguishing closely related taxa, the most effective approach would be to use a combined dataset of sequences from multiple DNA markers. The results showed discrepancies in the placement of several taxa (T. kaufmanniana, T. patens), likely due to introgression and natural spontaneous hybridization. The molecular phylogenetic analysis suggests the existence of a previously undescribed hybrid between T. patens and T. alberti. Further detailed population studies are needed to validate this hypothesis. [ABSTRACT FROM AUTHOR]

Gorgonzola is an Italian "erborinato" blue cheese from cow's milk, bearing blue-green "parsley-like" spots due to the spread of Penicillium roqueforti mycelium. Due to its pH, water activity, and high nutrient content, as well as the environmental conditions required for its maturation, Gorgonzola constitutes an optimal ecological niche supporting the growth of both yeasts and filamentous fungi. Therefore, exploring the abundant mycobiota present in this peculiar habitat is of great interest regarding the search for new probiotic strains. The present investigation aimed to characterize the Gorgonzola mycobiota using both phenotypic (macroscopic and microscopic morphological analyses) and genotypic (DNA barcoding) analyses to find possible putative probiotic strains to be used in veterinary medicine in feed supplements. Among the different isolated filamentous fungi (Mucor and Penicillium) and yeasts (Yarrowia, Debaryomyces, Saccharomyces, and Sporobolomyces), we selected a strain of Saccharomyces cerevisiae var. boulardii. We tested its adaptation to thermal stress and its stability in feed matrices. The overall results highlight that the selected strain is stable for three months and can be considered as a possible candidate for use as a probiotic in veterinary feed supplements. [ABSTRACT FROM AUTHOR]

Simple Summary: Chalcidoidea (chalcid wasps) are one of the most numerically, structurally, and biologically diverse groups in Hymenoptera. However, the conserved morphology coupled with a high intraspecific variability in certain key characters hinders research on chalcids. In this study, DNA barcoding with the COI gene was used to calibrate intra- and interspecific variation in diagnostic characters within the genus Dzhanokmenia. In addition, the nuclear gene, 28S D2, was used to infer a phylogeny to better understand the definition of Dzhanokmenia and its relationships to potentially close genera associated with Tetrastichus. A DNA barcode library that includes eight species was established for Dzhanokmenia. A new species, D. brevifunis Ganbaatar & Cao sp. nov., was described and illustrated. The results show that Dzhanokmenia is closely related to Baryscapus and suggest that the species diversity of Dzhanokmenia is underestimated. This study lays a foundation for further research on the diversity and classification of Dzhanokmenia. Although Chalcidoidea is one of the megadiverse superfamilies in Hymenoptera, numerous species are still being discovered and described. However, the difficulties in delimiting intra- and interspecific variation hinder this process. In this study, DNA barcoding methods using the COI gene were employed to investigate the morphological variation within Dzhanokmenia Kostjukov, 1977. The nuclear locus, 28S D2, was used to infer a phylogeny to gain an understanding of the relationship of Dzhanokmenia with other potentially close genera. Through a preliminary DNA barcode library established here, including eight species, we calibrated the intraspecific variation in certain diagnostic characters for the new species described here, D. brevifunis Ganbaatar & Cao sp. nov. Maximum likelihood results show that Dzhanokmenia is clustered with the genera associated with Tetrastichus, such as Chaenotetrastichus Graham, 1987, Baryscapus Förster, 1856, Tetrastichus Haliday, 1844, and Oomyzus Rondani, 1870 involved in this study. Our results indicate that the species diversity of Dzhanokmenia is understudied and tentatively confirm that Dzhanokmenia has a potential close relationship with Baryscapus. Along with the DNA barcode library, the referenced phylogeny datasets improve the understanding of the systematic position of Dzhanokmenia within the subfamily Tetrastichinae and the definition of this genus in terms of morphology, thereby facilitating species delimitation, discovery, and description within Dzhanokmenia. [ABSTRACT FROM AUTHOR]

Argonauts or paper nautiluses are pelagic octopod cephalopods with a cosmopolitan distribution in tropical and subtropical waters around the world. Unlike other species of octopus, these are characterized by the fact that the female has a shell that serves as the breeding chamber for the eggs. Over time, this structure has been used as a taxonomic diagnostic character, causing problems in the systematics of this genus, with around 50 synonymies reported. Only two species, Argonauta argo and A. nouryi, have been reported in the Northern Humboldt Current System; however, there is taxonomic uncertainty regarding these species, which is reflected in the paralarvae (the first stage of life after hatching). In the paralarvae, the chromatophore patterns are considered to be conservative and reliable taxonomic characteristics. The objective of this study is to demonstrate the extensive variability in the chromatophore arrangement of Argonauta paralarvae in the Northern Humboldt Current using DNA barcoding and five species delimitation models. Our results include up to 11 different paralarvae morphotypes according to the pattern of chromatophores (number and arrangement) and 2 shell morphotypes. Species delimitation methods divided the 13 Argonauta morphotypes into two consensus molecular taxonomic units (MOTUs), A. argo and A. nouryi. Additionally, the results revealed an extensive morphological variability in the paralarvae and female shells of A. nouryi, demonstrating the importance of molecular data in studies involving species with different life stages, especially when this extensive morphological variability obscures conventional analyses. [ABSTRACT FROM AUTHOR]

Numerous Cymbidium species have significant commercial value globally due to their exotic ornamental flowers. Identifying Cymbidium species is challenging due to their similar shapes, which hinders their rational use and the conservation of germplasm resources. In the present study, firstly, four plastid loci (matK, rbcL, psbA-trnH, and atpF-atpH) and a nuclear locus (internal transcribed spacer, ITS) were initially examined to identify Cymbidium species. Secondly, we inferred the interspecific phylogeny of Cymbidium species using ITS sequences. All of these DNA regions, with the exception of atpF-atpH, could be readily amplified from Cymbidium, and the corresponding DNA sequences can be successfully obtained by sequencing. Our research demonstrated that ITS exhibited the highest intra- and interspecific divergences, the greatest barcoding gap, and the highest proportion of species identification. The phylogenetic analysis of Cymbidium species based on the ITS regions primarily corroborated the results obtained using traditional morphological methods. A comparative analysis of candidate DNA barcodes has shown that the ITS can be used not only for barcoding Cymbidium species but also for the phylogenetic analysis of Cymbidium. [ABSTRACT FROM AUTHOR]

The genus Rorippa belongs to the family Brassicaceae, and its members usually have high medicinal value. The genus consists of approximately 75 species and mainly grows in the Northern Hemisphere, occurring in every continent except Antarctica. The taxonomy and phylogenetic relationships of Rorippa are still unsettled, largely due to complex morphological variations in Rorippa, which were caused by frequent hybridization events. Here, we sequenced four complete plastid genomes of Rorippa species by Illumina paired-end sequencing. The four new plastid genomes of Rorippa ranged in total size from 154,671 bp for R. palustris to 154,894 bp for R. sylvestris. There are 130 genes in the four plastomes, embodying 8 rRNA, 37 tRNA, and 85 protein-coding genes. Combining with six published plastid genomes, we carried on comparative and phylogenetic analyses. We found that the ten Rorippa plastid genomes were conservative in gene number and order, total size, genomic structure, codon usage, long repeat sequence, and SSR. Fourteen mutational hotspot regions could be selected as candidate DNA barcoding to distinguish Rorippa plants. The phylogenetic trees clearly identified that ten Rorippa species displayed monophyletic relationships within the tribe Cardamineae based on plastomes and nrDNA ITS sequences. However, there are significant cytonuclear discordances in the interspecific relationships within Rorippa, as well as the intergeneric relationships between Rorippa and its related genera. We inferred that the cytonuclear discordance is most likely a result of interspecific hybridization within Rorippa, as well as intergeneric hybridization with its related genera. These plastid genomes can offer precious information for studies of species authentication, evolutionary history, and the phylogeny of Rorippa. [ABSTRACT FROM AUTHOR]

The adoption of DNA-based assessments for biodiversity monitoring has been on the rise. However, the effectiveness of DNA-based taxonomic assignments heavily relies on the availability and reliability of DNA barcode libraries. There is growing demand for a comprehensive understanding of aquatic biodiversity and the critical role of Chironomidae, specifically Stenochironomus in freshwater ecosystems. Therefore, our objective is to develop a reference barcode library for Stenochironomus in China. From 2016 to 2021, we collected Stenochironomus specimens in diverse Chinese landscapes using malaise traps, light traps, and sweep nets. These specimens were carefully preserved for DNA extraction and barcode sequencing. Our analysis unveiled 36 unique operational taxonomic units from 180 COI barcode sequences through a Neighbor-Joining tree and Automatic Barcode Gap Discovery program, highlighting a significant diversity within the Stenochironomus species. The findings emphasize the constraints of conventional morphological identification methods, especially for species with ambiguous morphologies. It also underscores the effectiveness of DNA barcoding in revealing hidden species diversity, known as cryptic species. Consequently, this study advocates for an integrated taxonomic approach, combining morphological and molecular data, to refine species identification and conservation strategies. [ABSTRACT FROM AUTHOR]

Wood-decaying fungi are important study subjects for their ecological role as well as for their biotechnological applications. They break down lignin, cellulose, and hemicelluloses using enzymes that modify the chemical structure of these complex macromolecules. Due to their ability to degrade wood, these fungi can create structural damage to wooden structures and to trees, especially those with very low level of fitness. Previous studies on wood decay fungi in the Maltese Islands are limited to records and checklists described by a handful of authors. The aim of this study was to provide a comprehensive description of wood decay fungal diversity in the Maltese Islands including an updated checklist based on DNA barcoding, as well as to establish the first wood-decay fungal culture collection at the Biology Department Seed Bank of the University of Malta. Several surveys were carried out during the rainy season along wooded areas of the Maltese Islands as well as in historical gardens. Isolates were identified using macro- and micro-morphological features, dichotomous keys, as well as molecular data. Basidiomes were recorded growing on 14 different host plant species, 11 axenic cultures have been made and 9 species of wood decay fungi have been conclusively identified by DNA barcoding. The collection of the axenic isolates includes one of Aurificaria cf. euphoria, three of Ganoderma resinaceum sl., two of Laetiporus sulphureus, one of Inonotus sp., one of Inonotus rickii anamorph, one of Inocutis tamaricis, one of Stereum hirsutum, and one of Pleurotus eryngii. However, the mycelium of Coriolopsis gallica, though collected and identified, could not be isolated. [ABSTRACT FROM AUTHOR]

The genus Angelica comprises various species utilized for diverse medicinal purposes, with differences attributed to the varying levels or types of inherent chemical components in each species. This study employed DNA barcode analysis and HPLC analysis to genetically authenticate and chemically classify eight medicinal Angelica species (n = 106) as well as two non-medicinal species (n = 14) that have been misused. Nucleotide sequence analysis of the nuclear internal transcribed spacer (ITS) region revealed differences ranging from 11 to 117 bp, while psbA-trnH showed variances of 3 to 95 bp, respectively. Phylogenetic analysis grouped all samples except Angelica sinensis into the same cluster, with some counterfeits forming separate clusters. Verification using the NCBI database confirmed the feasibility of species identification. For chemical identification, a robust quantitative HPLC analysis method was developed for 46 marker compounds. Subsequently, two A. reflexa-specific and seven A. biserrata-specific marker compounds were identified, alongside non-specific markers. Moreover, chemometric clustering analysis reflecting differences in chemical content between species revealed that most samples formed distinct clusters according to the plant species. However, some samples formed mixed clusters containing different species. These findings offer crucial insights for the standardization and quality control of medicinal Angelica species. [ABSTRACT FROM AUTHOR]

This study aimed to provide an overview of the methodological approach used for the species determination of big cats. The molecular system described herein employs mitochondrial DNA control region (CR-mtDNA)-length polymorphism in combination with highly sensitive and precise capillary electrophoresis. We demonstrated that the described CR-mtDNA barcoding system can be utilized for species determination where the presence of biological material from big cats is expected or used as a confirmatory test alongside Sanger or massive parallel sequencing (MPS). We have also addressed the fact that species barcoding, when based on the analysis of mtDNA targets, can be biased by nuclear inserts of the mitochondrial genome (NUMTs). The CR-mtDNA barcoding system is suitable even for problematic and challenging samples, such as hair. CR-mtDNA-length polymorphisms can also distinguish hybrids from pure breeds. [ABSTRACT FROM AUTHOR]

Simple Summary: A striped dolphin was found dead in Maltese waters in July 2020, displaying a severe infestation of the mesoparasite Pennella balaenoptera. This parasite is the largest known mesoparasite to grow on cetaceans and the level of infestation is typically associated with the health status of its host. This incident emphasizes the importance of monitoring the presence of these parasites, to better understand the health of dolphins in the area and to contribute to conservation efforts. Here, we document the stranding of a striped dolphin Stenella coeruleoalba (Meyen, 1833) (Mammalia: Delphinidae), which was found dead in Maltese waters in July 2020. The stranded dolphin exhibited a severe infestation of the mesoparasitic copepod, Pennella balaenoptera Koren and Danielssen, 1877 (Copepoda: Pennelidae). Parasites of this genus represent the largest known mesoparasites to infest cetaceans. Under normal circumstances, cetaceans may have a few P. balaenoptera individuals attached to them, but cetaceans with compromised health are more prone to heavy infestations. The identification of the parasite was accomplished through morphological and genetic analyses. This incident highlights the significance of monitoring mesoparasitic infestations, offering valuable insights into the health of cetacean populations and emphasizing the potential implications for conservation efforts in the region. [ABSTRACT FROM AUTHOR]

Caulerpa J.V. Lamouroux is a genus of green macroalgae belonging to the family Caulerpaceae in the order Bryopsidales. The genus comprises 104 currently accepted species, of which 51 have been recorded from Pacific Islands. Among these islands, French Polynesia is found in the middle of the South Pacific Ocean and includes five archipelagos (i.e., the Austral, Gambier, Marquesas, Society, and Tuamotu Islands) where seaweed inventories have reported a total of 16 Caulerpa species so far based on morphology. Here, based on a sampling covering the five archipelagos of French Polynesia, we attempt to (i) verify the taxonomy of Caulerpa species present in these regions based on phylogeny, (ii) describe in more detail the specific diversity between the five archipelagos, and (iii) provide a morphological identification tool for these species. We successfully obtained 134 new tufA sequences for phylogenetic analyses, which corresponded to 13 species. We propose to resurrect C. pickeringii Harvey & Bailey for representatives of C. webbiana var. pickeringii and classify it in the Caulerpa subgenus Araucarioideae. We also transfer C. seuratii to C. pickeringii based on genetic results. A new morphological identification key is provided as well as an updated distribution of Caulerpa species across French Polynesia. [ABSTRACT FROM AUTHOR]

The Asian giant hornet, Vespa mandarinia, is an invasive species that could potentially destroy the local honeybee industry in North America. It has been observed to nest in the coastal regions of British Columbia in Canada and Washington State in the USA. What is the source population of the immigrant hornets? The identification of the source population can shed light not only on the route of immigration but also on the similarity between the native habitat and the potential new habitat in the Pacific Northwest. We analyzed mitochondrial COX1 sequences of specimens sampled from multiple populations in China, the Republic of Korea, Japan, and the Russian Far East. V. mandarinia exhibits phylogeographic patterns, forming monophyletic clades for 16 specimens from China, six specimens from the Republic of Korea, and two specimens from Japan. The two mitochondrial COX1 sequences from Nanaimo, British Columbia, are identical to the two sequences from Japan. The COX1 sequence from Blaine, Washington State, clustered with those from the Republic of Korea and is identical to one sequence from the Republic of Korea. Our geophylogeny, which allows visualization of genetic variation over time and space, provides evolutionary insights on the evolution and speciation of three closely related vespine species (V. tropica, V. soror, and V. mandarinia), with the speciation events associated with the expansion of the distribution to the north. [ABSTRACT FROM AUTHOR]

Simple Summary: The reduviid genus Sycanus Amyot & Serville, 1843, possesses higher aesthetic value and plays a crucial role as a natural enemy in the control of agricultural and forestry pests. However, Sycanus from China has not undergone a comprehensive review for over four decades. Based on both molecular data and morphological evidence, we conducted a systematic review of the 14 Chinese Sycanus species, including a description of three new species. Due to the variability of body coloration and morphological similarity among closely related species, unresolved issues and debates still persist in the taxonomic study of the genus Sycanus from China. In this study, we conducted phylogenetic analyses and species delimitation for Sycanus in China based on a COI DNA barcoding dataset comprising 81 samples. The results revealed that all the samples could be classified into 12 species by integrating molecular analyses with morphological comparison. This paper provides a comprehensive systematic review of the Sycanus species found in China, including descriptions of three new species: S. taiwanensis Zhao & Cai sp. nov., S. flavicorius Li & Cai sp. nov., and S. hainanensis Wang & Cai sp. nov. Furthermore, it is proposed that S. croceovittatus Dohrn, 1859, S. leucomesus Walker, 1873, and S. villicus Stål, 1863, are three synonyms of S. bifidus (Fabricius, 1787); S. bicolor Hsiao, 1979, is a synonym of S. versicolor Dohrn, 1859; and S. hsiaoi Maldonado-Capriles, 1990, is a synonym of S. marginellus Putshkov, 1987. Additionally, brief biological information is provided for two species, S. falleni Stål, 1863, and S. croceus Hsiao, 1979. [ABSTRACT FROM AUTHOR]

In this study, we investigate the phylogeographic patterns of Dolichophis species in the Aegean region, aiming to elucidate their genetic diversity and putative historical colonisation routes through mitochondrial and nuclear DNA data. Our findings revealed distinct phylogeographic patterns: D. caspius exhibited a higher level of haplotypes within two shallow mitochondrial lineages, contrasting with D. jugularis, which displayed lower genetic variability in the area. Additionally, we identified evidence showing possible human-mediated historical translocation of D. caspius populations to Karpathos from the Balkans mainland. The mitochondrial variability in D. jugularis remained relatively uniform across southwestern Anatolia and Dodecanese, except for Rhodes Island. The evidence from mitochondrial and nuclear data confirming the previously described morphological differentiation of the Rhodes snakes, and thus the name D. j. zinneri Cattaneo, 2012, described on the island, could be applied to this isolated population. This result addresses the first genetic view on the long-standing taxonomic uncertainties regarding the subspecies status of Rhodes D. jugularis. Our results also raise questions regarding possible historical hybridisations between D. caspius and D. jugularis in the Dodecanese islands, prompting the need for further investigation using extensive field studies and genomic approaches. Ultimately, the Aegean islands, particularly Kos and Rhodes, seem to be important sites for the evolution of these colubrid snakes and their historical dynamics. [ABSTRACT FROM AUTHOR]

Morphological stasis is a widespread characteristic of pseudoscorpions, suggesting that much cryptic diversity remains unexplored. Here, we revise the polytypic species Dactylochelifer latreillii in the framework of an integrative taxonomic approach, using DNA barcoding, multivariate ratio analysis, geometric morphometry of the male foretarsus, and genitalic morphology. The pattern of mitochondrial variation suggests three species-level entities in central Europe, which widely overlap in morphospace, but differentiate in the structure of the female genitalia, and by their ecology. Dactylochelifer latreillii (Leach) is a halobiont species, occurring exclusively in coastal habitats and in Pannonian salt steppes, and D. l. septentrionalis Beier syn. nov. is a junior synonym of the nominate species. Dactylochelifer degeerii (C. L. Koch) stat. rev. is the oldest available name for an inland species that has long been mistaken for the nominotypical subspecies of D. latreillii. New habitat information suggests a preference for higher shrub vegetation. Dactylochelifer ninnii (Canestrinii) stat. rev. is a halophilic Mediterranean species that extends to the northern limits of the Pannonian basin. The distinctiveness of the Mediterranean "form" was recognized by early naturalists in the 19th century, but was ignored by later authorities in the field. [ABSTRACT FROM AUTHOR]

Up to the present study, only 8.5% of the 522 macroalgal species reported at the Azores have sequences deposited in GenBank and BOLD public repositories. The sequences of four genetic markers (cox1, rbcL, UPA, tufA) were obtained for recently collected samples from two Azorean islands. DNA barcode-assisted identification was conducted on newly generated and unpublished sequences from public repositories. A literature review of recently published studies, including the molecular identifications of Azorean macroalgae, was also performed. The results confirm the occurrence of 51 species (including subspecific ranks) and provide four new records, namely, three cryptogenic species (Olokunia boudouresquei, Padina gymnospora, and Ulva lacinulata) and one introduced species (Ulva australis). This study contributes 23 DNA barcodes generated for the first time to the Azores, which now has 10.5% of its marine flora represented in public repositories. Additionally, UPA sequences were generated for the first time for the five taxa. [ABSTRACT FROM AUTHOR]

Simple Summary: The systematic status of Fuscozetes is not clear in the literature. Therefore, the morphological ontogeny of F. fuscipes, the type species of this genus, was investigated and compared with its congeners in this study, and a new diagnosis of Fuscozetes is given. The juveniles of F. fuscipes are light brown, with a brown prodorsum, sclerites, epimeres, and legs. In all juveniles, a humeral organ and a humeral macrosclerite are present. The gastronotum of the larva has 12 pairs of setae (h3 is present), while the nymphs have 15 pairs. In the larva, the gastronotal shield is weakly developed, most of the gastronotal setae are short and inserted on the microsclerites, and several other macrosclerites and many microsclerites are present on the hysterosoma. In the nymphs, the gastronotal shield is well developed, with 10 pairs of setae (d-, l-, and h-series, and p1), and setae p2 and p3 are located on a large posteroventral macrosclerite. In all the instars, femora I and II are oval in cross-section, without a large ventral carina. Mitochondrial COI sequence data revealed a deep split between the Nearctic and Palearctic populations of F. fuscipes, and a less, but significant, divergence within each continent. These strong geographical barriers were contrasted with multiple cases of shared haplotypes over long distances in the Palearctic, indicating high migration rates in modern times. The systematic status of Fuscozetes Sellnick, 1928, is not clear in the literature. Therefore, the morphological ontogeny of F. fuscipes (C.L. Koch, 1844), the type species of this genus, was investigated and compared with its congeners in this study, and a new diagnosis of Fuscozetes is given. The juveniles of F. fuscipes are light brown, with a brown prodorsum, sclerites, epimeres, and legs. In all juveniles, a humeral organ and a humeral macrosclerite are present. The gastronotum of the larva has 12 pairs of setae (h3 is present), whereas the nymphs have 15 pairs. In the larva, the gastronotal shield is weakly developed, and most gastronotal setae are short except for a slightly longer h2. Most of the gastronotal setae are inserted on the microsclerites except for h3, and several other macrosclerites and many microsclerites are present on the hysterosoma. In the nymphs, the gastronotal shield is well developed, with 10 pairs of setae (d-, l-, and h-series, and p1), and setae p2 and p3 are located on a large posteroventral macrosclerite. In all the instars, femora I and II are oval in cross-section, without a large ventral carina. Mitochondrial COI sequence data revealed a deep split between the Nearctic and Palearctic populations of F. fuscipes, and a less, but significant, divergence within each continent. These strong geographical barriers were contrasted with multiple cases of shared haplotypes over long distances in the Palearctic, indicating high migration rates in modern times. [ABSTRACT FROM AUTHOR]

In the last two decades, plant taxonomy has bloomed, following the development of a novel technique, namely, DNA barcoding. DNA barcodes are standardized sequences, ideally unique, coding or non-coding, either from the genome of the organism or from its organelles, that are used to identify/classify an organismal group; in short, the method includes amplification of the DNA barcode, sequencing and comparison with a reference database containing the relevant sequences from different species. In plants, the use a universal DNA barcode, such as COI, which is used in animals, has not been achieved so far. This review provides a comprehensive overview of the progress made in DNA barcoding within the field of plant taxonomy. It highlights the success of various barcode loci, the emergence of super barcodes from the chloroplast genome, and the overall impact of next-generation sequencing technologies on the field. The discussion of different approaches reflects the ongoing efforts to refine and optimize DNA barcoding techniques for plants, contributing to the advancement in our understanding of plant biodiversity. [ABSTRACT FROM AUTHOR]

The Siberian moth, Dendrolimus sibiricus Tschetv., 1908 (Lepidoptera: Lasiocampidae) is a conifer pest that causes unprecedented forest mortality in Northern Asia, leading to enormous ecological and economic losses. This is the first study summarizing data on the parasitoid diversity and parasitism of this pest over the last 118 years (1905–2022). Based on 860 specimens of freshly reared and archival parasitoids, 16 species from two orders (Hymenoptera and Diptera) were identified morphologically and/or with the use of DNA barcoding. For all of them, data on distribution and hosts and images of parasitoid adults are provided. Among them, the braconid species, Meteorus versicolor (Wesmael, 1835), was documented as a parasitoid of D. sibiricus for the first time. The eastern Palaearctic form, Aleiodes esenbeckii (Hartig, 1838) dendrolimi (Matsumura, 1926), status nov., was resurrected from synonymy as a valid subspecies, and a key for its differentiation from the western Palaearctic subspecies Aleiodes esenbeckii ssp. esenbecki is provided. DNA barcodes of 11 parasitoid species from Siberia, i.e., nine hymenopterans and two dipterans, represented novel records and can be used for accurate molecular genetic identification of species. An exhaustive checklist of parasitoids accounting for 93 species associated with D. sibirisus in northern Asia was compiled. Finally, the literature and original data on parasitism in D. sibiricus populations for the last 83 years (1940–2022) were analysed taking into account the pest population dynamics (i.e., growth, outbreak, decline, and depression phases). A gradual time-lagged increase in egg and pupal parasitism in D. sibiricus populations was detected, with a peak in the pest decline phase. According to long-term observations, the following species are able to cause significant mortality of D. sibiricus in Northern Asia: the hymenopteran egg parasitoids Telenomus tetratomus and Ooencyrtus pinicolus; the larval parasitoids Aleiodes esenbeckii sp. dendrolimi, Cotesia spp., and Glyptapanteles liparidis; and the dipteran pupal parasitoids Masicera sphingivora, Tachina sp., and Blepharipa sp. Their potential should be further explored in order to develop biocontrol programs for this important forest pest. [ABSTRACT FROM AUTHOR]

The updated checklist of the marine crabs of Guinea-Bissau presented in this work is the result of consolidating decades of research, ongoing systematic revisions of the regional carcinofauna, and the inclusion of new records. DNA markers and morphological analyses for accurate identifications are integrated. Sixty-one species are mentioned in this list after reviewing specimens from scientific collections and literature. Of these 61 species listed, the presence of 51 species in Guinea-Bissau is confirmed, and 21 are reported for the first time in the area. A total of 98 sequences were obtained from 41 species (51 of 16S for 38 species and 47 of COI for 35 species), and 48 sequences (24 of 16S and 24 of COI) were the first ones obtained for 29 species. The maximum or minimum bathymetric range is expanded by 16 species, and for other six species, a color description is provided for the first time. Merocryptus obsoletus is synonymized as Merocryptus boletifer. The present study will be a useful baseline for gathering further ecological information about globally important marine taxa, not only in Guinea-Bissau but about African brachyurans in general. [ABSTRACT FROM AUTHOR]

White Kwao Krua, a crucial Thai medicinal plant, contains various phytoestrogen substances used to alleviate menopausal symptoms in estrogen-deficient women. It originates from two species, namely Pueraria candollei Wall. ex Benth. and P. mirifica Airy Shaw & Suvat. However, there exists morphological variation, and the taxonomic status between both species is ambiguous, making discrimination challenging. In this study, we aim to clarify and differentiate the morphological characteristics, palynology, and DNA barcoding of both species. The morphological results showed the stipule size is a phenotypic marker for the differentiation of both species during the vegetative stage. The palynological results, however, exhibited similarity. Through an examination of nucleotide sequences and neighbor-joining tree analysis, it was determined that the DNA barcoding of the matK region has the capability to distinguish between P. candollei and P. mirifica at nucleotide position 702. Specifically, P. candollei manifested a G base, contrasting with the C base observed in P. mirifica. This study concludes that stipule size and the matK gene in DNA barcoding serve as a distinctive characteristic for distinguishing between P. candollei and P. mirifica. These methodologies prove valuable for ensuring the accurate identification of white Kwao Krua for horticulturists. [ABSTRACT FROM AUTHOR]

Red algae (Rhodophyta) are a heterogeneous group of marine algal species that have served as a source of high-value molecules, including antioxidants and scaffolds, for novel drug development. However, it is challenging to identify Rhodophytes through morphological features alone, and in most instances, that has been the prevailing approach to identification. Consequently, this study undertook the identification of red algae species in Kenton-on-Sea, South Africa, as a baseline for future research on red algae biodiversity and conservation. The identification was achieved by designing, analysing, and using a set of universal primers through DNA barcoding of the rbcL gene. The PCR products of the rbcL gene were sequenced, and 96% of the amplicons were successfully sequenced from this set and matched with sequences on BOLD, which led to these species being molecularly described. Amongst these species are medicinally essential species, such as Laurencia natalensis and Hypnea spinella, and potential cryptic species. This calls for further investigation into the biodiversity of the studied region. Meanwhile, the availability of these primers will ease the identification process of red algae species from other coastal regions. [ABSTRACT FROM AUTHOR]

Simple Summary: Morphology and barcode data were used to estimate the diversity and genetic variability of fourteen putative species of the genus Syringophiloidus Kethley, 1970. In most cases, both sources of information were consistent. The only exception was S. amazilia Skoracki, 2017, which according to our results is most likely a population of S. stawarczyki Skoracki, 2004, and probably should be treated as its junior synonym. The further findings of our study are six new-to-science species described herein. We indicate that both the host phylogeny and distribution can drive the evolution of quill mites. Our results increase the knowledge of quill mite diversity and provide some premises to formulate and further test evolutionary, ecological, and epidemiological inquiries. Quill mites (Acariformes: Syringophilidae) are poorly explored bird parasites. Syringophiloidus Kethley, 1970, is the most specious and widespread genus in this family. It is believed to contain mono-, steno- and poly-xenous parasites and thus seems to be an exemplary for studies on biodiversity and host associations. In this work, we applied the DNA barcode marker (mitochondrial cytochrome c oxidase subunit I gene fragment, COI) to analyze the species composition and host specificity of representatives of fifteen Syringophiloidus populations parasitizing fifteen bird species. The neighbor joining analyses distinguished thirteen monophyletic lineages, almost completely corresponding to seven previously known species recognized based on morphological features, and six new-to-science species. The only exception is S. amazilia Skoracki, 2017, which is most likely conspecific with Syringophiloidus stawarczyki Skoracki, 2004. The intraspecific distances of all species were not higher than 0.9%, whilst the interspecific diversity ranged from 5.9% to 19.2% and 6.3–22.4%, inferred as the distances p and K2P, respectively. Although all putative species (except S. amazilia) are highly supported, the relationships between them have not been fully resolved and only faintly indicate that both host phylogeny and distributions influence the phylogenetic structure of quill mite taxa. [ABSTRACT FROM AUTHOR]

Plants of the genus Physalis are of economic interest because of their fleshy edible fruits with high nutritional value. Some species have high medicinal value with a long history of ethno-medicinal use to treat diverse diseases. There is therefore a need to correctly discriminate the different species of Physalis for proper utilization. Although most Physalis species have unique morphologies, their vegetative stages are identical, making it difficult to accurately identify them based on morphological characteristics. DNA barcoding has the potential to discriminate species accurately. In this study, ribulose bisphosphate carboxylase large (rbcL) and internal transcribed spacer 2 (ITS2) regions were used to discriminate Physalis species and to reveal their phylogenetic relationships and genetic diversity. Physalis plant samples were collected from seven counties in Kenya based on the availability of the germplasm. The voucher specimens were identified using the botanical taxonomy method and were deposited in the University of Nairobi herbarium. Genomic DNA was isolated from leaf samples of 64 Physalis accessions and used for PCR amplification and the sequencing of rbcL and ITS2 barcode regions. The discriminatory ability of the barcodes was based on BLASTn comparison, phylogenetic reconstruction and cluster analysis, and the determination of inter- and intra-specific distances. The nucleotide polymorphism, genetic diversity and distance of the identified Physalis species were determined using DnaSP and MEGA 11.0 software. Species discrimination was more robust using ITS2 sequences. The species identified and discriminated by ITS2 sequences were Physalis purpurea, Physalis peruviana and Physalis cordata. The rbcL sequences were only able to identify Physalis to the genus level. There was high interspecific and low intraspecific divergence within the identified Physalis species based on ITS2 sequences. The ITS2 barcode is an ideal DNA barcode for use in the discrimination of species, as well as in genetic diversity studies of Physalis accessions in Kenya. [ABSTRACT FROM AUTHOR]

Simple Summary: The genus Colias Fabricius, 1807 is a taxonomically challenging group of butterflies. Many Colias taxa display a high level of intraspecific variation in wing pattern and are weakly differentiated with respect to genitalia structure; therefore, a conclusion on their status as a distinct species, subspecies or local form can be controversial. In such cases, it is crucial to conduct a comprehensive analysis based on various phylogenetic and biogeographical approaches and a large-scale sampling dataset in order to resolve existing taxonomic and nomenclatural problems. In the present study, we focused on two enigmatic Colias taxa of unclear taxonomic status, Colias mongola Alphéraky, 1897 and Colias tamerlana Staudinger, 1897, restricted in distribution to western Mongolia, northwestern China and the south Siberian part of Russia. Here, we conducted a DNA barcode-based analysis that revealed complicate genetic pattern with several differentiated haplotypes clustered in four distinct haplogroups. In addition, we found a strong correlation between a specific mitochondrial haplogroup and Wolbachia infection, suggesting that Wolbachia endosymbionts may have played an essential role in the biology and diversification of the taxa in question and the genus Colias as a whole. The genus Colias Fabricius, 1807 includes numerous taxa and forms with uncertain status and taxonomic position. Among such taxa are Colias mongola Alphéraky, 1897 and Colias tamerlana Staudinger, 1897, interpreted in the literature either as conspecific forms, as subspecies of different but morphologically somewhat similar Colias species or as distinct species-level taxa. Based on mitochondrial and nuclear DNA markers, we reconstructed a phylogeographic pattern of the taxa in question. We recover and include in our analysis DNA barcodes of the century-old type specimens, the lectotype of C. tamerlana deposited in the Natural History Museum (Museum für Naturkunde), Berlin, Germany (ZMHU) and the paralectotype of C. tamerlana and the lectotype of C. mongola deposited in the Zoological Institute, Russian Academy of Sciences, St. Petersburg, Russia (ZISP). Our analysis grouped all specimens within four (HP_I–HP_IV) deeply divergent but geographically poorly structured clades which did not support nonconspecifity of C. mongola–C. tamerlana. We also show that all studied females of the widely distributed haplogroup HP_II were infected with a single Wolbachia strain belonging to the supergroup B, while the males of this haplogroup, as well as all other investigated specimens of both sexes, were not infected. Our data highlight the relevance of large-scale sampling dataset analysis and the need for testing for Wolbachia infection to avoid erroneous phylogenetic reconstructions and species misidentification. [ABSTRACT FROM AUTHOR]

Simple Summary: The butterfly genus Parnassius attracts the attention of numerous researchers. However, species and populations from Central and Western Asia remain understudied compared to taxa from Western Europe and East Asia. In our study, using the analysis of DNA barcodes and morphology (wing color, male genitalia, and sphragis shape in females), we substantiate the species status for P. nubilosus stat. nov. from Turkmenistan and NE Iran. We demonstrate that the P. mnemosyne group includes three morphologically similar species, P. mnemosyne (Western Eurasia), P. turatii (southwestern Europe), and P. nubilosus, as well as P. ariadne (Altai). The latter species differs from the rest of the group in the presence of red spots on the wings. We also suggest that morphological evolution within the genus involved processes of loss and reappearance of red wing spotting. The pattern of these processes is consistent with the reinforcement theory, which predicts a higher level of divergence between sympatric and allopatric populations and species. In our study, using the analysis of DNA barcodes and morphology (wing color, male genitalia, and female sphragis shape), we show that the group of species close to P. mnemosyne comprises the western and eastern phylogenetic lineages. The eastern lineage includes P. stubbendorfii, P. glacialis, and P. hoenei. The western lineage includes three morphologically similar species: P. mnemosyne (Western Eurasia), P. turatii (southwestern Europe), and P. nubilosus stat. nov. (Turkmenistan and NE Iran), as well as the morphologically differentiated P. ariadne (Altai). The latter species differs from the rest of the group in the presence of red spots on the wings. Parnassius mnemosyne s.s. is represented by four differentiated mitochondrial clusters that show clear association with specific geographic regions. We propose to interpret them as subspecies: P. mnemosyne mnemosyne (Central and Eastern Europe, N Caucasus, N Turkey), P. mnemosyne adolphi (the Middle East), P. mnemosyne falsa (Tian Shan), and P. mnemosyne gigantea (Gissar-Alai in Central Asia). We demonstrate that in P. ariadne, the red spots on the wing evolved as a reversion to the ancestral wing pattern. This reversion is observed in Altai, where the distribution areas of the western lineage, represented by P. ariadne, and the eastern lineage, represented by P. stubbendorfii, overlap. These two species hybridize in Altai, and we hypothesize that the color change in P. ariadne is the result of reinforcement of prezygotic isolation in the contact zone. The lectotype of Parnassius mnemosyne var. nubilosus Christoph, 1873, is designated. [ABSTRACT FROM AUTHOR]

Physalis species are used as an indigenous food and medicine in Kenya. However, species identification and an analysis of the health-promoting bioactive compounds and antioxidant properties are lacking. In this study, we report the molecular identification and mineral and phytochemical profiling of wild Physalis accessions. Leaf samples of 10 Physalis accessions were collected and used for species identification using nuclear ITS2 and plastid rbcL barcodes. Ripe fruits were collected from the same accessions and analyzed for mineral, total phenolic, tannin, and flavonoid contents, and antioxidant activities. The Physalis species were discriminated based on the ITS2 barcode and identified as Physalis purpurea. The genetic diversity, distance, and polymorphism of the ITS2 region of Physalis accessions were high due to the high rate of singleton and parsimony mutations. No genetic diversity, distance, or polymorphism was observed based on the rbcL barcode. The mineral content was significantly different (p < 0.05) for calcium, zinc, nickel, copper, and lithium among the Physalis accessions. No significant variation (p > 0.05) was found for phenolic acids or flavonoids, but the tannic acid content varied significantly (p < 0.05). DPPH free radical scavenging varied significantly (p < 0.05) among Physalis accessions. In conclusion, nuclear ITS2 was used to successfully identify the Physalis species of all the accessions as Physalis purpurea. The present study confirmed that Physalis purpurea has a significantly high mineral and phytochemical content and antioxidant activity. The findings from this study can be used to facilitate exploitation of Physalis purpurea in genetic breeding, their application in pharmaceutical, cosmetic, and nutritional value as well as conservation and sustainable use. [ABSTRACT FROM AUTHOR]

Quinquelaophonte is a genus of laophontid harpacticoid, including 13 valid species around the world. Many of them are known to possess inter- and/or intra-individual variations in their swimming legs. During a survey of the meiofaunal biodiversity of Jeju Island in Korea, specimens of Quinquelaophonte were collected from an intertidal zone off the west coast of Jeju Island. This study examined the morphological characteristics and amplified partial sequence of four genes (18S rRNA, 28S rRNA, COX1, and CYTB). As with other Quinquelaophonte species, specimens of this new species showed variability in the chaetotaxy of their swimming legs in both sexes. There was a setal arrangement pattern in females that could be considered a standard, whereas male individuals showed two sets of the chaetotaxy on the fourth swimming legs. The molecular data confirmed that individuals belonged to a single species, regardless of morphological variations. The result of the morphological comparison showed that the new Quinquelaohponte species shared some characteristics with congeneric species but included some specific morphological characters different from its congeners. The new species was named Quinquelaophonte sominer sp. nov. and was presented with detailed descriptions, illustrations, and confocal micrographs. Furthermore, phylogenetic analyses were conducted using newly obtained data plus the sequences of other copepods and two Quinquelaophonte species. The result suggested that Q. sominer sp. nov. and two congeners were classified as branched lineages. This discovery brings the total number of species to three in the Northwest Pacific region. [ABSTRACT FROM AUTHOR]

Sea stars are a diverse and important component of the Southern Ocean benthos. However, scarce information is available regarding their diversity, distribution, evolution, and taxonomic uncertainties persist for multiple taxa. The Magellanic Region (south of Chile) remains under-sampled despite its pivotal location for species distribution and diversity, being located at the crossroad of three ocean basins. In this study, we assessed the biodiversity of coastal Magellanic sea stars and their affinities with other oceanic bioregions. An integrative approach combining morphological identification with DNA barcoding was implemented to highlight taxonomic discrepancies such as suspected synonymy and unrecognised diversity. Firstly, we identified a total of 15 species from the coastal Magellanic Region and reported the occurrence of Cycethra frigida Koehler, 1917 for the first time in this region. The distribution of these 15 species ranged from only in South America to circumpolar, bipolar, or possibly cosmopolitan. Secondly, we highlighted possible synonymy in two species pairs within the genera Anasterias and Odontaster. This preliminary biodiversity assessment forms an important baseline for monitoring and conservation purposes, especially in the face of distribution shifts as a response to climate change and the increased presence of invasive species. Developmental mode has previously been suggested to be important in shaping biogeographical patterns. However, developmental mode was insufficient to explain the observed patterns, and other factors (e.g., physiological constraints, competition, bathymetrical range, and the possibility of passively rafting on kelp) are suggested to be at least equally important. Finally, an increase in barcoding effort is needed to better capture phylogeographic patterns for each species, both by increasing the number of specimens investigated and by covering a broader geographical range. [ABSTRACT FROM AUTHOR]

Simple Summary: The species-rich subgenus Agrodiaetus Hübner, 1822 is a distinct monophyletic lineage within the diverse blue butterfly genus Polyommatus Latreille, 1804. Although the subgenus has attracted the attention of numerous researchers, a large number of unresolved taxonomic problems persist in Agrodiaetus. In our study, we analyzed the taxonomic structure of the subgenus via combining short mitochondrial DNA barcodes of several extremely rare species, for which multi-locus data were unavailable, with multi-gene mitochondrial and nuclear data for the other Agrodiaetus taxa. This approach resulted in a high phylogenetic resolution of the tree obtained, even for the clades, which were solely represented by DNA barcodes. The status and taxonomic position of the enigmatic species P. muellerae, P. afghanicus, P. frauvartianae, P. bogra and P. anticarmon from Afghanistan, Pakistan, Iran and Turkey are revealed and discussed. The complete list of species groups and species of the subgenus Agrodiaetus is presented. The need for multi-gene analysis in evolutionary and taxonomic studies is generally accepted. However, the sequencing of multiple genes is not always possible. For various reasons, short mitochondrial DNA barcodes are the only source of molecular information for some species in many genera, although multi-locus data are available for other species of the same genera. In particular, such situation exists in the species-rich butterfly subgenus Polyommatus (Agrodiaetus). Here, we analyzed the partitioning of this subgenus into species groups by using three data sets. The first data set was represented by short mitochondrial DNA barcodes for all analyzed samples. The second and third data sets were represented by a combination of short mitochondrial DNA barcodes for part of the taxa with longer mitochondrial sequences COI + tRNA-Leu + COII (data set 2) and with longer mitochondrial COI + tRNA-Leu + COII and nuclear 5.8S rDNA + ITS2 + 28S rDNA sequences (data set 3) for the remaining species. We showed that the DNA barcoding approach (data set 1) failed to reveal the phylogenetic structure, resulting in numerous polytomies in the tree obtained. Combined analysis of the mitochondrial and nuclear sequences (data sets 2 and 3) revealed the species groups and the position within these species groups, even for the taxa for which only short DNA barcodes were available. [ABSTRACT FROM AUTHOR]

Molecular data for 19 specimens of Elmis syriaca syriaca and E. s. zoufali from eight countries have been analysed in order to investigate the taxonomic status and the geographical distribution of these two subspecies. The nominative subspecies was previously thought to be endemic to the Levant (Israel, Lebanon, Syria), while E. s. zoufali was regarded as being widespread from the Balkans to eastern Anatolia and Afghanistan. The results of our molecular studies using DNA barcoding and nuclear DNA data reveal that the two taxa are in fact distinct species, which separated around 2 Mya. A distinction based on the external morphological characters of 354 specimens was found to be impossible due to the pronounced variability, especially of the pronotal microsculpture, which had hitherto been used as the main distinguishing feature. The two species can only be distinguished by the aedeagal parameres and by the geographical distribution, which deviates considerably from the concept of previous authors. Elmis zoufali is distributed in Romania, the Balkan Peninsula, some Aegean Islands and in western Anatolia, while E. syriaca occurs from the Caucasus region southwards to eastern Turkey, Iran (probably also Afghanistan) and the Levant. Geographically, both species are widely separated by the so-called Anatolian Diagonal. Elmis zoufali resp. E. syriaca are recorded for the first time from Croatia, Romania, Azerbaijan, Georgia, Iran and Turkey. In addition, we examined 13 specimens tentatively identified as Elmis quadricollis (Reitter, 1887), a closely related species from Central Asia; we sequenced one specimen from China, which was revealed to be a sister to E. zoufali and E. syriaca. [ABSTRACT FROM AUTHOR]

The charophytes in many regions of the world are still poorly understood. This hampers the synthesis of distributional and ecological datasets at worldwide and continental scales, as well as complicates the generalization of species concepts for widely distributed and local taxa. To fill in the blanks for charophytes in the Caucasus and to improve our knowledge of species distribution areas in Eurasia, a field survey and study of available specimens from Dagestan (North Caucasus, Russia) was conducted based on morphological observation using light and scanning electron microscopy and molecular genetic analyses allowing for the precise identification and testing of the presence of cryptic and undescribed taxa. Nineteen new localities for seven Chara species and one Tolypella species, seven new species, and one new genus were identified in the studied region, and one new species in the Caspian Sea region was found. Some species records changed the outline or filled in the gaps in species distribution data. The presence of species distributed mainly in central Eurasia (C. globata, C. neglecta) with mainly Mediterranean–Middle Eastern species (C. gymnophylla) is notable for this region, as well as for other studied regions of the Caucasus characterized by a mixture combination of species with different distribution patterns. Chara gymnophylla was frequent in Dagestan, similar to the Mediterranean and Middle Eastern regions. Small brackish waterbodies on the coast of the Caspian Sea, freshwater mountain rivers, small associated waterbodies, and water reservoirs are the main habitats of charophytes in the studied region. Based on habitat preference and distribution in the Caucasus, recommendations for the protection of some species were suggested. The lack of endemic species among charophytes from Dagestan and Caucasus contrasts with the flora of terrestrial magnoliophytes that is rich in species endemism. [ABSTRACT FROM AUTHOR]

This study aimed to explore the applicability of DNA barcoding for assessing the authenticity of caviar on the Chinese market. A set of universal COI primers and two sets of designed primers based on COI and D-loop genes were used to identify maternal species of samples from 21 batches of caviar. The results showed that the PCR products from three sets of primers had more than 98% similarity to the sequences in database. The COI gene could not distinguish sturgeons with closed genetic relationships, while D-loop gene could effectively improve the accuracy of DNA barcoding and was more suitable to the identification of interspecific sturgeon than the COI gene. The neighbor-joining dendrogram further confirmed the applicability and accuracy of COI and D-loop genes in identifying maternal relatives of caviar (Acipenser baerii/Acipenser gueldenstaedtii/Acipenser schrenckii/Huso dauricus/Huso huso). Despite the limitations of mitochondrial DNA in identifying hybrid sturgeon species, the presence of counterfeit caviar of non-sturgeon ingredients could be excluded. All the caviar samples were identified successfully as sturgeon species, but the mislabeling rate of species was 33.4%, indicating that there were illegal phenomena such as disorderly labeling, mislabeling, and adulteration on the market. [ABSTRACT FROM AUTHOR]

The identification of species present in an ecosystem and the assessment of a faunistic inventory is the first step in any ecological survey and conservation effort. Thanks to technological progress, DNA barcoding has sped up species identification and is a great support to morphological taxonomy. In this work, we used a "Reverse Taxonomy" approach, where molecular (DNA barcoding) analyses were followed by morphological (skeletal features) ones to determine the specific status of 70 echinoid and 22 crinoid specimens, collected during eight different expeditions in the Ross and Weddell Seas. Of a total of 13 species of sea urchins, 6 were from the Terra Nova Bay area (TNB, Ross Sea) and 4 crinoids were identified. Previous scientific literature reported only four species of sea urchins from TNB to which we added the first records of Abatus cordatus (Verrill, 1876), Abatus curvidens Mortensen, 1936 and Abatus ingens Koehler, 1926. Moreover, we found a previous misidentification of Abatus koehleri (Thiéry, 1909), erroneously reported as A. elongatus in a scientific publication for the area. All the crinoid records are new for the area as there was no previous faunistic inventory available for TNB. [ABSTRACT FROM AUTHOR]