Search Results (464)

The flowers of Panax ginseng C. A. Mey. (PG) and Panax notoginseng (Burkill) F. H. Chen ex C. H. Chow (PN) are morphologically indistinguishable after drying, leading to prevalent adulteration that compromises product quality and consumer safety. To address this issue, we developed a rapid, closed-tube molecular authentication method based on high-resolution melting (HRM) analysis. Species-specific primer pairs were designed to target the conserved ITS and rbcL-accD regions, with PNG-2 selected as the optimal candidate owing to its stable genotyping performance and moderate GC content. Our results established GC content, rather than amplicon length, as the primary determinant of the melting temperature (Tm). Notably, the experimentally measured Tm values were consistently 0.7–1.5 °C higher than theoretical predictions, a discrepancy attributable to the stabilizing effect of the saturated fluorescent dye. To ensure maximum diagnostic reliability, the HRM results were cross-validated through a three-tier system comprising ITS2 phylogenetic analysis, agarose gel electrophoresis, and Sanger sequencing. The practical utility and matrix robustness of the assay were further verified using a diversified validation cohort of 30 commercial samples, including 24 floral batches and 6 root-derived products (root slices and ultramicro powders). The HRM profiles demonstrated 100% concordance with DNA barcoding results, effectively identifying mislabeled products across different botanical matrices and processing forms. This methodology, which can be completed within 3 h, provides a significantly more cost-effective and rapid alternative to traditional sequencing-based methods for large-scale market surveillance and industrial quality control. Full article

Platynota stultana is a Nearctic moth of economic importance for many crops in North America. It is a quarantine pest in Europe, where Mediterranean regions, with warm climates similar to those of the moth’s native range, are at risk of invasion. To date, the species is established only in Spain. It has been reported sporadically in Italy, but it is unknown whether these were transient findings or the result of an establishment. In this study, the presence of P. stultana in the Campania region, Southern Italy, was recorded. Adults of both sexes were found in different locations and in two consecutive years, suggesting that the species is established. Sequencing the COI gene identified three haplotypes of P. stultana, suggesting possible multiple introductions. The two most numerous haplotypes were identical to haplotypes from Florida. Phylogenetic analysis showed that the P. stultana clade splits into two subclades. The Italian haplotypes are all grouped into the same subclade. Our data suggest that P. stultana is expanding its range of invasion into Southern Italy, where, due to global warming, it may find increasingly favorable conditions and become an economic pest. A monitoring plan is required to allow timely implementation of control measures. Full article

Limpets are marine gastropod molluscs well adapted to intertidal rocky environments, yet their taxonomic resolution remains challenging due to extensive morphological convergence and the presence of cryptic species. In this study, we applied an integrative taxonomic framework combining multi-locus DNA barcoding and fine-scale morphological characterization to clarify species boundaries within three families of limpets—Nacellidae, Lottiidae, and Siphonariidae. A total of 132 individuals collected from six coastal sites in Shenzhen and adjacent areas of southern China were analyzed using four markers Cytochrome c oxidase subunit I (COI), 16S ribosomal RNA (16S rRNA), Cytochrome b (Cytb) and 28S ribosomal RNA (28S rRNA), together with scanning electron microscopy (SEM) observations of radular morphology. Molecular analyses identified nine distinct species across five genera. Kimura two-parameter distance analyses revealed clear barcode gaps in 16S rRNA, Cytb, and 28S rRNA genes, particularly among Cellana and Nipponacmea, whereas COI exhibited stronger discriminatory power within Siphonaria. Moreover, our study provides newly 16S, 28S references for Nipponacmea formosa and Cytb references for Nipponacmea formosa, Lottia luchuana, Siphonaria atra, Siphonaria sirius, Siphonaria sp. and Siphonaria sirius, enriching the public references and explaining the lack of corresponding records in previous BLAST searches. In addition, we identified misannotated COI references in NCBI which were labelled as Nipponacema schrenckii but belong to Cellana toreuma, highlighting inconsistencies in existing reference data rather than issues with our samples. SEM-based radular features displayed consistent interspecific variation that corroborated molecularly defined clades, offering comprehensive search of the NCBI reliable morphological evidence for species delimitation. Collectively, our findings highlight the value of integrating lineage-specific molecular markers with detailed morphological analyses to resolve taxonomic ambiguities in morphologically conservative marine gastropods. Furthermore, this approach strengthens molecular reference resources essential for future biodiversity and evolutionary research on intertidal limpets. Full article

Arid and semi-arid ecosystems harbor a wealth of underexplored plant biodiversity with untapped ecological and pharmacological potential. This study integrates morphological and molecular barcoding (ITS and rbcL) to confirm the identity of eight wild plant species native to the Saudi Arabian desert: Calligonum crinitum, Tribulus terrestris, Cornulaca monacantha, Cleome pallida, Leptadenia pyrotechnica, Cyperus conglomeratus, Indigofera argentea, and Artemisia monosperma. High-resolution GC–MS analysis identified over 25 bioactive compounds across these taxa, grouped into functional classes including hydrocarbons, esters, fatty acids, quinones, terpenoids, and phenolics. Notable compounds such as n-hexadecanoic acid, 2,4-di-tert-butylphenol, lupeol, and D-limonene were linked to antioxidant activity, desiccation tolerance, and membrane protection under stress. L. pyrotechnica and A. monosperma emerged as chemical outliers with unique metabolite profiles, suggesting divergent strategies for climate resilience. Our results highlight the ecological and bioeconomic value of desert flora, positioning them as candidates for future research in metabolic engineering, dryland restoration, and plant-based pharmaceuticals. This integrative approach underscores the relevance of desert plants for sustainable development in the face of climate change. Full article

Thrips are cosmopolitan agricultural pests and important vectors of plant viruses, and the increasing coexistence of multiple morphologically similar species has intensified the demand for species-specific molecular identification. However, traditional morphological identification and PCR assays using universal primers are often inadequate for mixed-species samples and field-adaptable application. In this study, we developed a species-specific molecular identification framework targeting a polymorphism-rich region of the mitochondrial cytochrome c oxidase subunit I (COI) gene, which is more time-efficient than sequencing-based COI DNA barcoding, for four economically important thrips species in southern China, including the globally invasive Frankliniella occidentalis. By aligning COI sequences, polymorphism-rich regions were identified and used to design four species-specific primer pairs, each containing a diagnostic 3′-terminal nucleotide. These primers were combined with a PBS-based DNA extraction workflow optimized for single-insect samples that minimizes dependence on column-based purification. The assay achieved a practical detection limit of 1 ng per reaction, demonstrated species-specific amplification, and maintained reproducible amplification at DNA inputs of ≥1 ng per reaction. Notably, PCR inhibition caused by crude extracts was effectively alleviated by fivefold dilution. Although the chemical identities of the inhibitors remain unknown, interspecific variation in inhibition strength was observed, with T. hawaiiensis exhibiting the strongest suppression, possibly due to differences in lysate composition. This integrated framework balances target specificity, operational simplicity, and dilution-mitigated inhibition, providing a field-adaptable tool for thrips species identification and invasive species monitoring. Moreover, it provides a species-specific molecular foundation for downstream integration with visual nucleic acid detection platforms, such as the CRISPR/Cas12a system, thereby facilitating the future development of portable molecular identification workflows for small agricultural pests. Full article

Urban green spaces host complex arthropod communities, in which natural insect antagonists play a key role in regulating pest populations. The jumping plant-louse Cacopsylla pulchella is a sap-sucking pest widespread across Europe that attacks Cercis siliquastrum L., which is commonly used as an ornamental tree. Heavy infestations may contribute to host tree decline and cause indirect damage in urban environments by reducing aesthetic value and by extensive deposition of honeydew secretions on surrounding surfaces. As with many phytophagous insects occurring in urban contexts, information on the natural enemies of this species remains limited, particularly in Italy, and requires further documentation. Here, we investigated the parasitoids associated with C. pulchella in central and southern Italy based on surveys conducted between 2022 and 2025. Specimens were obtained from infested plant material and identified using an integrative taxonomic approach combining detailed morphological examination with DNA barcoding. Prionomitus mitratus was confirmed as the primary parasitoid of C. pulchella, while two species, Pachyneuron muscarum and Pachyneuron aphidis, were identified as hyperparasitoids. In addition, a single specimen of Anastatus bifasciatus was also recorded emerging from the psyllid as a hyperparasitoid. Molecular analyses generated the first publicly available mitochondrial and nuclear sequences for P. mitratus. For Pachyneuron, molecular results showed variable correspondence with available reference sequences, reflecting the uneven representation of species-level data for Pteromalidae in public databases. By integrating morphological and molecular evidence, this study clarifies trophic relationships within the C. pulchella parasitoid complex. It provides vouchered molecular references to support future taxonomic and ecological research in urban ecosystems. Full article

Ganoderma is a genus of medically significant fungi, that is used in traditional medicine and is increasingly incorporated into modern nutraceuticals and pharmaceuticals. Accurate species identification and product standardization remain major challenges due to morphological plasticity and cryptic diversity. This review articulates current advances in Ganoderma systematics, phylogenetics, and metabolomics, with an emphasis on molecular identification strategies and chemical profiling. Internal transcribed spacer (ITS) sequencing has substantially improved species delineation compared with morphology alone, but its resolving power is limited in closely related species complexes, necessitating complementary multilocus approaches. Advances in metabolomics, and LC-MS- and HPLC-based profiling of triterpenes and polysaccharides, have enhanced species discrimination, chemotaxonomic resolution, and quality control of commercial products. Integrating molecular barcoding with metabolomic fingerprints provides a more robust framework for classification, pharmacological evaluation, and standardization. This review also highlights significant geographic knowledge gaps, particularly in Africa, where molecular and metabolomic data remain scarce despite high species diversity. Full article

Modern sequencing technologies have transformed the identification of medicinal plant species and varieties, overcoming the limitations of traditional approaches. To address the challenge of discriminating Toona sinensis varieties, we sequenced and compared 15 complete chloroplast genomes from five varieties in northern China. Although these genomes exhibited a highly conserved structure, we identified eight variety-specific simple sequence repeats (SSRs), two unique tandem repeats, and several hypervariable regions with elevated nucleotide diversity. Phylogenetic analysis demonstrated that whole chloroplast genomes provided the highest resolution for variety identification, outperforming conventional barcodes. Furthermore, we developed 13 specific primer pairs targeting variable regions, and PCR validation confirmed their reliable amplification across varieties. In addition, sequence-level validation by Sanger sequencing of representative SSR and tandem repeat markers revealed stable, variety-specific repeat copy number differences. These results demonstrate that the identified chloroplast markers can effectively discriminate closely related T. sinensis varieties. This study confirms that despite overall conservation, the T. sinensis plastome contains sufficient variation for reliable identification, providing a robust framework for future germplasm conservation and molecular breeding. Full article

Freshwater ecosystems are among the most vulnerable habitats worldwide, and reliable biodiversity assessment is essential for their conservation. Baiyangdian Lake, the largest freshwater lake in northern China, has undergone severe ecological degradation but is now experiencing recovery through restoration efforts. To provide a molecular basis for monitoring biodiversity, we constructed a COI DNA barcode reference library of aquatic insects from Baiyangdian Lake. From January 2023 to May 2025, systematic sampling across representative habitats yielded 315 high-quality sequences covering 104 species, 74 genera, and 33 families within eight insect orders. Diptera, particularly Chironomidae, showed the highest diversity, followed by Odonata. Phylogenetic analysis using maximum likelihood resolved all orders and families as well-supported monophyletic groups, demonstrating strong congruence with morphological taxonomy. Genetic distance analysis revealed a pronounced barcode gap, with mean intraspecific divergence of 0.46% and nearest-neighbor divergence exceeding 15%, confirming the discriminatory power of COI for species identification. Accumulation curves indicated that genus-level diversity is largely captured, while species-level diversity, especially among Diptera, remains incompletely revealed. This study provides the first comprehensive DNA barcode reference library for Baiyangdian aquatic insects, supporting ecological restoration evaluation, eDNA applications, and regional biodiversity conservation strategies. Full article

(1) Background: The identification of Tropidothorax cruciger and T. sinensis is often complicated by the presence of the “intermediate form”. Due to the lack of molecular data, the taxonomic status of the “intermediate form” and the species boundaries between T. cruciger and T. sinensis remain uncertain; (2) Methods: In this study, we integrated morphological, molecular, and ecological data to delimit species boundaries of these two species using multiple species delimitation approaches; (3) Results: Most species delimitation analyses based on the cytochrome c oxidase subunit I (COI) fragment suggested that T. cruciger and the “intermediate form” comprised a single species, with T. sinensis representing a separate species. This delimitation result was also supported by the analyses of BFD* and genetic clustering based on genome-wide SNPs. Under this species delimitation scenario, a clear-cut barcode gap was discovered between the interspecific and intraspecific genetic distances. In addition, environmental-related analyses showed highly similar ecological requirements of T. cruciger and the “intermediate form”, supporting their recognition as a single species; (4) Conclusions: This study clarifies the taxonomic status of the “intermediate form” and the species boundaries between T. cruciger and T. sinensis, which is essential for further studies of ecology and evolution of these species. Full article

Yueqing Bay in China supports important marine resources that sustain local fisheries and food security, but increasing anthropogenic pressures and natural environmental changes threaten its biodiversity. This study used morphological identification, COI DNA barcoding, and 12S DNA metabarcoding to assess ichthyoplankton composition and fish spawning patterns. A total of 13,415 eggs and 17,291 larvae were collected using horizontal and vertical plankton nets. Morphological analysis identified 58 taxa, while molecular methods detected 21 species (COI) and 48 species (12S), with an overall total of 105 species from 78 genera and 42 families. Spawning activity showed clear seasonal and spatial patterns, with the highest abundance and diversity in spring and summer, particularly around Ximen Island and the Bay mouth. These areas function as key spawning and nursery grounds. 12S DNA metabarcoding contributed strongly to species detection, especially for cryptic and morphologically indistinguishable larvae, complementing traditional surveys. The findings provide essential baseline information for monitoring fish stocks, protecting critical habitats, and improving fishery management strategies under increasing anthropogenic and climate-related pressures in Yueqing Bay. Full article

This study presents the first comprehensive molecular analysis of the Lepidoptera fauna of Cyprus based on DNA barcoding. A total of 1859 DNA barcode sequences were generated, representing 701 Barcode Index Numbers (BINs) and thus putative species. Morphological examination enabled the assignment of 596 BINs to 580 Linnaean species. Based on this genetically validated species inventory—complemented by morphologically examined specimens and a critical review of the literature—a new checklist for the Lepidoptera of Cyprus is provided. In total, 1213 species are accepted as confirmed or considered likely based on published but unverified records. The checklist includes 57 genetically confirmed first records for Cyprus and 62 new records supported solely by morphology. Remarkably, 10 species are recorded as new to Europe: Alloclita deprinsi, Cochylimorpha diana, C. additana, Pammene avetianae, P. nannodes, Cydia alienana, Ephestia abnormalella, Hypsotropa paucipunctella, Dysauxes parvigutta, and Bryophilopsis roederi. In addition, 105 BINs could not be assigned to a species. Preliminary morphological assessment indicates that many of these represent cryptic taxa or belong to taxonomically unresolved species complexes. Furthermore, 35 morphology-based records could be identified at best to the genus level. The study also lists 158 previously published species that are now considered likely misidentifications and therefore excluded from the Cypriot fauna. Full article

DNA barcode sequences remain unavailable for many species of Mexican Coleoptera. This study presents the first COI barcode records for four saproxylophagous beetle species (Coleoptera: Buprestidae, Cerambycidae) from the Central Valleys of Oaxaca, Mexico: Acmaeodera scalaris, Placosternus erythropus, Parevander xanthomelas, and Stenaspis castaneipennis. The sequences, together with their associated metadata, were deposited in the Barcode of Life Data System (BOLD) under the project Oaxaca Central Valley Insect Diversity (OCVID). A. scalaris is newly barcoded for Mexico, while the other three species are newly represented in global barcode databases. The P. xanthomelas barcode differs by approximately 8% from a GenBank sequence labeled with the same name, suggesting either a misidentified reference or a cryptic mitochondrial lineage. These data expand the molecular reference coverage for Neotropical Buprestidae and Cerambycidae and highlight the need for additional taxonomic work to refine species boundaries within Cerambycidae. Full article

Bats (Chiroptera) represent nearly one-fifth of all mammalian species and play vital ecological roles as pollinators, pest controllers, and reservoirs of zoonotic pathogens. Accurate identification of bat species is essential for biodiversity monitoring, conservation, and disease surveillance. Traditional methods based on morphology or acoustic calls are often limited by overlapping features, while DNA barcoding using the cytochrome oxidase I (COI) gene can be hindered by sequence variability. In this study, we developed a simple, single-step PCR assay targeting a short, variable region of the mitochondrial 12S rRNA gene. Alignment of sequences from 232 bat species allowed the design of a single primer pair producing a 203–224 bp amplicon that successfully distinguished all species analyzed. The assay achieved 100% amplification success across 241 bat samples, with 97.2% concordance between molecular and morphological identification. Two samples showed sequence divergence suggestive of an undescribed species. Overall, ten bat species from six genera were identified, with Eptesicus fuscus being the most frequent. This assay offers a practical and robust approach for bat identification, supporting biodiversity assessment and pathogen surveillance in ecological and public health research. Full article

Beetles (Coleoptera) represent one of the most diverse insect groups and play vital ecological roles, yet their accurate identification is often challenging due to morphological similarities among taxa. DNA barcoding has emerged as a powerful and reliable tool for species-level identification and biodiversity monitoring. In this study, we established a local DNA barcode reference database for beetles in the Liancheng Nature Reserve, Gansu Province, China. From May to August 2024, beetle specimens were collected and identified using both morphological traits and DNA barcoding. Three species delimitation methods—Automatic Barcode Gap Discovery (ABGD), Assemble Species by Automatic Partitioning (ASAP), and Bayesian Poisson Tree Processes (bPTP)—were employed as complementary analytical tools, and phylogenetic relationships were inferred from cytochrome c oxidase subunit I (COI) sequences. A total of 164 COI sequences (650 bp) were obtained, representing 126 beetle species from 95 genera and 20 families. DNA barcoding successfully resolved morphologically ambiguous taxa, with many sequences reported here for the first time. Phylogenetic analysis revealed that species within the same genus formed cohesive clades before clustering at the family level, confirming the species-level discriminative power of the COI gene. Collectively, these findings demonstrate that COI-based DNA barcoding is a powerful complement to traditional taxonomy. The establishment of this preliminary reference database provides a valuable molecular resource for beetle identification and a practical tool to support biodiversity conservation, resource management, and long-term monitoring in the Liancheng Nature Reserve. Full article