Search Results (463)

The biodiversity of wild bees (Hymenoptera: Anthophila) in southern Italy—particularly in Calabria and Basilicata—remains poorly documented despite the region’s high species richness. This study addresses this gap by assessing bee fauna through field surveys conducted across diverse habitats, including national parks, reserves, and agroecosystems, between 2016 and 2023. Bees were collected using hand nets or traps, and DNA barcoding was applied to 63 specimens to support species identification. A total of 223 species were recorded, including 10 new records for Basilicata, 57 for Calabria, 20 for southern Italy, 3 for the Italian mainland, and 1 for Italy, Eucera (Eucera) colaris Dours, 1873 (Apidae). The results reveal the unexpected presence of several central European species in mountainous areas of southern Italy, co-occurring with Mediterranean taxa. These findings indicate that these regions act as important refugia for species from both Mediterranean and Central European biogeographic zones, contributing to high regional diversity. DNA barcoding also revealed notable genetic divergence in several species compared to other European populations, highlighting the uniqueness of southern Italian bee fauna. Continuous monitoring and habitat protection are urgently needed in light of agricultural intensification and climate change. Full article

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

Orbea is a morphologically diverse lineage within the subtribe Stapeliinae, yet plastome evolution in Arabian taxa remains insufficiently characterized. This study reports the first complete chloroplast genomes of Orbea sprengeri subsp. commutata and O. wissmannii var. eremastrum and investigates plastome structure, sequence variability, and phylogenetic relationships across tribe Ceropegieae. Chloroplast genomes were assembled, annotated, and compared with 13 published plastomes representing major Ceropegieae lineages. Both Arabian plastomes displayed the typical quadripartite structure and identical gene content of 114 unique genes, including 80 protein-coding genes, 30 transfer RNA genes, and four ribosomal RNA genes. However, O. wissmannii var. eremastrum exhibited pronounced structural divergence, possessing the largest plastome recorded for the tribe (170,054 bp), an 8.9 kb expansion of the inverted repeat regions, and an 8.4 kb inversion spanning the ndhG–ndhF region. Comparative analyses revealed conserved gene order across Ceropegieae but identified six highly variable loci (accD, clpP, ndhF, ycf1, psbM–trnD, and rpl32–trnL) as potential DNA barcodes. Selection pressure analyses indicated strong purifying selection across most genes, with localized adaptive signals in accD, ndhE, ycf1, and ycf2. Phylogenomic reconstruction consistently resolved the two Arabian Orbea taxa as a distinct clade separate from the African O. variegata. This study fills a gap in Ceropegieae plastid genomics and underscores the importance of sequencing additional Orbea species to capture the full extent of genomic variation within this diverse genus. Full article

Understanding patterns and mechanisms of species diversity is one fundamental issue in biogeography and ecology. As a critical region for biodiversity, the Qinghai-Xizang Plateau (QXP) still has unclear distribution patterns and drivers for cryptic, understudied taxa such as Curculionoidea. Here, we collected the distribution data of Curculionoidea on the QXP to analyze their diversity patterns and influencing factors, and compiled a DNA barcode dataset to uncover cryptic diversity. This comprehensive dataset encompasses 671 Curculionoidea species across 223 genera, demonstrating a level of diversity that surpasses that of certain vertebrate groups. We also observed an unbalanced biogeographic pattern of diversity, with a concentration of species in the eastern and southern regions and a scarcity in the northern and central areas of QXP. Further analysis showed that the elevation range is the most important factor influencing the diversity of Curculionoidea. In addition, based on 1147 COI-5′ barcode sequences from 217 species, we found that 11 morphological species may contain cryptic species based on DNA barcode datadset. Our findings significantly enhance the current understanding of cryptic biodiversity patterns among understudied taxa in the QXP, while simultaneously highlighting persistent knowledge gaps in characterizing the plateau’s full ecological complexity. 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

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

Echinoderms are marine invertebrates that play important roles in structuring marine benthic ecosystems. DNA barcoding has become a valuable tool for species identification; however, reference DNA barcode libraries for echinoderms remain incomplete. This study aims to: (i) develop a COI-5′ reference dataset for echinoderms from Vietnam by integrating DNA barcodes with morphological data; (ii) evaluate species resolution and barcode gaps using multiple analytical approaches; (iii) assess the consistency of species assignments from BOLD and GenBank for echinoderms collected in Vietnam; (iv) make barcode data publicly available to support global reference database development. Thirty-two echinoderm specimens representing 16 species were analyzed for COI-5′ sequences, and BLAST assignments were highly concordant with those from GenBank and BOLD. Integrative validation confirmed that all taxa were monophyletic in the Neighbor Joining Tree, formed single OTUs in Cluster Sequences, and exhibited clear barcode gaps greater than 3% to the nearest-neighbor species. These results provided species-level resolution for 75% and genus-level resolution for 90% of the records. The dataset, spanning four classes, eight orders, and eleven families, enhances barcode coverage and contributes records (ProcessIDs. BINs; GenBank accessions) to public repositories. This study delivers the first curated COI-5′ reference library, supporting regional baselines for taxonomy, conservation, and biodiversity assessment. 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

Accurate species identification in blackberries (Rubus spp.) is difficult because of morphological similarity and frequent hybridization. We studied 56 wild accessions from the Sirius Federal Territory (Russia), representing coastal and foothill ecosystems of the Black Sea region. Multilocus DNA barcoding with the plastid rbcL gene and nuclear ITS1 and ITS2 regions revealed signals of hybridization and hidden diversity. The rbcL marker showed low variation, grouping most accessions into two clusters with several singletons, which limited its use for distinguishing species. In contrast, ITS1 and ITS2 showed higher variation, forming six clusters and eight singletons, and allowed for clear separation of taxa such as Rubus caesius L., R. irritans Focke, and R. amabilis Focke. Accession 3 carried a raspberry (closely to R. corchorifolius L.fil) plastid haplotype, pointing to a hybrid origin. We also found groups of nearby plants with identical mutations, which likely reflect clonal spread with fixed somatic changes or the persistence of recent hybrid lineages. At the same time, accessions collected up to 140 km apart did not form separate clusters, showing weak geographic structuring along the coast. The results demonstrate that multilocus barcoding can reveal not only species boundaries but also evolutionary processes among Rubus such as hybridization, clonal propagation, and early stages of speciation. Full article

The paper assesses brown seaweed diversity following the catastrophic events of the 2010 Deepwater Horizon (DWH) oil spill in offshore deep bank habitats at 45–90 m depth in the northwestern Gulf of Mexico, and their potential regeneration and recovery in the region. Innovative approaches to expeditionary and exploratory research resulted in the discovery, identification, and classification of brown seaweed diversity associated with rhodoliths (free-living carbonate nodules predominantly accreted by crustose coralline algae). Whereas the rhodoliths collected in situ at our research sites pre-DWH were teeming with brown algae growing on their surface, post-DWH they looked dead, bare, and bleached. These post-DWH impacts appear long-lasting, with little macroalgal growth recovery in the field. However, these apparent “dead” rhodoliths collected post-DWH at banks offshore Louisiana showed macroalgal regeneration starting within three weeks when placed in microcosms in the laboratory, with 19 brown algal species emerging from the bare rhodoliths’ surface. Some taxa corresponded to new records for the GMx (genus Cutleria and Dictyota cymatophila). Padina vickersiae is resurrected from synonymy with P. gymnospora. Reproductive sori evidence is presented for Lobophora declerckii. A detailed nomenclatural list, morphological plates, and phylogenetic/barcoding trees of brown seaweed that emerged from rhodoliths’ surfaces in laboratory microcosms are provided. These findings provide key molecular and morphological insights that reinforce species boundaries and highlight the significance of mesophotic rhodolith beds as previously overlooked reservoirs of cryptic brown algal diversity. Full article

Bactrocera dorsalis (Hendel) is a major insect pest of commercial fruit and a quarantine priority in the European Union (EU). This tephritid species was previously recorded in Austria, France, and Italy, with more recent detections in Greece. In 2023 and 2024, B. dorsalis adult males were captured by the Federal Agency for the Safety of the Food Chain (FASFC) using traps placed in community gardens and produce markets in Belgium. Morphological identification confirmed the specimens as B. dorsalis. Genomes of trapped adults were sequenced, as well as a historical set of B. dorsalis larvae intercepted by FASFC from imported fruit. A nuclear single-nucleotide polymorphism (SNP) analysis revealed that three Belgian B. dorsalis adults originated from Africa, while eight others were of Asian origin. In contrast, almost all FASFC intercepted larvae had an African origin. A discriminant analysis of principal components (DAPC) of the “Folmer” region of the mitochondrial cytochrome c oxidase subunit I (COI) gene largely confirmed the SNP analysis but also indicated that, in a few cases, mitonuclear discordance may confound origin tracing. To enable geographic origin tracing in a laboratory setting, a diagnostic set of nuclear SNPs was developed. The DAPC was also implemented in a streamlined R-script, allowing origin assignment using a mitochondrial COI barcode. To conclude, our study reveals independent occurrences of B. dorsalis in Belgium and provides important tools for origin tracing of this quarantine pest. Full article

Background: Mycetia, a subshrub genus within the subfamily Rubioideae (Rubiaceae), is predominantly distributed in tropical Asia, lacking comprehensive plastid genomic resources. This study aimed to characterize the complete plastid genomes of two Mycetia species and explore their structural features and evolutionary relationships. Methods: The plastid genomes of Mycetia hirta and Mycetia sinensis were sequenced and assembled. We analyzed genome structure, simple sequence repeats (SSRs), long repeats, codon usage, nucleotide diversity (π), and Ka/Ks and conducted phylogenetic analysis. Results: Both genomes exhibited a typical quadripartite structure (153,989–154,588 bp; GC content 37.7–37.8%), encoding 126 genes (86 protein-coding, 8 rRNA, and 32 tRNA). Both chloroplast genomes contained 52–60 SSRs and three repeat types with minor interspecific differences. Junction regions and codon usage were highly conserved, with slight variations in RSCU values. The average π was 0.0096, and the non-coding trnE-trnT (π = 0.0817) emerged as a potential DNA barcode. The average Ka/Ks was 0.2900, indicating purifying selection. Phylogenetic analysis confirmed the monophyly of Mycetia within Argostemmateae. Conclusions: This study provides the first comparative plastid genomic analysis for Mycetia, enhancing our understanding of its genetic diversity and supporting future phylogenetic and taxonomic research on the genus. Full article

Despite extensive research into green microalgae belonging to the genus Chloroidium, their species diversity and biotechnological potential remain poorly characterized. The strain VKM Al-418, the subject of this study, was isolated from the soil of Duvannyi Yar (Russian Federation). The independent species status of this strain is supported by distinct morphological characteristics, robust phylogenetic placement based on the 18S-ITS1-5.8S-ITS2 fragment, and unique features in the secondary structures of both ITS1 and ITS2, including one compensatory base change (CBC) in the highly conserved helix III of ITS2. Additionally, the species delimitation was also confirmed using five independent algorithmic approaches analyzing four different DNA barcodes. The concatenated ITS1-5.8S-ITS2 fragment is more reliable for species discrimination than the individual ITS1 or ITS2 barcodes. Of the species delimitation methods evaluated, ASAP (Assemble Species by Automatic Partitioning) and GMYC (Generalized Mixed Yule Coalescent) performed best in distinguishing Chloroidium species across multiple barcode regions in our analysis. The fatty acid profile of strain VKM Al-418 was analyzed at 9 °C, 22 °C, and 27 °C and exhibited high plasticity in response to temperature, indicative of an adaptive strategy to its harsh environment. Using this integrative taxonomic approach, we describe Chloroidium pseudoellipsoideum sp. nov., a new species with a distinct phylogenetic positioning and promising biotechnological properties. Full article

Background: Orchids of the Masdevallia genus are characterized by their beautiful esthetic qualities. However, they are vulnerable to habitat destruction, illegal harvesting, tourism and climate change. These extinction threats have led to the listing of all Masdevallia species in Appendix II of CITES (Convention on International Trade in Endangered Species of Wild Fauna and Flora) and their selection by the IUCN (International Union for Conservation of Nature). Orchids are sold in various forms, e.g., stems or tubers, rendering them impossible to identify based on morphology alone. DNA barcoding is a method that enables reliable identification of organisms using DNA barcodes, and it offers an excellent solution to the need for efficient identification of Masdevallia orchids. The aim of this study was to determine the most effective locus for DNA barcode identification of Masdevallia orchids in order to develop a quick and practical identification method for this genus. Such a method can be used by CITES verification authorities to detect illegal trade. This is the first focused study to validate a DNA barcoding protocol for Masdevallia for CITES enforcement purposes. Methods: Three genetic regions were analyzed: matK, rbcL, and ITS. The effectiveness of identification was verified based on results obtained from the new version of the BOLD Systems v.5 reference database. Results: Although Masdevallia is not well represented in this database, successful identification to the genus level was achieved. Conclusions: The highest identification efficiency at the genus level was achieved for the ITS region (91%). Full article