Phakopsora pachyrhizi, the causal agent of soybean rust disease (SBR) on Glycine max (soybean), is considered one of the most globally devastating diseases of soybeans and is a particular problem in Brazil, China, Sub-Saharan Africa, and the southern United States. To better understand genetic diversity and epidemiological history of SBR in the United States, 49 P. pachyrhizi isolates collected from soybean fields in four Southeastern states (Alabama, Florida, Georgia, and Louisiana) from the 2008 to 2017 growing seasons were genotyped through restriction site-associated genotype by sequencing (GBS). Rarefaction analysis identified 54 informative SNPs among the P. pachyrhizi isolates. We found no evidence suggesting sexual or parasexual recombination, and measurements of genetic diversity were low to moderately low. Multiple different statistical approaches, including neighbor-joining trees, K-means hierarchical clustering, discriminant analysis of principal components, and principal coordinates analysis (PCoA) all identified two groups of P. pachyrhizi genotypes that associated with geographic location. One group was composed of isolates from south Georgia, and the other with isolates from Alabama, Florida, Georgia (excluding south Georgia), and Louisiana. Our results suggest that two genetically related but distinct genotypes were introduced to the continental United States in a two-phase introduction and overwinter in South Georgia and Florida. The first introduction of one genotype likely occurred in South Georgia in 2004 followed by a later introduction of a second genotype. One genotype remained in South Georgia while the other genotype became established through the Southeastern United States. Future studies are necessary to determine whether SBR in Brazil, China, or Sub-Saharan Africa shows similar patterns of genotype distribution and history or if the United States situation is unique.

Tall fescue (Festuca arundinacea Schreb.), adaptable across subtropical to temperate zones, is extensively cultivated in northern China and the Yangtze River basin (Richard et al. 2020, Yang et al. 2022). In September 2023, the symptoms of leaf blight were observed on F. arundinacea lawns of Linghu Park (117.07°E, 30.53°N, located in Anqing, Anhui Province, China), with 15～30% of the area being infected. The leaf tissues (5 × 5 mm) from the border of diseased and healthy areas were surface disinfested in a 1% sodium hypochlorite (NaOCl) for 5 min, washed three times with sterile water. After drying on sterile filter paper, leaf tissues were placed on potato dextrose agar (PDA) at 28°C for 3 days. Pure cultures were obtained by transferring hyphal tips from the edges of colonies onto PDA. Among the 22 isolates, 18 exhibited similar morphological characteristics, and one representative strain GYB-1 was selected for further identification. Colonies on PDA were pink with a white outer ring and a cottony surface, while the reverse side was light pink with some areas appearing red. Microconidia were hyaline, oval or ellipsoidal, had zero to two septa, and measured 4.3 to 14 × 1.8 to 4.2 μm (n = 50). Macroconidia were slightly curved with three to five septa, and measured 22 to 64 × 3.1 to 5.0 μm (n = 50). According to the morphological characteristics, the isolates were identified as Fusarium verticillioides. To further identify the species, rDNA internal transcribed spacer (ITS) regions, beta-tubulin (TUB2), translation elongation factor 1-alpha (TEF1), and RNA polymerase subunits RPB2 were amplified using the primer pairs, ITS1/ITS4, BT2A/BT2B, EF1/EF2 and 7cf/11ar (Lim et al. 2019, O′Donnell et al. 2022). The obtained sequences were deposited in GenBank with the accession numbers PP783464 for ITS, PQ351671 for TUB2, PQ557396 for RPB2, and PQ557397 for TEF1. Polyphasic identification of the TEF1, ITS, TUB2, and RPB2 sequences from GYB-1 showed 100% identity as the sequences of Fusarium verticillioides strain LC13655 in the FUSARIOID-ID database. To further assess the phylogenetic relationships, a neighbor-joining tree was constructed in MEGA-X, confirming that GYB-1 clusters with F. verticillioides. To confirm pathogenicity, five healthy F. arundinacea plants were inoculated by spraying the leaves with 10 ml of a spore suspension (1 × 106 macroconidia/ml), while five plants were sprayed with sterilized water as controls. All plants were placed in the same growth chamber with a 12-hour day/night cycle at 30/25°C, and 80% humidity. Ten days post-inoculation, typical lesions were observed on the treated plants. Pathogenesis involves three stages: initial light yellow spots on leaf margins/tips, progression to yellow, irregular lesions, and subsequent leaf curling and wilting. The fungi were re-isolated from the diseased leaves and confirmed to be the original pathogens through morphological and molecular identification. Thus, Koch’s postulates were fulfilled. The experiment was repeated three times with similar results. F. verticillioides is known to cause Fusarium blight on Artemisia argyi (Wang et al. 2024) and Schizonepeta tenuifolia (Li et al. 2024). This is the first report of F. verticillioides causing Fusarium blight on tall fescue in China. Identification of the pathogen is crucial for implementing management approaches to reduce lawn losses.

The Cassidinae family, comprising unique and beautiful leaf beetles, has been the subject of limited research regarding its diversity and richness in Malaysia. Consequently, this study aimed to perform DNA barcoding on the Cassidinae species collected from Peninsular Malaysia by using the cytochrome c oxidase subunit I (COI) gene. Prior to molecular work, each species was identified morphologically based on external morphological characteristics. This study reconfirmed the host plant record for only one species, Silana farinosa, which infests the curry leaf, Murraya koenigii. Notably, a total of ten species were morphologically identified, including those belonging to the tribe Aspidimorphini (Aspidimorpha assimilis, Aspidimorpha elevata, Aspidimorpha malaccana, Aspidimorpha miliaris, and Laccoptera nepalensis) and tribe Cassidini (Chiridopsis punctata, Cassida circumdata, Chiridopsis scalaris, Notosacantha taeniata, and Silana farinosa). In this study, only seven species were successfully barcoded, and the resulting data have been deposited in GenBank. Remarkably, the separation of species is clearly delineated within their respective lineages on the Neighbor-Joining tree, with the exception of several species that predominantly belong to the genus Aspidimorpha. The data gathered in this study are significant and contribute valuable information for genetic conservation and the preservation of plant species.

The species composition of bee and non-bee Hymenoptera in the Tengku Hassanal Wildlife Reserve (THWR) Forest, Pahang, Malaysia, was studied to document their diversity and to explore their relatedness using DNA barcoding of the cytochrome c oxidase subunit I (COI) region inferred by Neighbour-Joining tree. DNA of 56 specimens were successfully extracted from the trapping between February 2023 and January 2024 by using 12 malaise traps. The collection represents 34 species from 33 genera across 11 families; eight families identified as an effective pollinators (Apidae, Halictidae, Liopteridae, Tenthredinidae, Crabronidae, Pompilidae, Tiphiidae and Vespidae), while three families were classified as less effective pollinators (Braconidae, Evaniidae, and Ichneumonidae). Unique DNA sequences revealed limited or no matches in global databases, highlighting Malaysia’s underexplored hymenopteran diversity. This study provides essential baseline data on pollinator diversity, emphasizing the roles of effective pollinators (Apis cerana, Tetrigona apicalis) and less effective pollinators (e.g., parasitoid wasps) through flower visitation in sustaining ecosystem stability. The inclusion of genetic tools alongside morphological approaches offers valuable insights for biodiversity conservation and highlights the global importance of preserving tropical ecosystems.

Angiopteris fokiensis is an endangered fern with ecological and medicinal value, yet genetic studies to support its conservation have been scarce. We performed a genome survey using high-throughput sequencing, developed genomic SSR markers from a draft assembly, and genotyped 96 individuals from 10 populations in Guangdong Province. The genome size was ~4.44 Gb (1.89% heterozygosity). From a 3.58 Gb contig assembly, 4,327,181 SSR loci were identified, with 15 highly polymorphic SSR markers being developed. Genotyping showed high within-population genetic diversity, low inter-population differentiation, and 98.55% of variation within populations. Bayesian structure, principal coordinates analysis, and neighbor-joining tree analyses consistently indicated admixed genetic clusters without clear geographical division. Additionally, the analysis revealed no significant correlation between genetic and geographic distances. Conservation should prioritize intra-population diversity via in situ/ex situ strategies. This study provides the first genomic SSR resources for A. fokiensis and underscores the importance of conserving within-population genetic diversity through integrated in situ and ex situ strategies.

Macadamia nut (Macadamia integrifolia) is a crop with significant nutritional and economic value. In November 2024, one of leaf spot symptoms was observed in Dehong Dai and Jingpo Autonomous Prefecture, Yunnan Province, China (23°50′~25°20′ N, 97°31′~98°43′ E), with a disease incidence of 35% in field. The disease initially manifests along the main veins or leaf margins, forming irregular lesions. These lesions have grayish centers surrounded by dark brown to black margins. As the lesions expand and coalesce, they cover large areas of the leaf surface. In severe cases, the leaves become wilted and defoliated, with black acervuli visible on the affected spots. Single-spore isolation was used to isolate the pathogen and four strains were obtained. The fungal colonies on PDA exhibited a circular, white, cottony appearance with an undulated surface. After 18 days, black, globular acervuli emerged on the colony's surface. Conidia 18.59 to 25.96 × 4.50 to 6.54 µm (x̄ = 22.61 × 5.55 μm) (n = 50), fusiform, straight to slightly curved, 4-septate; basal cell obconic to conic, hyaline, thin and smooth-walled, 2.72 to 5.61 μm long (x̄ = 4.13 μm); second cell from base pale brown, 4.06 to 5.95 μm (x̄ = 4.81 μm); third cell darker brown, 3.57 to 5.76 μm (x̄ = 4.61 μm); fourth cell darker, 3.74 to 5.64 μm (x̄ = 4.70 μm); apical cell 2.23 to 4.37 μm long (x̄ = 3.34 μm), hyaline, conic to obconic; apical appendages 16.34 to 23.17 μm long (x̄ = 18.97 μm), 2–3, tubular, arising from the apex; basal appendage, 3.30 to 7.08 μm long (x̄ = 4.82 μm), filiform. Morphological characteristics were consistent with published descriptions of Neopestalotiopsis chrysea. The internal transcribed spacer (ITS) regions, large subunit (LSU), and translation elongation factor 1-alpha (TEF1-α) were amplified with the primers ITS1/ITS4, LR0R/LR5, and EF1-728F/EF-2, respectively. The obtained sequences from swfu06 showed similarity with those from Neopestalotiopsis chrysea, ITS (KT783664, 553/551, 99.82%), TEF1-α (OQ410712, 500/513, 99.80%), LSU (KM116261, 874/896, 100.00%) in GenBank，respectively. All the sequences were deposited in GenBank: ITS (PQ803208, PQ803209); LSU (PQ804966, PQ804967); TEF1-α (PQ867804, PQ867805). A neighbor-joining phylogenetic tree was constructed using MEGAX, incorporating Neopestalotiopsis sp. sequences from GenBank. The pathogen was found to cluster within the N. chrysea clade with 99% bootstrap support. To test pathogenicity, Five healthy 2-year-old seedlings were selected, and four leaves per seedling were inoculated with 200 μl of conidial suspension per leaf (106 conidia/ml) with syringe. Five control plants were inoculated with ddH2O. All inoculated seedlings were covered with plastic bags for 48 h then being placed in the nature. All the inoculated leaves showed symptoms similar to those observed in the field, whereas the control leaves were asymptomatic for 7 days. N. chrysea (swfu07) was reisolated from the lesions, whereas no fungus was isolated from the control leaves. This disease will lead to early leaf drop, affecting photosynthesis and fruit yield, and posing a threat to the macadamia nut industry. This is the first report that N. chrysea causing leaf spots on M. integrifolia in China. This is of great significance for ensuring the sustainable development of macadamia nut industry. To reduce economic losses, early monitoring and disease management should be strengthened in the field.

Cotton (Gossypium hirsutum L.), a primary fiber crop, is cultivated on approximately 1.0 million hectares in Xinjiang province, China. During late September 2021 and 2023, a peculiar boll and lint rot was observed in 80% of surveyed cotton fields (Dis 60/ Tol 75) in Xinjiang, and diseased incidence ranged from 0.09% to 3.75% (average 0.52%). The lint within the bolls failed to fluff upon opening and exhibited dark discoloration due to the presence of dark brown to black conidia on the surface, accompanied by lint degradation. Notably, the pathogen exclusively infected the lint, leaving the carpel wall unaffected. Symptomatic lint tissues from 138 bolls were cut into 4-5-mm pieces, surface disinfected with 3% NaClO for 1 min, rinsed three times with sterile water, dried on sterilized filter paper,and plated on potato dextrose agar (PDA). After incubation at 25℃, 27 single-spore isolates were obtained via conidial streaking. Colonies of five representative isolates were initially white, turned gray to black within 3 days, covered the entire 9 cm PDA plate at 25 ± 2°C, and produced abundant black conidia by day 5. Conidiophores mostly reduced to conidiogenous cells, which were monoblastic, solitary, discrete, determinate, hyaline to pale brown, with a doliiform to ampulliform shape, measuring 10.3 (6.5-16.1) ×6.4 (3.5–9.1) μm (n=30). Conidia were solitary, globose to oblate, dark brown to black, smooth-walled, shiny, with equatorial slits, averaged 12.9 (9.3–15.1 × 13.6 (10.9–17.8) μm (n=50). These morphological traits matched descriptions of Nigrospora gorlenkoana (Hao et al.2020;Wang et al. 2017). DNA was extracted from 27 isolates grown on PDA for 5 days. The ITS (ITS1/4), TEF-α (EF1-728F/EF-2) and β-tubulin (Bt-2a/2b) gene was amplified and sequenced (GenBank: PQ525293, PQ616136, PQ616135 (strain XJCN-1)). BLAST analysis revealed>99.3% identity with N. gorlenkoana reference sequences (GenBank: ITS KX986048 (499/499 bp), TEF1-α KY019420 (473/476 bp), TUB2 KY019456 (403/403 bp)). A neighbor-joining phylogenetic tree of concatenated sequences (MEGA 11 Software) confirmed the isolates as N.gorlenkoana. Pathogenicity tests were performed on 30–40-day-old cotton bolls, using both detached bolls (in vitro) and attached bolls (in planta). Twenty bolls pre group were inoculated by injecting 100 μL of a conidial suspension (10⁵ conidia/mL) into the rind, while controls received sterile water. Inoculated bolls were incubated at 25°C (experiment repeated twice). Within 4 days post-inoculation, the lint exhibited extensive black-gray rot, consistent with field symptoms. Controls remained asymptomatic. The same fungus was successfully re-isolated from the symptomatic plant tissues and was re-identified as N.gorlenkoana through morphological characteristics and ITS sequencing analysis, thereby fulfilling Koch’s postulates. While N. oryzae has been reported to cause cotton boll rot in Alabama,USA and Iran (Mirzaee et al. 2013; Palmateer et al. 2003), this is the first report of N.gorlenkoana causing boll and lint rot in Xinjiang, China. Given cotton’s economic importance in the region, further studies are needed to assess its spread, economic impact, and management strategies.

IntroductionKele pig (KLP) is a valuable Chinese indigenous pig breed, renowned for its strong adaptability, high intramuscular fat content, and excellent meat quality. However, the genomic characteristics of KLPs are still unknown. This study aims to investigate the genetic diversity, population structure, and trait-related selection signatures of KLPs based on whole-genome resequencing.MethodsThe genomes of 30 KLPs were resequenced and analyzed alongside genomic data from 90 pigs of three commercial breeds, comprising 30 Duroc (DUPs), 30 Landrace (LRPs), and 30 Yorkshire pigs (YRPs). To evaluate their genetic diversity, we calculated the expected heterozygosity, observed heterozygosity, polymorphic marker ratio, minor allele frequency, nucleotide diversity (π), runs of homozygosity (ROH), and inbreeding coefficient (FROH). Meanwhile, a neighbor-joining tree, principal component analysis, ADMIXTURE analysis, linkage disequilibrium (LD) analysis, genetic distance and relationship matrices were constructed to analyze the population structure. In addition, selection signatures between KLPs and DUPs, LRPs, and YRPs were detected using fixation index (Fst) and π ratio methods.Results and DiscussionA total of 66,204,339 autosomal single nucleotide polymorphisms (SNPs) were detected in the 120 pigs, and 21,738,497 SNPs were retained for further analysis after filtering. The results showed that KLPs had higher genetic diversity, along with the smallest FROH value compared to DUPs, LRPs, and YRPs. Moreover, KLPs displayed a relatively unique genetic structure with a higher LD decay, and the majority of individuals within the KLPs exhibited distant genetic distances and relationships. Totals of 688 selected regions were identified, including 723 published QTLs. Within the selected regions, 192 candidate genes were annotated, and seven genes were found to be functionally involved in coat color (KIT), immune response (JAK2 and SOCS1), heart development (NTRK3 and SRF), muscle growth and development (VDR), and fat deposition (KDR). These findings will provide valuable insights for the future conservation, breeding, and utilization of KLPs.

Novel genotyping and genetic heterogeneity of chicken parvovirus in central and Eastern China

Japan, an island nation comprising both continental and oceanic islands, is regarded as a biodiversity hotspot with a large number of endemic insect species. Among these, two spilomeline moth species - Duponchelia naitoi Sasaki, 2008, endemic to mainland Japan, and Metasia bilineatella Inoue, 1996, endemic to the Ogasawara Islands - have attracted our attention due to their uncertain generic placements and limited available information. In this study, we investigate the morphology and phylogenetic position of these two species, and accordingly establish a new genus, Micrometasiagen. nov., to accommodate them, providing redescriptions including the first description of the female of M. bilineatella. Additionally, two new congeners are described: Micrometasia ryukyuensissp. nov. from the Ryukyu Islands, and Micrometasia melanatellasp. nov. from the Ogasawara Islands, Japan. Photographs of the adults and genitalia, along with an identification key, are provided. Maximum likelihood (ML) phylogenetic analysis using one mitochondrial (COI) and three nuclear (CAD, EF1α and RpS5) genes places this genus within Steniini - a predominantly detritivorous tribe - yet specifically within a clade whose larval stages remain almost entirely unknown. In addition, intraspecific COI genetic distances and branching order in a neighbour-joining (NJ) tree, together with the distributional patterns of each species raise several biogeographic and evolutionary implications: (1) a case of small-scale radiation accompanied by remarkable morphological divergence on oceanic islands; and (2) at least two possible colonisation routes into the Japanese archipelago, along with the potential presence of undescribed diversity of this genus beyond Japan. ZooBank: urn:lsid:zoobank.org:pub:F4E35FA1-661F-4AE4-AD4B-4D7DB5CF1569.

The West Palearctic genus Heterotoma Lepeletier & Serville, 1825, and its included species H. planicornis (Pallas, 1772) (Hemiptera, Heteroptera, Miridae, Orthotylinae) are reported from the Korean Peninsula. Heterotoma planicornis is diagnosed, and its dorsal habitus and the genitalia of both sexes are illustrated. The possible introduction of this European species into the Korean Peninsula is discussed. The complete mitochondrial genome of the species is provided. The circular mitogenome of H. planicornis is 16,179 bp long, comprising 13 protein-coding genes, two ribosomal RNA genes, and 22 transfer RNAs, and shows the same gene order as the closely allied species Mecomma ambulans (Fallén, 1807). The A–T content of the total sequence was 71.54%. An analysis using a cytochrome oxidase I (COI) neighbor-joining (NJ) tree revealed that the Korean H. planicornis population formed two distinctive groups, with a 1.8% sequence difference between them.

The Lipizzan horse breed comprises 62 mare family lines. Mitochondrial DNA (mtDNA), especially the D-loop region, plays a vital role in tracing maternal lineage and studying the origin of these family lines. The primary aim of this study is to construct phylogenetic trees using the Neighbour-Joining and Median-Joining methods, thereby analysing the genetic diversity and evolutionary relationships among these mare family lines. This study included 13 horses from Serbia that belong to different mare family lines including Thais (Rebecca-Thais) (n=2), Drava-Dubovina (Djebrin) (n=2), Wera (Theodorosta) (n=4), Batosta (Africa) (n=3), and Zora (n=2). The D-loop region was sequenced, and the sequences were subjected to bioinformatics analyses. Multiple alignments of all obtained sequences and those downloaded from the GenBank were performed with the reference mtDNA sequence downloaded from the GenBank using the MEGA 11 software, and phylogenetic trees were constructed using the PopART 1.7 and iTOL v6 online tools for visualization. Results of this study indicate that the Zora family line, though not officially recognized as a Lipizzan mare family line by the LIF (Lipizzan International Federation), forms a cluster with classical Lipizzan mare lines, including Monteaura, Betalka, Wera, and Allegra. The Neighbour-Joining tree provided a hierarchical representation of genetic distances, while the Median-Joining network revealed multiple evolutionary pathways, illustrating intraspecific genetic variability.

Central Asia is a diversity hotspot of arid-adapted grasses from the genus Stipa, with approximately 100 taxa found in the region. Recent studies in the steppe areas of Kazakhstan revealed specimens displaying intermediate morphology, distinguishing them from other taxa that grow sympatrically. Using integrative taxonomy, we investigated whether these individuals resulted from natural speciation or hybridisation, and if so, we would like to know which species were involved in this process feathergrasses. Research conducted in steppes of central Kazakhstan (Kyzylorda region), revealed the existence of individuals morphologically intermediate between S. arabica and S. richteriana, suggesting that these are probably of hybrid origin. Morphology and SNP markers validated the specimens as F1 hybrid between the aforementioned species by cladding separately based on neighbor-joining phylogenetic tree. Moreover, genetic structure displayed a separate cluster and showed almost equal genetic admixture between S. arabica and S. richteriana. Additionally, fastStructure analysis detected two geographically separated cryptic genotypes within S. richteriana population and their involvement in the hybridisation resulted in occurrence of S. × heptapotamica, S. × czerepanovii and S. × korshinskyi which recently were suggested as hybrids. Based on these evidences, we described a new nothospecies S. × kyzylordensis, as F1 hybrid. Furthermore, morphologically, the nothospecies delimited with other hybrids in Kazakh steppe area, marking the first report of hybridisation between S. arabica and S. richteriana, along with molecular evidence for the origin of further species supposed to be hybrids. This finding is crucial to understanding species diversity and hybridisation process in morphologically and genetically distant Stipa species.Supplementary InformationThe online version contains supplementary material available at 10.1038/s41598-025-08934-y.

Hospital wastewater is a reservoir of antimicrobial resistance (AMR), yet the genetic diversity and resistance mechanisms of environmental Escherichia coli in such settings remain underexplored. This study aimed to investigate the genomic characteristics, resistance profiles, and virulence potential of E. coli isolates recovered from a hospital wastewater in Jakarta, Indonesia. Six Escherichia coli isolates from hospital wastewater were sequenced using short-read next-generation sequencing (NGS). Raw reads were quality-checked and assembled with SPAdes, with five genomes retained for downstream analysis. Antimicrobial resistance (AMR) genes were identified using Staramr and the CARD database, while virulence factors were predicted using Abricate against the Virulence Factors Database (VFDB). Plasmid replicons were detected with PlasmidFinder. Phylogroup assignment followed the Clermont typing method, and phylogenetic analysis was conducted using a neighbor-joining tree based on core genome MLST (cgMLST) generated with chewBBACA v3.3.10. Multilocus sequence typing (MLST) revealed five distinct sequence types (ST744, ST156, ST1196, ST38, and ST10) across three phylogroups (A, B1, and D). A total of 57 AMR genes were detected, including blaCTX-M-15, blaCMY-2, and blaOXA-1 along with plasmid-mediated and chromosomal mutations conferring resistance to fluoroquinolones, aminoglycosides, and tetracyclines. E. coli from hospital wastewater in Jakarta exhibited high genomic diversity, multidrug resistance, and variable virulence profiles. The findings support the role of untreated hospital effluents as a hotspot for AMR emergence and horizontal gene transfer. This showed the need for routine environmental surveillance to mitigate the public health risks associated with environmental reservoirs of resistant pathogens.

Aims: To develop cox1 based DNA barcodes for Heteropneustes fossilis and Mystus bleekeri, from Upper Lake, Bhopal, to support accurate species identification and molecular taxonomy. Study Design: The study involved the collection and morphological identification of samples followed by DNA isolation, PCR amplification, Sanger sequencing, and barcode generation. Place and Duration of Study: Laboratory of molecular Biology and Genomics, Department of Bioscience, Barkatullah University, Bhopal (M.P.), India, between March 2024 to July 2025. Methodology: The genomic DNA was extracted using Phenol:Chloroform:Isoamyl (25:24:1) method. The quality of extracted DNA was determined on a 1% agarose gel. The cox1 gene fragments were amplified using universal primer (FishF1 and FishR1). Sequenced through Sanger sequencing and analysed using BLAST, MEGA11, and BOLD systems for species identification, phylogenetic assessment and barcode development. Results: In this study, two samples were analysed to generate DNA barcodes. A 615 bases long fragment of the cox1 gene was sequenced, resulting sequences (658 bp for H. fossilis and 609 bp for M. bleekeri) were submitted to GenBank (accession number PP754237 and PX116877). Sequence composition analysis revealed a moderate AT-bias (A+T = 56.35%) with the highest GC content at the first codon position. BLAST analysis confirmed 99-100% similarity with reference sequences, validating species level identification. Phylogenetic analyses using Neighbor-Joining (NJ) trees highlighted contrasting patterns: H. fossilis sequences clustered into a single cohesive clade with low divergence across South Asia, while M. bleekeri grouped within a diverse genus level dataset showing clear interspecific separation but some overlap with congeners. Conclusion: These findings confirm the reliability of cox1 as a DNA barcode for both species. Overall, this study contributes validates barcodes for two freshwater catfishes of Upper Lake and emphasizes the value of DNA barcoding in fish taxonomy, biodiversity.

In terms of medicinal material market and seed sources, Schisandra sphenanthera Rehd.et Wils. is often misused as Schisandra chinensis (Turcz.) Baill. Although the Chinese Pharmacopoeia makes a distinction at the medicinal material level, there is no reliable identification method for seeds. The existing approach still relies on traditional morphology, which requires rich experience. This study aims to fill this gap and enable quality control at the source level. A neighbor-joining (NJ) tree based on the ITS2 gene was constructed to analyze species genetic relationships and find SNP site for S. chinensis. Primers targeting the SNP site were used in an allele-specific PCR system, whose sensitivity was tested with different amounts of DNA and further validated by commercial samples. The NJ tree showed that S. chinensis clustered into a single strand, clearly separated from other related species. When the primers were applied to the allele-specific PCR, a diagnostic 273 bp band was amplified specifically in S. chinensis seeds, with no cross-reactivity observed. This assay system exhibited high sensitivity, detecting as little as 0.5 ng of genomic DNA, and was validated by commercial samples, suggesting its accuracy, reproducibility, and practical applicability. This method is an ideal tool for large-scale seed authentication of S. chinensis due to its high efficiency, precision, and operational simplicity. Our research contributes to quality control throughout the supply chain by ensuring the purity of the seeds at the earliest stages of cultivation, and also promotes the standardized production of this valuable medicinal resource.

In this paper, a new species belonging to the Andrena pilipes/nigrospina complex, Andrena culucciaesp. nov., is described and illustrated from northern Sardinia (Italy). The species is distinguished by significant morphological characters, including the structure of the genital capsule and the translucent marginal zones of the tergites in males, as well as the coloration patterns of setae in females. These characters are discussed in detail and compared with those of closely related species. Morphological evidence is complemented by COI barcode data, which supports the distinctiveness of the new taxon, although they do not fully resolve the phylogenetic relationships within the A. pilipes/nigrospina complex. Both Neighbor-Joining and Maximum Likelihood trees consistently place A. culucciaesp. nov. in a well-supported monophyletic clade. The COI data also reveal p-distance values between A. culucciaesp. nov. and both A. nigrospina Thomson, 1872 and A. pilipes Fabricius, 1781 that are higher than those observed between A. nigrospina and A. pilipes themselves, and show distinct haplotypes for all three taxa. Furthermore, we argue that the Sardinian subspecies Andrena pilipes iliensis Alfken, 1938 is not valid, as it lacks significant morphological and molecular divergence from A. pilipes s.s. An identification key for the Andrena pilipes/nigrospina complex is provided. Ecological data on A. culucciaesp. nov. are also presented, suggesting a possible association with coastal dune habitats. This highlights important conservation implications, given the ecological vulnerability of these environments in the Mediterranean region.

In this study, three new species of the genus Argyresthia Hbner, [1825], A. (Blastotere) argentilucens Kim & Byun, sp. nov., A. (B.) koreana Kim & Byun, sp. nov., and A. (B.) maculofasciata Kim & Byun, sp. nov. are described from Korea, bringing the total number of Argyresthia species recorded in the country to 19 species. The adult habitus and genitalia of the species are illustrated. The COI barcode sequences are provided for the species, and a neighbor-joining (NJ) tree is used as supplementary evidence to support the distinctiveness of the new species from other Argyresthia species.

Simple SummaryThe family Entomobryidae Tömösvary, 1882, is the largest family in Collembola, with around 2500 species in the world. It is characterised by the reduced prothorax, long antennae and furcula, and 4th abdominal segment much longer than 3rd. The genus Homidia, belonging to Entomobryidae, is mainly distributed in China. To date, 60 species of Homidia have been reported from China and account for approximately 71% of all known species of the genus. Here, the sequences of COI for ten Homidia species are provided, a neighbour-joining tree of Homidia is presented, three new species are described from Chongqing, China, and the taxonomic statuses of some species are discussed.The genus Homidia contains 84 species of which 60 have been reported from China. The sequence of COI for ten Homidia species are provided and a neighbour-joining tree is presented. Three new species of Homidia are described from Chongqing Municipality, China. Homidia wuxiensis sp. nov. is characterised by its colour pattern and chaetotaxy of Abd. IV; Homidia pseudochroma sp. nov. by some expanded post-labial chaetae and chaetotaxy of dorsal head and Abd. II–IV and Homidia yangi sp. nov. by its colour pattern. Based on similarities in COI sequences and morphology, we designate Homidia linhaiensis (Shi, Pan & Qi), as a junior synonym of Homidia tiantaiensis (Chen & Li).

The molecular epidemiological methods are important in the study of transmission dynamics and population structure of Mycobacterium tuberculosis strains circulating in a geographical region. A total of 104 patients were selected who were diagnosed for the first time with pulmonary tuberculosis and were sputum smear positive. Drug sensitivity test, spoligotyping, and MIRU-VNTR typing were done with all strains. In our series, three isolates were resistant to all the four agents, while 13(12.5%) were classified as multi-drug resistant. Spoligotyping revealed the predominance of East African-India (EAI) lineage (66.3%) with EAI 3-Ind (44.2%) and EAI 5 (18.2%) making up the majority. Our study found no presence of Beijing strains, while 15 isolates exhibited 9 distinct spoligotyping patterns (orphan or novel) that are not listed in the spoligotyping database. Three strains belonging to CAS1-Delhi, EAI3-Ind, and Manu1 lineages were resistant to all four anti-tuberculosis agents. The discriminatory power of various MIRU-VNTR alleles was found to be lower for EAI family strains compared to Non-EAI family strains. The clustering rates of MIRU-VNTR and Spoligotyping were 0.52 and 0.711 respectively. The neighbor-joining tree, based on 24-loci MIRU-VNTR typing, revealed two main clusters: one group included the CAS1-Delhi, LAM6, Cameroon, and MANU families, while the other group contained the EAI family. The finding of half of the strains resistant to one or more anti-tuberculosis drugs and 12.5% MDR strains emphasizes the importance of susceptibility testing before initiation of treatment.EAI lineage is considered to be predominant in Southern part of India which was corroborated in our study. MIRU-VNTR should be used in addition to spoligotyping since it has got finer discriminatory power. Molecular epidemiological studies should be performed periodically to assess the circulating strains in a particular geographic area.