Search Results (47)

Background/Objectives: Changnienia amoena is a rare and endangered terrestrial orchid endemic to China, valued for its ornamental and medicinal properties. However, limited genomic resources hinder its effective conservation strategies and sustainable utilization. This study aimed to generate comprehensive plastome resources and develop molecular markers to support the phylogenetics, identification, and conservation management of C. amoena. Methods: Genome skimming was employed to assemble and annotate the complete plastomes of seven geographically distinct C. amoena accessions. Comparative analyses were conducted to assess structural features and sequence divergence within C. amoena and across related species in the Calypsoinae subtribe. Phylogenetic relationships were inferred from protein-coding genes. Simple sequence repeats (SSRs), dispersed repeats, and hypervariable regions were identified from the plastomes, while nuclear SSRs were developed from assembled nuclear sequences. Results: All seven plastomes exhibited a conserved quadripartite structure with identical gene content and order, showing only minor variations in genome size. Sequence divergence was mainly confined to non-coding regions. Across Calypsoinae species, mycoheterotrophic taxa exhibited reduced plastomes. Phylogenetic analyses resolved four well-supported intergeneric clades within Calypsoinae and revealed a notable divergence between the HuNGZ accession and other C. amoena accessions, which otherwise showed low plastome-level differentiation. We also identified 69–74 plastome-derived SSRs, 22–25 dispersed repeats, and three hypervariable regions that may serve as informative molecular markers for C. amoena. Additionally, 16 polymorphic nuclear SSRs were developed from assembled nuclear sequences. Conclusions: These findings significantly expand the genomic resources available for C. amoena and provide essential insights for its phylogeny, molecular identification, conservation management, and future breeding efforts. Full article

The improvement of carcass traits is a key focus in pig genetic breeding programs. To identify quantitative trait loci (QTLs) and genes linked to key carcass traits, we conducted a genome-wide association study (GWAS) using whole-genome sequencing data from 1118 commercial pigs (Duroc sires and Yorkshire/Landrace F1 dams). This study focused on six phenotypes: iodine value, belly firmness, belly side fat, total side thickness (belly SThK), belly subcutaneous fat (Subq), and belly seam. Phenotypes were measured using image analysis, DEXA, and fatty acid profiling, and genotyping was performed using low-pass sequencing (SkimSeq). After quality control, 18,911,793 single nucleotide polymorphisms (SNPs) were retained for further analysis. A GWAS was conducted using a linear mixed model implemented in GCTA. Key findings include a significant QTL on SSC15 (110.83–112.23 Mb), which is associated with the iodine value, containing genes such as COX15, CHUK, SCD, and HIF1AN, which have known roles in fatty acid metabolism. Additionally, PNKD, VIL1, and PRKAG3 (120.74–121.88 Mb on SSC15) were linked to belly firmness, influencing muscle structure and fat composition. Three QTLs for belly side fat were identified on SSC1, SSC2, and SSC3, highlighting genes like SLC22A18, PHLDA2, and OSBPL5, which regulate fat deposition and lipid metabolism. The results provide novel molecular markers that can be incorporated into selective breeding programs to improve pork quality, fat distribution, and meat composition. These findings enhance our understanding of the genetic mechanisms underlying carcass belly traits while offering tools to improve pork quality, optimize fat composition, and align with consumer preferences in the meat production industry. Full article

Fermentation is one of the oldest food processing techniques and is widely utilized in dairy product processing, during which nutrient availability and bioactive compounds are altered. However, the complete mode of action by which fermented dairy exerts beneficial effects on the host remains unknown. The present study investigated the effect of milk and yogurt ingestion alone or combined with prebiotic inulin on the transcriptome of colonic mucosa, liver, and femur in healthy rats. Young growing male rats were fed one of four experimental diets containing (1) skimmed milk, (2) skimmed milk supplemented with inulin (5% w/w), (3) yogurt, or (4) yogurt supplemented with inulin (5% w/w) for 6 weeks. Microarray results revealed that yogurt consumption resulted in 2195 upregulated differential expressed genes (DEGs) and 1474 downregulated DEGs in colonic mucosa as compared with milk consumption. According to Gene Ontology (GO) categories and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, tight junction-, immune system-related pathways in the colonic mucosa and metabolic pathways in the liver were enriched with yogurt consumption. No evident differences were identified in the bone transcriptome between the diet groups. In conclusion, the study found that the intake of fermented dairy exerts more pronounced effects on gene expression in the intestinal tissue than prebiotics supplementation. Full article

High-throughput sequencing has transformed molecular systematics. This study presents a semi-automated pipeline for genome skimming in Thysanoptera, an insect order known for challenging species identification and cryptic relationships. By efficiently obtaining mitochondrial genomes and nuclear genes from multiple thrips specimens, the study evaluates the limitations of traditional barcoding and the data required for accurate species delimitation. The results highlight the importance of the sequencing data volume and this pipeline in reconstructing Thysanoptera phylogeny. This research also showcases the potential of advanced sequencing techniques for species delimitation and phylogenetics. Full article

The allopolyploid complexes in Crocus series Verni represent taxonomic challenges due to their variable or mostly overlapping morphology with one parental species. Moreover, their diploid ancestors remain unidentified, even with genome-wide SNP data. One such case, collected from the southeasternmost point of the series’ geographical distribution, is herein characterised and described as a new species, C. bachofenii. This study integrates phylogenomics and cytogenetics to infer the parental origin of C. bachofenii and establish its diagnostic morphological characteristics. Genome skimming of C. bachofenii and 10 other C. ser. Verni species enabled the development of novel satellite repeats as cytogenetic markers and the assembly of their complete chloroplast genomes that were employed for phylogenetic analysis alongside GBS data. The allopolyploid origin of C. bachofenii (2n = 16) was confirmed with C. vernus as the maternal parent. The probably extinct paternal parent was affiliated with a clade comprising C. heuffelianus, C. tommasinianus, C. kosaninii, and C. bertiscensis. Morphologically, C. bachofenii is distinguished by larger flowers, perigone segment coloration, and a stigma–anther ratio from its close relatives. In conclusion, its phylogenetic affiliation, distinctive cytological status, and unique morphological features justified the description of this taxon as a new species. Full article

Naematelia aurantialba and its allies are important edible and medicinal mushrooms in China. They are usually called Jiner (金耳) and have been cultivated on a commercial scale. However, due to the lack of DNA sequences from the holotype of Naematelia aurantialba, the taxonomic issues of the species complex are unresolved. In this study, the authors successfully generated DNA sequences from the holotype of N. aurantialba by a genome skimming approach and additional allied species by Sanger sequencing. Based on morphological characteristics, molecular phylogenetic data, and geographic distribution patterns, four species, including three new ones, in the complex in southwestern China were uncovered. Naematelia aurantialba occurs at high altitudes (over 3000 m above sea level), with subalpine dead plants as its substrates, and has larger basidiospores, while the commonly cultivated species, described as N. sinensis in this work, is distributed in subtropical areas at altitudes between 1800 m and 2600 m on the dead wood of subtropical plants and has smaller basidiospores. The third species, namely N. nodulosa, has habitats similar to those of N. sinensis but differs from the latter in its basidiomata with an uneven nodulose surface, a loose context with small internal cavities, and numerous conidia. The fourth species, N. pedicellata, is easily distinguished from the others by its basidia, with long basal stalks and broadly ellipsoid basidiospores measuring 10.5–12.5 × 8.0–10.0 μm. All these species are parasitic on Stereum species. This study provides a solid basis for future guidance for the selection of new strains and cultivation practices of these valuable fungi. Full article

Within Polynoidae, a diverse aphroditiform family, the subfamily Macellicephalinae comprises anchialine cave-dwelling and deep-sea scaleworms. In this study, Lepidonotopodinae is synonymized with Macellicephalinae, and the tribe Lepidonotopodini is applied to a well-supported clade inhabiting deep-sea chemosynthetic-based ecosystems. Newly sequenced “genome skimming” data for 30 deep-sea polynoids and the comparatively shallow living Eulagisca gigantea is used to bioinformatically assemble their mitogenomes. When analyzed with existing scaleworm mitogenomes, deep-sea scaleworms exhibit increased gene order rearrangement events compared to shallow-water relatives. Additionally, comparative analyses of shallow-water vs. deep-sea polynoid substitution rates in mitochondrial protein-coding genes show an overall relaxed purifying selection and a positive selection of several amino acid sites in deep-sea species, indicating that polynoid mitogenomes have undergone selective pressure to evolve metabolic adaptations suited to deep-sea environments. Furthermore, the inclusion of skimming data for already known Lepidonotopodini species allowed for an increased coverage of DNA data and a representation of the taxa necessary to create a more robust phylogeny using 18 genes, as opposed to the six genes previously used. The phylogenetic results support the erection of Cladopolynoe gen. nov., Mamiwata gen. nov., Photinopolynoe gen. nov., Stratigos gen. nov., and Themis gen. nov., and emended diagnoses for Branchinotogluma, Branchipolynoe, Lepidonotopodium, and Levensteiniella. Full article

Genome skimming is a novel approach that enables obtaining large-scale genomic information based on high-copy DNA fractions from shallow whole-genome sequencing. The simplicity of this method, low analysis costs, and large amounts of generated data have made it widely used in plant research, including species identification, especially in the case of protected or endangered taxa. This task is particularly difficult in the case of closely related taxa. The Pinus mugo complex includes several dozen closely related taxa occurring in the most important mountain ranges in Europe. The taxonomic rank, origin, or distribution of many of these taxa have been debated for years. In this study, we used genome skimming and multilocus DNA barcoding approaches to obtain different sequence data sets and also to determine their genetic diversity and suitability for distinguishing closely related taxa in the Pinus mugo complex. We generated seven different data sets, which were then analyzed using three discrimination methods, i.e., tree based, distance based, and assembling species by automatic partitioning. Genetic diversity among populations and taxa was also investigated using haplotype network analysis and principal coordinate analysis. The proposed data set based on divergence hotspots is even twenty-times more variable than the other analyzed sets and improves the phylogenetic resolution of the Pinus mugo complex. In light of the obtained results, Pinus × rhaetica does not belong to the Pinus mugo complex and should not be identified with either Pinus uliginosa or Pinus rotundata. It seems to represent a fixed hybrid or introgressant between Pinus sylvestris and Pinus mugo. In turn, Pinus mugo and Pinus uncinata apparently played an important role in the origins of Pinus uliginosa and Pinus rotundata. Full article

Wastewater surveillance is crucial for the epidemiological monitoring of SARS-CoV-2. Various concentration techniques, such as skimmed milk flocculation (SMF) and polyethylene glycol (PEG) precipitation, are employed to isolate the virus effectively. This study aims to compare these two methods and determine the one with the superior recovery rates. From February to December 2021, 24-h wastewater samples were collected from the Ioannina Wastewater Treatment Plant’s inlet and processed using both techniques. Subsequent viral genome isolation and a real-time RT-qPCR detection of SARS-CoV-2 were performed. The quantitative analysis demonstrated a higher detection sensitivity with a PEG-based concentration than SMF. Moreover, when the samples were positive by both methods, PEG consistently yielded higher viral loads. These findings underscore the need for further research into concentration methodologies and the development of precise protocols to enhance epidemiological surveillance through wastewater analysis. Full article

Wastewater-based surveillance has emerged as an important method for monitoring the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). This study investigated the presence of SARS-CoV-2 in wastewater in Zambia. We conducted a longitudinal study in the Copperbelt and Eastern provinces of Zambia from October 2023 to December 2023 during which 155 wastewater samples were collected. The samples were subjected to three different concentration methods, namely bag-mediated filtration, skimmed milk flocculation, and polythene glycol-based concentration assays. Molecular detection of SARS-CoV-2 nucleic acid was conducted using real-time Polymerase Chain Reaction (PCR). Whole genome sequencing was conducted using Illumina COVIDSEQ assay. Of the 155 wastewater samples, 62 (40%) tested positive for SARS-CoV-2. Of these, 13 sequences of sufficient length to determine SARS-CoV-2 lineages were obtained and 2 sequences were phylogenetically analyzed. Various Omicron subvariants were detected in wastewater including BA.5, XBB.1.45, BA.2.86, and JN.1. Some of these subvariants have been detected in clinical cases in Zambia. Interestingly, phylogenetic analysis positioned a sequence from the Copperbelt Province in the B.1.1.529 clade, suggesting that earlier Omicron variants detected in late 2021 could still be circulating and may not have been wholly replaced by newer subvariants. This study stresses the need for integrating wastewater surveillance of SARS-CoV-2 into mainstream strategies for monitoring SARS-CoV-2 circulation in Zambia. Full article

Repeated sequences, especially transposable elements (TEs), are known to be abundant in some members of the important invertebrate class Gastropoda. TEs that do not have long terminal repeated sequences (non-LTR TEs) are frequently the most abundant type but have not been well characterised in any gastropod. Despite this, sequences in draft gastropod genomes are often described as non-LTR TEs, but without identification to family type. This study was conducted to characterise non-LTR TEs in neritimorph snails, using genomic skimming surveys of three species and the recently published draft genome of Theodoxus fluviatilis. Multiple families of non-LTR TEs from the I, Jockey, L1, R2 and RTE superfamilies were found, although there were notably few representatives of the first of these, which is nevertheless abundant in other Gastropoda. Phylogenetic analyses of amino acid sequences of the reverse transcriptase domain from the elements ORF2 regions found considerable interspersion of representatives of the four neritimorph taxa within non-LTR families and sub-families. In contrast, phylogenetic analyses of sequences from the elements’ ORF1 region resolved the representatives from individual species as monophyletic. However, using either region, members of the two species of the Neritidae were closely related, suggesting their potential for investigation of phyletic evolution at the family level. Full article

Enteric infections due to viral pathogens are a major public health concern. Detecting the risk areas requires a strong surveillance system for pathogenic viruses in sources such as wastewater. Towards building an environmental surveillance system in Zambia, we aimed to identify group A rotavirus (RVA) and human adenovirus (HAdV) in wastewater. Convenient sampling was conducted at four study sites every Tuesday for five consecutive weeks. The research team focused on three different methods of viral concentration to determine the suitability in terms of cost and applicability for a regular surveillance system: the bag-mediated filtration system (BMFS), polyethylene glycol-based (PEG) precipitation, and skimmed milk (SM) flocculation. We screened 20 wastewater samples for HAdV and RVA using quantitative polymerase chain reaction (qPCR) and conventional polymerase chain reaction (cPCR). Of the 20 samples tested using qPCR, 18/20 (90%) tested positive for HAdV and 14/20 (70%) tested positive for RVA. For the genetic sequencing, qPCR positives were subjected to cPCR, of which 12 positives were successfully amplified. The human adenovirus was identified with a nucleotide identity range of 98.48% to 99.53% compared with the reference genome from GenBank. The BMFS and SM flocculation were the most consistent viral concentration methods for HAdV and RVA, respectively. A statistical analysis of the positives showed that viral positivity differed by site (p < 0.001). SM and PEG may be the most appropriate options in resource-limited settings such as Zambia due to the lower costs associated with these concentration methods. The demonstration of HAdV and RVA detection in wastewater suggests the presence of the pathogens in the communities under study and the need to establish a routine wastewater surveillance system for the identification of pathogens. Full article

Cinnamomum verum (syn C. zeylanicum) is considered ‘true’ cinnamon. However, it is reported that less expensive sources of cinnamon from C. cassia (syn C. aromaticum), C. loureiroi, and C. burmannii (toxic coumarin) may be used in the place of C. verum. We lack the quality assurance tools that are required to differentiate C. verum from other cinnamon species when verifying that the correct species is sourced from ingredient suppliers. The current research on cinnamon species authentication using DNA tools is limited to a few species and the use of high-quality DNA extracted from raw leaf materials. The cinnamon bark traded in the supply chain contains much less DNA and poorer-quality DNA than leaves. Our research advances DNA methods to authenticate cinnamon, as we utilized full-length chloroplast genomes via a genome skimming approach for C. burmannii and C. cassia to facilitate the design of optimal mini DNA markers. Furthermore, we developed and validated the use of NMR fingerprints for several commercial cinnamon species, including the quantification of 16 molecules. NMR fingerprints provided additional data that were useful for quality assessment in cinnamon extract powders and product consistency. Both the new mini DNA markers and NMR fingerprints were tested on commercial cinnamon products. Full article

A new, facultatively anaerobic, light-yellow, and rod-shaped bacterium designated as 3B26^T isolated from Qi’ao Island’s tidal flat sediment was identified. Strain 3B26^T can hydrolyze gelatin, aesculin, and skim milk. The major cellular fatty acids were identified as iso-C_15:0, referred to as summed feature 3, and C_16:0; the polar lipids consisted of phosphatidylethanolamine, phosphatidylglycerol, and phospholipid; and the quinones contained Q-7, Q-8, MK-7, and MMK7. The genomic size of strain 3B26^T was 4,682,650 bp, and its genomic DNA G + C content was 54.8%. While a 16S rRNA gene-based phylogenetic analysis confirmed that strain 3B26^T belongs to the genus Shewanella, both phylogenomic inference and genomic comparison revealed that strain 3B26^T is distinguishable from its relatives, and digital DNA-DNA hybridization (dDDH) values of 24.4–62.6% and average nucleotide identities (ANIs) of 83.5–95.6% between them were below the 70% dDDH and 96% ANI thresholds for bacterial species delineation. Genomic functional analysis demonstrated that strain 3B26^T possesses complete gene clusters of eicosapentaenoic acid biosynthesis and denitrification. Based on the evidence above, strain 3B26^T is considered to represent a novel species of the genus Shewanella, and the name Shewanella zhuhaiensis sp. nov. (type strain 3B26^T = GDMCC 1.2057^T = KCTC 82339^T) is proposed. Full article

Aedes japonicus and Aedes koreicus are two invasive mosquitoes native to East Asia that are quickly establishing in temperate regions of Europe. Both species are vectors of arboviruses, but we currently lack a clear understanding of their evolution. Here, we present new short-read, shallow genome sequencing of A. japonicus and A. koreicus individuals from northern Italy, which we used for downstream phylogenetic and barcode analyses. We explored associated microbial DNA and found high occurrences of Delftia bacteria in both samples, but neither Asaia nor Wolbachia. We then assembled complete mitogenomes and used these data to infer divergence times estimating the split of A. japonicus from A. koreicus in the Oligocene, which was more recent than that previously reported using mitochondrial markers. We recover a younger age for most other nodes within Aedini and other Culicidae. COI barcoding and phylogenetic analyses indicate that A. japonicus yaeyamensis, A. japonicus amamiensis, and the two A. koreicus sampled from Europe should be considered as separate species within a monophyletic species complex. Our studies further clarify the evolution of A. japonicus and A. koreicus, and indicate the need to obtain whole-genome data from putative species in order to disentangle their complex patterns of evolution. Full article