Complete Sequence Research Articles

Molecular detection and sequencing of beet necrotic yellow vein virus and beet cryptic virus 2 in sugar beet from Kazakhstan

IntroductionBeet necrotic yellow vein virus (BNYVV) is a common viral pathogen that causes considerable economic loss globally. In the present study, a commercial realtime PCR test system and custom loop mediated amplification primers were used to detect the virus in asymptomatic sugar beet samples.MethodsA total of 107 of 124 samples tested positive for the presence of the A type BNYVV coat protein gene. Near complete sequences of RNA-3 and RNA-4 were obtained using reverse transcription, followed by nanopore sequencing of 14 samples.Results and discussionA comparison with available sequences, including previously published isolates Kas2 and Kas3 from Kazakhstan, identified RNA-3 as similar to such of the P-type isolates Puthiviers and Kas3. RNA-5 was not detected using real-time PCR or cDNA amplification. Unique variable sites were identified in the p25 protein sequence translated from RNA-3. Another virus, beet cryptic virus 2 (BCV2), was identified and sequenced in samples infected with BNYVV. With 85.28% genome coverage, the identified BCV2 samples were very similar to the previously reported isolates from Hungary and Germany

AI-enabled pipeline for virus detection, validation, and SNP discovery from next-generation sequencing data

Background and AimsThe rapid and accurate detection of viruses and the discovery of single nucleotide polymorphisms (SNPs) are critical for disease management and understanding viral evolution. This study presents a pipeline for virus detection, validation, and SNP discovery from next-generation sequencing (NGS) data. The pipeline processes raw sequencing data to identify viral sequences with high accuracy and sensitivity by integrating state-of-the-art bioinformatics tools with artificial intelligence.MethodsBefore aligning the reads to the reference genomes, quality control measures, and adapter trimming are performed to ensure the integrity of the data. Unmapped reads are subjected to de novo assembly to reveal novel viral sequences and genetic elements.ResultsThe effectiveness of the pipeline is demonstrated by the identification of virus sequences, illustrating its potential for detecting known and emerging pathogens. SNP discovery is performed using a custom Python script that compares the entire population of sequenced viral reads to a reference genome. This approach provides a comprehensive overview of viral genetic diversity and identifies dominant variants and a spectrum of genetic variations.ConclusionThe robustness of the pipeline is confirmed by the recovery of complete viral sequences, which improves our understanding of viral genomics. This research aims to develop an auto-bioinformatics pipeline for novel viral sequence discovery, in vitro validation, and SNPs using the Python (AI) language to understand viral evolution. This study highlights the synergy between traditional bioinformatics techniques and modern approaches, providing a robust tool for analyzing viral genomes and contributing to the broader field of viral genomics.

Phylogenomic insights into species relationships, reticulate evolution, and biogeographic diversification of the ginseng genus Panax (Araliaceae), with an emphasis on the diversification in the Himalayan‐Hengduan Mountains

AbstractPanax (Araliaceae) is a small genus containing several well known medicinally important species. It has a disjunct distribution between Eastern Asia and Eastern North America, with most species from eastern Asia, especially the Himalayan‐Hengduan Mountains (HHM). This study used the genomic target enrichment method to obtain 358 nuclear ortholog loci and complete plastome sequences from 59 accessions representing all 18 species of the genus. Divergence time estimation and biogeographic analyses suggest that Panax was probably widely distributed from North America to Asia during the middle Eocene. During the late Eocene to Oligocene Panax may have experienced extensive extinctions during global climate cooling. It survived and diverged early in the mountains of Southwest China and tropical Indochina, where some taxa migrated northwestward to the HHM, eastward to central and eastern China, and then onward toward Japan and North America. Gene flow is identified as the main contributor to phylogenetic discordance (33.46%) within Panax. We hypothesize that the common ancestors of the medicinally important P. ginseng + P. japonicus + P. quinquefolius clade had experienced allopolyploidization, which increased adaptability to cooler and drier environments. During the middle to late Miocene, several dispersals occurred from the region of the HHM to contiguous areas, suggesting that HHM acted as a refugium and also served as a secondary diversification center for Panax. Our findings highlight that the interplay of orographic uplift and climatic changes in the HHM greatly contributed to the species diversity of Panax.

EPCO-45. COMPLETE CHROMOSOME-SCALE DIPLOID-PHASED ASSEMBLY OF A NORMAL HUMAN ASTROCYTE CELL LINE

Abstract Genomic studies of the malignant brain tumor glioma invariably rely on aligning sequencing reads to a reference genome such as HG38, even though no single reference can capture the diversity of an individual patient or cell line. To begin addressing these biases in genomics research, we sought to construct a chromosome-scale diploid-phased assembly of a normal human astrocyte (NHA) cell line that is frequently used as a basis for developing isogenic glioma cell models. To achieve this, we generated 45x Pacific Biosciences HiFi data (N50: 16kb), 26x Oxford Nanopore Technologies ultra-long data (N50: 80kb), and 2 billion paired-end reads of Hi-C data. The assembler verkko2 was used to integrate these data and build a phased diploid assembly. We achieved an assembly size of 6.01 Gbp, with 17 gapless chromosomes. The two haplotypes contain 214 and 182 contigs, respectively. Our assembly includes 76 fully assembled telomeres and 20 fully assembled centromeres. The contiguity of our assembly is 130 Mbp, which vastly exceeds that of HG38 (N50: 68 Mbp) and approaches that of the recent telomere-to-telomere CHM13 assembly (N50: 155 Mbp). We ran BUSCO on each haplotype of the assembly using the metazoa database to quantify the single-copy genes that are expected to be present in every animal genome. The haplotypes of our assembly contain the complete sequence of 98.6% and 99.9% of these genes respectively. Finally, we used Merqury to perform a k-mer based evaluation of the two haplotypes and found quality values of 59.09 and 59.75 respectively. This diploid genome assembly of NHA will serve as a resource for researchers that wish to develop NHA-derived glioma models. It can be used to call indels or structural variants in a haplotype-specific manner which will further elucidate genotypes underlying glioma phenotypes. Most importantly, it sets the stage for personalized genome assembly and tackling the lack of diversity in genomic studies of glioma to date.

Assessment of Complete Plastid Genome Sequences of Tulipa alberti Regel and Tulipa greigii Regel Species from Kazakhstan

Background. Tulipa species are economically, culturally, scientifically, and ecologically important. Tulips present taxonomic complexities that cannot be adequately resolved by examining their morphological characteristics alone or by relying on a limited selection of genetic markers. Methods. In the present study, we assessed the complete plastid sequences of Tulipa alberti Regel and Tulipa greigii Regel collected from Kazakhstan. Additionally, 14 previously published plastomes were obtained from GenBank for comparison and phylogenetic analysis. Results. The plastid genome sizes of T. alberti and T. greigii were 152,359 bp and 152,242 bp, respectively. In the plastid genomes of T. alberti and T. greigii, 136 genes were annotated, 114 of which were unique. These unique genes comprised eighty protein-coding, thirty transfer RNA, and four ribosomal RNA genes. Additionally, 415 simple sequence repeats were identified, comprising 107 tandem, 40 forward, 49 palindromic, 8 reverse, and 1 complementary repeat. Notably, the region containing ycf1 exhibited high variability and may serve as an informative DNA barcode for this genus. Conclusion. Phylogenetic analysis showed strong support for the relationships among Tulipa species, indicating the utility of plastid genome data for further taxonomic studies within the genus.

Reducing amylose content in wheat (Triticum aestivum L.) using a novel Wx-D1 null allele generated by chemical mutagenesis.

Amylose has a major influence over starch properties and end-use quality in wheat. The granule-bound starch synthase I, encoded by Wx-1, is the single enzyme responsible for amylose synthesis. Natural null mutants of Wx-1 appear at extremely low frequencies, particularly in the Wx-D1 locus, where only four spontaneous null variants have been identified, with different geographic origins. The current study identified an induced Wx-D1 null mutant (M4-9484) from the M4 generation of an ethyl methanesulfonate-mutagenized population of wheat cv. 'SM126'. The sequencing showed that the complete Wx-D1 ORF sequences of 'SM126' and M4-9484 were 2862 bp long and that there was one SNP mutation between them. The mutation was located at the RNA splice site within the junction of exon 8 and intron 8, which led to abnormal transcription of Wx-D1, with five different aberrant transcripts being identified in the mutant. The Wx-D1 null allele resulted in amylose and total starch content being decreased in M4-9484 in comparison with the wild-type 'SM126', with higher swelling capacity and being fully pasted at higher temperatures than the wild-type parent. The mutation of the Wx-D1 null gene affects the formation of amylose directly, resulting in significantly altered starch properties. This discovery offers valuable insights for enhancing wheat starch quality and contributes to the diversification of starch characteristics. It also deepens our understanding of the genetic and molecular mechanisms underlying amylose synthesis, thereby supporting breeding programs. © 2024 Society of Chemical Industry.

De novo SCN1A missense variant in a patient with Parkinson’s disease

BackgroundVariants in a gene encoding sodium voltage-gated channel alpha subunit 1 (SCN1A) are known to cause a broad clinical spectrum of epilepsy and associated features, including Dravet syndrome (MIM 607208), non-Dravet developmental and epileptic encephalopathy (MIM 619317), familial febrile seizures (MIM 604403), familial hemiplegic migraine (MIM 609634), and generalized epilepsy with febrile seizures (MIM 604403).MethodsIn this study, we examined a patient with Parkinson’s disease (PD) without any clinical manifestations of epilepsy and associated features. Genomic nucleic acid was extracted, and a complete coding sequence of the human genome (whole-exome sequencing) was sequenced. Moreover, Sanger sequencing of variants of interest was performed to validate the exome-discovered variants.ResultsWe identified a heterozygous pathogenic missense mutation (c.1498C>T; p.Arg500Trp) in the SCN1A gene in the patient using the whole-exome sequencing approach. The onset of PD features in our patient occurred at the age of 30 years. Biochemical investigations were carried out to rule out any secondary cause of the disease, including Wilson's disease or another metabolic disorder. MRI of the brain and spinal images were unremarkable. Moreover, a dramatic response to carbidopa–levodopa treatment was also observed in the patient.ConclusionOur results suggest that the pathogenic variant in SCN1A may lead to PD features without epilepsy.

Characterizing practice-dependent motor learning after a stroke.

After stroke, patients must learn to use residual motor function correctly. Consistently, motor learning is crucial in stroke motor recovery. We assessed motor performance, practice-dependent on-line motor learning, and factors potentially affecting them in stroke patients. This is a cross-sectional observational study. Twenty-six patients with first brain stroke leading to upper limb motor deficit in the subacute or chronic timeframe were enrolled. They performed a Finger Tapping Task (FTT) with both the affected and unaffected limbs. We assessed how patients learn to perform motor tasks despite the motor deficit and the differences in performance between the unaffected and affected limbs. Furthermore, by randomizing the order, we evaluated the possible inter-limb transfer of motor learning (i.e. transfer of a motor skill learned in one limb to the opposite one). Moreover, sleep, attention, anxiety, and depression were assessed through specific tests and questionnaires. Improved FTT accuracy and completed sequences for the affected limb were observed, even if lower than for the unaffected one. Furthermore, when patients initially performed the FTT with the unaffected limb, they showed higher accuracy in subsequent task completion with the affected limb than subjects who started with the affected limb. Only anxiety and attentional abilities showed significant correlations with motor performance. This work provides relevant insights into motor learning in stroke. Practice-dependent on-line motor learning is preserved in stroke survivors, and an inter-limb transfer effect can be observed. Attentional abilities and anxiety can affect learning after stroke, even if the effect of other factors cannot be excluded.

Complete Dynamics From Ignition to Stabilization of a Lean Hydrogen Flame With Thickened Flame Model

Abstract In recent years, attention has been paid to hydrogen thanks to its carbon-free nature and its interesting characteristics as an energy vector. Despite the large number of numerical analyses regarding hydrocarbon combustion in all the steady and unsteady processes, few papers that cover all those aspects are available in the literature for hydrogen flames. Therefore, a numerical methodology to explore the complete ignition sequence from the spark release to the flame stabilization is validated on a single-sector hydrogen burner. In this context, a preliminary direct numerical simulation (DNS) investigation of laminar spherical expanding flames is performed using different diffusive transport models to isolate their impact. The present work, carried out within the European project HESTIA, investigates the atmospheric test rig installed at the Norwegian University of Science and Technology operating with a lean, perfectly premixed, hydrogen–air flame stabilized on a conical bluff body. Four simulations are performed adopting the thickened flame model with an energy deposition (ED) strategy to assess the impact of preferential and thermal diffusion, as well as grid resolution, on flame dynamics. Three-dimensional (3D) flame structure visualization coupled with detailed particle image velocimetry (PIV)/OH-planar laser induced fluorescence (OH-PLIF) measurements allows the investigation of the key mechanisms involved during the ignition. The dynamic response of the flame through axial fluctuations once the ignition transient is concluded is reconstructed by the numerical strategy employed. Although the overall behavior is almost unchanged by including or not thermal diffusion effects, their local impact on the flame is evident leading to a better agreement with experimental data.

Cultivation of deep-sea bacteria from the Northwest Pacific Ocean and characterization of Limnobacter profundi sp. nov., a phenol-degrading bacterium

Despite previous culture-independent studies highlighting the prevalence of the order Burkholderiales in deep-sea environments, the cultivation and characterization of deep-sea Burkholderiales have been infrequent. A total of 243 deep-sea bacterial strains were isolated from various depths in the Northwest Pacific Ocean, with 33 isolates (13.6%) from a depth of 4000 m classified into Burkholderiales. Herein, we report the isolation and genome characteristics of strain SAORIC-580T, from a depth of 4000 m in the Northwest Pacific Ocean. The strain showed a close phylogenetic relationship with Limnobacter thiooxidans CS-K2T, sharing 99.9% 16S rRNA gene sequence identity. The complete whole-genome sequence of strain SAORIC-580T comprised 3.3 Mbp with a DNA G+C content of 52.5%. Comparative genomic analysis revealed average nucleotide identities between 79.4–85.7% and digital DNA-DNA hybridization values of 19.9–29.5% when compared to other Limnobacter genomes, indicating that the strain represents a novel species within the genus. Genomic analysis revealed unique adaptations to deep-sea conditions, including genes associated with phenol degradation, stress responses, cold adaptation, heavy metal resistance, signal transduction, and carbohydrate metabolism. The SAORIC-580T genome was found to be more abundant in the deep sea than at the surface in the trenches of the Northwest Pacific Ocean, suggesting adaptations to the deep-sea environment. Phenotypic characterization highlighted distinct differences from other Limnobacter species, including variations in growth conditions, enzyme activities, and phenol degradation capabilities. Chemotaxonomic markers of the strain included ubiquinone-10, major fatty acids such as C16:0, C16:1, and C18:1, and major polar lipids including phosphatidylethanolamine, phosphatidylglycerol, and diphosphatidylglycerol. Based on the polyphasic taxonomic data, it is concluded that strain SAORIC-580T (= KACC 21440T = NBRC 114111T) represents a novel species, for which the name Limnobacter profundi sp. nov. is proposed.

Genome-Wide Identification and Analysis of Gene Family of Carbohydrate-Binding Modules in Ustilago crameri.

Ustilago crameri is a pathogenic basidiomycete fungus that causes foxtail millet kernel smut (FMKS), a devastating grain disease in most foxtail millet growing regions of the world. Carbohydrate-Binding Modules (CBMs) are one of the important families of carbohydrate-active enzymes (CAZymes) in fungi and play a crucial role in fungal growth and development, as well as in pathogen infection. However, there is little information about the CBM family in U. crameri. Here, 11 CBM members were identified based on complete sequence analysis and functional annotation of the genome of U. crameri. According to phylogenetic analysis, they were divided into six groups. Gene structure and sequence composition analysis showed that these 11 UcCBM genes exhibit differences in gene structure and protein motifs. Furthermore, several cis-regulatory elements involved in plant hormones were detected in the promoter regions of these UcCBM genes. Gene ontology (GO) enrichment and protein-protein interaction (PPI) analysis showed that UcCBM proteins were involved in carbohydrate metabolism, and multiple partner protein interactions with UcCBM were also detected. The expression of UcCBM genes during U. crameri infection is further clarified, and the results indicate that several UcCBM genes were induced by U. crameri infection. These results provide valuable information for elucidating the features of U. crameri CBMs' family proteins and lay a crucial foundation for further research into their roles in interactions between U. crameri and foxtail millet.

Features of the oropharyngeal microbiota of healthy children and those with acute respiratory infections. A prospective single-center randomized study

Background. The oropharyngeal microbiota is involved in the development of colonization resistance, affecting viral adhesion and metabolism. Any deviations in the stability of the environmental microbial niches of the oropharynx alter the local immune response and can trigger severe chronic somatic disorders. Aim. To compare the composition of the microbiota of children during an episode of respiratory infection and healthy volunteers examined in different periods of convalescence. The obtained data can be used to predict the course of the disease, analyze the risk of complications, and assess the frequency of the diseases in the future. Materials and methods. From 20.01.2022 to 23.12.2022, an open-label prospective single-center randomized comparative study was conducted, which included 120 children aged 5-10 who presented with symptoms of acute respiratory infection. The control group consisted of 15 asymptomatic children examined at different periods of convalescence. The study compared changes in the oral microbiota composition of patients and healthy children. The evaluation was performed using complete 16S rRNA gene sequencing on the Oxford Nanopore platform. Taxonometric analysis at the species and genera level was performed using the Knomics-Biota platform. The R programming language was used for statistical processing. In addition, the parameters of α- and β-diversity of the microbiota were assessed. The Chao1 and Shannon indices were calculated to assess α-diversity (diversity within one community). The Chao1 index is based on the number of species found in the sample and also rare species that are found only 1 or 2 times, thus preventing underestimation of diversity. The Shannon index includes both the number of species and their uniformity in the community. A change in the index indicates the dominance of one or more species. The following indicators were used to assess the β-diversity describing the differences between two microbiota samples at the species and genera level: Bray–Curtis dissimilarity (characterizing the ratios of common and different microorganisms between the samples) and the Aitchison distance (reflecting the differences in the proportions of microorganisms). In addition, the balance between the two groups of microorganisms was evaluated using the NearesBalance method, which characterizes the differences between the microbiota of the main and control groups. Results. The results of our study show that during respiratory infection, the state of the oropharyngeal microbiota is characterized by an increase in α-diversity, which is associated with an increase in the proportion of species of Streptococcus salivarius, Streptococcus pneumoniae, Streptococcus pseudopneumoniae, Streptococcus pyogenes, Streptococcus thermophilus and A12 versus the proportion of Streptococcus mitis, Streptococcus oralis, Streptococcus gwangjuense, Streptococcus sanguinis, Streptococcus gordonii and FDAARGOS_192. In the control group, the oropharyngeal microbiota was divided into two clusters. The dominance of streptococci was observed in the first group (control), while the second group had a more uniform representation. Analysis of the changes in microbial communities in the main and control groups can show the stages of oral microbiota recovery after an acute respiratory infection episode. Conclusion. More research is needed in the field of the oropharyngeal microbiome on a larger sample of healthy children, to standardize the communities of the oropharyngeal microbiota, to study the interaction within communities and with the body as a whole. Analysis of the impact of the microbiota on the frequency and course of respiratory infections and the rate of complications opens up new prospects in treating, rehabilitating, and preventing diseases. These research areas can contribute to improving children's health and quality of life and introducing new approaches into clinical practice.

CDC42 enhances phagocytic activity and exhibits antimicrobial functions through the recruitment of IQGAP1 in Apostichopus japonicus

Cell Division Cycle 42 protein (CDC42) is a pivotal member of the Rho family of small GTPases, integral to cell signaling and the regulation of cell polarity. CDC42 is extensively documented to participate in host immunity, particularly in processes such as phagocytosis and cell migration. Nevertheless, the precise mechanisms by which CDC42 affects host-bacteria interactions remain unclear in Echinoderms. In this study, we have isolated and characterized the CDC42 gene from Apostichopus japonicus, referred to as AjCDC42. The complete cDNA sequence of AjCDC42 extends 1282 base pairs, including a typical classical RHO domain, spanning amino acids from 6 to 179. Spatial expression analysis demonstrated that AjCDC42 is ubiquitously expressed across all examined tissues. Following infection with Vibrio splendidus and stimulation with LPS, the protein and mRNA levels of AjCDC42 were significantly upregulated in coelomocytes. Moreover, the knockdown of AjCDC42 significantly decreased V. splendidus-induced phagocytic activity in coelomocytes, leading to a reduced intracellular load of V. splendidus, as evidenced by flow cytometry and bacterial plate counting assays. Mechanistically, AjCDC42 was identified as an interacting partner of IQ Motif Containing GTPase Activating Protein 1 (AjIQGAP1) through the use of co-immunoprecipitation, immunofluorescence, and GST pull-down assays combined with mass spectrometry techniques. Additional functional analyses revealed that the knockdown of AjIQGAP1 similarly decreased the phagocytic activity and the intracellular load of V. splendidus in coelomocytes. Furthermore, we detected that decreased levels of AjCDC42 and AjIQGAP1 impaired cytoskeletal rearrangement and delayed lysosomal clearance of V. splendidus. In summary, our findings underscore the pivotal role of the AjCDC42-AjIQGAP1 axis in regulating coelomocytes phagocytosis in A. japonicus during V. splendidus infection.

Analysis of bZIP transcription factors in Rhododendron simsii and functional study of RsbZIP6 in regulating anthocyanin biosynthesis

The basic leucine zipper (bZIP) transcription factors play a critical role in various plant biological processes, including anthocyanin biosynthesis. This study focuses on Rhododendron simsii, a notable ornamental species with insufficiently explored bZIP transcription factors. We identified 66 bZIP transcription factors in the R. simsii genome and conducted comprehensive bioinformatics analyses to determine their gene localization, phylogenetic relationships, grouping, gene/protein structure, duplication events, synteny, and expression profiles. Our analysis identified RsbZIP6, a homolog of HY5 known to influence anthocyanin biosynthesis in many plants, as a potential regulator of this pathway. We cloned the complete coding sequence of RsbZIP6, which encodes a 170-amino acid protein spanning 510 bp. Subcellular localization analysis verified the nuclear presence of the RsbZIP6 protein. RT-qPCR analysis revealed the highest expression of RsbZIP6 in petals, which correlated with anthocyanin accumulation. Transgenic experiments indicated that overexpressing RsbZIP6 in Arabidopsis enhanced anthocyanin accumulation by upregulating genes involved in anthocyanin biosynthesis (4CL, CHS, CHI, DFR, F3H, F3’H, ANS and UF3GT). Our findings enhance understanding of the bZIP transcription factor family in R. simsii and underscore the vital role of RsbZIP6 in anthocyanin biosynthesis, providing insights for future genetic enhancement strategies.

The complete mitochondrial genome of the Rhacophorus chenfui Liu, 1945 and its phylogenetic analyses

We described the complete mitogenome sequence of Rhacophorus chenfui in this research. The circular mitogenome of R. chenfui is total length with 20,520 bp, encoded 39 genes (24 tRNA genes, 13 protein-coding genes, 2 rRNA genes) and two D-loop regions. The base composition of the mitogenome is 33.46% A, 30.80% T, 23.05% C, and 12.69% G. All tRNAs have the typical clover-leaf structure except for tRNASer1(AGN) which have a reduced DHU arm. The results showed that R. chenfui is closely related with R. schlegelii, R. arboreus, Zhangixalus omeimontis and Z. dugritei. This work enriches the library of Rhacophoridae mitoenomes and provides a valuable resource for understanding the evolutionary history of Rhacophorus.

Characterization of soil organic carbon at profile scale in two forest soils under pine and holm oak

Context It is essential to promote soil carbon sequestration as a means to mitigate climate change. Thus, it is fundamental to know the distribution of C in the soil at profile scale, the characteristics of humic substances as indicators of soil organic matter (SOM) turnover and their relationship with other soil properties. Aims Two forest ecosystems under pine and holm oak developed under a Meso-Mediterranean climate in Spain were used to characterize SOM through the complete sequence of layers of the soil profile. Methods General soil analysis, infrared spectroscopic analysis and soil color measurements were conducted for the characterization. Key results Humus form under oak was found to be Mull mesotrophic–Mull acid while humus under pine Moder oligotrophic. The infrared spectrum determined that oxidation of the humic acids was more complete in the deeper horizons. Relationships between intensities of the main spectral bands in both soils followed similar maximum and minimum sequence values. The total humic extract (THE) color measured by reflection was found inversely related to the THE color measured by transmission. In the same way, the color spectrum between 350 and 800 nm in the THE showed an inverse relation between hue/brightness and absorbance values. Conclusions Infrared analysis and color measurements provided evidence of a different level of stabilization of humic substances from each soil, and between the different horizons. Large spatial variability in soil organic carbon quantity and quality was observed. Implications Better understanding of carbon sequestration behavior in different soil ecosystems in its crucial role within the global carbon cycle

Complete genome sequence of Promicromonospora sp. strain Populi, an actinobacterium isolated from Populus trichocarpa rhizosphere.

Promicromonospora sp. strain Populi is an actinobacterium isolated from the rhizosphere of a black cottonwood tree, Populus trichocarpa. We completely sequenced its 5.2-Mbp chromosome using Oxford Nanopore long reads and predicted it to encode 4,685 proteins, 3 rRNA operons, and 54 tRNAs and noncoding RNAs.

The Complete Mitochondrial Genome of Nephropsis grandis: Insights into the Phylogeny of Nephropidae Mitochondrial Genome.

The systematic phylogeny of Pleocyemata species, particularly within the family Nephropidae, remains incomplete. In order to enhance the taxonomy and systematics of Nephropidae within the evolutionary context of Pleocyemata, we embarked upon a comprehensive study aiming to elucidate the phylogenetic position of Nephropsis grandis. Consequently, we determined the complete mitochondrial DNA sequence for N. grandis. The circular genome spans a length of 15,344bp and exhibits a gene composition analogous to that observed in other metazoans, encompassing a comprehensive set of 37 genes. Additionally, the genome features an AT-rich region. The rRNAs exhibited the highest AT content among the 37 genes (70.41%), followed by tRNAs (67.42%) and protein-coding genes (PCGs) (62.76%). The absence of a dihydrouracil arm in trnS1 prevented the formation of the canonical cloverleaf secondary structure. Selective pressure analysis indicated that the PCGs underwent purifying selection. The Ka/Ks ratios for cox1, cox2, cox3, and cob were considerably lower compared to other PCGs, implying strong purifying selection acting upon these particular genes. The mitochondrial gene order in N. grandis was consistent with the reported order in ancestral Pleocyemata. Phylogenetic revealed that N. grandis forms a cluster with the genus Metanephrops, and this cluster further groups with Homarus and the genus Nephrops within the Nephropidae family. These findings provide robust support for N. grandis as an ancestral member of the Nephropidae family. This study highlights the significance of employing complete mitochondrial genomes in phylogenetic analysis and deepens our understanding of the evolution of the Nephropidae family.

Plastid phylogenomics of Robinsonia (Senecioneae; Asteraceae), endemic to the Juan Fernández Islands: insights into structural organization and molecular evolution

BackgroundThe genus Robinsonia DC. (tribe Senecioneae, Asteraceae) endemic to the Juan Fernández Islands in Chile is one of the most conspicuous insular plant groups in the world. Unlike typical herbaceous Asteraceae plants, these plants demonstrate spectacular and unusual rosette tree growth forms as shown by the alpine giant senecios (genus Dendrosenecio, tribe Senecioneae) endemic to the East African mountains. However, monophyly of the genus and phylogenetic relationships among species of Robinsonia as well as their plastome evolution remain elusive. This study aims to explore their phylogeny, species diversification, and molecular evolution based on the complete plastome sequences in the context of adaptive radiation on oceanic islands.ResultsThe insular Robinsonia plastomes are highly conserved in their structures and organization of contents. Five divergence hotspots as potential chloroplast markers and five positively selected coding genes (accD, ndhF, rpoA, ycf1, and ycf2) are identified. Robinsonia plastomes has an overall nucleotide diversity higher than that of the sky island Dendrosenecio, but much lower than herbaceous Senecio. Phylogenetic analysis demonstrates the monophyly of Robinsonia and identifies two major infrageneric lineages. Both Robinsonia and Dendrosenecio are deeply nested within large genus Senecio.ConclusionsWhile plastid genomes of Robinsonia are highly conserved, their sequences strongly demonstrated the monophyly of the genus and inferred robust interspecific relationships, including herbaceous Senecio and woody Dendrosenecio. Different sets of positively selected chloroplast genes, five for Robinsonia and two for Dendrosenecio, may play an important role in the adaptation strategies of these fascinating woody species in insular and continental sky island habitats. Overall phylogenetic positions and sister lineages of Robinsonia and Dendrosenecio require additional study based on broader sampling of Senecio.

Numerical simulation on multi-well fracturing considering multiple thin layers in vertical direction

Multiwell fracturing is a key technology for developing shale gas and shale oil reservoirs. In this study, a multiple planar 3D (PL3D) fracture simulator that can capture multiple thin layers was developed to examine the propagation of multiple fractures during multicluster fracturing in multiple horizontal wells. The simulator considers multiple thin layers in the vertical direction. The results of the model are validated against the analytical solution of a single radial fracture and the implicit level set algorithm (ILSA). Using the simulator, a series of numerical simulations based on the field case are performed to investigate the fracture propagation mechanism of multiwell fracturing. The completion sequence, well placement pattern, well spacing, and cluster spacing are investigated to optimize the treatment parameters. The effective fracture area is used to quantitatively describe the stimulation effect. The adaptability of the completion sequence and well placement pattern is also analysed from the perspective of “frac hits”. The results show that the completion sequence has a critical influence on the stimulation effect and fracture geometry. From the perspective of avoiding “frac-hit” fractures, fracturing the low-stress layer can form an “artificial stress barrier”, which slightly protects the well from interference from other fractures. The staggered well pattern is better than the stacked well pattern. Compared with the stacked pattern, the staggered pattern can reduce the overlap area of fractures by 80 %, which greatly reduces the probability of “frac-hits”. With increasing well spacing from 200 m to 500 m, the fracture area increases by 25 %, and the degree of uneven stimulation between the two pay zones also increases by 6 %. Considering that a small well spacing is prone to “frac hits”, a large well spacing leads to an unstimulated area between two wells, and a 350 m well spacing is optimal. The effective fracture area decreases slightly with increasing perforation cluster spacing, but the fracture geometry becomes much more regular. The results can be helpful for the field design of multiwell fracturing.