Illumina High-throughput Sequencing Research Articles

Comprehensive transcriptome sequencing of silkworm Midguts: Uncovering extensive isoform diversity and alternative splicing in BmNPV-Sensitive and BmNPV-resistant strains

The silkworm, Bombyx mori, stands out as one of the few economically valuable insects within the realm of model organisms. However, Bombyx mori nucleopolyhedrovirus (BmNPV) poses a significant threat, decreasing the quality and quantity of silkworm cocoons. Over the past few decades, a multitude of researchers has delved into the mechanisms that underlie silkworm resistance to BmNPV, employing diverse methodologies and approaching the problem from various angles. Despite this extensive research, the role of alternative splicing (AS) in the silkworm's response to BmNPV infection has been largely unexplored. This study leveraged both third-generation (Oxford Nanopore Technologies) and second-generation (Illumina) high-throughput sequencing technologies to meticulously identify and analyze AS patterns in the context of BmNPV response, utilizing two distinct silkworm strains—the susceptible strain 306 and the resistant strain NB. Consequently, we identified five crucial genes (Dsclp, LOC692903, LOC101743583, LOC101742498, LOC101743809) that are linked to the response to BmNPV infection through AS and differential expression. Additionally, a thorough comparative analysis was conducted on their diverse transcriptomic expression profiles, including alternative polyadenylation, simple sequence repeats, and transcription factors.

Recruitment of beneficial cucumber rhizosphere microbes mediated by amino acid secretion induced by biocontrol Bacillus subtilis isolate 1JN2.

At present, the use of beneficial microorganisms to control cucumber Fusarium wilt is a widely used method, and the rhizosphere microecological reset is one of the mechanisms involved. However, how biocontrol strains reshape cucumber rhizosphere microecology remains to be further studied. The composition changes of cucumber root exudates induced by biocontrol strain 1JN2, the microbial ecology of cucumber rhizosphere and the colonization ability of biocontrol strain 1JN2 in cucumber rhizosphere were analyzed through UHPLC-MS/MS analysis, Illumina high-throughput sequencing and SEM, respectively. First, cucumber plants treated with biocontrol Bacillus 1JN2 reduced the disease severity of Fusarium wilt by 60%. Significant changes in cucumber root exudates were found after 1JN2 inoculation and the contents of four amino acids including glutamine, tryptophan, glycine and glutamic acid were significantly increased. Second, It was found that the bacterial diversity in the rhizosphere of cucumber was significantly increased in both the strain treatment group and the amino acid mixture treatment group, The number of Bacillus was the largest in all dominant populations, exceeded 20% in all treatment groups. The bacteria of Hydrogenispora and Vicinamibacteria were significantly increased after treatment. Overall, the results demonstrated that amino acid substances in cucumber root exudates induced by biocontrol strain 1JN2 can shift the cucumber root microenvironment and prevent the occurrence of Fusarium wilt disease.

Transcriptomic analysis of the response of Avena sativa to Bacillus amyloliquefaciens DGL1.

Bacillus amyloliquefaciens DGL1, isolated from the arid sandy areas in Dagler, Qinghai Province, China, promotes the growth of Avena sativa variety "Qing Yan 1". To elucidate the transcriptomic changes in the oat root system following interaction with DGL1 and to reveal the molecular mechanism by which DGL1 promotes oat growth, treatment and control groups of oat roots at 2, 4, 8, and 12 h after inoculation with a suspension of strain DGL1 were analyzed using Illumina high-throughput transcriptome sequencing technology. The differentially expressed genes were determined through Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses, and the metabolic pathways and key genes were analyzed. The results showed that 7874, 13,392, 13,169, and 19,026 differentially expressed genes were significantly enriched in the glycolysis/gluconeogenesis pathway, amino acid metabolism, nitrogen metabolism, plant hormone signal transduction, and other related metabolic pathways in the oat roots at 2, 4, 8, and 12 h after inoculation with a DGL1 suspension. The GO and KEGG enrichment analyses revealed that the genes encoding plasma membrane ATPase, phosphoglycerate kinase gene PGK, ammonium transporter protein gene AMT, cellulose synthase gene CSLF6, and growth hormone response family gene IAA18 were significantly upregulated. It is hypothesized that the pro-growth mechanism of strain DGL1 in oats is the result of the coordination of multiple pathways through the promotion of oat energy metabolism, phytohormone signaling, secondary metabolite synthesis, and amino acid metabolism.

Response of the microbial community structure to the environmental factors during the extreme flood season in Poyang Lake, the largest freshwater lake in China.

Poyang Lake is the largest freshwater lake in China, and there are several studies on the composition and diversity of bacteria in Poyang Lake, while few quantitative studies were carried out on the response of the bacterial community to environmental factors during the extreme flood season in Poyang Lake. The connected-lake heterogeneity of bacterial community composition (BCC) was investigated in Poyang Lake during the flood season in 2020. Illumina high-throughput sequencing technology was used in this study. The bacterial community structure in the water was different from that in the sediment of Poyang Lake during extreme flood seasons. The bacterial diversity in water was much lower than that in sediment. In the water column, the dominant phyla were Actinobacteriota, while the composition of bacteria in sediment was more complex than that in water, and the dominant phyla in sediment were Proteobacteria, Chloroflexi, Acidobacteriota, and Actinobacteriota. The bacterial diversity in the water of Poyang Lake showed seasonal dynamics, while no seasonal variation of bacterial communities in sediment was observed. The bacterial community structure in the sediment from the two bays and channel areas of Poyang Lake can be distinguished from each other. The microbial diversity in sediment gradually increased from the Sancha Bay to the Zhouxi Bay and then to the channel, but the total nitrogen (TN) concentration in sediment (STN) and the total phosphorus (TP) concentration in sediment (STP) showed opposite trends. This might be due to the anthropogenic disturbances from the extreme flood. The bacterial community structure in, water column was significantly correlated with WT, NH4-N, STP, SOM, Chl a, DO, TP, and Eh, while the bacterial community structure in sediment was significantly correlated with SOM and STP. The bacterial community structure in water was greatly different from that in sediment in Poyang Lake during extreme flood seasons. The bacterial community structure in the water column was not only sensitive to the geochemical characteristics of the water but also affected by some nutrient concentrations in the sediment. During the wet seasons, bacterial diversity was only affected by SOM and STP.

Paeonol attenuates nonalcoholic steatohepatitis by regulating intestinal flora and AhR/NLRP3/Caspase-1 metabolic pathway

Ethnopharmacological relevanceNon-alcoholic steatohepatitis (NASH) is a common metabolic liver injury disease that is closely associated with obesity and metabolic disorders. Paeonol, an active ingredient found in Moutan Cortex, a traditional Chinese medicine which exhibits significant therapeutic effect on liver protection, has shown promising effects in treating liver diseases, particularly NASH. However, the specific intervention mechanism of paeonol on NASH is still unknown. Aim of the studyOur objective is to elucidate the pharmacological mechanism of paeonol in intervening NASH at the in vivo level, focusing on the impact on intestinal flora, tryptophan-related targeted metabolome, and related Aryl hydrocarbon receptor (AhR) pathways. Materials and methodsHere, we explored the intervention effect of paeonol on NASH by utilizing the NASH mouse model. The Illumina highthroughput sequencing technology was preformed to determine the differences of gut microbiota of model and paeonol treatment group. The concentration of Indoleacetic acid is determined by ELISA. The intervention effect of NASH mouse and AhR/NLRP3/Caspase-1 metabolic pathway is analyzed by HE staining, oil red O staining, Immunohistochemistry, Immunofluorescence, Western blot and qRT-PCR assays. Fecal microbiota transplantation experiment also was performed to verify the intervention effect of paeonol on NASH by affecting gut microbiota. ResultsFirstly, we discovered that paeonol effectively reduced liver pathology and blood lipid levels in NASH mice, thereby intervening in the progression of NASH. Subsequently, through 16S meta-analysis, we identified that paeonol can effectively regulate the composition of intestinal flora in NASH mice, transforming it to resemble that of normal mice. Specifically, paeonol decreased the abundance of certain Gram-negative tryptophan-metabolizing bacteria. Moreover, we discovered that paeonol significantly increased the levels of metabolites Indoleacetic acid, subsequently enhancing the expression of AhR-related pathway proteins. This led to the inhibition of the NOD-like receptor protein 3 (NLRP3) inflammasome production and inflammation generation in NASH. Lastly, we verified the efficacy of paeonol in intervening NASH by conducting fecal microbiota transplantation experiments, which confirmed its role in promoting the AhR/NLRP3/cysteinyl aspartate specific proteinase (Caspase-1) pathway. ConclusionsOur findings suggest that paeonol can increase the production of Indoleacetic acid by regulating the gut flora, and promote the AhR/NLRP3/Caspase-1 metabolic pathway to intervene NASH.

Effects of Four-Year Oilseed Flax Rotations on the Soil Bacterial Community in a Semi-Arid Agroecosystem

Crop rotation aims to improve the sustainability and production efficiency of agricultural ecosystems, especially as demands for food and energy continue to increase. However, the regulation of soil microbial communities using crop rotation with oilseed flax and its relationship with key soil physicochemical driving factors are still not clear. In order to investigate this matter, we carried out a field study lasting four years involving various crop rotation sequences including FWPF, FPFW, PFWF, FWFP, ContF, and ContF1. In addition to evaluating soil physicochemical parameters, we employed Illumina high-throughput sequencing technology to explore the structure and variety of soil microbial communities. The findings indicated a notable rise in pH value with the FPFW treatment in contrast to other treatments, along with significant increases in AP, MBC, MBN, and qSMBC compared to ContF. The number of OTUs in the FPFW, WFPF, FPFW, and PFWF treatments was significantly increased by 4.10–11.11% compared to ContF (p < 0.05). The presence of Actinobacteria and Acidobacteria was greatly impacted by the FPFW treatment, whereas the presence of Actinobacteria and Chloroflexi was notably influenced by the ContF treatment. The soil bacterial community was primarily influenced by TC, pH, and NO3−-N according to correlation analysis. Specifically, the FPFW therapy notably raised the soil pH level while lowering the TC level. Furthermore, the FPFW therapy led to a notable rise in the proportion of Acidobacteria and a significant decline in the proportion of Actinobacteria. These findings provide important theoretical support for using FPFW rotation to regulate soil microbial communities and solve the problems of continuous cropping.

Influences of the Integrated Rice-Crayfish Farming System with Different Stocking Densities on the Paddy Soil Microbiomes.

Integrated rice-fish farming has emerged as a novel agricultural production pattern to address global food security challenges. Aiming to determine the optimal, scientifically sound, and sustainable stocking density of red claw crayfish (Cherax quadricarinatus) in an integrated rice-crayfish farming system, we employed Illumina high-throughput 16S rRNA gene sequencing to evaluate the impact of different stocking densities of red claw crayfish on the composition, diversity, function, and co-occurrence network patterns of soil bacterial communities. The high stocking density of red claw crayfish reduced the diversity and evenness of the soil bacterial community during the mid-culture stage. Proteobacteria, Actinobacteria, and Chloroflexi emerged as the most prevalent phyla throughout the experimental period. Low stocking densities initially boosted the relative abundance of Actinobacteria in the paddy soil, while high densities did so during the middle and final stages. There were 90 distinct functional groups identified across all the paddy soil samples, with chemoheterotrophy and aerobic chemoheterotrophy being the most abundant. Low stocking densities initially favored these functional groups, whereas high densities enhanced their relative abundances in the later stages of cultivation. Medium stocking density of red claw crayfish led to a more complex bacterial community during the mid- and final culture stages. The experimental period showed significant correlations with soil bacterial communities, with total nitrogen (TN) and total phosphorus (TP) concentrations emerging as primary factors contributing to the alterations in soil bacterial communities. In summary, our findings demonstrated that integrated rice-crayfish farming significantly impacted the soil microbiomes and environmental factors at varying stocking densities. Our study contributed to theoretical insights into the profound impact of integrated rice-crayfish farming with various stocking densities on bacterial communities in paddy soils.

Helicobacter pylori causes gastric dysbacteriosis in chronic gastritis patients.

Gastric mucosal samples were procured and underwent the sequencing of 16S ribosomal RNA (16S rRNA) via Illumina high-throughput sequencing technology to explore the impact of Helicobacter pylori (H. pylori) infection on the composition of gastric flora in chronic gastritis (CG) patients. In the results, the operational taxonomic unit (OTU) analysis revealed an overlap of 5706 OTUs shared between the two groups. The top 5 abundance ranking (TOP5) phyla comprised Bacteroidetes, Proteobacteria, Firmicutes, Actinobacteria, and Epsilonbacteraeota, while the TOP5 genus was Lachnospiraceae_NK4A136_group, Helicobacter, Bacteroides, Klebsiella, and Pseudomonas. In the metabolic pathways at the Kyoto Encyclopedia of Genes and Genomes (KEGG)_L3 level, conspicuous variations across seven functions were observed between the H. pylori-positive (HP_Pos) and H. pylori-negative (HP_Neg) groups. Subsequently, functional gene enrichment in KEGG pathways was further validated through animal experimentation. In contrast to the mice in the HP_Neg group, those infected with H. pylori manifested an infiltration of inflammatory cells, an augmentation in gastric acid secretion, and conspicuously elevated scores regarding gastric activity, along with heightened levels of malondialdehyde. In conclusion, CG patients infected with H. pylori displayed a disorder in gastric flora, furnishing a theoretical basis for the prophylaxis of H. pylori infection and its associated pathogenic ramifications.

Characterization of macadamia ringspot-associated virus, a novel Orthotospovirus associated with Macadamia integrifolia in South Africa

South Africa is the largest producer of macadamias in the world, producing an estimated 61,288 tons nut-in-shell in 2022. In order to ensure the sustainability of the industry, it is important that research focuses on the control and eradication of economically important pests and diseases. Macadamia trees in the Mpumalanga Lowveld of South Africa have for some time shown severe chlorosis, which coincides with a significant drop in production, with losses of up to 60% being recorded. The disease has since been coined Macadamia Chlorosis Disease (MCD). In an attempt to determine whether MCD may be associated with a virus, high-throughput Illumina sequencing was performed on RNA extracted from both diseased and healthy trees collected from farms in Mpumalanga. Subsequent data analyses could not link a specific virus to MCD, however, reads spanning the full genome of a novel virus belonging to the Orthotospovirus genus were obtained. An RT-PCR assay was optimized for the detection of this virus and subsequent surveys linked the virus to ringspot symptoms which are commonly observed on different macadamia cultivars. The virus has to date been identified from orchards in Mpumalanga, Limpopo and KwaZulu-Natal. Other viruses described in the genus are known to cause severe crop losses and it is therefore important that the virus, provisionally named macadamia ringspot-associated virus (MRSV), be further studied to determine whether association with this virus can lead to yield losses, and whether appropriate control strategies must be implemented to prevent the spread of MRSV.

Unlocking water potential in drylands: Quicklime and fly ash enhance soil microbiome structure, ecological networks and function in acid mine drainage water-irrigated agriculture

In water-stressed regions, treated acid mine drainage (AMD) water for irrigated agriculture is a potential solution to address freshwater scarcity. However, a significant knowledge gap exists on the short and long-term effects of treated AMD water on soil health. This study used high-throughput Illumina sequencing and predictive metagenomic profiling to investigate the impact of untreated AMD (AMD), quicklime- (A1Q and A2Q) and quicklime and fly ash-treated AMD water (AFQ) irrigation on soil bacterial diversity, co-occurrence networks and function. Results showed that untreated AMD water significantly increased soil acidity, electrical conductivity (EC), sulfate (SO42−), and heavy metals (HM), including reduced microbial diversity, disrupted interaction networks, and functional capacity. pH, EC, Cu, and Pb were identified as key environmental factors shaping soil microbial diversity and structure. Predominantly, Pseudomonas, Ralstonia picketti, Methylotenera KB913035, Brevundimonas vesicularis, and Methylobacteriumoryzae, known for their adaptability to acidic conditions and metal resistance, were abundant in AMD soils. However, soils irrigated with treated AMD water exhibited significantly reduced acidity (pH > 6.5), HM and SO42− levels, with an enrichment of a balanced bacterial taxa associated with diverse functions related to soil health and agricultural productivity. These taxa included Sphingomonas, Pseudoxanthomonas, Achromobacter, Microbacterium, Rhodobacter, Clostridium, Massillia, Rhizobium, Paenibacillus, and Hyphomicrobium. Moreover, treated AMD water contributed to higher connectivity and balance within soil bacterial co-occurrence networks compared to untreated AMD water. These results show that quicklime/fly ash treatments can help lessen impacts of AMD water on soil microbiome and health, suggesting its potential for irrigated agriculture in water-scarce regions.

Regulation and analysis of Simiao Yong'an Decoction fermentation by Bacillus subtilis on the diversity of intestinal microbiota in Sprague-Dawley rats

Background and Aim: Simiao Yong'an decoction (SYD) is a classic traditional Chinese medicine (TCM) prescription that has the effects of clearing heat, detoxifying, promoting blood circulation, and relieving pain. In this study, we investigated the effect of SYD on the diversity of intestinal microbiota after fermentation by Bacillus subtilis. Materials and Methods: SYD was fermented using B. subtilis. Female Sprague-Dawley rats were randomly divided into the following four groups with six rats in each group: Negative sample group (NS), water exaction non-fermentation group (WE), B. subtilis group (BS), and fermentation liquid group (FL). All rats were orally administered for 14 days. High-throughput Illumina sequencing was used to analyze 16S rRNA expression in rat fecal samples. Results: A total of 2782 operational taxonomical units (OTUs) were identified in this study, and 634 OTUs were shared among all samples. Bacteroidetes (28.17%-53.20%) and Firmicutes (48.35%-67.83%) were the most abundant phyla identified among the four groups. The abundance of Escherichia and Alistipes was lower in the FL group than in the NS group, whereas the abundance of Bifidobacteria and Lactobacillus was increased in the FL group (p < 0.05). The abundance of Bifidobacterium was significantly upregulated in the FL group compared with the WE and BS groups (p < 0.05). Conclusion: After fermentation, SYD had a significantly better effect than SYD or B. subtilis. SYD significantly promoted the growth of intestinal probiotics, inhibited the growth of pathogenic bacteria, and maintained the balance of intestinal microbiota in SD rats. This study provides new insights into the development and use of SYD. Keywords: Bacillus subtilis, fermentation, intestinal microbiota, probiotics, Simiao Yong'an Decoction.

MHC-I alleles mediate clearance and antibody response to the zoonotic Lassa virus in Mastomys rodent reservoirs.

West African Mastomys rodents are the primary reservoir of the zoonotic Lassa virus (LASV). The virus causes haemorrhagic Lassa fever and considerable mortality in humans. To date, the role of Mastomys immunogenetics in resistance to, and persistence of, LASV infections is largely unknown. Here, we investigated the role of Major Histocompatibility Complex class I (MHC-I) on LASV infection status (i.e., active vs. cleared infection, determined via PCR and an immunofluorescence assay on IgG antibodies, respectively) in Mastomys natalensis and M. erythroleucus sampled within southwestern Nigeria. We identified more than 190 and 90 MHC-I alleles by Illumina high throughput-sequencing in M. natalensis and M. erythroleucus, respectively, with different MHC allele compositions and frequencies between LASV endemic and non-endemic sites. In M. natalensis, the MHC allele ManaMHC-I*006 was negatively associated with active infections (PCR-positive) and positively associated with cleared infections (IgG-positive) simultaneously, suggesting efficient immune responses that facilitate LASV clearance in animals carrying this allele. Contrarily, alleles ManaMHC-I*008 and ManaMHC-I*021 in M. natalensis, and MaerMHC-I*008 in M. erythroleucus, were positively associated with active infection, implying susceptibility. Alleles associated with susceptibility shared a glutamic acid at the positively selected codon 57, while ManaMHC-I*006 featured an arginine. There was no link between number of MHC alleles per Mastomys individual and LASV prevalence. Thus, specific alleles, but not MHC diversity per se, seem to mediate antibody responses to viremia. We conclude that co-evolution with LASV likely shaped the MHC-I diversity of the main LASV reservoirs in southwestern Nigeria, and that information on reservoir immunogenetics may hold insights into transmission dynamics and zoonotic spillover risks.

Comparative profiling of the skin and gut microbiota during metamorphosis in Chinese spiny frogs (Quasipaa spinosa) highlights microbial communities involved in disease pathogenesis

The symbiotic microbiota inhabiting the skin and gut of amphibians exerts roles in maintaining host health, especially during the critical stage of complete metamorphosis where failure to remodel the microbiota can result in various diseases. However, the composition and alterations in the amphibian microbiota in diverse diseased populations, as well as the exact mechanisms underlying disruptions in skin and gut microbial communities leading to diseases, are not yet well established. Herein, skin and gut microbiota were sampled from healthy Chinese spiny frogs (CTL group) as well as individuals with ascites disease (AD group), cataract disease (CD group), and rotten-skin disease (RD group) for Illumina high-throughput sequencing analysis to comprehensively explore potential associations between microbiota composition and the health status of the host. Venn diagrams and UpSet analysis revealed that the CTL group exhibited the highest number of unique amplicon sequence variants (ASVs) in their skin, while the CD group displayed the lowest abundance of ASVs in their gut. Compared to the CTL group, the alpha-diversity of skin microbiota was dramatically depressed in diseased groups, and notable variations in the beta-diversity of both skin and gut microbiota were observed. Further compositional analysis identified Firmicutes, Proteobacteria, and Bacteroidota as the dominant bacterial phyla in both skin and gut microbiota, although their relative abundances varied widely among groups. The linear discriminant analysis effect size (LEfSe) analysis identified various bacteria species with substantial differences between skin and gut microbiota groups including beneficial populations as well as opportunistic ones. Functional predictions indicated significant differences between CTL and diseased groups mainly enriched in metabolism-relevant pathways. Furthermore, in vitro culture experiments confirmed the presence of a total of 19 strains of bacteria from the skin microbiota of RD group, including opportunistic bacteria population Acinetobacter closely associated with spiny frog rotten-skin disease. Taken together, this work provides insights into the composition dynamics of skin and gut microbiota under different health conditions in spiny frogs, highlighting potential applications for symbiotic microbiota modification in promoting healthy amphibian culture practices as well as preventing diseases.

Arsenic biomineralization and selenium nanoparticles biosynthesis by Halomonas boliviensis strain H-10 isolated from the high-altitude Salar de Huasco salt flat (Chile)

High-altitude Andean wetlands of the Atacama Desert (Chile), including the Salar de Huasco salt flat, are unusual environments characterized by a great variability of physical-chemical conditions, often combined with a high concentration of heavy metals, including arsenic (As) and semimetals, such as selenium (Se). Microorganisms present in these environments evolved adaptative strategies to cope with environmental stresses, with biotechnological potentials. The present work investigated the natural bacterial community composition in sediments collected from the Salar de Huasco salt flat, by high-throughput Illumina sequencing, and to isolate bacterial strains able to remove As and biosynthesize Se nanoparticles (SeNPs). Dominant taxonomic groups (abundance ≥1%) were affiliated to Proteobacteria (mainly consisting in Delta- and Alfa- Gamma-proteobacteria) Bacteriodetes, Firmicutes, Acidobacteria, Spirochaetes, Deinococcus-Thermus, Gemmatimonadetes and Verrucomicrobia. Halomonas boliviensis strain H-10 was able to both biomineralize As and biosynthesise SeNPs; therefore, the characteristics of H. boliviensis strain H-10 could be used for biotechnological purposes, including As bioremediation from water and bioproduction of SeNPs.

Molecular Characterization and Phylogenetic Analysis of Centipedegrass [Eremochloa ophiuroides (Munro) Hack.] Based on the Complete Chloroplast Genome Sequence.

Centipedegrass (Eremochloa ophiuroides) is an important warm-season grass plant used as a turfgrass as well as pasture grass in tropical and subtropical regions, with wide application in land surface greening and soil conservation in South China and southern United States. In this study, the complete cp genome of E. ophiuroides was assembled using high-throughput Illumina sequencing technology. The circle pseudomolecule for E. ophiuroides cp genome is 139,107 bp in length, with a quadripartite structure consisting of a large single copyregion of 82,081 bp and a small single copy region of 12,566 bp separated by a pair of inverted repeat regions of 22,230 bp each. The overall A + T content of the whole genome is 61.60%, showing an asymmetric nucleotide composition. The genome encodes a total of 131 gene species, composed of 20 duplicated genes within the IR regions and 111 unique genes comprising 77 protein-coding genes, 30 transfer RNA genes, and 4 ribosome RNA genes. The complete cp genome sequence contains 51 long repeats and 197 simple sequence repeats, and a high degree of collinearity among E. ophiuroide and other Gramineae plants was disclosed. Phylogenetic analysis showed E. ophiuroides, together with the other two Eremochloa species, is closely related to Mnesithea helferi within the subtribe Rottboelliinae. These findings will be beneficial for the classification and identification of the Eremochloa taxa, phylogenetic resolution, novel gene discovery, and functional genomic studies for the genus Eremochloa.

Impact of artisanal refining activities on bacterial diversity in a Niger Delta fallow land

Hydrocarbon pollution is a major ecological problem facing oil-producing countries, especially in the Niger Delta region of Nigeria. In this study, a site that had been previously polluted by artisanal refining activity was investigated using 16S rRNA Illumina high-throughput sequencing technology and bioinformatics tools. These were used to investigate the bacterial diversity in soil with varying degrees of contamination, determined with a gas chromatography-flame ionization detector (GC-FID). Soil samples were collected from a heavily polluted (HP), mildly polluted (MP), and unpolluted (control sample, CS) portion of the study site. DNA was extracted using the Zymo Research (ZR) Fungi/Bacteria DNA MiniPrep kit, followed by PCR amplification and agarose gel electrophoresis. The microbiome was characterized based on the V3 and V4 hypervariable regions of the 16S rRNA gene. QIIME (Quantitative Insights Into Microbial Ecology) 2 software was used to analyse the sequence data. The final data set covered 20,640 demultiplexed high-quality reads and a total of 160 filtered bacterial OTUs. Proteobacteria dominated samples HP and CS, while Actinobacteria dominated sample MP. Denitratisoma, Pseudorhodoplanes, and Spirilospora were the leading genera in samples HP, CS, and MP respectively. Diversity analysis indicated that CS [with 25.98 ppm of total petroleum hydrocarbon (TPH)] is more diverse than HP (with 490,630 ppm of TPH) and MP (with 5398 ppm of TPH). A functional prediction study revealed that six functional modules dominated the dataset, with metabolism covering up to 70%, and 11 metabolic pathways. This study demonstrates that a higher hydrocarbon concentration in soil adversely impacts microbial diversity, creating a narrow bacterial diversity dominated by hydrocarbon-degrading species, in addition to the obvious land and ecosystem degradation caused by artisanal refining activities. Overall, the artisanal refining business is significantly driving ecosystem services losses in the Niger Delta, which calls for urgent intervention, with focus on bioremediation.

Characterization of a New, Country Bean (Lablab purpureus) Lineage of Bean Common Mosaic Necrosis Virus.

Country bean (Lablab purpureus, family Fabaceae) is grown in subsistence agriculture in Bangladesh as a multipurpose crop for food, animal feed, and green manure. This study was undertaken to investigate the genetic diversity of bean common mosaic necrosis virus (BCMNV, genus Potyvirus, family Potyviridae) in country beans. Leaf samples from country beans showing yellowing, vein banding, and mosaic symptoms were collected during field surveys between 2015 and 2019 cropping seasons from farmers' fields in different geographic regions. These samples were tested by serological and molecular diagnostic assays for the presence of BCMNV. Virus-positive samples were subjected to high-throughput Illumina sequencing to generate near-complete genomes of BCMNV isolates. In pairwise comparisons, the polyprotein sequences of BCMNV isolates from Bangladesh showed greater than 98% identities among themselves and shared less than 84% sequence identity at the nucleotide level with virus isolates reported from other countries. In the phylogenetic analysis, BCMNV isolates from Bangladeshi country beans formed a separate clade from virus isolates reported from common beans in other countries in the Americas, Africa, Europe, and from East Timor. Grow-out studies showed seed-to-seedling transmission of BCMNV, implying a possible seedborne nature of the virus in country beans.

Whole-genome sequencing of Fusarium spp. causing sugarcane root rot on both chewing cane and sugar-making cane

Previously we isolated three Fusarium strains (a F. sacchari strain namely GXUF-1, and another two F. commune strains namely GXUF-2 and GXUF-3), and we verified that GXUF-3 was able to cause sugarcane root rot to the chewing cane cultivar Badila. Considering that Fusarium spp. are a group of widely distributed fungal pathogens, we tested whether these three Fusarium isolates were able to cause root rot to Badila as well as sugar-making cane cultivar (Guitang42), using a suitable inoculation method established based on infection assays using Badila. We found that the three Fusarium strains were able to cause root rot symptoms to both Badila and Guitang42, to different extents. To better investigate the potential pathogenicity mechanisms, we performed Illumina high-throughput sequencing and analyzed the whole genomic sequence data of these three Fusarium strains. The results reveal that the assembly sizes of the three Fusarium strains were in a range of 44.7–48.2 Mb, with G + C contents of 48.0–48.5%, and 14,154–15,175 coding genes. The coding genes were annotated by multiple public databases, and potential pathogenic genes were predicted using proprietary databases (such as PHI, DFVF, CAZy, etc.). Furthermore, based on evolutionary analysis of the coding sequence, we found that contraction and expansion of gene families occurred in the three Fusarium strains. Overall, our results suggest a potential risk that the root rot disease may occur to the sugar-making canes although it was initially spotted from fruit cane, and provide clues to understand the pathogenic mechanisms of Fusarium spp. causing sugarcane root rot.

Metabarcoding of zooplankton communities of Dianchi Lake based on the mitochondrial cytochrome oxidase subunit 1 gene.

Freshwater lakes as an essential component of the ecosystem, provide ecological resources in addition to economic source for humans. Under recent climate change scenario, preserving the biodiversity of freshwater ecosystems is crucial. This study aimed to characterize the diversity of zooplankton communities in Dianchi Lake, located in Kunming Municipality, Yunnan Province, China, using Illumina high-throughput sequencing of the cytochrome oxidase subunit 1 (COI) gene marker. A total of 18 water samples were collected including 16 from the outer sea area of Dianchi Lake: 4 from the east (E1-4), 4 from the west (W1-4), 4 from the south (S1-4), and 4 from the north (N1-4), and: 2 from the Caohai area (C1-2) as research sites. All environmental parameters including pH, ammonium (NH4+), total nitrogen (TN), total phosphorus (TP), chlorophyll a content (CHLA) were found to be insignificant (p > 0.05), except for chemical oxygen demand (COD) and transparency (T), which were found to be significant (p < 0.05). Alpha diversity indices including ACE, Chao1, Shannon, and Simpson showed non-significant differences (p > 0.05), indicating no variation in the richness of zooplankton communities at different locations of Dianchi Lake. However, principal coordinate analysis (PCoA) showed that most of the samples from East, West, and South groups were close to each other, showing more similarities among them, while Caohai and North group samples were distant from each other, showing more differences with other groups. Rotifera, Arthropoda, and Chordata were the top three phyla, while Keratella, Macrothrix, and Brachionus were the dominant genera. Mantel test analysis showed that COD and transparency were important environmental factors that shaped the Rotifera community structure of Dianchi Lake. In conclusion, this study provides insights on conserving the diversity of zooplankton communities in Dianchi Lake, especially by controlling COD and maintaining water transparency, in order to preserve its ecological resources and economic significance.

Paddy-Lilium Crop Rotation Improves Potential Beneficial Soil Fungi and Alleviates Soil Acidification in Lilium Cropping Soil

Lilium growth is severely impeded by continuous cropping, and crop rotation is essential to reducing the detrimental effects of monocultures. Soil (0–20 cm) was collected in three Lilium cropping patterns in Longshan County, Hunan Province, including continuous Lilium cropping (Lilium), corn upland rotation with Lilium (Corn), and paddy rotation with Lilium (Rice). Using Illumina high-throughput sequencing technology, the fungal ribosomal DNA internal-transcribed spacer 1 (ITS1) was examined to evaluate the features of soil fungi communities among three cropping patterns. Crop rotation has an impact on soil properties and the microbial community. Rice soil has a significantly higher pH than Lilium and corn soil, while corn and rice soil have a greater total nitrogen and total phosphorus content than Lilium soil. Rotation cropping clearly shifted the fungi community diversity based on the results of principal coordinate analysis (PCoA) and nonmetric multidimensional scaling (NMDS). Ascomycota was the most prevalent phylum, with the highest levels in Lilium soil. Genetic analysis revealed that paddy rotation led to a clear reduction in or non-detection of eight potentially pathogenic fungal genera and a noticeable accumulation of eight beneficial fungal genera compared to Lilium continuous cropping. Fungi communities and their abundant taxa were correlated with soil pH and nutrients. Altogether, we propose that rice rotation, with its ability to mitigate soil acidification, reducing pathogenic and accumulating beneficial communities, may be an effective strategy for alleviating the continuous cropping barrier.