MiSeq Sequencing Research Articles

Response of Soil Nitrogen Components and nirK- and nirS-Type Denitrifying Bacterial Community Structures to Drip Irrigation Systems in the Semi-Arid Area of Northeast China

Denitrification is a key process in soil available nitrogen (N) loss. However, the effects of different water-saving irrigation systems on soil N components and denitrifying bacterial communities are still unclear. In this study, quantitative fluorescence PCR and Illumina MiSeq sequencing were used to investigate the effects of three main irrigation systems, conventional flooding irrigation (FP), shallow buried drip irrigation (DI), and mulched drip irrigation (MF), on the abundance, community composition, and diversity of soil nirK- and nirS-type denitrifying bacteria in the semi-arid area of Northeast China, and to clarify the driving factors of nirK- and nirS-type denitrifying bacterial community variations. The results showed that, compared with FP, MF significantly increased soil moisture, alkaline hydrolyzed nitrogen (AHN), nitrate nitrogen (NO3−-N), non-acid hydrolyzed nitrogen (AIN), and amino sugar nitrogen (ASN), but significantly decreased the contents of ammonium nitrogen (NH4+-N) and acid hydrolyzed ammonium nitrogen (AN). The irrigation system changed the relative abundance of the dominant genera of denitrifying bacteria, DI and MF significantly increased nitrate reductase (NR) and nitrite reductase (NiR) activities, and MF significantly increased the diversity of nirK- and nirS-type denitrifying bacteria but significantly decreased the richness. The community structure of nirK- and nirS-type denitrifying bacteria was significantly different among the three irrigation systems. NO3−-N was the main driving factor affecting the community structure of nirS-type denitrifying bacteria, and moisture significantly affected the community structure of nirK-type denitrifying bacteria. DI and MF significantly increased the abundance of nirK- and nirS-type denitrifying bacteria and also increased the abundance ratio of nirS/nirK genes. Therefore, although DI and MF significantly increased the abundance of denitrifying microorganisms, they did not lead to an increase in the N2O emission potential.

Microbiomic Analysis of Bacteria Associated with Rock Tripe Lichens from Alpine Areas in Eastern Alps and Equatorial Africa

The diversity of bacteria associated with alpine lichens was profiled. Lichen samples belonging to the Umbilicariaceae family, commonly known as rock tripe lichens, were gathered from two distinct alpine fellfields: one situated on Mt. Brennkogel located in the Eastern European Alps (Austria), and the other on Mt. Stanley located in the Rwenzori mountains of equatorial Africa (Uganda). The primary aim of this research was to undertake a comparative investigation into the bacterial compositions, and diversities, identifying potential indicators and exploring their potential metabolisms, of these lichen samples. Bulk genomic DNA was extracted from the lichen samples, which was used to amplify the 18S rRNA gene by Sanger sequencing and the V3-V4 region of the 16S rRNA gene by Illumina Miseq sequencing. Examination of the fungal partner was carried out through the analysis of 18S rRNA gene sequences, belonging to the genus Umbilicaria (Ascomycota), and the algal partner affiliated with the lineage Trebouxia (Chlorophyta), constituted the symbiotic components. Analyzing the MiSeq datasets by using bioinformatics methods, operational taxonomic units (OTUs) were established based on a predetermined similarity threshold for the V3-V4 sequences, which were assigned to a total of 26 bacterial phyla that were found in both areas. Eight of the 26 phyla, i.e. Acidobacteriota, Actinomycota, Armatimonadota, Bacteroidota, Chloroflexota, Deinococcota, Planctomycetota, and Pseudomonadota, were consistently present in all samples, each accounting for more than 1% of the total read count. Distinct differences in bacterial composition emerged between lichen samples from Austria and Uganda, with the OTU frequency-based regional indicator phyla, Pseudomonadota and Armatimonadota, respectively. Despite the considerable geographic separation of approximately 5430 km between the two regions, the prediction of potential metabolic pathways based on OTU analysis revealed similar relative abundances. This similarity is possibly influenced by comparable alpine climatic conditions prevailing in both areas.

Research on carbon and nitrogen removal of tetramethylammonium hydroxide containing wastewater by combined anaerobic/integrated fixed film activated sludge process

Tetramethylammonium hydroxide (TMAH) is widely used as a developer and etchant in the thin-film transistor liquid crystal display industry, which is the main component of developer wastewater with low C/N ratio. This study investigated TMAH degradation by combined anaerobic/integrated fixed film activated sludge (A/IFAS) process, especially for nitrogen removal. Effects of process condition on the TMAH degradation were studied, including dissolved oxygen concentration in IFAS reactor and the temperature of anaerobic reactor. Especially, the nitrogen removal was studied through the monitoring of intermediate products during TMAH biodegradation. The results indicated that lower the anaerobic treatment temperature can provide more available organic matters to enhance the denitrification in the subsequent IFAS reactor. Less oxygen supply in the IFAS reactor contributed to simultaneous nitrification and denitrification. Removal efficiency of total organic carbon and total nitrogen was up to 95.8% and 80.7%, when the temperature of anaerobic treatment was controlled at 30 °C with the DO kept at 0.7 mg/L. It indicated that A/IFAS process was efficient in carbon and nitrogen removal for TMAH degradation. The results also confirmed intermediate products of TMAH biodegradation can be used as the electron donor during denitrification, including trimethylamine, dimethylamine and methylamine. Illumina MiSeq sequencing showed that Proteobacteria was the dominant phylum contribute to nitrogen removal. Compared to sludge flocs in IFAS reactor, richer community and higher microbial diversity were observed in the biofilm.

Characteristics of intestinal microbiota in preterm infants and the effects of probiotic supplementation on the microbiota.

In this study, we investigated the characteristics of the intestinal microbiota of preterm infants, and then analyzed the effects of probiotics supplementation on intestinal microbiota in preterm infants. This study enrolled 64 infants born between 26 and 32 weeks gestational age (GA) and 22 full-term infants. 34 premature infants received oral probiotic supplementation for 28 days. Stool samples were obtained on the first day (D1) and the 28th day (D28) after birth for each infant. Total bacterial DNA was extracted and sequenced using the Illumina MiSeq Sequencing System, specifically targeting the V3-V4 hyper-variable regions of the 16S rDNA gene. The sequencing results were then used to compare and analyze the composition and diversity index of the intestinal microbiota. There was no significant difference in meconium bacterial colonization rate between premature and full-term infants after birth (p > 0.05). At D1, the relative abundance of Bifidobacterium, Bacteroides, and Lactobacillus in the stool of preterm infants was lower than that of full-term infants, and the relative abundance of Acinetobacter was higher than that of full-term infants. The Shannon index and Chao1 index of intestinal microbiota in preterm infants are lower than those in full-term infants (p < 0.05). Supplementation of probiotics can increase the relative abundance of Enterococcus and Enterobacter, and reduce the relative abundance of Escherichia and Clostridium in premature infants. The Chao1 index of intestinal microbiota decreased in preterm infants after probiotic supplementation (p < 0.05). The characteristics of intestinal microbiota in preterm infants differ from those in full-term infants. Probiotic supplementation can reduce the relative abundance of potential pathogenic bacteria and increase the abundance of beneficial microbiota in premature infants.

Investigation of gut microbiota diversity according to infectious agent in pediatric infectious acute gastroenteritis in a Korean university hospital

BackgroundAcute gastroenteritis (AGE) is a common cause of pediatric morbidity and mortality worldwide. AGE can cause an imbalance in the intestinal microbiota. This study aimed to investigate the diversity of the gut microbiome in Korean children hospitalized for infectious AGE at a university hospital. MethodsA total of 23 stool samples from patients aged 5 months to 11 years with AGE were analyzed. Thirteen convalescent stool samples were collected 1 month after discharge. Multiplex polymerase chain reaction (PCR) for the five viruses and 16 bacteria-specific AGE pathogens (PowerChek Multiplex Real time PCR Kit, Seoul, Korea), and 16s rRNA sequencing (Illumina MiSeq Sequencing system, Illumina, USA) were performed. ResultsAccording to the results of multiplex PCR for causative pathogens, the microbiome taxonomic profile (MTP) of the gut microbiome in three groups of AGE, norovirus AGE (n = 11), Campylobacter AGE (n = 7) and Salmonella AGE (n = 5) was compared. The phylum Actinobacteria was significantly more abundant in the norovirus AGE (P = 0.011), whereas the phylum Proteobacteria was significantly more abundant in Salmonella AGE (P = 0.012). Alpha diversity, which indicates species richness and diversity, showed no statistical differences. However, beta diversity, representing the similarity in MTP between norovirus, Campylobacter, and Salmonella AGE, was significantly different (P = 0.007). In convalescence, compared with their corresponding AGE samples, the phylum Firmicutes; and the lower taxa Christensenellaceae (P = 0.0152) and Lachnospiraceae (P = 0.0327) were significantly increased. ConclusionsIn pediatric AGE, the type of infectious agent can affect the diversity and dominance of gut microbiota in pediatric patients. Furthermore, healthy gut bacteria increased during the period of 1 month after infection, allowing a return to a healthy state without causing long-term dysbiosis.

Seasonal changes of prokaryotic microbial community structure in Zhangjiayan Reservoir and its response to environmental factors

As a typical sub-deep reservoir in the upper reaches of the Yangtze River in the southwest region, Zhangjiayan Reservoir is also an important source of drinking water. Exploring the role of microorganisms in the material cycle of water bodies is of great significance for preventing the exacerbation of eutrophication in the reservoir. In this study, water samples from the overlying water of five points in the reservoir were collected four times in spring (April), summer (July), autumn (November), and winter (January) of 2022–2023 using a gas-tight water sampler. Physicochemical factors were measured, and the microbial community structure was analyzed by high-throughput MiSeq sequencing of the V3–V4 hypervariable region of 16S rRNA gene in order to explore the relationship between physicochemical factors and microbial community structure and the dominant microbial populations that affect eutrophication of the reservoir. The following results were obtained through analysis. Among the 20 overlying water samples from Zhangjiayan Reservoir, a total of 66 phyla, 202 classes, 499 orders, 835 families, 1716 genera, and 27,904 ASVs of the bacterial domain were detected. The phyla Proteobacteria and Actinobacteria were dominant in the microbial community of the overlying water in Zhangjiayan Reservoir. At the genus level, hgcI_clade and Actinobacteria had the highest abundance and was the dominant population. The microbial community in the water of Zhangjiayan Reservoir has a high level of diversity. The diversity index ranked by numerical order was winter > autumn > summer > spring. Significant differences were found in the composition and structure of the microbial community between the spring/summer and autumn/winter seasons (p < 0.05). Total phosphorus, dissolved total phosphorus, soluble reactive phosphorus, and dissolved oxygen have a significant impact on the composition and structure of the microbial community (p < 0.01). The bacterial community in the overlying water of Zhangjiayan Reservoir showed a mainly positive correlation. Sphingomonas, Brevundimonas, and Blastomonas were the central populations of the bacterial community in the overlying water of Zhangjiayan Reservoir. This study indicates that environmental factors, such as phosphorus and other nutrients, have a significant impact on the formation of the microbial community structure in different seasons. Sphingomonas, Brevundimonas, and Blastomonas are key populations that may have a significant impact on eutrophication in Zhangjiayan Reservoir.

Somatic CAG repeat instability in intermediate alleles of the HTT gene and its potential association with a clinical phenotype.

Huntington disease (HD) is a neurodegenerative disorder caused by ≥36 CAGs in the HTT gene. Intermediate alleles (IAs) (27-35 CAGs) are not considered HD-causing, but their potential association with neurocognitive symptoms remains controversial. As HTT somatic CAG expansion influences HD onset, we hypothesised that IAs are somatically unstable, and that somatic CAG expansion may drive phenotypic presentation in some IA carriers. We quantified HTT somatic CAG expansions by MiSeq sequencing in the blood DNA of 164 HD subjects and 191 IA (symptomatic and control) carriers, and in the brain DNA of a symptomatic 33 CAG carrier. We also performed genotype-phenotype analysis. The phenotype of symptomatic IA carriers was characterised by motor (85%), cognitive (27%) and/or behavioural (29%) signs, with a late (58.7 ± 18.6 years), but not CAG-dependent, age at onset. IAs displayed somatic expansion that were CAG and age-dependent in blood DNA, with 0.4% and 0.01% of DNA molecules expanding by CAG and year, respectively. Somatic expansions of +1 and +2 CAGs were detected in the brain of the individual with 33 CAGs, with the highest expansion frequency in the putamen (10.3%) and the lowest in the cerebellum (4.8%). Somatic expansion in blood DNA was not different in symptomatic vs. control IA carriers. In conclusion, we show that HTT IAs are somatically unstable, but we found no association with HD-like phenotypes. It is plausible, however, that some IAs, close to the HD pathological threshold and with a predisposing genetic background, could manifest with neurocognitive symptoms.

Variation of Soil Bacterial Communities in Forest Soil Contaminated with Chainsaw Lubricants.

Pollutants can exist in the soil for a long time and alter the bacterial community. Using lubricants to prevent the wear of chainsaw blades is necessary for thinning activities and wood harvesting. We investigated the influences of soil contamination with chainsaw lubricants on soil bacterial communities. Bio-oil, mineral oil, and recycled oil were scattered on each treatment to investigate variations in soil bacterial structure during treated periods using the Illumina MiSeq sequencing platform. The results obtained were 5943 ASVs, 5112 ASVs, and 6136 ASVs after treatment at one month, six months, and twelve months, respectively. There was a significant difference in Shannon and Simpson indices between treatments and controls. A total of 46 bacterial genera with an average relative abundance of more than 1.0% were detected in all soil samples. Massilia was the most common genus detected in control at one month, with an average relative abundance of 14.99%, while Chthoniobacter was the most abundant genus detected in bio-oil, mineral oil, and recycled oil treatments at one month, with an average relative abundance of 13.39%, 14.32%, and 10.47%, respectively. Among the three chainsaw lubricants, bio-oil and mineral oil had fewer impacts than recycled oil. The abundances of several functional bacteria groups in the bio-oil treatment were higher than in other treatments and controls. Our results indicated that different chainsaw lubricants and their time of application affected the soil bacterial community composition.

Unraveling the correlations between microbial communities and metabolic profiles of strong-flavor Jinhui Daqu with different storage periods

Daqu is a saccharification agent required for fermenting Baijiu, a popular Chinese liquor. Our objective was to investigate the relationships between physicochemical indices, microbial community diversity, and metabolite profiles of strong-flavor Jinhui Daqu during different storage periods. During different storage periods of Jinhui Daqu, we combined Illumina MiSeq sequencing and non-target sequencing techniques to analyze dynamic changes of the microbial community and metabolite composition, established a symbiotic network and explored the correlation between dominant microorganisms and differential metabolites in Daqu. Fungal community diversity in 8d_Daqu was higher than that in 45d_Daqu and 90d_Daqu, whereas bacterial community diversity was higher in 90d_Daqu. Twelve bacterial and four fungal genera were dominant during storage of Daqu. Bacillus, Leuconostoc, Kroppenstedtia, Lactococcus, Thermomyces and Wickerhamomyces decreased as the storage period increased. Differences of microbiota structure led to various metabolic pathways, and 993 differential metabolites were found in all Daqu samples. Differential microorganisms were significantly related to key metabolites. Major metabolic pathways involved in the formation of amino acids and lipids, such as l-arogenate and hydroxyproline, were identified. Interactions between moisture, acidity, and microbes may drive the succession of the microbial community, which further affects the formation of metabolites.

Soil Microbiome of Abandoned Plaggic Podzol of Different-Aged Fallow Lands and Native Podzol in South Taiga (Leningrad Region)

The soil microbiome is composed of various communities that play an important role in the existence of ecosystem services and the sustainable functioning of ecosystems under high anthropogenic loads. The transition of soils to a fallow state and their subsequent transformation lead to a notable alteration in the taxonomic composition of the soil microbiome, impacting the biochemical processes within the soil and its fertility levels. The object of this study comprised different-aged fallow soils of the southern taiga in the vicinity of Ban’kovo village, Leningrad region. The method comprising the high-throughput sequencing of 16S rRNA gene fragments using an Illumina MiSEQ sequencer was used to analyze the microbial community. The general processing of sequences was carried out with the dada2 (v1.14.1) package. It was found that the morphological organization of fallow soils has significant differences from the native podzol. In fallow soils, there are signs of leaching expressed in the accumulation of leached mineral particles, which indicates the degradation of the fallow–arable horizon. At the same time, there is a decrease in the content of P2O5 and K2O and an increase in the content of N-NH4 and N-NO3 in fallow soil. The analysis of alpha diversity index values showed that the highest level of alpha diversity in the microbial community is characteristic of 40-year-old soil, the alpha diversity index decreased with the increasing time of the fallow state, and the lowest alpha diversity index was observed in the native podzol. According to the values of the beta diversity index, a high correlation between the soil microbiome and the physicochemical characteristics of the soil was revealed, which indicates the formation of functional specialization in the studied microbial communities. As a result of the study of the taxonomic composition of microbial communities in fallow soils, it was found that the most represented microbial communities in fallow soils belong to Nitrosomonadaceae (Pseudomonadota), Mycobacterium (Actinobacteria), Nitrospira (Nitrospirota), and Luteolibacter (Verrucomicrobiota). The duration of post-agrogenic transformation is the leading factor influencing the changes in microbial communities; so, with an increase in the time that soils were in a fallow state, an increase in the oligotrophic microbial community was observed.

Isolation and characterization of polymorphic microsatellite loci for the three Iberian vipers, Vipera aspis, V. latastei and V. seoanei by Illumina MiSeq sequencing

BackgroundEuropean vipers (genus Vipera) are a well-studied taxonomic group, but the low resolution of nuclear sanger-sequenced regions has precluded thorough studies at systematic, ecological, evolutionary and conservation levels. In this study, we developed novel microsatellite markers for the three Iberian vipers, Vipera aspis, V. latastei and V. seoanei, and assessed their polymorphism in north-central Iberian populations.Methods and resultsGenomic libraries were developed for each species using an Illumina Miseq sequencing approach. From the 70 primer pairs initially tested, 48 amplified reliably and were polymorphic within species. Cross-species transferability was achieved for 31 microsatellites loci in the three target species and four additional loci that were transferable to one species only. The 48 loci amplified in average seven alleles, and detected average expected and observed heterozygosities of 0.7 and 0.55, in the three genotyped populations/species (26 V. aspis, 20 V. latastei and 10 V. seoanei).ConclusionsOur study provides a selection of 48 polymorphic microsatellite markers that will contribute significantly to current knowledge on genetic diversity, gene flow, population structure, demographic dynamics, systematics, reproduction and heritability in these species, and potentially in other congeneric taxa.

Peanut-based intercropping systems altered soil bacterial communities, potential functions, and crop yield.

Intercropping is an efficient land use and sustainable agricultural practice widely adopted worldwide. However, how intercropping influences the structure and function of soil bacterial communities is not fully understood. Here, the effects of five cropping systems (sole sorghum, sole millet, sole peanut, sorghum/peanut intercropping, and millet/peanut intercropping) on soil bacterial community structure and function were investigated using Illumina MiSeq sequencing. The results showed that integrating peanut into intercropping systems increased soil available nitrogen (AN) and total nitrogen (TN) content. The alpha diversity index, including Shannon and Chao1 indices, did not differ between the five cropping systems. Non-metric multidimensional scaling (NMDS) and analysis of similarities (ANOSIM) illustrated a distinct separation in soil microbial communities among five cropping systems. Bacterial phyla, including Actinobacteria, Proteobacteria, Acidobacteria, and Chloroflexi, were dominant across all cropping systems. Sorghum/peanut intercropping enhanced the relative abundance of phyla Actinobacteriota and Chloroflexi compared to the corresponding monocultures. Millet/peanut intercropping increased the relative abundance of Proteobacteria, Acidobacteriota, and Nitrospirota. The redundancy analysis (RDA) indicated that bacterial community structures were primarily shaped by soil organic carbon (SOC). The land equivalent ratio (LER) values for the two intercropping systems were all greater than one. Partial least squares path modeling analysis (PLS-PM) showed that soil bacterial community had a direct effect on yield and indirectly affected yield by altering soil properties. Our findings demonstrated that different intercropping systems formed different bacterial community structures despite sharing the same climate, reflecting changes in soil ecosystems caused by interspecific interactions. These results will provide a theoretical basis for understanding the microbial communities of peanut-based intercropping and guide agricultural practice.

Phylogeny and Fungal Community Structures of Helotiales Associated with Sclerotial Disease of Mulberry Fruits in China.

Mulberry fruit sclerotiniose is a prevalent disease caused by the fungal species Ciboria shiraiana, C. carunculoides, and Scleromitrula shiraiana of the order Helotiales, and severely affects the production of mulberry. However, these species have only been identified using morphological and rDNA-ITS sequence analyses, and their genetic variation is unclear. To address this, morphological and two-locus (ITS and RPB2) phylogenetic analyses were conducted using culture-dependent and independent methods for 49 samples from 31 orchards across four provinces in China. Illumina MiSeq sequencing was used to assess the fungal communities obtained from fruits varying in disease severity and color from an orchard in Wuhan. Conidial suspensions of C. shiraiana and C. carunculoides isolated from diseased fruits, diseased fruits affected with hypertrophy and pellet sorosis sclerotiniose, and mycelia of Sclerotinia sclerotiorum were determined to be pathogenic to the mulberry cultivar YSD10. However, fruits inoculated with S. sclerotiorum mycelia exhibited nontypical disease symptoms, and mycelia and conidia obtained from C. carunculoides and S. shiraiana strains were not pathogenic. Maximum parsimony and Bayesian analyses using the sequences of the assessed loci indicated species variability with no evidence of geographic specialization. Metagenomic analysis revealed that the diversity of fungal communities was reduced with disease progression. Furthermore, within a single fruit, the presence of two Ciboria spp. was detected. These results provide novel insights into Ciboria spp., revealing the secondary infections caused by conidia in diseased fruits, genetic variations of the pathogens, and the occurrence of coinfection. This improved understanding of fungal pathogens will aid in developing effective disease control strategies.

An improved and extended dual-index multiplexed 16S rRNA sequencing for the Illumina HiSeq and MiSeq platform.

Recent advancements in next-generation sequencing (NGS) technology have ushered in significant improvements in sequencing speed and data throughput, thereby enabling the simultaneous analysis of a greater number of samples within a single sequencing run. This technology has proven particularly valuable in the context of microbial community profiling, offering a powerful tool for characterizing the microbial composition at the species level within a given sample. This profiling process typically involves the sequencing of 16S ribosomal RNA (rRNA) gene fragments. By scaling up the analysis to accommodate a substantial number of samples, sometimes as many as 2,000, it becomes possible to achieve cost-efficiency and minimize the introduction of potential batch effects. Our study was designed with the primary objective of devising an approach capable of facilitating the comprehensive analysis of 1,711 samples sourced from diverse origins, including oropharyngeal swabs, mouth cavity swabs, dental swabs, and human fecal samples. This analysis was based on data obtained from 16S rRNA metagenomic sequencing conducted on the Illumina MiSeq and HiSeq sequencing platforms. We have designed a custom set of 10-base pair indices specifically tailored for the preparation of libraries from amplicons derived from the V3-V4 region of the 16S rRNA gene. These indices are instrumental in the analysis of the microbial composition in clinical samples through sequencing on the Illumina MiSeq and HiSeq platforms. The utilization of our custom index set enables the consolidation of a significant number of libraries, enabling the efficient sequencing of these libraries in a single run. The unique array of 10-base pair indices that we have developed, in conjunction with our sequencing methodology, will prove highly valuable to laboratories engaged in sequencing on Illumina platforms or utilizing Illumina-compatible kits.

Comparative Analysis of Rhizosphere and Endosphere Fungal Communities in Healthy and Diseased Faba Bean Plants.

This study used the ITS approach based on Illumina MiSeq sequencing to assess the endosphere and rhizosphere fungal communities in healthy and diseased faba bean plants. The findings indicate that the most predominant phyla in all samples were Ascomycota (49.89-99.56%) and Basidiomycota (0.33-25.78%). In healthy endosphere samples, Glomeromycota (0.08-1.17%) was the only predominant phylum. In diseased endosphere samples, Olpidiomycota (0.04-1.75%) was the only predominant phylum. At the genus level, Penicillium (0.47-35.21%) was more abundant in rhizosphere soil, while Paraphoma (3.48-91.16%) was predominant in the endosphere roots of faba bean plants. Significant differences were observed in the alpha diversity of rhizosphere samples from different germplasm resources (p < 0.05). The fungal community structures were clearly distinguished between rhizosphere and endosphere samples and between healthy and diseased endosphere samples (p < 0.05). Saccharomyces was significantly enriched in diseased endosphere samples, whereas Apiotrichum was enriched in healthy endosphere samples. Vishniacozyma and Phialophora were enriched in diseased rhizosphere samples, while Pseudogymnoascus was enriched in healthy rhizosphere samples. Diseased samples displayed more strongly correlated genera than healthy samples. Saprotrophs accounted for a larger proportion of the fungal microbes in rhizosphere soil than in endosphere roots. This study provides a better understanding of the composition and diversity of fungal communities in the rhizosphere and endosphere of faba bean plants as well as a theoretical guidance for future research on the prevention or control of faba bean root rot disease.

Operating performance of sequential batch system with calcium-modified basalt fiber bio-nest exposed to two typical perfluoroalkyl compounds (PFOA and PFBA)

The effects of per- and polyfluoroalkyl substances (PFASs) on the application of sequential batch system with basalt fiber as a biological carrier material have been rarely reported. This study explored the decontamination performance, extracellular polymeric substances (EPS) and microbial community structure of sequencing batch system with calcium-modified basalt fiber (Ca-MBF) bio-nest exposed to perfluorooctanoic acid (PFOA) and perfluorobutyric acid (PFBA). The results showed that 0.5 mg/L PFOA and PFBA both inhibited nitrification by 24.67 % and 19.47 %, while promoted denitrification by 16.54 % and 11.15 %, respectively. Overall, compared to PFBA, PFOA inhibited the removal of total nitrogen (TN) and total phosphorus (TP) by 11.27 % and 9.49 %, respectively. Further analysis of EPS showed that under PFOA exposure, the total EPS and polysaccharide (PS) content decreased by 34.57 % and 40.78 %, respectively. In addition, PFOA and PFBA exhibited a phenomenon of initially promoting by 89.21 % and 55.29 %, and subsequently inhibiting on protein (PN) content by 50.22 % and 51.37 %, respectively. Analyses of Illumina Miseq sequencing showed that PFOA and PFBA exposure for 50 days both inhibited the richness and diversity of microbial communities, and the inhibitory effect of PFOA was greater than that of PFBA. In addition, PFOA and PFBA respectively domesticated the resistant bacteria Meganema and Acinetobacter. These results may provide valuable information for ecological restoration and biodegradation of PFOA and PFBA.

Stochastic processes dominated the soil bacterial community assemblages along an altitudinal gradient in boreal forests

Soil bacteria, as integral components of the soil microbial community, play pivotal roles in biogeochemical cycles and ecosystem functions in boreal forests. The altitudinal patterns of soil bacteria have been widely reported, but their community assembly is uncertain. Here, we investigated the soil bacterial community attributes (diversity, taxonomic and functional composition, and bacterial interactions) and ecological processes associated with community assembly on Mt. Oakley in the northern Greater Khingan Mountains via Illumina MiSeq sequencing and functional annotation tools. The alpha diversity indices of the soil bacteria exhibited a progressively decreasing trend with increasing altitude across seasons. The relative abundance of the dominant bacterial taxa was more sensitive to altitude than to season. More complex (more nodes and links) bacterial interactions were detected at the lowest and highest altitudinal sites, as well as in September. The variations in the taxonomic and functional compositions of the soil bacteria induced by altitude were mainly driven by the variations in soil pH and extracellular enzyme activities. Stochastic (dispersal limitation and drift) processes largely controlled the soil bacterial community assemblages across spatiotemporal scales. The community assemblages of soil bacteria were affected by altitude-induced changes in the taxonomic composition and functional groups. Collectively, our results have significant implications for understanding bacterial biogeography and community assembly mechanisms along altitudinal gradients in boreal forest ecosystems.

Response of the rhizosphere soil microbial diversity to different nitrogen and phosphorus application rates in a hulless barley and pea mixed-cropping system

The mixed-cropping mode of hulless barley (Hordeum vulgare L) and peas (Pisum sativum L) can optimize the planting system, effectively enhancing crop nitrogen and phosphorus utilization and increasing yield. This mode has been widely applied in Qinghai Province, China. In this study, a split-plot experimental design was employed to investigate the bacterial and fungal communities in the rhizosphere soil of monocropping and mixed-cropping treatments under different fertilization levels. The Illumina MiSeq sequencing technology was used for the research, and the physicochemical properties of the soil were measured. The results indicated that with the increase in the application of nitrogen and phosphorus, the bacterial α-diversity in the rhizosphere of the mixed-cropping system initially increased and then decreased after reaching a peak. The α-diversity index of soil bacteria was higher under the mixed-cropping mode than under the monocropping mode. For the rhizosphere soil fungi in the mixed-cropping system, the Shannon index initially increased and then decreased, while the Chao1 and ACE indices decreased initially and then increased. Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium, Sphingomonas, and Pseudomonas comprised the predominant bacterial genera in the rhizosphere soil of the hulless barley–pea intercropping treatment, whereas Fusarium, Metarhizium, and Cladosporium dominated among the fungal genera across the 12 treatments. Under the same fertilization levels, the relative abundances of Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium in the rhizosphere soil of the mixed-cropping treatment increased by 8.4 %, 7.1 %, 5.7 %, and 5.6 %, respectively, compared to the monoculture of hulless barley, with a decreasing trend as nitrogen and phosphorus levels increased. Compared to the monoculture of hulless barley, the relative abundance of Fusarium in the mixed-cropping treatment decreased by 37.3 %, 43.8 %, 56.8 %, and 40.3 %, respectively. Redundancy and correlation analyses suggested that the variation in bacterial and fungal diversity and community composition in the mixed-cropping zone due to the interaction between hulless barley and peas under different nitrogen and phosphorus fertilizer application levels was significantly correlated with changes in soil organic matter, available phosphorus, and alkaline hydrolyzable nitrogen.

Depth effects on bacterial community altitudinal patterns and assembly processes in the warm-temperate montane forests of China

Soil bacterial communities are essential for ecosystem function, yet their response along altitudinal gradients in different soil strata remains unclear. Understanding bacterial community co-occurrence networks and assembly patterns in mountain ecosystems is crucial for comprehending microbial ecosystem functions. We utilized Illumina MiSeq sequencing to study bacterial diversity and assembly patterns of surface and subsurface soils across a range of elevations (700 to 2100 m) on Dongling Mountain. Our results showed significant altitudinal distribution patterns concerning bacterial diversity and structure in the surface soil. The bacterial diversity exhibited a consistent decrease, while specific taxa demonstrated unique patterns along the altitudinal gradient. However, no altitudinal dependence was observed for bacterial diversity and community structure in the subsurface soil. Additionally, a shift in bacterial ecological groups is evident with changing soil depth. Copiotrophic taxa thrive in surface soils characterized by higher carbon and nutrient content, while oligotrophic taxa dominate in subsurface soils with more limited resources. Bacterial community characteristics exhibited strong correlations with soil organic carbon in both soil layers, followed by pH in the surface soil and soil moisture in the subsurface soil. With increasing depth, there is an observable increase in taxa-taxa interaction complexity and network structure within bacterial communities. The surface soil exhibits greater sensitivity to environmental perturbations, leading to increased modularity and an abundance of positive relationships in its community networks compared to the subsurface soil. Furthermore, the bacterial community at different depths was influenced by combining deterministic and stochastic processes, with stochasticity (homogenizing dispersal and undominated) decreasing and determinism (heterogeneous selection) increasing with soil depth.

Oral microbiome characteristics in patients with pediatric solid tumor.

Pediatric solid tumor, the abnormal proliferation of solid tissues in children resulting in the formation of tumors, represent a prevailing malignant ailment among the younger population. Extensive literature highlights the inseparable association linking oral microbiome and adult tumors, but due to differences in age of onset, characteristics of onset, etc., there are many differences between Pediatric solid tumors and adult tumors, and therefore, studying the relationship between Pediatric solid tumor and the oral microbiota is also essential. To unravel the distinct characteristics of the oral microbiota within Pediatric solid tumor patients, 43 saliva samples, encompassing 23 Pediatric solid tumor patients and 20 healthy controls, were diligently procured. A meticulous screening process ensued, and conducted microbial MiSeq sequencing after screening. We documented the oral microbiome attributes among pediatric diagnosed with solid tumors (PST), and meanwhile, we observed a significant trend of decreased oral microbiota diversity in the pediatric solid tumor group. There were notable disparities in microbial communities observed between the two groups, 18 genera including Veillonellaceae, Firmicutes unclassified, Coriobacteriia, Atopobiaceae, Negativicutes, were significantly enriched in PST patients, while 29 genera, including Gammaproteobacteria, Proteobacteria, Burkholderiales, Neisseriaceae, were dominant in the HCs group. It was found that PST group had 16 gene functions, including Amino acid metabolism, Cysteine and methionine metabolism, Photosynthesis antenna proteins, Arginine and proline metabolism, and Aminoacyl tRNA biosynthesi, were significantly dominant, while 29 gene functions that prevailed in HCs. This study characterized the oral microbiota of Pediatric solid tumor patients for the first time, and importantly, targeted biomarkers of oral microbiota may serve as powerful and non-invasive diagnostic tools for pediatric solid tumor patients.