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• The commonly used MEA medium proved to be the least suitable for fungal capture. • Cultivation strategies to capture a wider range of fungi are still insufficient. • The need to use both culture-dependent and -independent approaches was highlighted. • Environmental conditions, not material composition, impact material colonisation. The biodeterioration of cultural heritage objects is often attributed to microscopic fungi and their metabolic activity. These fungi often produce pigments that result in aesthetic alterations. Furthermore, through physical mechanisms like hyphal penetration and chemical mechanisms such as the production of acids or other harmful substances, they can degrade the integrity and structure of a wide range of materials. It is therefore necessary to identify these microorganisms as an initial step towards preserving cultural heritage objects for future generations. This study aims to identify the fungi present on audio-visual materials stored in two Czech Republic archives. Employing both a culture-dependent approach, using four different cultivation media, and a culture-independent approach with Illumina MiSeq sequencing, comprehensive fungal identification was conducted. The culture-dependent method yielded the identification of only 11 isolates from 7 genera, highlighting the necessity to enhance fungal capture efficiency, with the MEA medium proving the least suitable. In contrast, 75 genera were identified by the culture-independent approach. However, minimal overlap in results between the approaches, with only the Aspergillus and Bjerkandera genera in common, emphasises the significance of employing both methods. Next, analysis in two different archives suggested a potential association of 16 genera directly with audio-visual materials rather than storage conditions, including the genera Filobasidium, Naganishia, Sporobolomyces , or Wallemia. Furthermore, factors influencing the composition of fungal communities on audio-visual materials were investigated, of which locality proved to be statistically significant. [Display omitted] [ABSTRACT FROM AUTHOR]

Litter decomposition is a key process governing carbon and nutrient cycles in forest ecosystems that is expected to be impacted by increasing atmospheric carbon dioxide (CO2) concentrations.We conducted two complementary field studies to assess the effects of elevated CO2 on Eucalyptus tereticornis litter decomposition processes. First, we used bags of two different mesh sizes to assess the effect of macrofauna and elevated CO2 over 24 months on mass loss of litter grown under ambient CO2. We then assessed the effect of elevated CO2 during the decomposition of litter grown under each combination of (i) ambient CO2 or elevated CO2 and (ii) during a psyllid outbreak that triggered significant canopy loss or later in canopy developing when psyllid densities were low.Both macrofauna and elevated CO2 enhanced mass loss at late decay stages in the first study, with no interactive effect. Again, mass loss was greater at elevated CO2 at late decay stages in the second study, particularly for non‐psyllid‐impacted litter grown at elevated CO2. In both studies, CO2 concentration during decomposition influenced fungal assemblages and these effects were observed before any effects on decomposition were observed, with some fungi linked to saprotrophic guilds being found with higher frequency under elevated CO2. CO2 concentrations under which leaves developed and whether leaves were psyllid‐impacted was also important in shaping fungal assemblages.Synthesis. The positive effect on mass loss at late decay stages is contrary to previous findings where elevated CO2 generally reduced decomposition rates. Our results show that elevated CO2 effects on decay rates are context‐specific. Further research is required to establish the mechanisms through which this occurs to better model elevated CO2 effects on global carbon dynamics. Read the free Plain Language Summary for this article on the Journal blog. [ABSTRACT FROM AUTHOR]

The study of microalgal communities is critical for understanding aquatic ecosystems. These communities primarily comprise diatoms (Heterokontophyta), with two methods commonly used to study them: Microscopy and metabarcoding. However, these two methods often deliver different results; thus, their suitability for analyzing diatom communities is frequently debated and evaluated. This study used these two methods to analyze the diatom communities in identical water samples and compare the results. The taxonomy of the species constituting the diatom communities was confirmed, and both methods showed that species belonging to the orders Bacillariales and Naviculales (class Bacillariophyceae) are the most diverse. In the lower taxonomic levels (family, genus, and species), microscopy tended to show a bias toward detecting diatom species (Nitzschia frustulum, Nitzschia inconspicua, Nitzschia intermedia, Navicula gregaria, Navicula perminuta, Navicula recens, Navicula sp.) belonging to the Bacillariaceae and Naviculaceae families. The results of the two methods differed in identifying diatom species in the communities and analyzing their structural characteristics. These results are consistent with the fact that diatoms belonging to the genera Nitzschia and Navicula are abundant in the communities; furthermore, only the Illumina MiSeq data showed the abundance of the Melosira and Entomoneis genera. The results obtained from microscopy were superior to those of Illumina MiSeq regarding species-level identification. Based on the results obtained via microscopy and Illumina MiSeq, it was revealed that neither method is perfect and that each has clear strengths and weaknesses. Therefore, to analyze diatom communities effectively and accurately, these two methods should be combined. [ABSTRACT FROM AUTHOR]

Infectious salmon anemia (ISA) is an infectious disease primarily affecting farmed Atlantic salmon, Salmo salar, which is caused by the ISA virus (ISAV). ISAV belongs to the Orthomyxoviridae family. The disease is a serious condition resulting in reduced fish welfare and high mortality. In this study, we designed an amplicon-based sequencing protocol for whole genome sequencing of ISAV. The method consists of 80 ISAV-specific primers that cover 92% of the virus genome and was designed to be used on an Illumina MiSeq platform. The sequencing accuracy was investigated by comparing sequences with previously published Sanger sequences. The sequences obtained were nearly identical to those obtained by Sanger sequencing, thus demonstrating that sequences produced by this amplicon sequencing protocol had an acceptable accuracy. The amplicon-based sequencingmethod was used to obtain the whole genome sequence of 12 different ISAV isolates froma small local epidemic in the northern part of Norway. Analysis of the whole genome sequences revealed that segment reassortment took place between some of the isolates and could identify which segments that had been reassorted. [ABSTRACT FROM AUTHOR]

The formation of oxygen-depleted zones in the bottom waters is one of the most widespread phenomena in coastal areas. Upwelling episodes along India's west coast due to the southwest monsoon increase biological productivity, further lowering the dissolved oxygen in the upwelled waters, which intensifies annually between June and October. Here, we have determined the changes in the microbial community in response to the varying oxygen levels and other physicochemical parameters at the Candolim Time Series Station using highthroughput sequencing. Amplicon Sequence Variants across all the samples collected in different seasons were mostly affiliated to the phyla Proteobacteria, Actinobacteria, Bacteroidetes, Verrucomicrobia, Chloroflexi, Firmicutes and Planctomycetes, with the most dominant being Proteobacteria (21-41%). Statistical analysis revealed that microbial diversity differed significantly with changing DO, ammonia, nitrate and nitrite concentrations during different seasons. The microbial community shift due to seasonal hypoxia results in the differential biogeochemical cycling of essential nutrients, with certain years seeing redox conditions up to sulphate reduction while certain years seeing only nitrogen loss. Future global warming scenarios will serve as a big challenge for understanding the role of microbial diversity and its implications in the cycling of natural elements. [ABSTRACT FROM AUTHOR]

We identified full-length sequence of a potexvirus infecting Nerine plants in the UK. By sequencing viruses in historical collections we have been able to demonstrate that the identified virus, for which we propose the name nerine potexvirus 1, is a potentially novel species in the genus Potexvirus. Analysis of the potexviruses in the historic collections has also enabled us to generate the first full-length sequence of nerine virus X (NVX) to be isolated from Nerine; and to clarify the taxonomy of NVX isolates infecting Nerine and Agapanthus. Analysis of isolates from the historical collections has enabled us to link biological data gathered in the pre-genomic era to specific isolate sequences. [ABSTRACT FROM AUTHOR]

Straw return is an effective agricultural management practice for alleviating soil sickness, but only a few studies have focused on the incorporation of straw with deep plowing and rotary tillage practices in vegetable production. To determine the effects of rice straw return on Chinese cabbage clubroot, a field experiment for three consecutive years in the same area was performed. Soil microbial high-throughput sequencing, quantitative real-time polymerase chain reaction (PCR) and other methods were used to detect Chinese cabbage plant growth, clubroot occurrence, soil chemical properties and soil microbial diversity and abundance. The results showed that straw addition could significantly reduce the clubroot disease incidence. Through Illumina Miseq sequencing, the diversity of the fungi decreased obviously. The relative abundance of the phyla Proteobacteria and Firmicutes was strikingly reduced, while that of Chloroflexi was significantly increased. Redundancy analysis suggests that soil properties may also affect the soil microbial composition; changes in the microbial structure of bacteria and fungi were associated with the available phosphorus. In conclusion, the continuous addition of rice straw can promote the growth and control the occurrence of clubroot, which is closely related to the microbial composition, and the inhibition effect is proportional to the age of addition. [ABSTRACT FROM AUTHOR]

Introduction: Numerous factors are known to influence reproductive efficiency in ewes, but few studies have investigated the potential role of vaginal microbiota in sheep reproductive success. The objective of this study was to thoroughly characterize the ewe vaginal microbiota throughout the course of pregnancy. Methods: Vaginal samples were collected from 31 pregnant Hampshire and Hampshire X Suffolk crossbred ewes on a weekly basis from pre-breeding to pregnancy testing and then biweekly until just after lambing. To characterize the vaginal microbial communities, DNA was extracted and 16S rRNA gene Illumina MiSeq amplicon sequencing was performed. Results and Discussion: Alpha diversity metrics indicated an increase in species richness, evenness, and overall diversity throughout gestation. Distinct shifts in the bacterial communities were observed during gestation and were segregated into three periods: early gestation, a transitional period and mid/late gestation. During early gestation, Actinobacillus, Histophilus, and unclassified Leptotrichiaceae were found in greater relative abundance. During the transitional period, a population shift occurred characterized by increasing relative abundance of Streptococcus and Staphylococcus. During mid/late gestation, Staphylococcus, Streptococcus, and Ureaplasma had the greatest relative abundance. These shifts in the microbial population throughout the ewe's gestation are likely related to hormonal changes triggered by the growing conceptus, specifically increasing blood concentration of progesterone. The transitional period shift in vaginal microbial communities potentially aligns with the placental take-over of progesterone production from the corpus luteum at approximately day 50 after conception (gestational week 7). Understanding the observed variability of the vaginal microbiota throughout pregnancy will allow for future comparison of ewes that did not become pregnant or had abnormal pregnancies, which could lead to the discovery of potential bacterial biomarkers for pregnancy outcome; this understanding could also lead to development of probiotics to improve sheep reproductive success. [ABSTRACT FROM AUTHOR]

The diversity, composition, and abundance of soil fungi from three sacred groves in Kerala, namely Iringole kavu of Ernakulam District, Kollakal Thapovanam of Alappuzha District, and Poyilkavu of Kozhikode District were analysed using Metagenomics analysis and Illumina sequencing. A total of 30,584, 78,323, and 55,640 reads were obtained from these groves, respectively. Ascomycota constitutes over 96% of the total fungi, making it the most abundant phylum, followed by Mortierellomycota, Basidiomycota, Chytridiomycota, and Rozellomycota. These phyla were subdivided into 20 classes, 40 orders, 83 families, 119 genera, and 135 species, while 1269 OTUs remained unidentified at the species level. Eurotiomycetes predominates the class, while the genus Talaromyces from the family Trichomaceae dominates the genera. Neocarmospora falciformis, Trichoderma lixii, and Candida ethanolic are the most abundant fungal species. Diversity analysis shows that Kollakal Thapovanam is rich in fungal species, while Poyilkavu is rich in biodiversity, with a high degree of dominance. Several species were found only in a particular grove and were absent in others and vice-versa, indicating high fungal specificity. Therefore, fungi have to be preserved in their original habitat. The Principal Coordinate Analysis revealed that each grove is distinct highlighting the importance of preserving the unique diversity of each sacred grove. In conclusion, this research provides valuable information about the soil fungal genera in their natural habitat. It emphasizes the need for more systematic research to understand the actual diversity and ecological role of fungi in sacred groves. This study is the first of its kind to analyse and compare soil fungal diversity in sacred groves using the metagenomics approach. [ABSTRACT FROM AUTHOR]

Serpentoviruses are a subfamily of positive sense RNA viruses in the order Nidovirales, family Tobaniviridae, associated with respiratory disease in multiple clades of reptiles. While the broadest viral diversity is reported from captive pythons, other reptiles, including colubrid snakes, turtles, and lizards of captive and free-ranging origin are also known hosts. To better define serpentoviral diversity, eleven novel serpentovirus genomes were sequenced with an Illumina MiSeq and, when necessary, completed with other Sanger sequencing methods. The novel serpentoviral genomes, along with 57 other previously published serpentovirus genomes, were analyzed alongside four outgroup genomes. Genomic analyses included identifying unique genome templates for each serpentovirus clade, as well as analysis of coded protein composition, potential protein function, protein glycosylation sites, differences in phylogenetic history between open-reading frames, and recombination. Serpentoviral genomes contained diverse protein compositions. In addition to the fundamental structural spike, matrix, and nucleoprotein proteins required for virion formation, serpentovirus genomes also included 20 previously uncharacterized proteins. The uncharacterized proteins were homologous to a number of previously characterized proteins, including enzymes, transcription factors, scaffolding, viral resistance, and apoptosis-related proteins. Evidence for recombination was detected in multiple instances in genomes from both captive and free-ranging snakes. These results show serpentovirus as a diverse clade of viruses with genomes that code for a wide diversity of proteins potentially enhanced by recombination events. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

The seasonal changes in environmental conditions can alter the growth states of host plants, thereby affecting the living environment of endophytes and forming different endophytic communities. This study employs Illumina MiSeq next-generation sequencing to analyze the 16SrRNA and ITS rDNA of endophytes in 24 samples of Actinidia arguta stem tissues across different seasons. The results revealed a high richness and diversity of endophytes in Actinidia arguta, with significant seasonal variations in microbial community richness. This study identified 897 genera across 36 phyla for bacteria and 251 genera across 8 phyla for fungi. Notably, 69 bacterial genera and 19 fungal genera significantly contributed to the differences in community structure across seasons. A distinctive feature of coexistence in the endophytic community, both specific and conservative across different seasons, was observed. The bacterial community in winter demonstrated significantly higher richness and diversity compared to the other seasons. Environmental factors likely influence the optimal timing for endophyte colonization. Solar radiation, temperature, precipitation, and relative humidity significantly impact the diversity of endophytic bacteria and fungi. In addition, seasonal variations show significant differences in the nutritional modes of fungal endophytes and the degradation, ligninolysis, and ureolysis functions of bacterial endophytes. This study elucidates the potential role of endophytes in assisting Actinidia arguta in adapting to seasonal changes and provides a theoretical basis for further exploration of functional microbial strains. [ABSTRACT FROM AUTHOR]

Infectious salmon anemia (ISA) is an infectious disease primarily affecting farmed Atlantic salmon, Salmo salar, which is caused by the ISA virus (ISAV). ISAV belongs to the Orthomyxoviridae family. The disease is a serious condition resulting in reduced fish welfare and high mortality. In this study, we designed an amplicon-based sequencing protocol for whole genome sequencing of ISAV. The method consists of 80 ISAV-specific primers that cover 92% of the virus genome and was designed to be used on an Illumina MiSeq platform. The sequencing accuracy was investigated by comparing sequences with previously published Sanger sequences. The sequences obtained were nearly identical to those obtained by Sanger sequencing, thus demonstrating that sequences produced by this amplicon sequencing protocol had an acceptable accuracy. The amplicon-based sequencing method was used to obtain the whole genome sequence of 12 different ISAV isolates from a small local epidemic in the northern part of Norway. Analysis of the whole genome sequences revealed that segment reassortment took place between some of the isolates and could identify which segments that had been reassorted.

The formation of oxygen-depleted zones in the bottom waters is one of the most widespread phenomena in coastal areas. Upwelling episodes along India’s west coast due to the southwest monsoon increase biological productivity, further lowering the dissolved oxygen in the upwelled waters, which intensifies annually between June and October. Here, we have determined the changes in the microbial community in response to the varying oxygen levels and other physicochemical parameters at the Candolim Time Series Station using high-throughput sequencing. Amplicon Sequence Variants across all the samples collected in different seasons were mostly affiliated to the phyla Proteobacteria, Actinobacteria, Bacteroidetes, Verrucomicrobia, Chloroflexi, Firmicutes and Planctomycetes, with the most dominant being Proteobacteria (21-41%). Statistical analysis revealed that microbial diversity differed significantly with changing DO, ammonia, nitrate and nitrite concentrations during different seasons. The microbial community shift due to seasonal hypoxia results in the differential biogeochemical cycling of essential nutrients, with certain years seeing redox conditions up to sulphate reduction while certain years seeing only nitrogen loss. Future global warming scenarios will serve as a big challenge for understanding the role of microbial diversity and its implications in the cycling of natural elements.

Despite the diligent efforts of libraries, archives, and similar institutions to preserve cultural monuments, biodeterioration continues to pose a significant threat to these objects. One of the main sources of microorganisms responsible for the biodeterioration process is the presence of airborne microorganisms. Therefore, this research aims to monitor and compare outcomes of both culture-dependent (utilising various cultivation strategies) and culture-independent approaches (RNA-based sequencing) to identifying metabolically active airborne microorganisms in archives in the Czech Republic. Through this study, several species that have the potential to pose risks to both cultural heritage objects and the health of institution employees were found. Additionally, the efficacy of different cultivation media was demonstrated to be varied across archive rooms, highlighting the necessity of employing multiple cultivation media for comprehensive analyses. Of noteworthy importance, the resuscitating-promoting factor (Rpf) proved to be a pivotal tool, increasing bacterial culturability by up to 30% when synergistically employed Reasoner's 2A agar (R2A) and R2A + Rpf media. Next, the study emphasises the importance of integrating both culture-dependent and culture-independent approaches. The overlap between genera identified by the culture-dependent approach and those identified also by the culture-independent approach varied from 33% to surpassing 94%, with the maximum alignment exceeding 94% in only one case. Our results highlight the importance of actively monitoring and assessing levels of microbial air contamination in archives to prevent further deterioration of cultural heritage objects and to promote improved conditions for employees in archives and similar institutions.

Anguina tritici is the first plant-parasitic nematode described in literature, dating back to the year 1743. It is responsible for causing earcockle (seed gall) and tundu diseases in wheat and rye. Notably, this nematode has been observed to survive in an anhydrobiotic state for up to 32 years within wheat seed galls. These exceptional characteristics have inspired the sequencing of the A. tritici genome. In this study, we present the initial draft genome of A. tritici, obtained using the Illumina MiSeq platform with coverage of 60-fold. The genome is estimated to have a size of 164 Mb and comprises 39,965 protein-coding genes, exhibiting a GC content of 39.1%. The availability of this genome data will serve as a foundation for future functional biological investigations, particularly for genes whose functions remain unknown to this day.

Metabarcoding is an important tool for understanding fungal communities. The internal transcribed spacer (ITS) rDNA is the accepted fungal barcode but has known problems. The large subunit (LSU) rDNA has also been used to investigate fungal communities but available LSU metabarcoding primers were mostly designed to target Dikarya (Ascomycota + Basidiomycota) with little attention to early diverging fungi (EDF). However, evidence from multiple studies suggests that EDF comprise a large portion of unknown diversity in community sampling. Here, we investigate how DNA marker choice and methodological biases impact recovery of EDF from environmental samples. We focused on one EDF lineage, Zoopagomycota, as an example. We evaluated three primer sets (ITS1F/ITS2, LROR/LR3, and LR3 paired with new primer LR22F) to amplify and sequence a Zoopagomycota mock community and a set of 146 environmental samples with Illumina MiSeq. We compared two taxonomy assignment methods and created an LSU reference database compatible with AMPtk software. The two taxonomy assignment methods recovered strikingly different communities of fungi and EDF. Target fragment length variation exacerbated PCR amplification biases and influenced downstream taxonomic assignments, but this effect was greater for EDF than Dikarya. To improve identification of LSU amplicons we performed phylogenetic reconstruction and illustrate the advantages of this critical tool for investigating identified and unidentified sequences. Our results suggest much of the EDF community may be missed or misidentified with "standard" metabarcoding approaches and modified techniques are needed to understand the role of these taxa in a broader ecological context.

Some insect species have gained attention as efficient bioconverters of low-value organic substrates (i.e., residual streams) into high-value biomass. Black soldier fly (BSF) (Hermetia illucens) larvae are particularly interesting for bioconversion due to their ability to grow on a wide range of substrates, including low-value industrial residual streams. This is in part due to the plasticity of the gut microbiota of polyphagous insects, like BSF. Gut microbiota composition varies depending on rearing substrates, via a mechanism that might support the recruitment of microorganisms that facilitate digestion of a specific substrate. At the same time, specific microbial genera do persist on different substrates via unknown mechanisms. This study aimed to offer insights on this microbial plasticity by investigating how the composition of the bacterial community present in the gut of BSF larvae responds to two industrial residual streams: swill (a mixture of catering and supermarket leftovers) and distiller's dried grains with solubles. The bacterial biota composition of substrates, whole larvae at the beginning of the rearing period and at harvest, rearing residues, and larval gut regions were investigated through 16S rRNA gene sequencing. It was observed that both substrate and insect development influenced the bacterial composition of the whole larvae. Zooming in on the gut regions, there was a clear shift in community composition from a higher to a lower diversity between the anterior/middle midgut and the posterior midgut/hindgut, indicating a selective pressure occurring in the middle midgut region. Additionally, the abundance of the bacterial biota was always high in the hindgut, while its diversity was relatively low. Even more, the bacterial community in the hindgut was found to be relatively more conserved over the different substrates, harboring members of the BSF core microbiota. We postulate a potential role of the hindgut as a reservoir for insect-associated microbes. This warrants further research on that underexplored region of the intestinal tract. Overall, these findings contribute to our understanding of the bacterial biota structure and dynamics along the intestinal tract, which can aid microbiome engineering efforts to enhance larval performance on (industrial) residual streams. [ABSTRACT FROM AUTHOR]

Introduction: The soil microbial community plays an important role in modulating cotton soil fertility. However, the effects of chemical fertilizer combined with organic fertilizer on soil chemical properties, microbial community structure, and crop yield and quality in arid areas are still unclear. This study aimed to explore the effects of different organic fertilizers on soil microbial community structure and diversity and cotton growth and yield. Methods: High-throughput sequencing was used to study the soil bacteria and fungi in different growth stages of cotton. The field fertilization experiment had five treatments. Results: The results indicated that the treatments of chemical fertilizer reduction combined with organic fertilizer significantly increased soil available nitrogen and phosphorus in cotton field. There were significant differences in the abundance of the bacterial and fungal communities in the dominant phyla among the treatments. At the phyla level, there were not significantly different in the diversity of bacteria and fungi among treatments. There were significant differences in the composition and diversity of bacterial and fungal communities during the entire cotton growth period (p = 0.001). The rhizosphere bacterial and fungal community structure was significantly affected by soil TK, NH4 +, AK, TP, AN, and NO3 -. The different fertilization treatments strongly influenced the modular structure of the soil bacterial and fungal community co-occurrence network. A reduction in chemical fertilizer combined with organic fertilizer significantly improved cotton stem diameter and seed yield, and the effect of the biological organic fertilizer on plant growth and yield formation was greater than that of ordinary organic fertilizer. Discussion: This study provide a scientific and technical basis for the establishment of environmentally friendly green fertilization technology for cotton in arid areas and the promotion of sustainable development of cotton industry. [ABSTRACT FROM AUTHOR]

Aims Approximately 10% of children are born prematurely, and bacterial vaginosis during pregnancy is associated with preterm delivery. Highly accurate species-level vaginal microflora analysis helps control bacteria-induced preterm birth. Therefore, we aimed to conduct a bioinformatic analysis of gene sequences using 16S databases and compare their efficacy in comprehensively identifying potentially pathogenic vaginal microbiota in Japanese women. Methods and results The 16 s rRNA databases, Silva, Greengenes, and the basic local alignment search tool (BLAST) were compared to determine whether the classification quality could be improved using the V3–V4 region next-generation sequencing (NGS) sequences. It was found that NGS data were aligned using the BLAST database with the QIIME 2 platform, whose classification quality was higher than that of Silva, and the combined Silva and Greengenes databases based on the mutual complementarity of the two databases. Conclusions The reference database selected during the bioinformatic processing influenced the recognized sequence percentage, taxonomic rankings, and accuracy. This study showed that the BLAST database was the best choice for NGS data analysis of Japanese women's vaginal microbiota. [ABSTRACT FROM AUTHOR]

Understanding prey consumption patterns is critical to understanding the ways in which seabirds cope with a changing ocean. However, characterizing the dietary habitats of seabirds can be challenging. In this study, we investigated the diet of the Marbled Murrelet Brachyramphus marmoratus population that lives in waters off California, Oregon, and Washington, USA, using fecal DNA, custom metabarcoding, and high-throughput sequencing. Murrelets were captured at sea by dip-netting at night. Across this region, murrelets consumed highly diverse prey types including 17 fish species and 10 invertebrate species, in accord with previous work indicating the species' forage on a wide range of prey. Pacific Herring Clupea pallasii was the most common prey in Washington and Oregon (frequency of occurrence = 0.84 and 0.69, respectively), replaced by Northern Anchovy Engraulis mordax in California (frequency of occurrence = 0.77). In Oregon, where our sample size was sufficient, diet composition differed between the 2017 and 2018 breeding seasons, with an apparent decline in the proportional consumption of energy-dense prey. Common and energy-dense prey were consumed in equal proportions by males and females, perhaps because of foraging in the same habitat. Diet did not vary between breeders and non-breeders. Our study offers the first detailed report on the diet of adult Marbled Murrelets in waters where they are listed as Threatened by the US federal government. This indicates that managing fisheries and conserving spawning habitat for high-occurrence prey species could benefit murrelet populations. [ABSTRACT FROM AUTHOR]

IntroductionNumerous factors are known to influence reproductive efficiency in ewes, but few studies have investigated the potential role of vaginal microbiota in sheep reproductive success. The objective of this study was to thoroughly characterize the ewe vaginal microbiota throughout the course of pregnancy.MethodsVaginal samples were collected from 31 pregnant Hampshire and Hampshire X Suffolk crossbred ewes on a weekly basis from pre-breeding to pregnancy testing and then biweekly until just after lambing. To characterize the vaginal microbial communities, DNA was extracted and 16S rRNA gene Illumina MiSeq amplicon sequencing was performed.Results and DiscussionAlpha diversity metrics indicated an increase in species richness, evenness, and overall diversity throughout gestation. Distinct shifts in the bacterial communities were observed during gestation and were segregated into three periods: early gestation, a transitional period and mid/late gestation. During early gestation, Actinobacillus, Histophilus, and unclassified Leptotrichiaceae were found in greater relative abundance. During the transitional period, a population shift occurred characterized by increasing relative abundance of Streptococcus and Staphylococcus. During mid/late gestation, Staphylococcus, Streptococcus, and Ureaplasma had the greatest relative abundance. These shifts in the microbial population throughout the ewe’s gestation are likely related to hormonal changes triggered by the growing conceptus, specifically increasing blood concentration of progesterone. The transitional period shift in vaginal microbial communities potentially aligns with the placental take-over of progesterone production from the corpus luteum at approximately day 50 after conception (gestational week 7). Understanding the observed variability of the vaginal microbiota throughout pregnancy will allow for future comparison of ewes that did not become pregnant or had abnormal pregnancies, which could lead to the discovery of potential bacterial biomarkers for pregnancy outcome; this understanding could also lead to development of probiotics to improve sheep reproductive success.

Straw return is an effective agricultural management practice for alleviating soil sickness, but only a few studies have focused on the incorporation of straw with deep plowing and rotary tillage practices in vegetable production. To determine the effects of rice straw return on Chinese cabbage clubroot, a field experiment for three consecutive years in the same area was performed. Soil microbial high-throughput sequencing, quantitative real-time polymerase chain reaction (PCR) and other methods were used to detect Chinese cabbage plant growth, clubroot occurrence, soil chemical properties and soil microbial diversity and abundance. The results showed that straw addition could significantly reduce the clubroot disease incidence. Through Illumina Miseq sequencing, the diversity of the fungi decreased obviously. The relative abundance of the phyla Proteobacteria and Firmicutes was strikingly reduced, while that of Chloroflexi was significantly increased. Redundancy analysis suggests that soil properties may also affect the soil microbial composition; changes in the microbial structure of bacteria and fungi were associated with the available phosphorus. In conclusion, the continuous addition of rice straw can promote the growth and control the occurrence of clubroot, which is closely related to the microbial composition, and the inhibition effect is proportional to the age of addition.

Some insect species have gained attention as efficient bioconverters of low-value organic substrates (i.e., residual streams) into high-value biomass. Black soldier fly (BSF) (Hermetia illucens) larvae are particularly interesting for bioconversion due to their ability to grow on a wide range of substrates, including low-value industrial residual streams. This is in part due to the plasticity of the gut microbiota of polyphagous insects, like BSF. Gut microbiota composition varies depending on rearing substrates, via a mechanism that might support the recruitment of microorganisms that facilitate digestion of a specific substrate. At the same time, specific microbial genera do persist on different substrates via unknown mechanisms. This study aimed to offer insights on this microbial plasticity by investigating how the composition of the bacterial community present in the gut of BSF larvae responds to two industrial residual streams: swill (a mixture of catering and supermarket leftovers) and distiller’s dried grains with solubles. The bacterial biota composition of substrates, whole larvae at the beginning of the rearing period and at harvest, rearing residues, and larval gut regions were investigated through 16S rRNA gene sequencing. It was observed that both substrate and insect development influenced the bacterial composition of the whole larvae. Zooming in on the gut regions, there was a clear shift in community composition from a higher to a lower diversity between the anterior/middle midgut and the posterior midgut/hindgut, indicating a selective pressure occurring in the middle midgut region. Additionally, the abundance of the bacterial biota was always high in the hindgut, while its diversity was relatively low. Even more, the bacterial community in the hindgut was found to be relatively more conserved over the different substrates, harboring members of the BSF core microbiota. We postulate a potential role of the hindgut as a reservoir for insect-associated microbes. This warrants further research on that underexplored region of the intestinal tract. Overall, these findings contribute to our understanding of the bacterial biota structure and dynamics along the intestinal tract, which can aid microbiome engineering efforts to enhance larval performance on (industrial) residual streams.

Simple Summary: In this study, the researchers investigated the role of gut bacteria in an insect pest called Megalurothrips usitatus and its resistance to an insecticide and an entomopathogenic fungus. They compared the gut bacterial communities of an insecticide-susceptible population and an insecticide-resistant population of Megalurothrips usitatus. Through High-throughput sequencing, they found that the insecticide-resistant population exhibited significantly higher resistance to the insecticide and the Beauveria brongniartii. The sequencing also revealed changes in the abundance of specific bacterial species in response to the insecticide and the Beauveria brongniartii. The researchers observed that the gut bacteria of Megalurothrips usitatus are primarily involved in various metabolic activities, including the degradation of the Beauveria brongniartii. Some of these bacteria may even reduce the virulence of the fungus, thereby weakening its biocontrol ability. The results of this study provide a basis for future elucidation of the role of intestinal bacteria in the symbiotic relationship of insecticide resistance and the design of new management strategies for Megalurothrips usitatus using Beauveria brongniartii for biological control. The gut bacterial microbiota of insects plays a crucial role in physiological, metabolic, and innate immune processes. In the current study, the gut bacterial communities of an insecticide-susceptible (IS), and a resistant (IR) population of a major legume pest, Megalurothrips usitatus (Bagnall), were evaluated. The 16S rDNA V3 + V4 regions of M. usitatus infected with Beauveria brongniartii along with the intestinal flora of both populations were sequenced based on a High-throughput sequencing platform. Toxicological bioassays revealed that the IR population exhibited resistance to acetamiprid and B. brongniartii isolate SB010 at levels of 138.0-fold and 55.6-fold higher, respectively, compared to the IS population. Through 16S High-throughput sequencing, the results indicate that both resistant populations, as well as B. brongniartii infestation, reduce the number of species of M. usitatus gut microbes. Using KEGG function prediction, it was found that most intestinal bacteria were involved in various metabolic activities, and the abundance of resistant populations was higher than that of sensitive populations. The bacteria in the gut of M. usitatus are mainly involved in various metabolic activities to achieve the degradation of B. brongniartii. This study provides valuable insights into the interaction between gut bacteria, insecticide resistance, and Beauveria. brongniartii infection in Megalurothrips usitatus, which can help inform future pest control strategies. [ABSTRACT FROM AUTHOR]

To ensure long-term food production in a changing world, it is critical to identify field management practices that increase crop yields and maintain soil health. Additionally, sustainable agriculture needs to provide experimental evidence to support the use of traditional agricultural practices. In this study, a 20-year investigation of the effects of different combinations of fertilizer types (control, chemical fertilizer, organic fertilizer, and bio-fertilizer) and fertilization amount (conventional dosages and high dosages) on wheat yield and soil health, including soil enzyme activity and microbial biomass, soil microbial diversity, and crop yield. Our long-term study indicates that the use of high dosages of bio-fertilizer can increase the fertilizer yield contribution rate by a minimum of 76.7% compared to other management combinations. Furthermore, this practice can improve soil biological quality, including the concentration of soil microbial biomass carbon, promote bacterial biodiversity, and enhance the soil health index. The effect of high dosages fertilizer was greater than that of conventional dosages fertilizer. The highest soil health index was 0.88 in high dosage bio-fertilizer, and the lowest was 0.12 in chemical fertilizer. In summary, these results suggested that the use of bio-fertilizer can help maintain soil health and crop productivity in the long term. [ABSTRACT FROM AUTHOR]

During reclamation organisms like fungi directly interact at the soil–plant interface and can potentially impact plant growth, health, and overall revegetation success. Therefore, a study was conducted to determine how topsoil amendments (mycorrhizae, biochar, compost, manure) influence root‐associated fungal communities and growth of fast growing Populus balsamifera trees on a phosphogypsum (PG) reclamation site in Alberta, Canada. Tree height, diameter, biomass, and diversity of Ascomycota and Basidiomycota were assessed using Illumina MiSeq sequencing targeting the fungal internal transcribed spacer region 1 (ITS1) and ITS2. Topsoil amendments and root location (topsoil vs. PG within a treatment), greatly affected the root fungal community. Basidiomycota preferentially grew in PG, and had a high treatment specific root fungal community with highest diversity of ectomycorrhizal fungi and basidiomycete yeast in biochar PG and compost PG. Populus balsamifera roots were associated with a high diversity of Ascomycota, found equally in roots in topsoil and PG, with a trend towards higher diversity and abundance in PG. The core genus found across all samples was Fusarium, a potential plant pathogen. All topsoil amendments strongly reduced abundance of Fusarium and other Ascomycota in poplar roots. Biochar and compost reduced number and abundance of potential fungal pathogens and increased number of potential plant growth promoting Basidiomycota, indicating favorable growth conditions for P. balsamifera on a phosphygypsum reclamation site by application of these amendments. This study indicates compost and biochar can be effective soil amendments to support P. balsamifera establishment and growth on a PG reclamation site. [ABSTRACT FROM AUTHOR]

Abstract Isolation of heavy metals-resistant bacteria from their original habitat is a crucial step in bioremediation. Six lead (Pb) resistant bacterial strains were isolated and identified utilizing 16S rRNA to be Enterobacter ludwigii FACU 4, Shigella flexneri FACU, Microbacterium paraoxydans FACU, Klebsiella pneumoniae subsp. pneumonia FACU, Raoultella planticola FACU 3 and Staphylococcus xylosus FACU. It was determined that all these strains had their Minimum inhibitory concentration (MIC) to be 2500 ppm except R. planticola FACU 3 has a higher maximum tolerance concentration (MTC) up to 2700 ppm. We evaluated the survival of all six strains on lead stress, the efficiency of biosorption and lead uptake. It was found that R. planticola FACU 3 is the highest MTC and S. xylosus FACU was the lowest MTC in this evaluation. Therefore, transmission electron microscopy (TEM) confirmed the difference between the morphological responses of these two strains to lead stress. These findings led to explore more about the genome of R. planticola FACU 3 using illumine Miseq technology. Draft genome sequence analysis revealed the genome size of 5,648,460 bp and G + C content 55.8% and identified 5526 CDS, 75 tRNA and 4 rRNA. Sequencing technology facilitated the identification of about 47 genes related to resistance to many heavy metals including lead, arsenic, zinc, mercury, nickel, silver and chromium of R. planticola FACU 3 strain. Moreover, genome sequencing identified plant growth-promoting genes (PGPGs) including indole acetic acid (IAA) production, phosphate solubilization, phenazine production, trehalose metabolism and 4-hydroxybenzoate production genes and a lot of antibiotic-resistant genes.

Serpentoviruses are a subfamily of positive sense RNA viruses in the order Nidovirales, family Tobaniviridae, associated with respiratory disease in multiple clades of reptiles. While the broadest viral diversity is reported from captive pythons, other reptiles, including colubrid snakes, turtles, and lizards of captive and free-ranging origin are also known hosts. To better define serpentoviral diversity, eleven novel serpentovirus genomes were sequenced with an Illumina MiSeq and, when necessary, completed with other Sanger sequencing methods. The novel serpentoviral genomes, along with 57 other previously published serpentovirus genomes, were analyzed alongside four outgroup genomes. Genomic analyses included identifying unique genome templates for each serpentovirus clade, as well as analysis of coded protein composition, potential protein function, protein glycosylation sites, differences in phylogenetic history between open-reading frames, and recombination. Serpentoviral genomes contained diverse protein compositions. In addition to the fundamental structural spike, matrix, and nucleoprotein proteins required for virion formation, serpentovirus genomes also included 20 previously uncharacterized proteins. The uncharacterized proteins were homologous to a number of previously characterized proteins, including enzymes, transcription factors, scaffolding, viral resistance, and apoptosis-related proteins. Evidence for recombination was detected in multiple instances in genomes from both captive and free-ranging snakes. These results show serpentovirus as a diverse clade of viruses with genomes that code for a wide diversity of proteins potentially enhanced by recombination events.

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.

The seasonal changes in environmental conditions can alter the growth states of host plants, thereby affecting the living environment of endophytes and forming different endophytic communities. This study employs Illumina MiSeq next-generation sequencing to analyze the 16SrRNA and ITS rDNA of endophytes in 24 samples of Actinidia arguta stem tissues across different seasons. The results revealed a high richness and diversity of endophytes in Actinidia arguta, with significant seasonal variations in microbial community richness. This study identified 897 genera across 36 phyla for bacteria and 251 genera across 8 phyla for fungi. Notably, 69 bacterial genera and 19 fungal genera significantly contributed to the differences in community structure across seasons. A distinctive feature of coexistence in the endophytic community, both specific and conservative across different seasons, was observed. The bacterial community in winter demonstrated significantly higher richness and diversity compared to the other seasons. Environmental factors likely influence the optimal timing for endophyte colonization. Solar radiation, temperature, precipitation, and relative humidity significantly impact the diversity of endophytic bacteria and fungi. In addition, seasonal variations show significant differences in the nutritional modes of fungal endophytes and the degradation, ligninolysis, and ureolysis functions of bacterial endophytes. This study elucidates the potential role of endophytes in assisting Actinidia arguta in adapting to seasonal changes and provides a theoretical basis for further exploration of functional microbial strains.

ABSTRACT Microbial contamination in cultural heritage storage facilities is undoubtedly still a huge problem and leads to the biodeterioration of historical objects and thus the loss of information for future generations. Most studies focus on fungi that colonize materials, which are the primary agents of biodeterioration. However, bacteria also play crucial roles in this process. Therefore, this study focuses on identifying bacteria that colonize audio-visual materials and those present in the air in the archives of the Czech Republic. For our purposes, the Illumina MiSeq amplicon sequencing method was used. Using this method, 18 bacterial genera with an abundance of higher than 1% were identified on audio-visual materials and in the air. We also evaluated some factors that were assumed to possibly influence the composition of bacterial communities on audio-visual materials, of which locality was shown to be significant. Locality also explained most of the variability in bacterial community structure. Furthermore, an association between genera colonizing materials and genera present in the air was demonstrated, and indicator genera were evaluated for each locality. IMPORTANCE The existing literature on microbial contamination of audio-visual materials has predominantly used culture-based methods to evaluate contamination and has overlooked the potential impact of environmental factors and material composition on microbial communities. Furthermore, previous studies have mainly focused on contamination by microscopic fungi, neglecting other potentially harmful microorganisms. To address these gaps in knowledge, our study is the first to provide a comprehensive analysis of bacterial communities present on historical audio-visual materials. Our statistical analyses demonstrate the critical importance of including air analysis in such studies, as airborne microorganisms can significantly contribute to the contamination of these materials. The insights gained from this study are not only valuable in developing effective preventive measures to mitigate contamination but also valuable in identifying targeted disinfection methods for specific types of microorganisms. Overall, our findings highlight the need for a more holistic approach to understanding microbial contamination in cultural heritage materials.

IntroductionThe native Atlantic rock crab (Cancer irrotatus) and the invasive European green crab (Carcinus maenas) are commercially and ecologically important crustacean species in Atlantic Canada. The importance of microbiomes for host health and ecology has been recognized in many species, although very few studies have focused on crustaceans or their external shell microbiome. This is the first-ever study to characterize and analyze the microbial communities associated with the external carapace of C. irrotatus and C. maenas.MethodsMicrobiome samples were collected from three locations in Atlantic Canada, processed using standard 16S Illumina MiSeq PE250 sequencing and analyzed with the open-access QIIME2 software.ResultsTaxonomic classification of the microbial compositions, as well as alpha- and beta diversities, reveal that the shell microbiome differs by host species between C. irrotatus and C. maenas sampled from the same location and between C. irrotatus sampled from different locations. Interestingly, the differences are greater between species at the same location than between locations for the same species.DiscussionThese are the first-ever results showing that the crustacean shell microbiome not only depends on geographical factors but also on intrinsic factors specific to the host species. This implies that crustaceans exert some impact on their shell microbiome, potentially selecting beneficial taxa. These are important findings that could elucidate contributing factors of crustacean shell diseases that are still poorly understood.

Pulse crop rotation in rice cultivation is a widely accepted agronomic practice. Depending upon the water regime, rice cultivation has been classified into wetland and aerobic practices. However, no studies have been conducted so far to understand the impact of pulse crop rotation and rice mono-cropping on fungal diversity, particularly in aerobic soil. A targeted metagenomic study was conducted to compare the effects of crop rotations (rice-rice and rice-pulse) on fungal diversity in wetland and aerobic rice soils. Out of 445 OTUs, 41.80% was unknown and 58.20% were assigned to six phyla, namely Ascomycota (56.57%), Basidiomycota (1.32%), Zygomycota (0.22%), Chytridiomycota (0.04%), Glomeromycota (0.03%), and Blastocladiomycota (0.02%). Functional trait analysis found wetland rice-pulse rotation increased symbiotrophs (36.7%) and saprotrophs (62.1%) population, whereas higher pathotrophs were found in aerobic rice–rice (62.8%) and rice-pulse (61.4%) cropping system. Certain soil nutrients played a major role in shaping the fungal community; Ca had significant (p < 0.05) positive impact on saprotroph, symbiotroph and endophytes, whereas Cu had significant (p < 0.05) negative impact on pathotrophs. This study showed that rice-pulse crop rotation could enhance the saprophytic and symbiotic fungal diversity in wetland and reduce the population of pathogens in aerobic rice cultivation. [ABSTRACT FROM AUTHOR]

Aim Stormwater is a major source of many contaminants of emerging concern, which can be toxic to both aquatic and terrestrial organisms. This project aimed to identify novel biodegraders of toxic tire wear particle (TWP) contaminants associated with coho salmon mortality. Methods and results This study has (i) characterized the prokaryotic communities of stormwater in both urban and rural settings; (ii) evaluated the ability of stormwater isolates to degrade two model TWP contaminants, hexa(methoxymethyl)melamine and 1,3-diphenylguanidine; and (iii) evaluated the toxicological impact of these model contaminants on the growth of six model bacteria. Rural stormwater possessed a diverse microbiome dominated by Oxalobacteraceae, Microbacteriaceae, Cellulomonadaceae, and Pseudomonadaceae taxa, while urban stormwater showed much less microbial diversity overall. Additionally, multiple stormwater isolates appeared capable of using model TWP contaminants as their sole carbon source. Each model contaminant was also found to alter growth patterns of model environmental bacteria including, with 1,3-DPG appearing more acutely toxic at high concentrations. Conclusion This study identified several stormwater isolates that have the potential to be used as a sustainable solution to stormwater quality management. [ABSTRACT FROM AUTHOR]

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The rise in wildfire frequency and severity across the globe has increased interest in secondary succession. However, despite the role of soil microbial communities in controlling biogeochemical cycling and their role in the regeneration of post‐fire vegetation, the lack of measurements immediately post‐fire and at high temporal resolution has limited understanding of microbial secondary succession. To fill this knowledge gap, we sampled soils at 17, 25, 34, 67, 95, 131, 187, 286, and 376 days after a southern California wildfire in fire‐adapted chaparral shrublands. We assessed bacterial and fungal biomass with qPCR of 16S and 18S and richness and composition with Illumina MiSeq sequencing of 16S and ITS2 amplicons. Fire severely reduced bacterial biomass by 47%, bacterial richness by 46%, fungal biomass by 86%, and fungal richness by 68%. The burned bacterial and fungal communities experienced rapid succession, with 5–6 compositional turnover periods. Analogous to plants, turnover was driven by "fire‐loving" pyrophilous microbes, many of which have been previously found in forests worldwide and changed markedly in abundance over time. Fungal secondary succession was initiated by the Basidiomycete yeast Geminibasidium, which traded off against the filamentous Ascomycetes Pyronema, Aspergillus, and Penicillium. For bacteria, the Proteobacteria Massilia dominated all year, but the Firmicute Bacillus and Proteobacteria Noviherbaspirillum increased in abundance over time. Our high‐resolution temporal sampling allowed us to capture post‐fire microbial secondary successional dynamics and suggest that putative tradeoffs in thermotolerance, colonization, and competition among dominant pyrophilous microbes control microbial succession with possible implications for ecosystem function. [ABSTRACT FROM AUTHOR]

Background: The genus Ternstroemia is associated with the vulnerable tropical montane cloud forest in Mexico and with other relevant vegetation types worldwide. It contains threatened and pharmacologically important species and has taxonomic issues regarding its species limits. This study describes 38 microsatellite markers generated using a genomic-based approach. Methods and results: We tested 23 of these markers in a natural population of Ternstroemia lineata. These markers are highly polymorphic (all loci polymorphic with 3–14 alleles per locus and expected heterozygosity between 0.202 and 0.908), most of them (19 out of 23) are in Hardy-Weinberg Equilibrium and free of null alleles (18 out of 23). Also we found no evidence of linkage among them. Finally, we tested the transferability to six other American species of Ternstroemia, two other Pentaphylacaceae species, and four species from different families within the order Ericales. Conclusions: These molecular resources are promising tools to investigate genetic diversity loss and as barcodes for ethnopharmacological applications and species delimitation in the family Pentaphylacaceae and some Ericales, among other applications. [ABSTRACT FROM AUTHOR]

Abstract Background Biofumigation is a non-chemical sustainable approach that reshapes soil microbiota to overcome challenges in way of continuous cultivation. However, the type and quantity of substrate have a significant impact on microbiota shifts and the subsequent success of biofumigation. Moreover, studies on the effects of biofumigant concentration in combination with fumigation duration on soil microbiota dynamics are very rare. Research methods We performed microcosm experiments to investigate how a biofumigant (Korean canola cultivar, HanRa) at various concentrations (0.5%, 1%, 2–4% w/w: biofumigant/soil) and fumigation periods (2–4 weeks) affects the soil bacterial and fungal communities. Subsequently, pot experiments employing two Korean canola cultivars (HanRa and YongSan) at 1% (w/w) were carried out. Results Illumina MiSeq analysis revealed that 2–4% biofumigant, regardless of incubation period, had a significant negative impact on microbial diversity and network complexity. In contrast, 1% biofumigant transformed the bacterial, fungal, and inter-kingdom networks into a highly connected and complex network without affecting microbial diversity. Bacillus, Clostridium, and Pseudomonas were the most highly stimulated bacterial genera in the biofumigated soils, whereas the abundance of Acidobacteria members was greatly reduced. The 2–4% amendments had substantially and more differentially abundant Fusarium than the 1%. Soil nutrition (e.g., pH, nitrate, ammonium, and exchangeable potassium), fruit yield, and weed suppression were enhanced in subsequent pot experiments. Of the nine soil chemical properties, phosphate and exchangeable potassium were the main factors influencing the microbial community assembly. Conclusions Optimized biofumigation-mediated increase in nitrate, ammonium, and potassium availability in the soil without causing any negative effects on soil microbial diversity indicates its potential as a preplant to improve crop productivity. This study contributes significantly to our understanding of how an optimal biofumigant can help ameliorate obstacles in continuous cropping. Graphical Abstract

AbstractIn this study, the roots and rhizosphere soil of Abies koreana and Taxus cuspidata were collected from sites at two different altitudes on Mt. Halla. Ectomycorrhizal fungi (EMF) were identified by Illumina MiSeq sequencing. The proportion of EMF from the roots was 89% in A. koreana and 69% in T. cuspidata. Among EMF in rhizosphere soils, the genus Russula was the most abundant in roots of A. koreana (p

In this study, pepper and tomato sour soup，the raw materials of red sour soup were taken as the research objects. Illumina MiSeq, total acid titration, gas chromatography-mass spectrometry and other equipment or methods are used to analyze the dynamic trend of microflora, total acid, pH value and volatile substances in the fermentation process of two kinds of sour soup. The results showed that Lactobacillus namurensis (27.67%) and Lactobacillus pentosus (41.75%) were the dominant lactic acid bacteria in pepper and tomato sour soup, respectively; Debaryomyces hansenii (76.05%，pepper sour soup) and Pichia membranifaciens (80.88%，tomato sour soup) were the dominant fungi in the later stage of fermentation. In terms of volatile substances, 156 and 92 compounds such as alcohols, ketones, aldehydes and esters were detected in pepper and tomato sour soup respectively. Among them, esters (24.99%), terpenes (20.21%) and ketones (17.60%) were several compounds with high content in chili acid soup in the early stage of fermentation; In tomato sour soup, aldehydes (41.30%) and ketones (20.28%) were the main compounds. With the change of fermentation, the main flavor compounds of chili acid soup were alcohols and esters, represented by linalool (10.43%) and ethyl hexadecanoate (12.12%); In tomato sour soup, alcohols, esters and phenols were the advantages. Among them, phenylethanol (31.93%) and 4-ethylphenol (8.71%) played an important role in the flavor of tomato sour soup. In general, the dominant functional microorganisms in the naturally fermented sour soup are Lactobacillus and yeast, and the total acid content higher than 25 g/kg can be obtained within 21~28 days after fermentation. The volatile substances vary greatly with the fermentation period. The relevant data can provide a reference for the development of directed vat set for sour soup and the optimization of processing technology.

Coffee is one of the most valuable agricultural commodities worldwide, second only to oil in terms of international trade. Coffea arabica L. is a widely cultivated and economically important crop that is responsible for about 90% of the global production of coffee. In this study, we selected five C. arabica cultivation sites at different altitudes to clarify the effects of altitude on rhizospheric soil physical–chemical characteristics and microbial communities. The samples collected at low altitudes were more nutrient-deficient and acidic than the soil samples collected at medium–high altitudes. The Proteobacteria-to-Acidobacteria ratio increased from lower altitudes to medium–high altitudes. Additionally, although Ascomycota was the dominant fungal phylum, it was unaffected by the altitude. Furthermore, the alpha richness and diversity of the bacterial and fungal communities were higher at medium–high altitudes than at low altitudes. Moreover, the redundancy analysis indicated that microbial phyla were closely associated with pH. These findings suggest that C. arabica should be cultivated at medium–high altitudes, which is conducive to sustainable management and the production of high-quality C. arabica beans. [ABSTRACT FROM AUTHOR]

Isolation of heavy metals-resistant bacteria from their original habitat is a crucial step in bioremediation. Six lead (Pb) resistant bacterial strains were isolated and identified utilizing 16S rRNA to be Enterobacter ludwigii FACU 4, Shigella flexneri FACU, Microbacterium paraoxydans FACU, Klebsiella pneumoniae subsp. pneumonia FACU, Raoultella planticola FACU 3 and Staphylococcus xylosus FACU. It was determined that all these strains had their Minimum inhibitory concentration (MIC) to be 2500 ppm except R. planticola FACU 3 has a higher maximum tolerance concentration (MTC) up to 2700 ppm. We evaluated the survival of all six strains on lead stress, the efficiency of biosorption and lead uptake. It was found that R. planticola FACU 3 is the highest MTC and S. xylosus FACU was the lowest MTC in this evaluation. Therefore, transmission electron microscopy (TEM) confirmed the difference between the morphological responses of these two strains to lead stress. These findings led to explore more about the genome of R. planticola FACU 3 using illumine Miseq technology. Draft genome sequence analysis revealed the genome size of 5,648,460 bp and G + C content 55.8% and identified 5526 CDS, 75 tRNA and 4 rRNA. Sequencing technology facilitated the identification of about 47 genes related to resistance to many heavy metals including lead, arsenic, zinc, mercury, nickel, silver and chromium of R. planticola FACU 3 strain. Moreover, genome sequencing identified plant growth-promoting genes (PGPGs) including indole acetic acid (IAA) production, phosphate solubilization, phenazine production, trehalose metabolism and 4-hydroxybenzoate production genes and a lot of antibiotic-resistant genes. [ABSTRACT FROM AUTHOR]

Ginseng is a profitable crop worldwide; however, the ginseng replanting problem (GRP) is a major threat to its production. Soil amendment is a non-chemical method that is gaining popularity for alleviating continuous cropping obstacles, such as GRP. However, the impact of soil amendment with either cow dung or canola on GRP reduction and the associated soil microbiota remains unclear. In the present study, we evaluated the effect of soil amendment with cow dung, canola seed powder, and without amendment (control), on the survival of ginseng seedling transplants, the soil bacterial and fungal communities, and their associated metabolic functions. The results showed that cow dung increased ginseng seedling survival rate by 100 percent and had a remarkable positive effect on ginseng plant growth compared to control, whereas canola did not. Cow dung improved soil nutritional status in terms of pH, electrical conductivity, NO3−, total carbon, total phosphorus, and available phosphorus. The amplicon sequencing results using Illumina MiSeq showed that canola had the strongest negative effect in reducing soil bacterial and fungal diversity. On the other hand, cow dung stimulated beneficial soil microbes, including Bacillus, Rhodanobacter, Streptomyces, and Chaetomium, while suppressing Acidobacteriota. Community-level physiological profiling analysis using Biolog Ecoplates containing 31 different carbon sources showed that cow dung soil had a different metabolic activity with higher utilization rates of carbohydrates and polymer carbon sources, mainly Tween 40 and beta-methyl-d-glucoside. These carbon sources were most highly associated with Bacillota. Furthermore, predicted ecological function analyses of bacterial and fungal communities showed that cow dung had a higher predicted function of fermentation and fewer functions related to plant pathogens and fungal parasites, signifying its potential to enhance soil suppressiveness. Co-occurrence network analysis based on random matrix theory (RMT) revealed that cow dung transformed the soil microbial network into a highly connected and complex network. This study is the first to report the alleviation of GRP using cow dung as a soil amendment, and the study contributes significantly to our understanding of how the soil microbiota and metabolic alterations via cow dung can aid in GRP alleviation. [ABSTRACT FROM AUTHOR]

Introduction: Anaerobic ammonium oxidation (anammox) plays a vital role in the global nitrogen cycle by oxidizing ammonium to nitrogen under anaerobic environments. However, the existence, abundance, and diversity of anammox bacteria between different temperatures are less studied, particularly in purple paddy soils. Methods: 13C-DNA stable-isotope probe combined with Illumina MiSeq high-throughput sequencing was employed to explore soil abundance and diversity of anammox bacteria. In doing so, 40-60 cm depth soils from typical purple paddy soils in Chongqing, southwest China, were cultured under 12CO2-labeled and 13CO2-labeled at 35°C, 25°C, 15°C, and 5°C for 56 days. Results and Discussion: Anammox bacteria were not labeled at all by 13CO2 at 5°C. The highest abundance of anammox bacteria was found at 25°C (3.52 x 106 -3.66 x 106 copies·g-1 dry soil), followed by 35°C and 15°C (2.01 x 106 -2.37 x 106 copies·g-1 dry soil) and almost no increase at 5°C. The relative abundance of Candidatus Jettenia sp. was higher at 25°C and 15°C, while Candidatus Brocadia sp. was higher at 35°C and 5°C. Our results revealed differences in anammox bacteria at different temperatures in purple paddy soils, which could provide a better understanding of soil N cycling regulated by anammox bacteria. [ABSTRACT FROM AUTHOR]

Bacteria have a fundamental role in determining the fitness of grapevine, the composition of grapes and the features of wines but at present, little information is available. In this work, the bacteria colonizing the different portions of grapevine (bark, leaves and grapes) were explored in the vineyards of the Alpine region of Trentino, considering the impact of different environmental and agronomical variables. The vineyards included in the work were selected based on their different geographical positions (altitude) and grapevine training systems in order to explore the whole variability of the grapevine ecosystem. Moreover, the surface amount of copper was measured on grapes and leaves during the vegetative growth. Bacterial analysis, performed using plate counts and Illumina MiSeq, revealed an increase in the concentration of grape bacteria proportional to the progress of the ripening stage. Conversely, the peak of bacterial concentration onto leaf and bark samples occurred in August, probably due to the more favourable environmental conditions. In bark samples, the bacterial microbiota reached the 7 log CFU/cm2, while 6 log UFC/g were measured in grape samples. A remarkable biodiversity was observed, with 13 phyla, 35 classes, 55 orders, 78 families and 95 genera of bacteria present. The presence of some taxa (Alphaproteobacteria, Desulfovibrionaceae, Clostriadiales, Oscillospira, Lachnospiraceae and Bacteroidales) was ubiquitous in all vineyards, but differences in terms of relative abundance were observed according to the vegetative stage, altitude of the vineyard and training system. Bacteria having oenological implication (Lactobacillus, Pediococcus and Oenococcus) were detected in grape samples collected in August, in low abundance. The data revealed a complex bacterial ecosystem inside the vineyard that, while maintaining common traits, evolves according to environmental and agronomical inputs. This study contributes to define the role of bacteria in the complex balance established in each vineyard between human actions and agricultural environment, known as terroir. [ABSTRACT FROM AUTHOR]

Continuous cropping and soilborne diseases affect soil properties and soil microbial diversity and structure, which are the main factors posing obstacles to the continuous cropping of ginseng. This study explored the response of the physicochemical properties, enzyme activity, and microbial community of ginseng rhizosphere soil to continuous cropping and soilborne disease (root rot of ginseng). We used woodland soil without ginseng planting as a control to study these changes. The results showed that continuous cropping and soilborne disease significantly affected soil physicochemical properties, enzyme activities, and microbial communities. The levels of total nitrogen, hydrolyzable nitrogen, organic matter, and soil pH decreased significantly, while available phosphorus, available potassium, total phosphorus, and total potassium showed significant accumulation after continuous cropping. The activities of urease, catalase, sucrase, acid phosphatase, alkaline phosphatase, and polyphenol oxidase decreased significantly after continuous cropping. Using MiSeq high-throughput sequencing, we found that the alpha diversity and the number of bacterial and fungal communities significantly changed after continuous cropping and soilborne disease. A redundancy analysis suggests that soil physicochemical properties and enzyme activities also affect soil microbial communities. In summary, this study revealed the effects of continuous cropping and soilborne disease on soil and provides a theoretical basis for alleviating soilborne disease in ginseng. [ABSTRACT FROM AUTHOR]