Decoding the chromosome-scale genome of the nutrient-rich Agaricus subrufescens: a resource for fungal biology and biotechnology.

Agaricus subrufescens, also known as the "sun mushroom," has significant nutritional and medicinal value. However, its short shelf life due to the browning process results in post-harvest losses unless it's quickly dehydrated. This restricts its availability to consumers in the form of capsules. A genome sequence of A. subrufescens may lead to new cultivation alternatives or the application of gene editing strategies to delay the browning process. We assembled a chromosome-scale genome using a hybrid approach combining Illumina and Nanopore sequencing. The genome was assembled into 13 chromosomes and 31 unplaced scaffolds, totaling 44.5 Mb with 96.5% completeness and 47.24% GC content. 14,332 protein-coding genes were identified, with 64.6% of the genome covered by genes and 23.41% transposable elements. The mitogenome was circularized and encoded fourteen typical mitochondrial genes. Four polyphenol oxidase (PPO) genes and the Mating-type locus were identified. Phylogenomic analysis supports the placement of A. subrufescens in the Agaricomycetes clade. This is the first available genome sequence of a strain of the "sun mushroom." Results are available through a Genome Browser (https://plantgenomics.ncc.unesp.br/gen.php?id=Asub) and can support further fungal biological and genomic studies.

A comprehensive study on diesel oil bioremediation under microcosm conditions using a combined microbiological, enzymatic, mass spectrometry, and metabarcoding approach.

This study aims at the application of a marine fungal consortium (Aspergillus sclerotiorum CRM 348 and Cryptococcus laurentii CRM 707) for the bioremediation of diesel oil-contaminated soil under microcosm conditions. The impact of biostimulation (BS) and/or bioaugmentation (BA) treatments on diesel-oil biodegradation, soil quality, and the structure of the microbial community were studied. The use of the fungal consortium together with nutrients (BA/BS) resulted in a TPH (Total Petroleum Hydrocarbon) degradation 42% higher than that obtained by natural attenuation (NA) within 120 days. For the same period, a 72 to 92% removal of short-chain alkanes (C12 to C19) was obtained by BA/BS, while only 3 to 65% removal was achieved by NA. BA/BS also showed high degradation efficiency of long-chain alkanes (C20 to C24) at 120 days, reaching 90 and 92% of degradation of icosane and heneicosane, respectively. In contrast, an increase in the levels of cyclosiloxanes (characterized as bacterial bioemulsifiers and biosurfactants) was observed in the soil treated by the consortium. Conversely, the NA presented a maximum of 37% of degradation of these alkane fractions. The 5-ringed PAH benzo(a)pyrene, was removed significantly better with the BA/BS treatment than with the NA (48 vs. 38 % of biodegradation, respectively). Metabarcoding analysis revealed that BA/BS caused a decrease in the soil microbial diversity with a concomitant increase in the abundance of specific microbial groups, including hydrocarbon-degrading (bacteria and fungi) and also an enhancement in soil microbial activity. Our results highlight the great potential of this consortium for soil treatment after diesel spills, as well as the relevance of the massive sequencing, enzymatic, microbiological and GC-HRMS analyses for a better understanding of diesel bioremediation.

Methanogenic communities and methane emissions from enrichments of Brazilian Amazonia soils under land-use change.

Amazonian forest conversion into agricultural and livestock areas is considered one of the activities that contribute most to the emission of greenhouse gases, including methane. Biogenic methane production is mainly performed by methanogenic Archaea, which underscores the importance of understanding the drivers shaping microbial communities involved in the methane cycling and changes in methane metabolism. Here, we aimed to investigate the composition and structure of bacterial and archaeal communities in tropical soils in response to land-use changes, emphasizing the methanogenic communities. We collected soil samples from primary forest, pasture, and secondary forest of the Amazonian region and used a strategy based on the enrichment of the methanogenic community with three different methanogenic substrates followed by measurements of methane emission, quantification of mcrA gene copies by qPCR, and total 16 S rRNA gene sequencing (metataxonomics). We observed variations in the structure of bacterial and archaeal communities of soils under different uses. The richness of methanogenic communities was higher in pasture than forest soils and this richness remained during the incubation period, and as a consequence, the enrichment induced earlier methane emission in pastures-derived samples. Furthermore, pastures enrichments exhibited methanogenic archaea networks more complex than primary and secondary forests. In conclusion, pastures harbor a richer and more responsive methanogenic community than forest samples, suggesting that conversion of forest areas to pasture may boost methane emission.

Interactions between Soil Bacterial Diversity and Plant-Parasitic Nematodes in Soybean Plants.

Plant-parasitic nematodes are an important group of pests causing economic losses in agriculture worldwide. Among the plant-parasitic nematodes, the root-knot (Meloidogyne spp.) and root-lesion nematodes (Pratylenchus spp.) are considered the two most important ones affecting soybeans. In general, they damage soybean roots, causing a reduction of about one-third in productivity. The soil microbial community can exert a suppressive effect on the parasitism of plant-parasitic nematodes. Here, we investigated the effects of soil bacterial diversity on Meloidogyne javanica (Meloidogyne-assay) and Pratylenchus brachyurus (Pratylenchus-assay) suppression by manipulating microbial diversity using the dilution-to-extinction approach in two independent experiments under controlled conditions. Furthermore, we recorded the changes in the soil microbial community induced by plant-parasitic nematode infection. In Meloidogyne-assay, microbial diversity reduced the population density of M. javanica and improved plant performance. In Pratylenchus-assay, microbial diversity sustained the performance of soybean plants even at high levels of P. brachyurus parasitism. Each nematode population affected the relative abundance of different bacterial genera and altered the core microbiome of key groups within the bacterial community. Our findings provide fundamental insights into the interactions between soil bacterial diversity and plant-parasitic nematodes in soybean plants. IMPORTANCE Root-knot and root-lesion nematodes cause losses of billions of dollars every year to agriculture worldwide. Traditionally, they are controlled by using chemical nematicides, which in general have a negative impact on the environment and human health. Fortunately, the soil microbial community may suppress these pests, acting as an environmentally friendly alternative to control nematodes. However, the effects of soil microbial diversity on the parasitism of plant-parasitic nematodes still poorly understood. In this study, we provide fundamental insight into the interactions between soil bacterial diversity and plant-parasitic nematodes in soybean plants, which may be useful for the development of new strategies to control these phytopathogens.

Maintaining grass coverage increases methane uptake in Amazonian pastures, with a reduction of methanogenic archaea in the rhizosphere.

Cattle ranching is the largest driver of deforestation in the Brazilian Amazon. The rainforest-to-pasture conversion affects the methane cycle in upland soils, changing it from sink to source of atmospheric methane. However, it remains unknown if management practices could reduce the impact of land-use on methane cycling. In this work, we evaluated how pasture management can regulate the soil methane cycle either by maintaining continuous grass coverage on pasture soils, or by liming the soil to amend acidity. Methane fluxes from forest and pasture soils were evaluated in moisture-controlled greenhouse experiments with and without grass cover (Urochloa brizantha cv. Marandu) or liming. We also assessed changes in the soil microbial community structure of both bare (bulk) and rhizospheric pasture soils through high throughput sequencing of the 16S rRNA gene, and quantified the methane cycling microbiota by their respective marker genes related to methane generation (mcrA) or oxidation (pmoA). The experiments used soils from eastern and western Amazonia, and concurrent field studies allowed us to confirm greenhouse data. The presence of a grass cover not only increased methane uptake by up to 35% in pasture soils, but also reduced the abundance of the methane-producing community. In the grass rhizosphere this reduction was up to 10-fold. Methane-producing archaea belonged to the genera Methanosarcina sp., Methanocella sp., Methanobacterium sp., and Rice Cluster I. Further, we showed that soil liming to increasing pH compromised the capacity of forest and pasture soils to be a sink for methane, and instead converted formerly methane-consuming forest soils to become methane sources in only 40-80 days. Liming reduced the relative abundance of Beijerinckiacea family in forest soils, which account for many known methanotrophs. Our results demonstrate that pasture management that maintains grass coverage can mitigate soil methane emissions, compared to bare (bulk) pasture soil.

Land use and roles of soil bacterial community in the dissipation of atrazine.

Atrazine (ATZ) is one of the most widely used herbicides in the world even though it is classified as a carcinogenic endocrine disruptor. This study focused on how land use (grazing versus cultivation in parallel soils, the latter under no-till with a seven-year history of ATZ application) and bacterial community diversity affected ATZ dissipation. Proteobacteria, Bacteroidetes, Firmicutes, Actinobacteria, Acidobacteria, Verrucomicrobia, Planctomycetes, and Gemmatimonadetes were the dominant phyla in both soils. The mineralization of ATZ was much higher in soils under cultivation up to the onset of moderate diversity depletion (dilution =10-3), corresponding to 44-52% of the amount applied (< 5% in the grazed soil). This was attributed to the higher diversity and complexity of the soils´ bacterial communities which consist of microbial groups that were more adapted as a result of previous exposure to ATZ. In these cases, ATZ dissipation was attributed mainly to mineralization (DT50 = 4-11 d). However, formation of non-extractable ATZ residues was exceptionally important in the other cases (DT50 = 17-44 d). The cultivated soils also presented a higher number of bacterial genera correlated with ATZ dissipation, in which Acidothermus, Aquicela, Arenimonas, Candidatus_Koribacter, Hirschia, MND1, Nitrospira, Occallatibacter, OM27_clade, and Ralstonia are suggested as potential ATZ-degraders. Finally, ATZ dissipation was mostly associated with an abundance of microbial functions related to energy supply and N-metabolism, suggesting co-metabolism is its first biodegradation step.

Mucilaginibacter sp. Strain Metal(loid) and Antibiotic Resistance Isolated from Estuarine Soil Contaminated Mine Tailing from the Fundão Dam.

In 2015 a mine dam with Mn-Fe-rich tailings collapsed releasing million tons of sediments over an estuary, in the Southwest of Brazil. The tailings have a high concentration of metals that contaminated soil until the present day. The high contaminant concentrations possibly caused a selection for microorganisms able to strive in such harsh conditions. Here, we isolated metal(loid) and anti-biotic resistance bacteria from the contaminated estuarine soil. After 16S rDNA sequencing to identify the strains, we selected the Mucilaginibacter sp. strain for a whole-genome sequence due to the bioprospective potential of the genus and the high resistance profile. We obtained a complete genome and a genome-guided characterization. Our finding suggests that the 21p strain is possibly a new species of the genus. The species presented genes for resistance for metals (i.e., As, Zn, Co, Cd, and Mn) beyond resistance and cross-resistance for antibiotics (i.e., quinolone, aminoglycoside, ß-lactamase, sulphonamide, tetracycline). The Mucilaginibacter sp. 21p description as new species should be further explored, as their extracellular polymeric substances and the potential of this strain as bioremediation and as a growth promoter in high met-al(loid) contaminated soil.

Experimental validation under controlled conditions of real time PCR to quantify arbuscular mycorrhizal colonization in root.

Mycorrhizal colonization of roots is traditionally evaluated by empirical methods, such as root microscopy. We compared this method with data from using a real time PCR technique, and determined the correlation between methods, indicating particularities of a promising system for a quick and accurate molecular diagnostic of arbuscular mycorrhization.

Microorganisms: the secret agents of the biosphere, and their key roles in biotechnology / Micro-organismos: os agentes secretos da biosfera, e seus papéis determinantes na biotecnologia

Abstract We present a survey of projects that have been funded by FAPESP under the BIOTA-Microorganisms program. These projects generated a wide variety of results, including the identification of novel antibacterial-producing microorganisms, the characterization of novel microbial enzymes for industrial applications, taxonomic classification of novel microorganisms in several environments, investigation of the soil and mangrove microbial ecosystems and its influence on endangered plant species, and the sequencing of novel metagenome-assembled genomes. The results surveyed demonstrate the importance of microorganisms in environments that play important roles in human activities as well as the potential that many of these microorganisms have in contributing to biotechnological applications crucial for human survival in the 21st century.

Resumo Apresentamos um levantamento comentado de projetos financiados pelo programa BIOTA-Micro-organismos. Estes projetos geraram uma variada gama de resultados, incluindo a identificação de novos micro-organismos produtores de compostos antibacterianos, a caracterização de novas enzimas microbianas para usos industriais, classificação taxonômica de novos micro-organismos presentes em diversos ambientes, investigação de ecossistemas microbianos em solos e mangues e sua influência sobre plantas ameaçadas, e o sequenciamento de vários novos genomas microbianos derivados de metagenomas. Os resultados descritos demonstram o papel-chave de micro-organismos em ecossistemas importantes para atividades humanas, assim como o potencial que vários desses micro-organismos tem de contribuir para aplicações biotecnológicas cruciais para a sobrevivência humana no século 21.

Complete Genome Sequence of a Mucilaginibacter sp. Strain Isolated from Estuarine Soil Contaminated with Mine Tailings from the Samarco Disaster at Fundão Dam.

We report the complete genome sequence of Mucilaginibacter strain 21P, which was isolated from estuarine soil contaminated with mine tailings from the Samarco disaster, which occurred in 2015 in Brazil. The genome sequence comprised 4,739,655 bp, with a G+C content of 43.2%, and harbors multiple antibiotic and metal resistance genes.

Complete Genome Sequence of Bacillus safensis Strain 3A, a Heavy Metal-Resistant Bacterium Isolated from Contaminated Estuarine Sediment in Brazil.

Bacillus safensis 3A was isolated from a contaminated estuarine sediment sample with mine tailing from the Samarco dam disaster, which occurred in 2015 in Minas Gerais State, Brazil. We report here a draft genome sequence (3.6 Mb) of this bacterial strain. B. safensis exhibited strong resistance to heavy metals.

Genomic analysis reveals the potential for hydrocarbon degradation of Rhodopirellula sp. MGV isolated from a polluted Brazilian mangrove.

Planctomycetes are bacteria found in several environments, such as mangroves. In the coastline of the State of Sao Paulo (Brazilian Southeast), mangroves occur in different stages of environmental contamination, promoted by the proximity to the city and industrial activities. One of these mangroves (located in the city of Bertioga) is characterized by the high impact due to past petroleum and ongoing urban contamination. We isolated five bacteria affiliated to Planctomycetes from this mangrove and further subjected them to phenotypical and genetic analysis. The tolerance for salinity was demonstrated by the cultivation under distinct concentrations of NaCl. The ability of this bacterium to use diverse carbon sources was revealed by the use of 30 C-sources from a total of 31 tests. We found the isolate Rhodopirellula sp. MGV very closely affiliated to species of the genus Rhodopirellula, harboring a genome with 7.16 Mbp and 55.3% of GC. The annotation of the 77 contigs resulted in 6.284 CDS, with a remarkable occurrence of sequences associated with aromatic carbon metabolism. In conclusion, we present the isolation and characterization of a Planctomycetes from mangroves, suggesting its participation in the degradation of hydrocarbons present in the contaminated mangroves studied.

Sulfadiazine dissipation as a function of soil bacterial diversity.

Antibiotic residues in the environment are concerning since results in dispersion of resistance genes. Their degradation is often closely related to microbial metabolism. However, the impacts of soil bacterial community on sulfadiazine (SDZ) dissipation remains unclear, mainly in tropical soils. Our main goals were to evaluate effects of long-term swine manure application on soil bacterial structure as well as effects of soil microbial diversity depletion on SDZ dissipation, using "extinction dilution approach" and 14C-SDZ. Manure application affected several soil attributes, such as pH, organic carbon (OC), and macronutrient contents as well as bacterial community structure and diversity. Even minor bacterial diversity depletion impacted SDZ mineralization and non-extractible residue (NER) formation rates, but NER recovered along 42 d likely due to soil diversity recovery. However, this period may be enough to spread resistance genes into the environment. Surprisingly, the non-manured natural soil (NS-S1) showed faster SDZ dissipation rate (DT90 = 2.0 versus 21 d) and had a great number of bacterial families involved in major SDZ dissipation pathways (mineralization and mainly NER), such as Isosphaeraceae, Ktedonobacteraceae, Acidobacteriaceae_(Subgroup_1), Micromonosporaceae, and Sphingobacteriaceae. This result is unique and contrasts our hypothesis that long-term manured soils would present adaptive advantages and, consequently, have higher SDZ dissipation rates. The literature suggests instantaneous chemical degradation of SDZ in acidic soils responsible to the fast formation of NER. Our results show that if chemical degradation happens, it is soon followed by microbial metabolism (biodegradation) performed by a pool of bacteria and the newly formed metabolites should favors NER formation since SDZ presented low sorption. It also showed that SDZ mineralization is a low redundancy function.

Application of extracellular polymers on soil communities exposed to oil and nickel contamination.

The petrochemical industry is responsible for many accidental releases of pollutants in soil such as hydrocarbons and toxic metals. This co-contamination is responsible for a delay in the degradation of the organic pollution. Many successful technologies to remove these metals apply extracellular polymeric substances (EPS). In this study, we tested the application of an EPS from a Paenibacillus sp. to aid the bioremediation of soils contaminated with crude oil and nickel. We conducted a microcosm experiment to soils containing combinations of oil, nickel, and EPS. The final concentration of oil was evaluated with an infrared spectrometer. Also, we sequenced the metagenomes of the samples in an ion torrent sequencer. The application of EPS did not aid the removal of hydrocarbons with or without the presence of nickel. However, it led to a smaller decrease in the diversity indexes. EPS decreased the abundance of Actinobacteria and increased that of Proteobacteria. The EPS also decreased the connectivity among Actinobacteria in the network analysis. The results indicated that the addition of EPS had a higher effect on the community structure than nickel. Altogether, our results indicate that this approach did not aid the bioremediation of hydrocarbons likely due to its effect in the community structure that affected hydrocarbonoclastic microorganisms.

Genetic diversity of N-fixing and plant growth-promoting bacterial community in different sugarcane genotypes, association habitat and phenological phase of the crop.

This study aimed to evaluate the genetic diversity of bacterial community associated to different sugarcane genotypes, association habitat and phenological phase of the culture, as well as to isolate, to identify and to characterize your potential for plant growth-promoting. Root and rhizospheric soil samples from RB 92579 and RB 867515 varieties were collected at 120 and 300 days after regrowth (DAR). The diversity of bacterial was evaluated through of the 16S rRNA and nifH genes. We found greater genetic diversity in the root endophytic habitat at 120 DAR. We identify the genera Burkholderia sp., Pantoea sp., Erwinia sp., Stenotrophomonas sp., Enterobacter sp. and Pseudomonas sp. The genera Bacillus sp. and Dyella sp. were only identified in the variety RB 92579. We found indices above 50% for biological nitrogen fixation, production of indole acetic acid and phosphate solubilization, showing that the use of these bacteria in biotechnological products is very promising.

Soil Microbial Diversity Affects the Plant-Root Colonization by Arbuscular Mycorrhizal Fungi.

Terrestrial plants establish symbiosis with arbuscular mycorrhizal fungi (AMF) to exchange water and nutrients. However, the extent to which soil biodiversity influences such association remains still unclear. Here, we manipulated the soil microbial diversity using a "dilution-to-extinction" approach in a controlled pot microcosm system and quantified the root length colonization of maize plants by the AMF Rhizophagus clarus. The experiment was performed by manipulating the soil microbiome within a native and foreign soil having distinct physicochemical properties. Overall, our data revealed significant positive correlations between the soil microbial diversity and AMF colonization. Most importantly, this finding opposes the diversity-invasibility hypothesis and highlights for a potential overall helper effect of the soil biodiversity on plant-AMF symbiosis.

The bacterial and fungal communities associated with Anthurium ssp. leaves: Insights into plant endemism and microbe association.

It is known that different plant species select specific microbes to live inside their tissues in a process determined by the host genotype, phenotype and geographic location, which can introduce discussion on plant endemism and the assembly of specific microbial communities. Herein, we report the results of an investigation relating the geographic distribution of plant species and the composition of microbial communities associated with plant hosts. The bacterial and fungal community associated with Anthurium plant leaves was mapped to assess the diversity and ecology of the endophytic community associated with Anthurium spp. collected on islands and on the Brazilian mainland. Twenty-six Anthurium specimens were surveyed, distributed throughout the São Paulo state coastline, including Alcatrazes Island, some coastal islands and distinct mainland environments. Bacterial and fungal endophytes were obtained from the leaves of A. alcatrazense, A. loefgrenii, A. penthaphyllum, A. urvellianum and A. intermedium and subjected to massive bacterial 16S rRNA and fungal ITS sequencing. The results indicated that A. alcatrazense, endemic to Alcatrazes Island, hosted a specific bacterial community structure, while its fungal community was similar to that of Anthurium species from other locations. Betaproteobacteria showed a high differential occurrence in A. alcatrazense. Some groups of fungi were found mainly inhabiting A. loefgrenii plants. While Alphaproteobacteria, Gammaproteobacteria, Actinobacteria and Sordariomycetes, Dothiodeomycetes and Tremellomycetes composed the core microbial community among Anthurium plants. The results suggest crucial role for the bacterial communities to endemic plants, while endophytic fungal diversity is less specifically distributed among endemic and nonendemic plant species.

Bacteria from tropical semiarid temporary ponds promote maize growth under hydric stress.

World climate change has triggered soil water stress and imposed limitations on agricultural production. Plant growth-promoting bacteria (PGPBs) have been an efficient strategy to improve the biological supply and growth of plants under distinct abiotic stress conditions. We hypothesized that the soils from a temporary pond may harbor PGPBs with potential strains which increase maize tolerance to water deficit. We studied rhizosphere and bulk soil of Mimosa bimucronata in a temporary pond from semiarid Northeast Brazil to access strains with characteristics to promote plant growth and mitigate abiotic stress for maize crop. We isolated 355 bacterial isolates, from which 96 were selected based on the morphophysiological characterization to assess IAA production (42 % produced over 50 µg mL-1 of IAA), calcium phosphate solubilization (with one isolate achieving medium IS), biofilm and exopolysaccharides production (66 % and 98 % of isolates, respectively). Based on these mechanisms, the 30 most promising bacterial isolates were selected to assess biological nitrogen fixation (74 % of the isolates showed nitrogenase activity greater than 20 C2H4.h-1.mg-1), ACC deaminase activity (80 % of isolates) and growth in medium with reduced water activity (8 % of isolates grew in medium with water activity (Aw) of 0.844). We sequenced the 16S rRNA gene from the seven most promising isolates in in vitro and in vivo assays, which were identified as Staphylococcus edaphicus, Bacillus wiedmannii, Micrococcus yunnanensis, Streptomyces alboflavus, Streptomyces alboflavus, Bacillus wiedmanni and Bacillus cereus. In vivo, eleven isolates and three bacterial consortia did not differ from the control with nutrient solution, for total leaf area and root dry mass of maize. S. alboflavus (BS43) had the best in vivo results, not differing from the control with nutrient solution. We highlight the unpublished potential of Staphylococcus edaphicus and Streptomyces alboflavus in promoting the growth of plants under water stress. In addition, it is the first report of bacteria isolated from a temporary pond in the Brazilian semiarid which promoting plant growth attributes and development.

The natural recovery of soil microbial community and nitrogen functions after pasture abandonment in the Amazon region.

We assessed the impacts of forest-to-pasture conversion on the dynamic of soil microbial communities, especially those involved in the N-cycle, and their potential functions, using DNA-metagenomic sequencing coupled with the quantification of marker genes for N-cycling. We also evaluated whether the community's dynamic was reestablished with secondary forest growth. In general, the microbial community structure was influenced by changes in soil chemical properties. Aluminum and nitrate significantly correlated to community structure and with 12 out of 21 microbial phyla. The N-related microbial groups and their potential functions were also affected by land-use change, with pasture being clearly different from primary and secondary forest systems. The microbial community analysis demonstrated that forest-to-pasture conversion increased the abundance of different microbial groups related to nitrogen fixation, including Bacteroidetes, Chloroflexi and Firmicutes. In contrast, after pasture abandonment and with the secondary forest regeneration, there was an increase in the abundance of Proteobacteria taxa and denitrification genes. Our multi-analytical approach indicated that the secondary forest presented some signs of resilience, suggesting that the N-related microbial groups and their potential functions can be recovered over time with implications for future ecological restoration programs.