Microbial central carbon metabolism in a tidal freshwater marsh and an upland mixed conifer soil under oxic and anoxic conditions.

The central carbon (C) metabolic network is responsible for most of the production of energy and biosynthesis in microorganisms and is therefore key to a mechanistic understanding of microbial life in soil communities. Many upland soil communities have shown a relatively high C flux through the pentose phosphate (PP) or the Entner-Doudoroff (ED) pathway, thought to be related to oxidative damage control. We tested the hypothesis that the metabolic organization of the central C metabolic network differed between two ecosystems, an anoxic marsh soil and oxic upland soil, and would be affected by altering oxygen concentrations. We expected there to be high PP/ED pathway activity under high oxygen concentrations and in oxic soils and low PP/ED activity in reduced oxygen concentrations and in marsh soil. Although we found high PP/ED activity in the upland soil and low activity in the marsh soil, lowering the oxygen concentration for the upland soil did not reduce the relative PP/ED pathway activity as hypothesized, nor did increasing the oxygen concentration in the marsh soil increase the PP/ED pathway activity. We speculate that the high PP/ED activity in the upland soil, even when exposed to low oxygen concentrations, was related to a high demand for NADPH for biosynthesis, thus reflecting higher microbial growth rates in C-rich soils than in C-poor sediments. Further studies are needed to explain the observed metabolic diversity among soil ecosystems and determine whether it is related to microbial growth rates.IMPORTANCEWe observed that the organization of the central carbon (C) metabolic processes differed between oxic and anoxic soil. However, we also found that the pentose phosphate pathway/Entner-Doudoroff (PP/ED) pathway activity remained high after reducing the oxygen concentration for the upland soil and did not increase in response to an increase in oxygen concentration in the marsh soil. These observations contradicted the hypothesis that oxidative stress is a main driver for high PP/ED activity in soil communities. We suggest that the high PP/ED activity and NADPH production reflect higher anabolic activities and growth rates in the upland soil compared to the anaerobic marsh soil. A greater understanding of the molecular and biochemical processes in soil communities is needed to develop a mechanistic perspective on microbial activities and their relationship to soil C and nutrient cycling. Such an increased mechanistic perspective is ecologically relevant, given that the central carbon metabolic network is intimately tied to the energy metabolism of microbes, the efficiency of new microbial biomass production, and soil organic matter formation.

Microbial carbohydrate active enzyme (CAZyme) genes and diversity from Menagesha Suba natural forest soils of Ethiopia as revealed by shotgun metagenomic sequencing.

BACKGROUND: The global over-reliance on non-renewable fossil fuels has led to the emission of greenhouse gases, creating a critical global environmental challenge. There is an urgent need for alternative solutions like biofuels. Advanced biofuel is a renewable sustainable energy generated from lignocellulosic plant materials, which can significantly contribute to mitigating CO2 emissions. Microbial Carbohydrate Active Enzymes (CAZymes) are the most crucial enzymes for the generation of sustainable biofuel energy. The present study designed shotgun metagenomics approaches to assemble, predict, and annotate, aiming to gain an insight into the taxonomic diversity, annotate CAZymes, and identify carbohydrate hydrolyzing CAZymes from microbiomes in Menagesha suba forest soil for the first time. RESULTS: The microbial diversity based on small subunit (SSU) rRNA analysis revealed the dominance of the bacterial domain representing 81.82% and 92.31% in the studied samples. Furthermore, the phylum composition result indicated the dominance of the phyla Proteobacteria (23.08%, 27.27%), Actinobacteria (11.36%, 20.51%), and Acidobacteria (10.26%, 15.91%). The study also identified unassigned bacteria which might have a unique potential for biopolymer hydrolysis. The metagenomic study revealed that 100,244 and 65,356 genes were predicted from the two distinct samples. A total number of 1806 CAZyme genes were identified, among annotated CAZymes, 758 had a known enzyme assigned to CAZymes. Glycoside hydrolases (GHs) CAZyme family contained most of the CAZyme genes with known enzymes such as ß-glucosidase, endo-ß-1,4-mannanase, exo-ß-1,4-glucanase, α-L-arabinofuranosidase and oligoxyloglucan reducing end-specific cellobiohydrolase. On the other hand, 1048 of the identified CAZyme genes were putative CAZyme genes with unknown enzymatical activity and the majority of which belong to the GHs family. CONCLUSIONS: In general, the identified putative CAZymes genes open up an opportunity for the discovery of new enzymes responsible for hydrolyzing biopolymers utilized for biofuel energy generation. This finding is used as a first-hand piece of evidence to serve as a benchmark for further and comprehensive studies to unveil novel classes of bio-economically valuable genes and their encoded products.

Metatranscriptomics sheds light on the links between the functional traits of fungal guilds and ecological processes in forest soil ecosystems.

Soil fungi belonging to different functional guilds, such as saprotrophs, pathogens, and mycorrhizal symbionts, play key roles in forest ecosystems. To date, no study has compared the actual gene expression of these guilds in different forest soils. We used metatranscriptomics to study the competition for organic resources by these fungal groups in boreal, temperate, and Mediterranean forest soils. Using a dedicated mRNA annotation pipeline combined with the JGI MycoCosm database, we compared the transcripts of these three fungal guilds, targeting enzymes involved in C- and N mobilization from plant and microbial cell walls. Genes encoding enzymes involved in the degradation of plant cell walls were expressed at a higher level in saprotrophic fungi than in ectomycorrhizal and pathogenic fungi. However, ectomycorrhizal and saprotrophic fungi showed similarly high expression levels of genes encoding enzymes involved in fungal cell wall degradation. Transcripts for N-related transporters were more highly expressed in ectomycorrhizal fungi than in other groups. We showed that ectomycorrhizal and saprotrophic fungi compete for N in soil organic matter, suggesting that their interactions could decelerate C cycling. Metatranscriptomics provides a unique tool to test controversial ecological hypotheses and to better understand the underlying ecological processes involved in soil functioning and carbon stabilization.

Collimonas rhizosphaerae sp. nov., a novel species isolated from the beech rhizosphere.

Bacterial strain H4R21T was isolated from beech rhizosphere soil sampled in the forest experimental site of Montiers (Meuse, France). It effectively weathers minerals, hydrolyses chitin and produces quorum sensing signal molecules. The strain is aerobic and Gram-stain-negative. Phylogenetic analysis based on its 16S rRNA gene sequence indicated that strain H4R21T belongs to the genus Collimonas with high sequence similarity to C. arenae Ter10T (99.38 %), C. fungivorans Ter6T(98.97 %), C. pratensis Ter91T (98.76 %), C. humicola RLT1W51T (98.46 %) and C. silvisoli RXD178 T (98.46 %), but less than 98 % similarity to other strains of the genus Collimonas. The predominant quinone in H4R21T is ubiquinone-8 (Q8). The major polar lipids are diphosphatidylglycerol, phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol and lipid. The major fatty acids identified were C12 : 0, C12:0 3-OH, C16  :  0 and C17:0 cyclo. The digital DNA G+C content of the genomic DNA was 59.5 mol%. Furthermore, the strain could be clearly distinguished from its closely related type strains by a combination of phylogenomic and in silico DNA-DNA hybridization results, and phenotypic characteristics. Therefore, strain H4R21T represents a novel species within the genus Collimonas, for which the name Collimonas rhizosphaerae sp. nov. is proposed, with strain H4R21T (=CFBP 9203T=DSM 117599T) as the type strain.

Eastern Canadian boreal forest soil and foliar chemistry show evidence of resilience to long-term nitrogen addition.

The boreal forest is one of the world's largest terrestrial biome and plays crucial roles in global biogeochemical cycles, such as carbon (C) sequestration in vegetation and soil. However, the impacts of decades of N deposition on N-limited ecosystems, like the eastern Canadian boreal forest, remain unclear. For 13 years, N deposition was simulated by periodically adding ammonium nitrate on soils of two boreal coniferous forests (i.e., balsam fir and black spruce) of eastern Canada, at low (LN) and high (HN) rates, corresponding to 3 and 10 times the ambient N deposition, respectively. We show that more than a decade of N addition had no strong effects on mineral soil C, N, P, and cation concentrations and on foliar total Ca, K, Mg, and Mn concentrations. In organic soil, C stock was not affected by N addition while N stock increased, and exchangeable Ca2+ and Mg2+ decreased at the balsam fir site under HN treatment. At both sites, LN treatment had nearly no impact on foliage and soil chemistry but foliar N and N:P significantly increased under HN treatment, potentially leading to foliar nutrient imbalance. Overall, our work indicates that, in the eastern Canadian boreal forest, soil and foliar nutrient concentrations and stocks are resilient to increasing N deposition potentially because, in the context of N limitation, extra N would be rapidly immobilized by soil micro-organisms and vegetation. These findings could improve modeling future boreal forest soil C stocks and biomass growth and could help in planning forest management strategies in eastern Canada.

Occurrence and fate of current-use pesticides in Chinese forest soils.

Pesticides play a crucial role in securing global food production to meet increasing demands. However, because of their pervasive use, they are now ubiquitous environmental pollutants that have adverse effects on both ecosystems and human health. In this study, the environmental occurrence and fate of 16 current-use pesticides (CUPs) were investigated in 93 forest soil samples obtained from 11 distinct mountains in China. The concentrations of the target pesticides ranged from 0.36 to 55 ng/g dry weight. Cypermethrin, dicofol, chlorpyrifos, chlorothalonil, and trifluralin were the most frequently detected CUPs. The CUP concentrations were generally higher in the O-horizon than in the A-horizon. Chlorpyrifos, chlorothalonil, and dicofol were detected in most deep layers in soil profiles from three mountains selected to represent distinct climate zones. No clear altitudinal trend in organic carbon-normalized concentrations of CUPs was observed in the O- or A-horizons within individual mountains. A negative correlation was noted between the CUP concentrations and the altitudes across all sampling sites. This indicated that proximity to emission sources was a key factor affecting the spatial distribution of CUPs in mountain forest soil on a national scale. The ecological risk assessment showed that dicofol and cypermethrin pose potential risks to earthworms. This study emphasizes the importance of source control when setting management strategies for CUPs.

Paenibacillus silvisoli sp. nov. and Paenibacillus humicola sp. nov., isolated from forest soil.

During the study of microbial ecology of forest soil, two circular, white-colored bacterial colonies were isolated and labeled as strains TW38T and TW40T. Both strains were catalase positive and oxidase negative. Strains TW38T and TW40T demonstrated growth within a temperature range of 10-37 °C and 18-37 °C, respectively, and thrived within a pH range of 5.5-9.0 and 6.0-8.0, respectively. Both strains grew at 0-2.0% (w/v) NaCl concentrations. The phylogenetic analysis indicated that strains TW38T and TW40T affiliated to the genus Paenibacillus, with the closest neighbors being Paenibacillus montanisoli RA17T (98.6%) and Paenibacillus arachidis E3T (95.4%), respectively. In both strains, the sole respiratory quinone was MK-7, the signature fatty acid was antiso-C15:0, and the major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, and phosphatidylcholine. The digital DNA-DNA hybridization and the average nucleotide identity values between TW38T, TW40T, and closest reference strains were < 29.0% and < 85.0%, respectively. The DNA G+C content of TW38T and TW40T was 54.5% and 57.1%, respectively. In general, the phylogenetic, genomics, chemotaxonomic, and phenotypic data support the differentiation of TW38T and TW40T from other closest members of the genus Paenibacillus. Thus, we conclude both strains TW38T and TW40T represent novel species of the genus Paenibacillus, for which the name Paenibacillus silvisoli sp. nov. and Paenibacillus humicola sp. nov. are proposed, respectively. The type strain of Paenibacillus silvisoli is TW38T (= KCTC 43468T = NBRC 116015T) and type strain of Paenibacillus humicola is TW40T (= KCTC 43469T = NBRC 116016T).

Chitinophaga nivalis sp. nov., isolated from forest soil in Pyeongchang, Republic of Korea.

Rod-shaped Gram-stain-negative, aerobic bacterial strains, designated PC14 and PC15T, were isolated from a forest soil sample collected in Pyeongchang county, Gangwon-do, Republic of Korea. Strains PC14 and PC15T grew at 15-37 °C (optimum, 28-30 °C in tryptone soya agar and Mueller-Hinton agar), hydrolysed chitin and casein, and tolerated pH 8.5 and 2 % (w/v) NaCl. The strains were most closely related to members of the genus Chitinophaga, namely Chitinophaga arvensicola DSM 3695T (98.4 %), Chitinophaga longshanensis Z29T (98.3 %), Chitinophaga ginsengisegetis Gsoil 040T (97.8 %), Chitinophaga polysaccharea MRP-15T (97.8 %) and Chitinophaga niastensis JS16-4T (97.7 %). The type strain grew well on conventional commercial media in the laboratory, including tryptone soya agar, Mueller-Hinton agar, Reasoner's 2A agar, nutrient agar and Luria-Bertani agar. The major polar lipid profile comprised phosphatidylethanolamine, an unidentified aminolipid and unidentified polar lipids. The major respiratory quinone was menaquinone-7. The main fatty acids were iso-C15:0, C16:1 ω5c, C16:0 3-OH, iso-C15:0 3-OH and iso-C17:0 3-OH. The DNA G+C content of the isolated strain based on the whole genome sequence was 46.6 mol%. The average nucleotide identity and digital DNA-DNA hybridization values between strains PC14 and PC15T and the reference type strains ranged from 71.0 to 76.5 %, and from 20.3 to 20.7 %, respectively. Based on phenotypic, chemotypic and genotypic evidence, strain PC15T could be differentiated phylogenetically and phenotypically from the recognized species of the genus Chitinophaga. Therefore, strain PC15T is considered to represent a novel species, for which the name Chitinophaga nivalis sp. nov. is proposed. The type strain is PC15T (=KACC 22893T=JCM 35788T).

Scleromatobacter humisilvae gen. nov., sp. nov., a novel bacterium isolated from oak forest soil.

A novel bacterial strain, designated BS-T2-15T, isolated from forest soil in close proximity to decaying oak wood, was characterized using a polyphasic taxonomic approach. Phylogenetic analyses based on 16S rRNA gene sequences as well as phylogenomic analyses based on coding sequences of 340 concatenated core proteins indicated that strain BS-T2-15T forms a distinct and robust lineage in the Rubrivivax-Roseateles -Leptothrix-Azohydromonas -Aquincola-Ideonella branch of the order Burkholderiales. The amino acid identity and the percentage of conserved proteins between the genome of strain BS-T2-15T and genomes of closely related type strains ranged from 64.27 to 66.57% and from 40.89 to 49.27 %, respectively, providing genomic evidence that strain BS-T2-15T represents a new genus. Its cells are Gram-stain-negative, aerobic, motile by a polar flagellum, rod-shaped and form incrusted white to ivory colonies. Optimal growth is observed at 20-22 °C, pH 6 and 0% NaCl. The predominant fatty acids of strain BS-T2-15T are C16 : 1 ω7c, C16 : 0 and C14 : 0 2-OH. Its polar lipid profile consists of a mixture of phosphatidylethanolamine, diphosphatidylglycerol and phosphatidylglycerol and its main respiratory quinone is ubiquinone 8. The estimated size of its genome is 6.28 Mb with a DNA G+C content of 69.56 mol%. Therefore, on the basis of phenotypic and genotypic properties, the new strain BS-T2-15T represents a novel genus and species for which the name Scleromatobacter humisilvae gen. nov., sp. nov., is proposed. The type strain is BS-T2-15T (DSM 113115T=UBOCC-M-3373T).

Paenibacillus caseinilyticus sp. nov., isolated forest soil.

A milky-white-coloured, aerobic, Gram-stain-positive, rod-shaped and motile bacterial strain (GW78T) was isolated from forest soil. GW78T was catalase-positive and oxidase-negative. The strain was able to grow optimally at 37 °C and at pH 7.0 in Reasoner's 2A media. The phylogenetic and 16S rRNA gene sequence analysis of GW78T showed its affiliation with the genus Paenibacillus. The 16S rRNA gene sequence of GW78T revealed 98.3 % similarity to its nearest neighbour Paenibacillus mucilaginosus VKPM B-7519T. Its chemotaxonomic properties included MK-7 as the sole menaquinone, phosphatidylglycerol, diphosphatidylglycerol, phosphatidylmonomethylethanolamine and phosphatidylethanolamine as major polar lipids, and anteiso-C15 : 0, C16 : 1 ω11c and anteiso-C17 : 0 as predominant fatty acids. Digital DNA-DNA hybridization and average nucleotide identity results with its closest relatives were <74.0 % and <14.0 %, respectively. Overall, 16S rRNA gene sequence comparisons, phylogenetic and genomic evidence, and phenotypic and chemotaxonomic data allow the differentiation of GW78T from other members of the genus Paenibacillus. Thus, we propose that strain GW78T represents a novel species of the genus Paenibacillus, with the name Paenibacillus caseinilyticus sp. nov. The type strain is GW78T (=KCTC 43430T=NBRC 116023T).

Streptantibioticus silvisoli sp. nov., acidotolerant actinomycetes from pine litter, reclassification of Streptomyces cocklensis, Streptomyces ferralitis, Streptomyces parmotrematis and Streptomyces rubrisoli as Actinacidiphila cocklensis comb. nov., Streptantibioticus ferralitis comb. nov., Streptantibioticus parmotrematis comb. nov. and Streptantibioticus rubrisoli comb. nov., and emended descriptions of the genus Streptantibioticus, the family Streptomycetaceae and Streptomyces iconiensis.

Filamentous actinomycetes, designated SL13 and SL54T, were isolated from pine litter and their taxonomic status resolved using a polyphasic approach. The isolates exhibit chemotaxonomic and morphological properties consistent with their classification in the family Streptomycetaceae. They form extensively branched substrate mycelia bearing aerial hyphae that differentiate into straight chains of cylindrical spores. The whole-organism hydrolysates contain ll-diaminopimelic acid, glucose, mannose and ribose, the predominant isoprenologue is MK-9(H8), the polar lipids are diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, phosphatidylglycerol and glycophospholipids, and the major fatty acids are anteiso-C15 : 0, iso-C15 : 0, iso-C16 : 0 and anteiso-C17 : 0. Phylogenetic trees based on 16S rRNA gene sequences and multilocus gene sequences of conserved housekeeping genes show that the isolates form a well-supported lineage that is most closely related to Streptomyces parmotrematis NBRC 115203T. All of these strains form a well-defined clade in the multilocus sequence analysis tree together with Streptantibioticus cattleyicolor DSM 46488T, Streptomyces ferralitis DSM 41836T and Streptomyces rubrisoli DSM 42083T. Draft genomes assemblies of the isolates are rich in biosynthetic gene clusters predicted to produce novel specialized metabolites and stress-related genes which provide an insight into how they have adapted to the harsh conditions that prevail in pine litter. Phylogenomically, both isolates belong to the same lineage as the type strains of S. cattleyicolor, S. ferralitis, S. parmotrematis and S. rubrisoli; these relationships are underpinned by high average amino acid identity, average nucleotide identity and genomic DNA-DNA hybridization values. These metrics confirm that isolates SL13 and SL54T belong to a novel species that is most closely related to S. parmotrematis NBRC 115203T and that these strains together with S. ferralitis DSM 41836T, S. rubrisoli DSM 42083T belong to the genus Streptantibioticus. Consequently, it is proposed that the isolates be recognized as a new Streptantibioticus species, Streptantibioticus silvisoli comb. nov., with isolate SL54T (=DSM 111111T=PCM3044T) as the type strain, and that S. ferralitis, S. parmotrematis and S. rubrisoli be transferred to the genus Streptantibioticus as Streptantibioticus ferralitis comb. nov., Streptantibioticus parmotrematis comb. nov. and Streptantibioticus rubrisoli comb. nov. Emended descriptions are given for the genus Streptantibioticus, the family Streptomycetaceae and for Streptomyces iconiensis which was found to be a close relative of the isolates in the 16S rRNA gene sequence analyses. It is also proposed that Streptomyces cocklensis be transferred to the genus Actinacidiphila as Actinacidiphila cocklensis comb. nov based on its position in the MLSA and phylogenomic trees and associated genomic data.

Dyella humicola sp. nov., Dyella subtropica sp. nov., Dyella silvatica sp. nov. and Dyella silvae sp. nov., isolated from subtropical forest soil.

Four novel bacterial strains, designated RBB1W86T, RXD159T, RBB189T and RLT163T, were isolated from subtropical forest soil of the Nanling National Nature Reserve located in Guangdong Province, PR China. 16S rRNA gene phylogeny indicated their affiliation to the genus Dyella, among which strains RBB1W86T and RXD159T were closely related to Dyella halodurans CGMCC 1.15435T with 16S rRNA gene sequence similarities of 98.8 and 99.5 %, respectively, and strains RBB189T and RLT163T were closely related to Dyella tabacisoli CGMCC 1.16273T (98.8 %) and Dyella japonica JCM 21530T (99.4 %), respectively. Phylogenomic analysis based on 92 core genes showed consistent phylogeny with the 16S rRNA gene phylogeny for strains RBB1W86T, RBB189T and RLT163T, while strain RXD159T showed a closer relationship with D. tabacisoli CGMCC 1.16273T and strain RBB189T. The genome-derived average nucleotide identity (ANI) values between the newly isolated strains and their closely related species were 70.18‒90.20 %, and the corresponding digital DNA-DNA hybridization (dDDH) values were 20.80‒40.30 %. Meanwhile, the ANI and dDDH values between each pair of the newly isolated strains were 75.80‒79.77 % and 21.30‒23.30 %, respectively. They all took iso-C15 : 0 and summed feature 9 (10-methyl C16  : 0 and/or iso-C17  : 1 ω9c) as the major fatty acids. Moreover, C16 : 0, iso-C16 : 0, iso-C17 : 0 and summed feature 3 (C16 : 1 ω7c and/or C16 : 1 ω6c) were also variously distributed as major components. They all took ubiquinone 8 as the only predominant respiratory quinone and phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine and an unidentified phospholipid as the major polar lipids. Phosphatidylmethylethanolamine was only present in strain RBB189T as another major component. Based on the results of phenotypic, genotypic and chemotaxonomic analyses, the newly isolated strains could be clearly distinguished from their closely related species and should represent four distinct novel species of the genus Dyella, for which the names Dyella humicola sp. nov. (type strain RBB1W86T=GDMCC 1.1901T=KACC 21988T), Dyella subtropica sp. nov. (type strain RXD159T=GDMCC 1.1902T=KACC 21989T), Dyella silvatica sp. nov. (type strain RBB189T=GDMCC 1.1900T=KACC 21990 T) and Dyella silvae sp. nov. (type strain RLT163T=GDMCC 1.1916T=KACC 21991T) are proposed.

Prolonged Effect of Forest Soil Compaction on Methanogen and Methanotroph Seasonal Dynamics.

Methane (CH4) oxidation by methanotrophic bacteria in forest soils is the largest biological sink for this greenhouse gas on earth. However, the compaction of forest soils by logging traffic has previously been shown to reduce the potential rate of CH4 uptake. This change could be due to not only a decrease of methanotrophs but also an increase in methanogen activity. In this study, we investigated whether the decrease in CH4 uptake by forest soils, subjected to compaction by heavy machinery 7 years earlier, can be explained by quantitative and qualitative changes in methanogenic and methanotrophic communities. We measured the functional gene abundance and polymorphism of CH4 microbial oxidizers (pmoA) and producers (mcrA) at different depths and during different seasons. Our results revealed that the soil compaction effect on the abundance of both genes depended on season and soil depth, contrary to the effect on gene polymorphism. Bacterial pmoA abundance was significantly lower in the compacted soil than in the controls across all seasons, except in winter in the 0-10 cm depth interval and in summer in the 10-20 cm depth interval. In contrast, archaeal mcrA abundance was higher in compacted than control soil in winter and autumn in the two soil depths investigated. This study shows the usefulness of using pmoA and mcrA genes simultaneously in order to better understand the spatial and temporal variations of soil CH4 fluxes and the potential effect of physical disturbances.

Distinct Nitrification Rates and Nitrifiers in Needleleaf and Evergreen Broadleaf Forest Soils.

Research on niche specialization in the microbial communities of ammonia oxidizers is important for assessing the consequences of vegetation shift on nitrogen (N) cycling. In this study, soils were sampled from three tree stands (needleleaf, mixed, and evergreen broadleaf) from the Hannam experimental forest in South Korea in spring (May 2019), summer (August 2019), autumn (November 2019), and winter (January 2020). Quantitative polymerase chain reaction (qPCR) and high-throughput sequencing were used to measure the abundance and community structure of various nitrifiers: ammonia-oxidizing archaea and bacteria (AOA and AOB, respectively) as well as complete ammonia oxidizers (comammox). Nitrification rates and total ammonia oxidizer abundance were significantly higher in needleleaf forest soil than those in other forest stands, and they were lowest in evergreen broadleaf forest soil. Comammox clade B was most abundant in needleleaf and evergreen broadleaf forest soils, while AOA were significantly more abundant in mixed forest soil. The abundances of comammox clade B and AOA were negatively correlated with dissolved organic carbon. Phylogenetic analysis showed that NT-alpha and NS-gamma-2.3.2 were the most abundant AOA lineages in all the samples. The seasonal of AOA, AOB, and comammox varied with the sites, suggesting the need to examine the combinations of environmental factors when considering the effects of seasonal changes in the environment. Overall, the results suggest that potential vegetation shifts in forest ecosystems might affect nitrification activities by regulating the abundance and community structure of ammonia oxidizers.

Holm oak (Quercus ilex L.) cover: A key soil-forming force in controlling C and nutrient stocks in long-time coppice-managed forests.

In forest ecosystems, soil-plant interactions drive the physical, chemical, and biological soil properties and, through soil organic matter cycling, control the dynamics of nutrient cycles. Parent material also plays a fundamental role in determining soil's chemical properties and nutrient availability. In this study, eight long-time coppice-managed Holm oak forests under conversion to high forest, located under similar climatic conditions in Tuscany and Sardinia Regions (Italy), and grown on soils developed from three different lithologies (limestone, biotite granite, and granite with quartz veins) were evaluated. The research aimed to a) estimate the amount of C and nutrients (total N and potentially available P, Ca, Mg, and K) stored both in the organic, organo-mineral, and mineral horizons and at fixed depth intervals (0-0.3 and 0.3-0.5 m), and b) assess the dominant pedological variables driving elemental accumulation. The soils were described and sampled by genetic horizons and each sample was analyzed for its C and nutrient concentration in both the fine earth and skeleton fractions. Despite the different parent materials from which the soils had evolved, the physicochemical properties and the C and nutrient stocks for the 0-0.3 and 0.3-0.5 m layers did not show substantial differences among the eight soils. Conversely, some differences were observed in the stocks of potentially available P and Ca per 0.01 m of mineral horizons. The findings show that over time, plant-induced pedogenic processes (acidification, mineral weathering, organic matter addition, and nutrient cycling) almost obliterated the influence of parent materials on soil properties. This resulted in the upper soil horizons that showed similar characteristics, even though derived from different lithologies. However, among the study sites, some differences occurred due to lithology, as in the case of the soils derived from calcareous parent materials that had high concentrations of exchangeable Ca in the mineral horizons and, likely, to environmental variables (e.g., exposure), which possibly influenced litter degradation and the release of nutrients such as N and available P.

Labile carbon-induced soil organic matter turnover in a subtropical forest under different redox conditions.

Labile organic carbon (LOC) input strongly affects soil organic matter (SOM) dynamics, including gains and losses. However, it is unclear how redox fluctuations regulate these processes of SOM decomposition and formation induced by LOC input. The objective of this study was to explore the impacts of LOC input on SOM turnover under different redox conditions. Soil samples were collected in a subtropical forest. A single pulse of 13C-labeled glucose (i.e., LOC) was applied to the soil. Soil samples were incubated for 40 days under three redox treatments, including aerobic, anoxic, and 10-day aerobic followed by 10-day anoxic conditions. Results showed that LOC input affected soil priming and 13C-SOM accumulation differently under distinct redox conditions by altering the activities of various microorganisms. 13C-PLFAs (phospholipid fatty acids) were analyzed to determine the role of microbial groups in SOM turnover. Increased activities of fungi and gram-positive bacteria (i.e., the K-strategists) by LOC input could ingest metabolites or residues of the r-strategists (e.g., gram-negative bacteria) to result in positive priming. Fungi could use gram-negative bacteria to stimulate priming intensity via microbial turnover in aerobic conditions first. Reduced activities of K-strategists as a result of the aerobic to anoxic transition decreased priming intensity. The difference in LOC retention in SOM under different redox conditions was mainly attributable to 13C-particulate organic carbon (13C-POC) accumulation. Under aerobic conditions, fungi and gram-positive bacteria used derivatives from gram-negative bacteria to reduce newly formed POC. However, anoxic conditions were not conducive to the uptake of gram-negative bacteria by fungi and gram-positive bacteria, favoring SOM retention. This work indicated that redox-regulated microbial activities can control SOM decomposition and formation induced by LOC input. It is extremely valuable for understanding the contribution of soil affected by redox fluctuations to the carbon cycle.

Microvirga terricola sp. nov. and Microvirga solisilvae sp. nov, isolated from forest soil.

Two Gram-staining-negative, aerobic and rod-shaped strains, designated c23x22T and sex2T, were isolated from forest soil collected from Chebaling National Nature Reserve in Guangdong Province and Limu Mountain National Forest Park in Hainan Province, P. R. China, respectively. Phylogenetic analyses based on 16S rRNA gene sequences revealed that they belonged to the genus Microvirga, and strain c23 x22T was most closely related to 'Microvirga alba' KCTC 72385, while strain sex2T showed close relationship with Microvirga guangxiensis CGMCC 1.7666T. The average nucleotide identity and digital DNA-DNA hybridization values between strains c23 x22T and sex2T and their close relatives, 'M. alba' KCTC 72385 and M. guangxiensis CGMCC 1.7666T, were all below the threshold values for species delimitation. The predominant quinones of the two novel strains were ubiquinone 10, and the major fatty acids contained C19:0 cyclo ω8c and summed feature 8 (C18:1 ω7c and/or C18:1 ω6c). Their predominant polar lipids were phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylcholine. The phenotypic, genotypic and chemotaxonomic analyses clearly supported that strains c23 x 22T and sex2T represent two novel species of the genus Microvirga, for which the name Microvirga terricola sp. nov. (type strain c23 x 22T = GDMCC 1.1700T = KCTC 62432T) and Microvirga solisilvae sp. nov. (type strain sex2T = GDMCC 1.1651T = KACC 21311T) are proposed, respectively.

Corallococcus silvisoli sp. nov., a novel myxobacterium isolated from subtropical forest soil.

An orange-pigmented myxobacterium, designated strain c25j21T, was isolated from subtropical forest soil collected from the Chebaling National Nature Reserve in Guangdong Province, China. Phylogenetic analysis based on the 16S rRNA gene and core genes clearly showed that the novel strain was affiliated within the genus Corallococcus and most closely related to Corallococcus aberystwythensis DSM 108846T (99.3% 16S rRNA gene sequence similarity), while C. exercitus DSM 108849T (99.2%) and C. carmarthensis DSM 108842T (99.0%) were the next most closely related type strains. The draft genome sequence of strain c25j21T was 9.23 Mb in length with a G + C content of 70.7 mol%. The average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values between strain c25j21T and its closely related type strains were 88.1-89.1 and 34.1-36.3%, respectively. The major fatty acids contained iso-C15:0, iso-C17:0, iso-C17:1ω5c and iso-C17:0 2-OH. The predominant respiratory quinone was menaquinone 7. Based on phylogenetic, phenotypic and chemotaxonomic analysis, strain c25j21T represents a novel species of the genus Corallococcus, for which the name Corallococcus silvisoli sp. nov. is proposed. The type strain is c25j21T (= GDMCC 1.1387T = KCTC 62437T).

Effect of coppice conversion into high forest on soil organic C and nutrients stock in a Turkey oak (Quercus cerris L.) forest in Italy.

In forest ecosystems, a variety of abiotic and biotic soil forming factors drives soil organic matter (SOM) and nutrients cycling with a profitable outcome on climate change mitigation. As a consequence, type and intensity of forest management, through its impact on carbon (C) and nutrient soil stocks, can be considered as an additional soil forming force. In this study, we investigated the influence of the coppice conversion into high forest on pedogenesis and on soil C and nutrient (N, P, Ca, Mg, and K) stocks, fifty years later the beginning of the conversion-cycle. The trial was established in a Turkey oak forest historically managed under the coppice system in central Italy. Specifically, we considered tree population density (natural evolution - control, moderate thinning, heavy thinning) where soil samples were collected according to genetic horizon to estimate C, N, and P stocks both in the forest floor and at fixed depth intervals (0-30, 30-50 and 50-75 cm). Further, the stocks of exchangeable Ca, Mg, and K were also assessed for the mineral layers. The results showed that litter and the upper layer of mineral soil (0-30 cm) contained a similar quantity of C (about 74-83 Mg ha-1), independently of the trials and no differences were observed also in the whole soil stocks (about 192-213 Mg ha-1). The comparison of the mean stocks calculated per 1-cm of thickness of organic (O), organo-mineral (OM), and mineral (M) layers, although it did not display any difference among trials (excepted for P and Mg), showed a similar capability of the organo-mineral horizons to store C and nutrients compared with the organic ones (e.g., about 6-12 Mg ha-1, 0.3-0.5 Mg ha-1 and 0.5-1.5 kg ha-1 for C, N and P, respectively). Our findings showed that thinning operated on Turkey oak coppice did not affect soil capacity to store C and nutrients. These results suggested that the forest ecosystem itself is the main soil forming force and this is consistent with the target of adopting forest management able to control the global C cycle through the storage of SOM in the mineral soil rather than in forest floor, where SOM turnover is faster.

Biological nitrogen fixation in logging residue piles of different tree species after final felling.

Logging residues influence the nitrogen cycling processes that play a key role in risks for nitrogen losses from the ecosystem after the clear cut. Therefore, our aim was to identify the potential ability of logging residues to gain external nitrogen via biological nitrogen fixation. We measured biological nitrogen fixation as nitrogenase activity in logging residues of three different tree species (Norway spruce (Picea abies (L.) Karst.), Scots pine (Pinus sylvestris L.), and silver birch (Betula pendula Roth.). The study site was located in south-eastern Finland and was clear cut in 2014 and piles of logging residues were established. Sampling was performed in June 2016, September 2018, and August 2019 and nitrogenase activity in branches and needles or leaves was measured using the acetylene (C2H2) reduction assay. Nitrogenase activity (ethylene production) was shown in all residue types. Nitrogenase activity tended to be higher in branches than in needles or leaves and in coniferous residues than in birch. C-to-N ratios were higher in branches than in needles/leaves and in coniferous residues than in birch. Our results indicate that logging residues can acquire external nitrogen from the atmosphere via biological nitrogen fixation and can thus bring nitrogen to the forest ecosystem and substitute some part of the N losses occurring when residues are retained at the site after clear cutting.