Impacts of phosphoenolpyruvate carboxylase gene silencing on photosynthetic efficiency and carbon fixation in Skeletonema costatum.

Diatoms play a crucial role in marine primary productivity through carbon fixation, which is essential for understanding the operation of marine biological pumps and carbon sinks. This study focuses on the phosphoenolpyruvate carboxylase (PEPC) gene, a key enzyme in the carbon assimilation pathway of diatoms, by investigating the consequences of its silencing in Skeletonema costatum. Through this approach, we aimed to clarify the distinct contributions of PEPC to the overall carbon fixation process. The mutant strains of S. costatum were subjected to thorough analysis to identify any shifts in physiological behavior, alterations in the gene expression of key carbon-fixing enzymes, and changes in the associated enzyme activities. Notably, the inhibition of the PEPC gene did not significantly affect the growth rate of S. costatum; however, it did have a notable impact on the photosynthetic apparatus, as evidenced by a reduction in the maximal electron transport rate and a decline in light utilization efficiency. A significant decrease was observed in both the enzymatic activity and gene expression of PEPCase. This down-regulation also affected other enzymes integral to the carbon fixation pathway, such as phosphoenolpyruvate carboxykinase and pyruvate-phosphate dikinase, indicating a wider metabolic perturbation. In contrast, the expression and activity of the Rubisco enzyme suggested that some facets of carbon fixation remained resilient. Furthermore, the substantial upregulation of carbonic anhydrase expression and activity probably represented an adaptive mechanism to sustain the inorganic carbon supply necessary for the carboxylation process of Rubisco. This research not only underscores the pivotal role of the PEPC gene in the carbon fixation of S. costatum but also expands our comprehension of carbon fixation mechanisms in diatoms.

Reduced primary productivity and notable resilience of phytoplankton community in the coastal water of southern China under a marine heatwave.

Increasing frequency, intensity and duration of marine heatwaves (MHWs) are supposed to affect coastal biological production in different regions to different extents. To understand how MHWs impact coastal primary productivity and community succession of phytoplankton and assess the changes in resilience of phytoplankton communities, we conducted a mesoscale enclosure experiment simulating a marine MHW in the coastal water of southern China. After 8 days of the MHW (+3 oC) treatment, community biomass was significantly lower than the control's, and primary productivity per volume of water was reduced by about 56%. Nevertheless, the phytoplankton community retrieved its biomass and primary productivity after the temperature was subsequently reset to that of the control. Although the MHW treatment decreased the abundance of diatom and increased the percentages of Synechococcus and Prasinophytes, the main phytoplankton functional types showed positive resilience that allowed the recovery of the phytoplankton community after the MHW. Our results indicate that key phytoplankton functional types in the southern coastal waters of China exhibited significant resilience, recovery, and temporal stability under the influence of the marine MHW by 3 oC rise. However, reduced primary productivity during the MHW period, along with decreased biomass density, might significantly influence secondary producers. In addition, the altered phytoplankton community structure may affect coastal food web processes at least during the MHW period.

Atmospheric Trace Gas Oxidizers Contribute to Soil Carbon Fixation Driven by Key Soil Conditions in Terrestrial Ecosystems.

Microbial oxidizers of trace gases such as hydrogen (H2) and carbon monoxide (CO) are widely distributed in soil microbial communities and play a vital role in modulating biogeochemical cycles. However, the contribution of trace gas oxidizers to soil carbon fixation and the driving environmental factors remain unclear, especially on large scales. Here, we utilized biogeochemical and genome-resolved metagenomic profiling, assisted by machine learning analysis, to estimate the contributions of trace gas oxidizers to soil carbon fixation and to predict the key environmental factors driving this process in soils from five distinct ecosystems. The results showed that phylogenetically and physiologically diverse H2 and CO oxidizers and chemosynthetic carbon-fixing microbes are present in the soil in different terrestrial ecosystems. The large-scale variations in soil carbon fixation were highly positively correlated with both the abundance and the activity of H2 and CO oxidizers (p < 0.05-0.001). Furthermore, soil pH and moisture-induced shifts in the abundance of H2 and CO oxidizers partially explained the variation in soil carbon fixation (55%). The contributions of trace gas oxidizers to soil carbon fixation in the different terrestrial ecosystems were estimated to range from 1.1% to 35.0%. The estimated rate of trace gas carbon fixation varied from 0.04 to 1.56 mg kg-1 d-1. These findings reveal that atmospheric trace gas oxidizers may contribute to soil carbon fixation driven by key soil environmental factors, highlighting the non-negligible contribution of these microbes to terrestrial carbon cycling.

Diatom pyrenoids are encased in a protein shell that enables efficient CO2 fixation.

Pyrenoids are subcompartments of algal chloroplasts that increase the efficiency of Rubisco-driven CO2 fixation. Diatoms fix up to 20% of global CO2, but their pyrenoids remain poorly characterized. Here, we used in vivo photo-crosslinking to identify pyrenoid shell (PyShell) proteins, which we localized to the pyrenoid periphery of model pennate and centric diatoms, Phaeodactylum tricornutum and Thalassiosira pseudonana. In situ cryo-electron tomography revealed that pyrenoids of both diatom species are encased in a lattice-like protein sheath. Single-particle cryo-EM yielded a 2.4-Å-resolution structure of an in vitro TpPyShell1 lattice, which showed how protein subunits interlock. T. pseudonana TpPyShell1/2 knockout mutants had no PyShell sheath, altered pyrenoid morphology, and a high-CO2 requiring phenotype, with reduced photosynthetic efficiency and impaired growth under standard atmospheric conditions. The structure and function of the diatom PyShell provide a molecular view of how CO2 is assimilated in the ocean, a critical ecosystem undergoing rapid change.

Effects of manure and nitrogen fertilization on soil microbial carbon fixation genes and associated communities in the Loess Plateau of China.

The effects of long-term fertilization on soil carbon (C) cycling have been a key focus of agricultural sustainable development research. However, the influences of different fertilization treatments on soil microbial C fixation profiles are still unclear. Metagenomics technology and multivariate analysis were employed to inquire changes in soil properties, soil microbial C fixation genes and associated bacterial communities, and the influence of dominant soil properties on C fixation genes. The contents of soil C and nitrogen fractions were signicficantly higher in manure or combined with nitrogen fertilization (NM) than other treatments. The composition of soil microbial C fixation genes and associated bacterial communities varied among different fertilization treatments. Compared with other treatments, the total abundance of microbial C fixation genes and the abundance of Proteobacteria were significantly higher in NM than in other treatments, as well as the abundances of C fixation genes involved in dicarboxylate/4-hydroxybutyrate cycle and reductive citrate cycle. Key functional genes and main bacterial communities presented in the middle of the co-occurrence network. Soil organic carbon, total nitrogen, and microbial biomass nitrogen were the dominant soil properties influencing microbial C fixation genes and associated bacterial communitis. Fertilization increased the abundance of C fixation genes by affecting the changes in bacterial communities abundance mediated by soil properties. Overall, elucidating the responses of soil microbial C fixation genes and associated communities to different fertilization will enhance our understanding of the processes of soil C fixation in farmland.

Exploring the leaf regeneration cycles response of Zostera japonica to ocean acidification.

Ocean acidification is one of the major global environmental problems facing humankind today, and it has far-reaching impacts on marine organisms and the entire marine ecosystem. Zostera japonica, an important supporting species of intertidal seagrass beds, exhibits high photosynthetic productivity and plays an important role in the carbon cycle of nearshore waters. However, little is known about the characteristics, processes, and mechanisms of its response to ocean acidification. In this study, we conducted a 120-day acidification experiment in Z. japonica; here, plants underwent four leaf regeneration cycles to reveal the response mechanism of Z. japonica to ocean acidification (OA). We found that acidification significantly affected the seedling stage of Z. japonica, impacting leaf regeneration cycles by altering physiological and molecular responses. In one leaf regeneration cycle, the short-term exposure to CO2 affected the seagrass parameters, such as the regulation of inorganic carbon uptake modes and the regulation of photosynthesis between the dark and light reactions, with the potential to affect the carbon sinks of the marine organisms. The long-term effects on the regulation of antioxidant enzymes and antioxidant metabolites, caused an improvement in the marine life adaptation to OA. In a comparison of the different leaf regeneration cycles, the response pattern of Z. japonica showed an offset of the acidification during the short cycles and an adaption to the acidification during the long cycles. This study revealed the response mechanism of Z. japonica to OA at different time scales and could provide a theoretical basis for accurately assessing the impact of OA on seagrass and the entire seagrass bed ecosystem.

A robust synthetic biology toolkit to advance carboxysome study and redesign.

Carboxysomes are polyhedral protein organelles that microorganisms use to facilitate carbon dioxide assimilation. They are composed of a modular protein shell which envelops an enzymatic core mainly comprised of physically coupled Rubisco and carbonic anhydrase. While the modular construction principles of carboxysomes make them attractive targets as customizable metabolic platforms, their size and complexity can be a hinderance. In this work, we design and validate a plasmid set - the pXpressome toolkit - in which α-carboxysomes are robustly expressed and remain intact and functional after purification. We tested this toolkit by introducing mutations which influence carboxysome structure and performance. We find that deletion of vertex-capping genes results in formation of larger carboxysomes while deletion of facet forming genes produces smaller particles, suggesting that adjusting the ratio of these proteins can rationally affect morphology. Through a series of fluorescently labeled constructs, we observe this toolkit leads to more uniform expression and better cell health than previously published carboxysome expression systems. Overall, the pXpressome toolkit facilitates the study and redesign of carboxysomes with robust performance and improved phenotype uniformity. The pXpressome toolkit will support efforts to remodel carboxysomes for enhanced carbon fixation or serve as a platform for other nanoencapsulation goals.

Crystal Structure and Molecular Mechanism of Isocitrate Lyase from Chloroflexus aurantiacus.

Chloroflexus aurantiacus is a green, nonsulfur bacterium that employs the 3-hydroxypropionate cycle to grow, using carbon dioxide/bicarbonate as its primary carbon source. Like most bacteria, it possesses the glyoxylate cycle, facilitated by malate synthase and isocitrate lyase (ICL), allowing a "tricarboxylic acid cycle" bypass. C. aurantiacus also harbors ICL, an enzyme that catalyzes reversible isocitrate cleavage into glyoxylate and succinate. This study presents the crystal structures of C. aurantiacus-derived ICL (CaICL), in its Mg2+-bound and Mn2+ and isocitrate-bound forms, elucidating its substrate-binding mechanism and catalytic loop dynamics. CaICL forms a homotetramer and interacts with isocitrate via critical active-site residues, revealing its catalytic mechanism. The stabilization of the catalytic loop and adjacent terminal regions upon isocitrate binding underscores its functional significance. These findings advance our understanding regarding ICL enzymes, offering a basis for future investigations into their biological roles and potential applications.

Rubisco packaging and stoichiometric composition of a native ß-carboxysome.

Carboxysomes are anabolic bacterial microcompartments that play an essential role in carbon fixation in cyanobacteria. This self-assembling proteinaceous organelle encapsulates the key CO2-fixing enzymes, Rubisco and carbonic anhydrase, using a polyhedral shell constructed by hundreds of shell protein paralogs. Deciphering the precise arrangement and structural organization of Rubisco enzymes within carboxysomes is crucial for understanding the formation process and overall functionality of carboxysomes. Here, we employed cryo-electron tomography and subtomogram averaging to delineate the three-dimensional packaging of Rubiscos within ß-carboxysomes in the freshwater cyanobacterium Synechococcus elongatus PCC7942 that were grown under low light. Our results revealed that Rubiscos are arranged in multiple concentric layers parallel to the shell within the ß-carboxysome lumen. We also identified the binding of Rubisco with the scaffolding protein CcmM in ß-carboxysomes, which is instrumental for Rubisco encapsulation and ß-carboxysome assembly. Using QconCAT-based quantitative mass spectrometry, we further determined the absolute stoichiometric composition of the entire ß-carboxysome. This study and recent findings on the ß-carboxysome structure provide insights into the assembly principles and structural variation of ß-carboxysomes, which will aid in the rational design and repurposing of carboxysome nanostructures for diverse bioengineering applications.

Zinc oxide nanoparticles reduce cadmium accumulation in hydroponic lettuce (Lactuca sativa L.) by increasing photosynthetic capacity and regulating phenylpropane metabolism.

Due to the continuous production of industrial wastes and the excessive use of chemical fertilizers and pesticides, severe cadmium (Cd) pollution in soil has occurred globally. This study investigated the impacts of incorporating zinc oxide nanoparticles (ZnONPs) into hydroponically grown lettuce (Lactuca sativa) under cadmium stress conditions, to seek effective methods to minimize Cd buildup in green leafy vegetables. The results showed that 1 mg/L of Cd significantly inhibited lettuce growth, decreasing in leaves (29 %) and roots (33 %) biomass. However, when lettuce was exposed to 2.5 mg/L ZnONPs under cadmium stress, the growth, chlorophyll content, net photosynthetic rate (Pn), stomatal conductance (Gs), actual photochemical efficiency of PSII (φPSII), and activity of key enzymes in photosynthesis were all significantly enhanced. Furthermore, ZnONPs significantly decreased the accumulation of Cd in lettuce leaves (36 %) and roots (13 %). They altered the subcellular distribution and chemical morphology of Cd in lettuce by modifying the composition of cell walls (such as pectin content) and the levels of phenolic compounds, resulting in a reduction of 27 % in Cd translocation from roots to leaves. RNA sequencing yielded 45.9 × 107 and 53.4 × 107 clean reads from plant leaves and roots in control (T0), Cd (T1), Cd+ZnONPs (T2), and ZnONPs (T3) treatment groups respectively, and 3614 and 1873 differentially expressed genes (DEGs) were identified. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis identified photosynthesis, carbon fixation, and phenylpropanoid metabolism as the main causes of ZnONPs-mediated alleviation of Cd stress in lettuce. Specifically, the DEGs identified included 12 associated with photosystem I, 13 with photosystem II and 23 DEGs with the carbon fixation pathway of photosynthesis. Additionally, DEGs related to phenylalanine ammonia-lyase, caffeoyl CoA 3-O-methyltransferase, peroxidase, 4-coumarate-CoA ligase, hydroxycinnamoyl transferase, and cytochrome P450 proteins were also identified. Therefore, further research is recommended to elucidate the molecular mechanisms by which ZnONPs reduce Cd absorption in lettuce through phenolic acid components in the phenylpropanoid metabolism pathway. Overall, treatments with ZnONPs are recommended to effectively reduce Cd accumulation in the edible portion of lettuce.

Electrogenic performance and carbon sequestration potential of biophotovoltaics.

Biophotovoltaics (BPV) is a clean and sustainable solar energy generation technology that operates by utilizing photosynthetic autotrophic microorganisms to capture light energy and generate electricity. However, a major challenge faced by BPV systems is the relatively low electron transfer efficiency from the photosystem to the extracellular electrode, which limits its electrical output. Additionally, the transfer mechanisms of photosynthetic microorganism metabolites in the entire system are still not fully clear. In response to this, this article briefly introduces the basic BPV principles, reviews its development history, and summarizes measures to optimize its electrogenic efficiency. Furthermore, recent studies have found that constructing photosynthetic-electrogenic microbial consortia can achieve high power density and stability in BPV systems. Therefore, the article discusses the potential application of constructing photosynthetic-electrogenic microbial aggregates in BPV systems. Since photosynthetic-electrogenic microbial communities can also exist in natural ecosystems, their potential contribution to the carbon cycle is worth further attention.

Genomic synergistic efficient carbon fixation and nitrogen removal induced by excessive inorganic carbon in the anammox-centered coupling system.

Given the significance of HCO3- for autotrophic anammox bacteria (AnAOB), excessive HCO3- was always provided in anammox-related systems and engineering applications. However, its impact mechanism on anammox process at genome-level remains unknown. This study firstly established an anammox-centered coupling system that entails heterotrophic partial denitrification (PD) and hydrolytic acidification (A-PDHA) fed mainly with inorganic carbon (high HCO3- concentration and low C/N ratio). Metagenomic binning and metatranscriptomics analyses indicated that high HCO3- concentration enhanced expression of natural most efficient phosphoenolpyruvate (PEP) carboxylase within AnAOB, by up to 30.59 folds. This further induced AnAOB to achieve high-speed carbon-fixing reaction through cross-feeding of phosphate and PEP precursors with heterotrophs. Additionally, the enhanced activity of transporters and catalytic enzymes (up to 4949-fold) induced by low C/N ratio enabled heterotrophs to eliminate extracellular accumulated energy precursors mainly derived from carbon fixation products of AnAOB. This maintained high-speed carbon-fixing reaction within AnAOB and supplemented heterotrophs with organics. Moreover, assimilated energy precursors stimulated nitrogen metabolism enzymes, especially NO2- reductase (968.14 times), in heterotrophs. This established an energy-saving PD-A process mediated by interspecies NO shuttle. These variation resulted in efficient nitrogen removal (>95 %) and reduced external organic carbon demand (67 %) in A-PDHA system. This study unveils the great potential of an anammox-centered autotrophic-heterotrophic coupling system for achieving cost-effective nitrogen removal and enhancing carbon fixation under excessive HCO3- doses.

Root-based inorganic carbon uptake increases the growth of Arabidopsis thaliana and changes transporter expression and nitrogen and sulfur metabolism.

Root-based uptake of inorganic carbon has been suggested as an additional carbon source. Our study aimed to characterize and understand the root-based uptake and fixation mechanisms and their impact on plant growth. 13C-labeled bicarbonate fed to Arabidopsis roots was assimilated into aspartic acid but mainly into sucrose, indicating that the added inorganic carbon was transported to the leaves. A hydroponic treatment was also established for A. thaliana using 2 mM NaHCO3 at pH 5.6, which enhanced the photosynthetic and growth parameters. According to transcriptome sequencing data, the observed enhancement in growth may be orchestrated by trehalose-6-phosphate signaling and supported by augmented nitrogen and sulfur assimilation. The analysis also revealed regulatory and transporter activities, including several nitrate (NRT2.1), and sulfate transporter (SULTR1;1 and SULTR1;2) candidates that could participate in bicarbonate uptake. Different transporters and carbon fixation mutants were assessed. Arabidopsis homologs of SLOW-TYPE ANION CHANNEL 1 (slah3) CARBONIC ANHYDRASE (ßca4), and SULFATE TRANSPORTER (sultr1;2) mutants were shown to be inferior to the bicarbonate-treated wild types in several growth and root ultrastructural parameters. Besides, aquaporin genes PIP1;3 and PIP2;6 could play a negative role in the carbon uptake by venting carbon dioxide out of the plant. The findings support the hypothesis that the inorganic carbon is taken up by the root anion channels, mostly transported up to the shoots by the xylem, and fixed there by RuBisCo after the conversion to CO2 by carbonic anhydrases. The process boosts photosynthesis and growth by providing an extra carbon supply.

Machine learning reveals the transcriptional regulatory network and circadian dynamics of Synechococcus elongatus PCC 7942.

Synechococcus elongatus is an important cyanobacterium that serves as a versatile and robust model for studying circadian biology and photosynthetic metabolism. Its transcriptional regulatory network (TRN) is of fundamental interest, as it orchestrates the cell's adaptation to the environment, including its response to sunlight. Despite the previous characterization of constituent parts of the S. elongatus TRN, a comprehensive layout of its topology remains to be established. Here, we decomposed a compendium of 300 high-quality RNA sequencing datasets of the model strain PCC 7942 using independent component analysis. We obtained 57 independently modulated gene sets, or iModulons, that explain 67% of the variance in the transcriptional response and 1) accurately reflect the activity of known transcriptional regulations, 2) capture functional components of photosynthesis, 3) provide hypotheses for regulon structures and functional annotations of poorly characterized genes, and 4) describe the transcriptional shifts under dynamic light conditions. This transcriptome-wide analysis of S. elongatus provides a quantitative reconstruction of the TRN and presents a knowledge base that can guide future investigations. Our systems-level analysis also provides a global TRN structure for S. elongatus PCC 7942.

Synthetic biology of metabolic cycles for Enhanced CO2 capture and Sequestration.

In most organisms, the tri-carboxylic acid cycle (TCA cycle) is an essential metabolic system that is involved in both energy generation and carbon metabolism. Its uni-directionality, however, restricts its use in synthetic biology and carbon fixation. Here, it is describing the use of the modified TCA cycle, called the Tri-carboxylic acid Hooked to Ethylene by Enzyme Reactions and Amino acid Synthesis, the reductive tricarboxylic acid branch/4-hydroxybutyryl-CoA/ethylmalonyl-CoA/acetyl-CoA (THETA) cycle, in Escherichia coli for the purposes of carbon fixation and amino acid synthesis. Three modules make up the THETA cycle: (1) pyruvate to succinate transformation, (2) succinate to crotonyl-CoA change, and (3) crotonyl-CoA to acetyl-CoA and pyruvate change. It is presenting each module's viability in vivo and showing how it integrates into the E. coli metabolic network to support growth on minimal medium without the need for outside supplementation. Enzyme optimization, route redesign, and heterologous expression were used to get over metabolic roadblocks and produce functional modules. Furthermore, the THETA cycle may be improved by including components of the Carbon-Efficient Tri-Carboxylic Acid Cycle (CETCH cycle) to improve carbon fixation. THETA cycle's promise as a platform for applications in synthetic biology and carbon fixation.

Bifunctional Phenylalanine/Tyrosine Ammonia-Lyase (PTAL) Enhances Lignin Biosynthesis: Implications in Carbon Fixation in Plants by Genetic Engineering.

Lignin is a key metabolite for terrestrial plants. Two types of aromatic amino acids, phenylalanine (Phe) and tyrosine (Tyr), serve as the precursors for lignin biosynthesis. In most plant species, Phe is deaminated by Phe ammonia-lyase (PAL) to initiate lignin biosynthesis, but in grass species, Phe and Tyr are deaminated by Phe/Tyr ammonia-lyase (PTAL). To understand the efficiency of PAL and PTAL, we used transgenic and non-transgenic Arabidopsis with PAL and crop-weedy rice hybrids (CWRH) with PTAL to analyze lignin-biosynthesis-associated metabolites. The transgenic plants overexpressed the exogenous 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene, whereas the non-transgenic plants normally expressed the endogenous EPSPS gene. Our results show significantly increased Phe/Tyr contents in transgenic Arabidopsis and CWRH plants, leading to substantially increased lignin and biomass. In addition, the PTAL pathway promotes a much greater proportion of increased lignin and biomass in transgenic CWRH than in transgenic Arabidopsis lineages. Evidently, more efficient lignin biosynthesis characterized the grass species possessing the PTAL pathway. These findings are important for a better understanding of the PAL and PTAL's functions in the phenylpropanoid metabolic pathways in the evolution of plant species. These findings also have great value for implications such as effective carbon fixation by enhancing lignin biosynthesis through genetic engineering of their key genes in appropriately selected plant species.

Identification of carbon fixation microorganisms and pathways in an aquifer contaminated with long-chain petroleum hydrocarbons.

Petroleum hydrocarbons (PHCs) can be biodegraded into CO2, and PHC-contaminated aquifers are always deemed as carbon sources. Fortunately, some carbon fixation microorganisms have been found in PHC-contaminated sites. However, most of the studies are related to volatile short-chain PHC, and few studies focus on long-chain PHC-contaminated sites. To reveal the carbon fixation microorganisms in these sites, in the study, a long-chain PHC polluted site in North China was selected. Through hydrochemical and metagenomics analysis, the structure and capacity of carbon fixing microorganisms in the site were revealed. Results showed that there were many kinds of carbon fixed microorganisms that were identified such as Flavobacterium, Pseudomonas. HP/4HB, rTCA, and DC/4HB cycles were dominated carbon fixation pathways. The long-chain PHC were weakly correlated with carbon fixation microorganisms, but it may stimulate the growth of some carbon fixation microorganisms, such as microorganisms involved in rTCA cycle. PRACTITIONER POINTS: The microorganisms with carbon fixation gene exist in the aquifer contaminated by long-chain petroleum hydrocarbon. Microorganisms that have the ability to degrade petroleum also have the ability to carbon fixation. Long-chain petroleum hydrocarbon may promote the growth of carbon fixation microorganisms.

Metabolic adaptations underpin high productivity rates in relict subsurface water.

Groundwater aquifers are ecological hotspots with diverse microbes essential for biogeochemical cycles. Their ecophysiology has seldom been studied on a basin scale. In particular, our knowledge of chemosynthesis in the deep aquifers where temperatures reach 60 °C, is limited. Here, we investigated the diversity, activity, and metabolic potential of microbial communities from nine wells reaching ancient groundwater beneath Israel's Negev Desert, spanning two significant, deep (up to 1.5 km) aquifers, the Judea Group carbonate and Kurnub Group Nubian sandstone that contain fresh to brackish, hypoxic to anoxic water. We estimated chemosynthetic productivity rates ranging from 0.55 ± 0.06 to 0.82 ± 0.07 µg C L-1 d-1 (mean ± SD), suggesting that aquifer productivity may be underestimated. We showed that 60% of MAGs harbored genes for autotrophic pathways, mainly the Calvin-Benson-Bassham cycle and the Wood-Ljungdahl pathway, indicating a substantial chemosynthetic capacity within these microbial communities. We emphasize the potential metabolic versatility in the deep subsurface, enabling efficient carbon and energy use. This study set a precedent for global aquifer exploration, like the Nubian Sandstone Aquifer System in the Arabian and Western Deserts, and reconsiders their role as carbon sinks.

Aquibium pacificus sp. nov., a Novel Mixotrophic Bacterium from Bathypelagic Seawater in the Western Pacific Ocean.

A novel Gram-stain-negative, facultatively anaerobic, and mixotrophic bacterium, designated as strain LZ166T, was isolated from the bathypelagic seawater in the western Pacific Ocean. The cells were short rod-shaped, oxidase- and catalase-positive, and motile by means of lateral flagella. The growth of strain LZ166T was observed at 10-45 °C (optimum 34-37 °C), at pH 5-10 (optimum 6-8), and in the presence of 0-5% NaCl (optimum 1-3%). A phylogenetic analysis based on the 16S rRNA gene showed that strain LZ166T shared the highest similarity (98.58%) with Aquibium oceanicum B7T and formed a distinct branch within the Aquibium genus. The genomic characterization, including average nucleotide identity (ANI, 90.73-76.79%), average amino identity (AAI, 88.50-79.03%), and digital DNA-DNA hybridization (dDDH, 36.1-22.2%) values between LZ166T and other species within the Aquibium genus, further substantiated its novelty. The genome of strain LZ166T was 6,119,659 bp in size with a 64.7 mol% DNA G+C content. The predominant fatty acid was summed feature 8 (C18:1ω7c and/or C18:1ω6c). The major polar lipids identified were diphosphatidylglycerol (DPG), phosphatidylethanolamine (PE), glycolipid (GL), and phosphatidylglycerol (PG), with ubiquinone-10 (Q-10) as the predominant respiratory quinone. The genomic annotation indicated the presence of genes for a diverse metabolic profile, including pathways for carbon fixation via the Calvin-Benson-Bassham cycle and inorganic sulfur oxidation. Based on the polyphasic taxonomic results, strain LZ166T represented a novel species of the genus Aquibium, for which the name Aquibium pacificus sp. nov. is proposed, with the type strain LZ166T (=MCCC M28807T = KACC 23148T = KCTC 82889T).

Stratified Effects of Tillage and Crop Rotations on Soil Microbes in Carbon and Nitrogen Cycles at Different Soil Depths in Long-Term Corn, Soybean, and Wheat Cultivation.

Understanding the soil bacterial communities involved in carbon (C) and nitrogen (N) cycling can inform beneficial tillage and crop rotation practices for sustainability and crop production. This study evaluated soil bacterial diversity, compositional structure, and functions associated with C-N cycling at two soil depths (0-15 cm and 15-30 cm) under long-term tillage (conventional tillage [CT] and no-till [NT]) and crop rotation (monocultures of corn, soybean, and wheat and corn-soybean-wheat rotation) systems. The soil microbial communities were characterized by metabarcoding the 16S rRNA gene V4-V5 regions using Illumina MiSeq. The results showed that long-term NT reduced the soil bacterial diversity at 15-30 cm compared to CT, while no significant differences were found at 0-15 cm. The bacterial communities differed significantly at the two soil depths under NT but not under CT. Notably, over 70% of the tillage-responding KEGG orthologs (KOs) associated with C fixation (primarily in the reductive citric acid cycle) were more abundant under NT than under CT at both depths. The tillage practices significantly affected bacteria involved in biological nitrogen (N2) fixation at the 0-15 cm soil depth, as well as bacteria involved in denitrification at both soil depths. The crop type and rotation regimes had limited effects on bacterial diversity and structure but significantly affected specific C-N-cycling genes. For instance, three KOs associated with the Calvin-Benson cycle for C fixation and four KOs related to various N-cycling processes were more abundant in the soil of wheat than in that of corn or soybean. These findings indicate that the long-term tillage practices had a greater influence than crop rotation on the soil bacterial communities, particularly in the C- and N-cycling processes. Integrated management practices that consider the combined effects of tillage, crop rotation, and crop types on soil bacterial functional groups are essential for sustainable agriculture.