Biosynthesis Genes Research Articles

SMPD4 mediated sphingolipid metabolism regulates brain and primary cilia development.

Genetic variants in multiple sphingolipid biosynthesis genes cause human brain disorders. A recent study collected patients from twelve unrelated families with variants in the gene SMPD4, a neutral sphingomyelinase which metabolizes sphingomyelin into ceramide at an early stage of the biosynthesis pathway. These patients have severe developmental brain malformations including microcephaly and cerebellar hypoplasia. The disease mechanism of SMPD4 was not known and we pursued a new mouse model. We hypothesized that the role of SMPD4 in producing ceramide is important for making primary cilia, a crucial organelle mediating cellular signaling. We found that the mouse model has cerebellar hypoplasia due to failure of Purkinje cell development. Human induced pluripotent stem cells exhibit neural progenitor cell death and have shortened primary cilia which is rescued by adding exogenous ceramide. SMPD4 production of ceramide is crucial for human brain development.

Whole-genome sequence of Bacillus subtilis TR6 strain isolated from tomato (Solanum lycopersicum L.) roots.

Here, we announce the sequence of endophyte Bacillus subtilis TR6 strain isolated from tomato roots growing in a greenhouse in St. Petersburg, Russia. With a length of 4,071,622 bp, this strain contained 4,073 potential coding sequences and 10 predicted secondary metabolite biosynthesis gene clusters.

Varying levels of natural light intensity affect the phyto-biochemical compounds, antioxidant indices and genes involved in the monoterpene biosynthetic pathway of Origanum majorana L.

BackgroundLight is a critical environmental factor in plants, encompassing two vital aspects: intensity and quality. To assess the influence of different light intensities on Origanum majorana L., pots containing the herb were subjected to four levels of light intensity: 20, 50, 70, and 100% natural light. After a 60-day treatment period, the plants were evaluated for metabolite production, including total sugar content, protein, dry weight, antioxidant indices, expression of monoterpenes biosynthesis genes, and essential oil compounds. The experimental design followed a randomized complete blocks format, and statistical analysis of variance was conducted.ResultsThe results indicated a correlation between increased light intensity and elevated total sugar and protein content, which contributed to improved plant dry weight. The highest levels of hydrogen peroxide and malondialdehyde (MDA) were observed under 100% light intensity. Catalase and superoxide dismutase enzymes exhibited increased activity, with a 4.23-fold and 2.14-fold increase, respectively, under full light. In contrast, peroxidase and polyphenol oxidase enzyme activities decreased by 3.29-fold and 3.24-fold, respectively. As light intensity increases, the expression level of the 1-deoxy-D-xylulose 5-phosphate reductoisomerase (DXR) gene increases. However, beyond a light intensity of 70%, the DXR gene expression level decreased. Furthermore, the expression levels of the cytochrome P450 genes CYP71D178 and CYP71D179 exhibited an increasing trend in response to elevated light intensity. Essential oil content increased from 0.02 to 0.5% until reaching 70% light intensity. However, with further increases in light intensity, the essential oil content decreased by 54 to 0.23%.ConclusionsThese findings emphasize the importance of balancing plant growth promotion and stress management under different light conditions. The research suggests that sweet marjoram plants thrive best in unshaded open spaces, resulting in maximum biomass. However, essential oil production decreases under the same conditions. For farmers in areas with an average light intensity of approximately 1700 µmol m−2s−1, it is recommended to cultivate sweet marjoram in shade-free fields to optimize biomass and essential oil production. Towards the end of the growth cycle, it is advisable to use shades that allow 70% of light to pass through. The specific duration of shade implementation can be further explored in future research.

The Characterization of R2R3-MYB Genes in Water Lily Nymphaea colorata Reveals the Involvement of NcMYB25 in Regulating Anthocyanin Synthesis

Nymphaea colorata, valued for its diverse flower colors and attractive shapes, is a popular ornamental aquatic plant. Anthocyanins provide color to flowers, and their biosynthesis is regulated by the R2R3-MYB transcription factor. In this study, we identified and analyzed the R2R3-MYB genes in N. colorata, focusing on their structure, evolution, expression patterns, regulatory mechanisms, and biological functions. We also investigated the role of the NcMYB25 gene in anthocyanin biosynthesis. There were 59 R2R3-MYB genes in N. colorata, distributed across 14 chromosomes. Among these, 14 genes were involved in segmental duplications and 6 in tandem duplications. Multiple R2R3-MYB transcription factors appeared to play a role in biological processes in N. colorata, including NcMYB48 in flavonoid synthesis, NcMYB33 in lignin synthesis, NcMYB23 in cold stress response, and NcMYB54 in osmotic stress response. Additionally, we identified 92 miRNAs in N. colorata, with 43 interacting with 35 R2R3-MYB genes. The NcMYB25 protein is localized in the nucleus and possesses transcriptional activation activity. Overexpression of the NcMYB25 gene in an apple pericarp resulted in anthocyanin accumulation. These findings provide insight into the evolutionary trajectory of the R2R3-MYB genes in N. colorata and highlight the regulatory function of the NcMYB25 gene in anthocyanin biosynthesis.

Functions of exogenous strigolactone application and strigolactone biosynthesis genes GhMAX3/GhMAX4b in response to drought tolerance in cotton (Gossypium hirsutum L.)

BackgroundDrought stress markedly constrains plant growth and diminishes crop productivity. Strigolactones (SLs) exert a beneficial influence on plant resilience to drought conditions. Nevertheless, the specific function of SLs in modulating cotton’s response to drought stress remains to be elucidated.ResultsIn this study, we assess the impact of exogenous SL (rac-GR24) administration at various concentrations (0, 1, 5, 10, 20 µM) on cotton growth during drought stress. The findings reveal that cotton seedlings treated with 5 µM exogenous SL exhibit optimal mitigation of growth suppression induced by drought stress. Treatment with 5 µM exogenous SL under drought stress conditions enhances drought tolerance in cotton seedlings by augmenting photosynthetic efficiency, facilitating stomatal closure, diminishing reactive oxygen species (ROS) generation, alleviating membrane lipid peroxidation, enhancing the activity of antioxidant enzymes, elevating the levels of osmoregulatory compounds, and upregulating the expression of drought-responsive genes. The suppression of cotton SL biosynthesis genes, MORE AXILLARY GROWTH 3 (GhMAX3) and GhMAX4b, impairs the drought tolerance of cotton. Conversely, overexpression of GhMAX3 and GhMAX4b in respective Arabidopsis mutants ameliorates the drought-sensitive phenotype in these mutants.ConclusionThese observations underscore that SLs significantly bolster cotton’s resistance to drought stress.

Integrated Analysis of Metabolome and Transcriptome Reveals the Effect of Burdock Fructooligosaccharide on the Quality of Chinese Cabbage (Brassica rapa L. ssp. Pekinensis)

Burdock fructooligosaccharide (BFO) is fructose with a low polymerization degree, which could improve the immunity to pathogens, quality, and stress resistance of vegetables. Still, there are no studies on applying BFO in Chinese cabbage. In this study, the effects of exogenous BFO sprayed with different concentrations (0, 5, 10, 20, 30 g·L−1) on the growth and soluble sugar content of Chinese cabbage seedlings were determined. The result showed that 10 g·L−1 was the appropriate spraying concentration. Based on metabolome analysis, a total of 220 differentially accumulated metabolites (DAMs) were found, among which flavonoid metabolites, glucosinolate metabolites, and soluble sugar-related metabolites were the key metabolites involved in improving the quality of Chinese cabbage caused by BFO. Further combination analysis with transcriptome, trans-cinnamate 4-monooxygenase (CYP73A5), and chalcone synthase 1 (CHS1) were more closely associated with the DAMs of flavonoid biosynthesis. Sulfotransferases 18 (SOT18), Branched-chain amino acid amino transferases 6 (BCAT6), and cytochrome P450 monooxygenase (CYP83A1) were the key genes in glucosinolate biosynthesis. Hexokinase (HxK1), beta-glucosidase 8 (BGL08), invertase 3 (INV3), beta-glucosidase 3B (BGL3B), and sucrose phosphate synthase 1 (SPS1) were significantly upregulated, potentially playing crucial roles in the soluble sugar metabolism. In conclusion, these results provided an understanding of the effects of BFO on the expression of genes and the accumulation of metabolites related to quality formation in Chinese cabbage.

Comparative transcriptome analysis of cucumber fruit tissues reveals novel regulatory genes in ascorbic acid biosynthesis.

Ascorbic acid (AsA) is one of the most abundant natural antioxidants, and it is an important indicator of the nutritional value of cucumber fruit. The aim of this study was to elucidate the regulatory mechanism affecting AsA metabolism in cucumber fruit. In this study, the AsA content in the fruit of two cucumber cultivars (H28 and H105) was significantly higher in the exocarp and endocarp than in the mesocarp. To clarify the regulation of AsA in cucumber fruit, the transcriptomes of three fruit tissues (i.e., the exocarp, mesocarp, and endocarp) of two cucumber cultivars (H28 and H105) were sequenced. Transcriptomic profiling combined with transcription factors (TFs) and correlation analysis were performed to reveal that three genes, including CsaV3_5G014110 (phosphomannomutase, PMM), CsaV3_2G004170 (GDP-mannose-3', 5'-epimerase, GME) and CsaV3_5G006680 (dehydroascorbate reductase, DHAR), were expressed at higher level in the exocarp and endocarp than in the mesocarp. In both two cultivars, CsaV3_4G028360 (ethylene-responsive transcription factor, ERF) was negatively correlated with PMM and GME, and positively correlated with DHAR. CsaV3_6G042110 (ethylene-responsive transcription factor, ERF) was positively correlated with PMM and GME, and negatively correlated with DHAR. CsaV3_6G032360 (mitogen-activated protein kinase, MAPK) as positively correlated with PMM, GME and DHAR. These six genes are considered the key candidate genes for further research. This study provides insight for further study on the regulation of AsA biosynthesis in cucumber fruit and provide potential candidate genes for future genetic improvement of cucumber germplasm with enhanced AsA accumulation.

Growth of soil ammonia-oxidizing archaea on air-exposed solid surface

Abstract Soil microorganisms often thrive as microcolonies or biofilms within pores of soil aggregates exposed to the soil atmosphere. However, previous studies on the physiology of soil ammonia-oxidizing microorganisms (AOM), which play a critical role in the nitrogen cycle, were primarily conducted using freely suspended AOM cells (planktonic cells) in liquid media. In this study, we examined the growth of two representative soil ammonia-oxidizing archaea (AOA), Nitrososphaera viennensis EN76 and “Nitrosotenuis chungbukensis” MY2, and a soil ammonia-oxidizing bacterium, Nitrosomonas europaea ATCC 19718 on polycarbonate membrane filters floated on liquid media to observe their adaptation to air-exposed solid surfaces. Interestingly, ammonia oxidation activities of N. viennensis EN76 and “N. chungbukensis” MY2 were significantly repressed on floating filters compared to the freely suspended cells in liquid media. Conversely, the ammonia oxidation activity of N. europaea ATCC 19718 was comparable on floating filters and liquid media. N. viennensis EN76 and N. europaea ATCC 19718 developed microcolonies on floating filters. Transcriptome analysis of N. viennensis EN76 floating filter-grown cells revealed upregulation of unique sets of genes for cell wall and extracellular polymeric substance biosynthesis, ammonia oxidation (including ammonia monooxygenase subunit C (amoC3) and multicopper oxidases), and defense against H2O2-induced oxidative stress. These genes may play a pivotal role in adapting AOA to air-exposed solid surfaces. Furthermore, the floating filter technique resulted in the enrichment of distinct soil AOA communities dominated by the “Ca. Nitrosocosmicus” clade. Overall, this study sheds light on distinct adaptive mechanisms governing AOA growth on air-exposed solid surfaces.

Histological, Transcriptomic, and Functional Analyses Reveal the Role of Gibberellin in Bulbil Development in Lilium lancifolium

Lily bulbils, advantageous axillary organs used for asexual reproduction, have an underexplored developmental mechanism. Gibberellins are known to participate in bulbil development, but the regulatory mechanisms remain unclear. In this study, exogenous gibberellin (GA3) significantly increased the bulbil length, width, and weight by raising the endogenous gibberellin levels and elongating the scale cells. Transcriptomic analysis identified LlGA20ox2, a key gibberellin biosynthesis gene, which was upregulated during bulbil development and significantly responsive to GA3 treatment. Given the similarities in bulbil and bulblet development, we determined the roles of LlGA20ox2 using a bulblet system. Silencing LlGA20ox2 in bulblets inhibited development by reducing the cell length, while overexpression increased the bulblet length and width. In the gibberellin signaling pathway, we identified two key genes, LlGID1C and LlCIGR2. Silencing these genes resulted in phenotypes similar to LlGA20ox2, inhibiting bulblet development. Further transcriptomic analysis revealed that gibberellin-responsive genes were enriched in the glucuronate pathway, pentose phosphate pathway and galactose metabolism pathways. Most of these differentially expressed genes responded to gibberellin and were highly expressed in later stages of bulbil development, suggesting their involvement in gibberellin-regulated bulbil growth. In conclusion, we preliminarily explored the mechanisms of gibberellin regulation in bulbil development, offering significant commercial potential for new lily reproductive organs.

Acquisition and evolution of the neurotoxin domoic acid biosynthesis gene cluster in Pseudo-nitzschia species

Of the hitherto over 60 taxonomically identified species in the genus of Pseudo-nitzschia, 26 have been confirmed to be toxigenic. Nevertheless, the acquisition and evolution of the toxin biosynthesis (dab) genes by this extensive group of Pseudo-nitzschia species remains unclear. Through constructing chromosome-level genomes of three Pseudo-nitzschia species and draft genomes of ten additional Pseudo-nitzschia species, putative genomic integration sites for the dab genes in Pseudo-nitzschia species were explored. A putative breakpoint was observed in syntenic regions in the dab gene cluster-lacking Pseudo-nitzschia species, suggesting potential independent losses of dab genes. The breakpoints between this pair of conserved genes were also identified in some dab genes-possessing Pseudo-nitzschia species, suggesting that the dab gene clusters transposed to other loci after the initial integration. A “single acquisition, multiple independent losses (SAMIL)” model is proposed to explain the acquisition and evolution of the dab gene cluster in Pseudo-nitzschia species.

Genome-Wide Identification of COMT Gene Family in Maize and its Function in Response to Light.

Maize is a major crop, feed, and industrial material. Caffeic acid-O-methyltransferase (COMT) is a methylase closely associated with lignin biosynthesis and plant growth and resistance. In this study, we identified the COMT gene (ZmCOMT) family in maize and further analyzed its phylogenetic evolution, subcellular localization, and its function in response to light. Thirty-one ZmCOMT genes were identified in the maize genome, which were distributed across eight chromosomes and mainly clustered on chromosome 4. Most ZmCOMT proteins were predicted to localize in the cytoplasm. Ten different conserved motifs were present in most ZmCOMT proteins, and motif1, motif6, and motif7 were highly conserved and present in all ZmCOMT proteins. The photoresponsivity elements were conserved among all members, and ZmCOMT22 and ZmCOMT10 genes responsive to light. This result suggests a potential function for these two genes in lignin biosynthesis which a previous study had linked to light regulation. Jasmonic acid responsive and abscisic acid cis-acting elements were present in the promoter regions of family members, thus the family may be regulated by hormone signaling pathways of maize. In summary, ZmCOMT genes are ancient, and the highly conserved motifs may be significant in survival and evolution of maize. Furthermore, light may influence lignin biosynthesis and photosynthesis through ZmCOMT genes. This research provided theoretical basis for lignin biosynthesis of maize and the potential value of ZmCOMT22 and ZmCOMT10 genes to enhance plant photosynthesis for facing global warming.

Tomato SlARF5 participate in the flower organ initiation process and control plant height

Plant height is a critical agronomic trait closely linked to yield, primarily regulated by Gibberellins (GA) and auxins, which interact in complex ways. However, the mechanism underlying their interactions remain incompletely understood. In this study, we identified a tomato mutant exhibiting significantly reduced plant height. Through gene cloning and bulked segregant analysis (BSA) sequencing, we found that the mutant gene corresponds to the tomato auxin response factor gene SlARF5/MP. Here, we show that overexpression of SlARF5/MP significantly enhances plant height. Additionally, treatment with GA3 restored the plant height of the mutant to wild-type (WT) levels, indicating that GA content is a key factor influencing plant height. We also observed significant upregulation of GA-biosynthesis genes, including GA2-oxidases GA20ox3 and GA20ox4, as well as the GA3 biosynthesis gene GA3ox1, in SlARF5-overexpressing plants. Furthermore, we demonstrated that SlARF5 directly binds to SlGA2ox3, which mediates the conversion of GA3 to inactive GA, therebyregulating its expression. Our findings suggest that SlARF5 modulates GA3 metabolism by regulating GA synthesis genes, ultimately leading to alterations in plant height.

Maize DLR1/NHX7 Is Required for Root Development Under Potassium Deficiency.

Root System Architecture (RSA) is a crucial plant trait that governs a plant's ability to absorb water and nutrients. In this study, we describe a mutant with nutrient-dependent defects in root development, affecting both the primary root and lateral roots (LRs). This mutant, identified through a screen for defects in LR development, has been designated dlr1-1. The dlr1-1 mutant exhibits impaired LR emergence rather than defects in the LR primordium (LRP) formation, particularly under potassium (K+)-deprivation conditions. This impairment likely stems from inhibited cell proliferation caused by the dlr1-1 mutation. K+ deprivation specifically leads to the accumulation of salicylic acid (SA) in the dlr1-1 mutant, consistent with the upregulation of SA biosynthesis genes. Moreover, exogenous application of SA to wild-type plants (B73) mimics the dlr1-1 phenotype. Conversely, treatment of the dlr1-1 mutant with 2-aminoindane-2-phosphonic acid, an SA biosynthesis inhibitor, partially restores LR emergence, indicating that elevated SA levels may be responsible for the mutant's developmental defects. MutMap analysis and allelism tests confirmed that the phenotypes of the dlr1-1 mutant results from the loss of the Na+/H+ antiporter, ZmNHX7. Additionally, the application of NaCl exacerbates the dlr1-1 mutant phenotype, suggesting that the root defects in dlr1-1 mutant depend on ion homoeostasis. In conclusion, our findings demonstrate that maize DLR1/NHX7 is essential for root development under potassium deprivation.

De Novo Synthesis of Anticholinergic Hyoscyamine and Scopolamine in Nicotiana benthamiana Based on Elucidating Tropane Alkaloid Biosynthetic Pathway of Anisodus luridus

Anisodus luridus, a perennial herb belonging to the genus Anisodus of the Solanaceae family, is an important Tibetan medicinal plant that produces pharmaceutical tropane alkaloids (TAs) including hyoscyamine and scopolamine. Its high yield of hyoscyamine makes A. luridus a valuable plant source for commercially producing TAs. In this study, we conduct homologous gene research across transcriptome data of different tissues together with functionally tested sequences in Atropa belladonna as a reference and identify 13 candidate genes for TAs biosynthesis in A. luridus. The results show that these 13 TAs biosynthesis genes identified in A. luridus were highly conserved in terms of sequence similarity and gene expression patterns compared to A. belladonna, suggesting that the two species may share the same biosynthetic pathway for TAs biosynthesis. Furthermore, scopolamine was detected in Nicotiana benthamiana leaves when these 13 enzymes were co-expressed in N. benthamiana, which confirmed that these 13 TAs biosynthesis genes are involved in the biosynthesis of TAs. The results of our study not only systematically elucidate the tropane alkaloid biosynthetic pathway of A. luridus, but also realize the de novo synthesis of TAs in N. benthamiana for the first time. It is now possible to make N. benthamiana a potential source for TAs production through synthetic biology techniques.

The effect of spermidine supplementation on polyamine metabolism and Fusarium infection in Linum usitatissimum

Flax (Linum usitatissimum) is a valuable industrial crop in temperate climate zones. However, the prevalence of fungal diseases, particularly those caused by Fusarium oxysporum sp. linii, poses a substantial threat, leading to significant crop losses and diminished interest in flax cultivation. In this study, flax plants were treated with spermidine and subsequently infected with Fusarium oxysporum to investigate multiple aspects: (1) the uptake of exogenous spermidine by the plants, (2) the impact of this uptake on the levels of other polyamines and the expression of polyamine biosynthesis genes, and (3) the effects of fungal infection on these parameters. The results demonstrated that flax plants effectively absorb spermidine, leading to substantial changes in the levels of polyamines and the expression of related genes in both roots and shoots. Notably, spermidine treatment resulted in significant alterations in the levels of putrescine and spermine, as well as in the transcript levels of key genes involved in polyamine biosynthesis. Moreover, Fusarium oxysporum infection itself induced changes in polyamine content and gene expression, which varied between root and shoot tissues. When spermidine-treated plants were infected with Fusarium oxysporum, a more complex response was observed, characterized by modifications in polyamine levels and gene expression that suggest an enhanced defense mechanism against the pathogen. These findings indicate that polyamine treatment not only affects the infection process but also modulates the plant's internal polyamine metabolism and gene expression, contributing to an improved resistance to fungal attack. This study highlights the potential of spermidine as a protective agent in flax and provides a foundation for further exploration of polyamine-related defense mechanisms.

De novo transcriptome analysis and gene expression profiling of Conticribra weissflogii during low salinity reveals regulation of fatty acid biosynthesis genes

De novo transcriptome analysis and gene expression profiling of Conticribra weissflogii during low salinity reveals regulation of fatty acid biosynthesis genes

Chemical and Transcriptomic Analyses Provide New Insights into Key Genes for Ginsenoside Biosynthesis in the Rhizome of Panax japonicus C. A. Meyer.

Panax japonicus C. A. Meyer is renowned for its significant therapeutic effects and is commonly used worldwide. Its active ingredients, triterpenoid saponins, show variation in content among different tissues. The tissue-specific distribution of saponins is potentially related to the expression of vital genes in the biosynthesis pathway. In this study, the contents of five saponins (ginsenoside Ro, chikusetsusaponin IV, chikusetsusaponin IVa, ginsenoside Rg1, and ginsenoside Rb1) in three different tissues were determined by HPLC. Transcriptome sequencing analysis identified differentially expressed genes (DEGs) involved in triterpenoid saponin biosynthesis, highlighting significant correlations between saponin contents and the expression levels of 10 cytochrome p450 monooxygenase (CYP) and 3 UDP-glycosyltransferase (UGT) genes. Cloning, sequencing, and prokaryotic expression of UGT genes confirmed the molecular weights of UGT proteins. Gene sequence alignment and phylogenetic analysis provided preliminary insights into UGT gene functions. Meanwhile, the function of one UGT gene was characterized in the yeast. These findings advance our understanding of the triterpenoid saponin biosynthesis in P. japonicus and support future research in traditional Chinese medicine (TCM) and synthetic biology.

A transcription factor, PbWRKY24, contributes to russet skin formation in pear fruits by modulating lignin accumulation

Abstract Skin color is one of the major traits of fruit appearance quality in pear (Pyrus), which affects the fruit commodity value. Russet skin protects pear fruits from environmental stresses and its formation process is closely linked to lignin accumulation. However, the molecular regulatory networks underlying russet skin formation in pear fruits involve complex secondary metabolic pathways and remain elusive. Here, we explored the regulatory mechanisms underlying lignin accumulation in pear skin based on transcriptome sequencing, co-expression network analysis, and gene expression profiling. We identified a WRKY transcription factor gene, PbWRKY24, that regulates russet skin formation in pear fruits. The relative expression of PbWRKY24 in russet pear skin was significantly correlated with lignin content. We then verified the function of PbWRKY24 in lignin accumulation via genetic transformation. DNA affinity purification sequencing revealed that PbWRKY24 directly binds to the promoter of a lignin biosynthesis gene, PbPRX4. This binding was confirmed by yeast one-hybrid, dual-luciferase, and electrophoretic mobility shift assays. Overexpression of PbPRX4 in pear skin stimulated lignin accumulation and consequently promoted russet skin formation. This study provides a glimpse into the intricate lignin biosynthesis mechanisms during russet skin formation in pear fruits, which is of practical significance to pear breeding for fruit quality.

Diverse and specialized metabolic capabilities of microbes in oligotrophic built environments

BackgroundBuilt environments (BEs) are typically considered to be oligotrophic and harsh environments for microbial communities under normal, non-damp conditions. However, the metabolic functions of microbial inhabitants in BEs remain poorly understood. This study aimed to shed light on the functional capabilities of microbes in BEs by analyzing 860 representative metagenome-assembled genomes (rMAGs) reconstructed from 738 samples collected from BEs across the city of Hong Kong and from the skin surfaces of human occupants. The study specifically focused on the metabolic functions of rMAGs that are either phylogenetically novel or prevalent in BEs.ResultsThe diversity and composition of BE microbiomes were primarily shaped by the sample type, with Micrococcus luteus and Cutibacterium acnes being prevalent. The metabolic functions of rMAGs varied significantly based on taxonomy, even at the strain level. A novel strain affiliated with the Candidatus class Xenobia in the Candidatus phylum Eremiobacterota and two novel strains affiliated with the superphylum Patescibacteria exhibited unique functions compared with their close relatives, potentially aiding their survival in BEs and on human skins. The novel strains in the class Xenobia possessed genes for transporting nitrate and nitrite as nitrogen sources and nitrosative stress mitigation induced by nitric oxide during denitrification. The two novel Patescibacteria strains both possessed a broad array of genes for amino acid and trace element transport, while one of them carried genes for carotenoid and ubiquinone biosynthesis. The globally prevalent M. luteus in BEs displayed a large and open pangenome, with high infraspecific genomic diversity contributed by 11 conspecific strains recovered from BEs in a single geographic region. The versatile metabolic functions encoded in the large accessory genomes of M. luteus may contribute to its global ubiquity and specialization in BEs.ConclusionsThis study illustrates that the microbial inhabitants of BEs possess metabolic potentials that enable them to tolerate and counter different biotic and abiotic conditions. Additionally, these microbes can efficiently utilize various limited residual resources from occupant activities, potentially enhancing their survival and persistence within BEs. A better understanding of the metabolic functions of BE microbes will ultimately facilitate the development of strategies to create a healthy indoor microbiome.Bh9ATT9ssZEtNaJHv1wg7GVideo

Reconfiguring the Escherichia coli Electron Transport Chain to Enhance trans-2-Decenoic Acid Production.

trans-2-Decenoic acid is a pivotal α,β-medium-chain unsaturated fatty acid that serves as an essential intermediary in the synthesis of 10-hydroxy-2-decenoic acid and various pharmaceutical compounds. Biosynthesis yield of trans-2-decenoic acid by decanoic acid has significantly improved in recent years; however, the oxidative stress of Escherichia coli at high fatty acid concentrations restricts the conversion rate. Here, we introduced a combination of rational design and metabolic rewiring of the E. coli electron transport chain (ETC) to improve trans-2-decenoic acid production. Overexpressing ubiquinone (UbQ) biosynthesis genes enhanced the expression of ETC complex III: UbQ to reduce reactive oxygen species (ROS) accumulation. Furthermore, applying rotenone to inhibit ETC complex I improved the electron transfer efficiency of complex II. The integration of Vitamin B5 and B2 into the fermentation process increased the activities of fatty acyl-CoA synthetase (MaMACS) and fatty acyl-CoA dehydrogenase (PpfadE). Finally, the constructed E. coli BL21(DE3)(ΔfadBJR/pCDFDuet-1-PpfadE-MaMACS/pRSFDuet-1-sumo-CtydiI-ubiI) strain exhibited a 51.50% decrease in ROS and a 93.33% enhancement in trans-2-decenoic acid yield, reaching 1.45 g/L after 66 h, which is the highest yield reported for flask fermentation. This study reports the feasibility of rewiring the ETC regulation and energy metabolism to improve α,β-UCA biosynthesis efficiency.