E3 ligase DECREASED GRAIN SIZE 1 promotes degradation of a G-protein subunit and positively regulates grain size in rice.

Grain size is one of the most important traits determining crop yield. However, the mechanism controlling grain size remains unclear. Here, we confirmed the E3 ligase activity of DECREASED GRAIN SIZE 1 (DGS1) in positive regulation of grain size in rice (Oryza sativa) suggested in a previous study. Rice G-protein subunit gamma 2 (RGG2), which negatively regulates grain size, was identified as an interacting protein of DGS1. Biochemical analysis suggested that DGS1 specifically interacts with canonical Gγ subunits (rice G-protein subunit gamma 1 [RGG1] and rice G-protein subunit gamma 2 [RGG2]) rather than non-canonical Gγ subunits (DENSE AND ERECT PANICLE 1 [DEP1], rice G-protein gamma subunit type C 2 [GCC2], GRAIN SIZE 3 [GS3]). We also identified the necessary domains for interaction between DGS1 and RGG2. As an E3 ligase, DGS1 ubiquitinated and degraded RGG2 via a proteasome pathway in several experiments. DGS1 also ubiquitinated RGG2 by its K140, K145 and S147 residues. Thus, this work identified a substrate of the E3 ligase DGS1 and elucidated the post transcriptional regulatory mechanism of the G-protein signalling pathway in the control of grain size.

Key roles of autophagosome/endosome maturation mediated by Syntaxin17 in methamphetamine-induced neuronal damage in mice.

BACKGROUND: Autophagic defects are involved in Methamphetamine (Meth)-induced neurotoxicity. Syntaxin 17 (Stx17), a member of the SNARE protein family, participating in several stages of autophagy, including autophagosome-late endosome/lysosome fusion. However, the role of Stx17 and potential mechanisms in autophagic defects induced by Meth remain poorly understood. METHODS: To address the mechanism of Meth-induced cognitive impairment, the adenovirus (AV) and adeno-associated virus (AAV) were injected into the hippocampus for stereotaxis to overexpress Stx17 in vivo to examine the cognitive ability via morris water maze and novel object recognition. In molecular level, the synaptic injury and autophagic defects were evaluated. To address the Meth induced neuronal damage, the epidermal growth factor receptor (EGFR) degradation assay was performed to evaluate the degradability of the "cargos" mediated by Meth, and mechanistically, the maturation of the vesicles, including autophagosomes and endosomes, were validated by the Co-IP and the GTP-agarose affinity isolation assays. RESULTS: Overexpression of Stx17 in the hippocampus markedly rescued the Meth-induced cognitive impairment and synaptic loss. For endosomes, Meth exposure upregulated Rab5 expression and its guanine-nucleotide exchange factor (GEF) (immature endosome), with a commensurate decreased active form of Rab7 (Rab7-GTP) and impeded the binding of Rab7 to CCZ1 (mature endosome); for autophagosomes, Meth treatment elicited a dramatic reduction in the overlap between Stx17 and autophagosomes but increased the colocalization of ATG5 and autophagosomes (immature autophagosomes). After Stx17 overexpression, the Rab7-GTP levels in purified late endosomes were substantially increased in parallel with the elevated mature autophagosomes, facilitating cargo (Aß42, p-tau, and EGFR) degradation in the vesicles, which finally ameliorated Meth-induced synaptic loss and memory deficits in mice. CONCLUSION: Stx17 decrease mediated by Meth contributes to vesicle fusion defects which may ascribe to the immature autophagosomes and endosomes, leading to autophagic dysfunction and finalizes neuronal damage and cognitive impairments. Therefore, targeting Stx17 may be a novel therapeutic strategy for Meth-induced neuronal injury.

Biocompatible Silica-Coated Europium-Doped CsPbBr3 Nanoparticles with Luminescence in Water for Zebrafish Bioimaging.

Cesium lead halide (CsPbX3, X = Br, Cl, and I) nanocrystals (NCs) are widely concerned and applied in many fields due to the excellent photoelectric performance. However, the toxicity of Pb and the loss of luminescence in water limit its application in vivo. A stable perovskite nanomaterial with good bioimaging properties is developed by incorporating europium (Eu) in CsPbX3 NCs followed with the surface coating of silica (SiO2) shell (CsPbX3:Eu@SiO2). Through the surface coating of SiO2, the luminescence stability of CsPbBr3 in water is improved and the leakage of Pb2+ is significantly reduced. In particular, Eu doping inhibits the photoluminescence quantum yield reduction of CsPbBr3 caused by SiO2 coating, and further reduces the release of Pb2+. CsPbBr3:Eu@SiO2 nanoparticles (NPs) show efficient luminescence in water and good biocompatibility to achieve cell imaging. More importantly, CsPb(ClBr)3:Eu@SiO2 NPs are obtained by adjusting the halogen components, and green light and blue light are realized in zebrafish imaging, showing good imaging effect and biosafety. The work provides a strategy for advanced perovskite nanomaterials toward biological practical application.

The Protective Role of Transcript-Induced in Spermiogenesis 40 in Cerebral Ischemia-Reperfusion Injury.

Prompt reperfusion after cerebral ischemia is important to maintain neuronal survival and reduce permanent disability and death. However, the resupply of blood can induce oxidative stress, inflammatory response and apoptosis, further leading to tissue damage. Here, we report the versatile biological roles of transcript-induced in spermiogenesis 40 (Tisp40) in ischemic stroke. We found that the expression of Tisp40 was upregulated in ischemia/reperfusion-induced brain tissues and oxygen glucose deprivation/returned -stimulated neurons. Tisp40 deficiency increased the infarct size and neurological deficit score, and promoted inflammation and apoptosis. Tisp40 overexpression played the opposite role. In vitro, the oxygen glucose deprivation/returned model was established in Tisp40 knockdown and overexpression primary cultured cortical neurons. Tisp40 knockdown can aggravate the process of inflammation and apoptosis, and Tisp40 overexpression ameliorated the aforementioned processes. Mechanistically, Tisp40 protected against ischemic stroke via activating the AKT signaling pathway. Tisp40 may be a new therapeutic target in brain ischemia/reperfusion injury.

Genome-wide identification and expression analysis of C-type lectins in discus fish (Symphysodon aequifasciatus) during parental care.

Discus fish (Symphysodon aequifasciatus) exhibit a unique parental care behavior: adult discus produces secretion through their skin, on which the larvae live after birth. The immune components in the skin mucus of parental discus would change during different parental care. C-type lectins (CTLs) could identify and eliminate pathogenic microorganisms and play important roles in innate immunity. Studies on CTLs of discus fish especially during parental care, however, are scarce. Here, we identified 186 CTL genes that distributed in 27 linkage groups based on discus genome. Phylogenetic analysis showed that S. aequifasciatus CTL (SaCTL) members were grouped into 14 subfamilies. A total of 80 gene replication events occurred, of which 15 pairs were subjected to segmental duplication and 65 pairs underwent tandem duplication. Ka/Ks ranged from 0.11 (SaCTL25/SaCTL158) to 0.68 (SaCTL36/SaCTL69), all undergoing purifying selection. RNA-seq analysis revealed that SaCTL members, including duplicated genes, in the skin of parental discus show distinct expression patterns in different care stages and between male and female parents. The SaCTL11 was differentially expressed in most care stages and reached the maximum after eggs spawned, but the expression of its paired SaCTL14 was low in each stage. The SaCTL39 increased first and then decreased, reaching a peak in eggs spawned, while paired SaCTL48 first decreased and then increased, reaching a peak in hatched eggs. The SaCTL50 was differentially expressed only in female fish during care, but not in male fish. These results provide new insights into the evolution and potential functional differentiation of CTLs in discus fish during parental care.

TOPK promotes the development of psoriasis and worenine alleviates psoriasiform dermatitis by inhibiting TOPK activity.

BACKGROUND: Psoriasis is an inflammatory skin disease. The pathogenesis of psoriasis has not been fully elucidated. T-lymphokine-activated killer cell-originated protein kinase (TOPK) activity increases in a proinflammatory environment, and inhibiting TOPK blocks inflammation. However, whether TOPK is involved in the pathogenesis of psoriasis remains to be identified. OBJECTIVES: We aimed to study the role of TOPK in psoriasis and attempted to find a drug targeting TOPK for the prevention and treatment of psoriasis. METHOD: Firstly, the expressions of TOPK in psoriatic patients, psoriatic cell and animal model were analysed by Gene Expression Omnibus database, immunohistochemistry (IHC) staining and western blot (WB). After inhibiting TOPK by chemical or gene knockout, the effect of TOPK on the development of psoriasis was verified in cell and animal model by WB, qRT-PCR, ELISA, haematoxylin-eosin (H&E) and IHC staining. Moreover, phosphoproteomic analysis was performed to explore the signalling pathways regulated by TOPK in the occurrence and development of psoriasis. Then, an in vitro kinase assay was performed to prove TOPK kinase activity was inhibited by worenine. Ultimately, WB, qRT-PCR, ELISA, H&E and IHC staining were used to verify the anti-psoriasis effect of worenine by inhibiting TOPK was in cell and animal model. RESULTS: In this study, we found that TOPK was highly expressed in psoriasis patients, psoriatic cell and animal model, which suggests that TOPK might be associated with psoriasis pathogenesis. Interestingly, chemical or genetic inhibition of TOPK alleviated M5- and imiquimod (IMQ)-induced psoriasis-like dermatitis, which further confirmed the role of TOPK in promoting the development of psoriasis. Moreover, we determined that worenine inhibited TOPK kinase activity. In addition, worenine relieved M5- and IMQ-induced psoriasiform dermatitis by inhibiting TOPK activity. CONCLUSIONS: T-lymphokine-activated killer cell-originated protein kinase promotes the development of psoriasis. Therefore, TOPK might be a promising drug target for the prevention and treatment of psoriasis. Worenine alleviates psoriasiform dermatitis by inhibiting TOPK activity, providing new strategies for clinical intervention.

Removal of proteins and lipids affects structure, in vitro digestion and physicochemical properties of rice flour modified by heat-moisture treatment.

BACKGROUND: The objective of this experiment was to investigate the role of endogenous proteins and lipids in the structural and physicochemical properties of starch in heat-moisture treatment (HMT) rice flour and to reveal their effect on starch digestibility under heat. RESULTS: The findings indicate that, in the absence of endogenous proteins and lipids acting as a physical barrier, especially proteins, the interaction between rice flour and endogenous proteins and lipids diminished. This reduction led to fewer starch-protein inclusion complexes and starch-lipid complexes, altering the granule aggregation structure of rice flour. It resulted in a decrease in particle size, an increase in agglomeration between starch granules, and more surface cracking on rice granules. Under HMT conditions with a moisture content of 30%, slight gelatinization of the starch granules occurred, contributing to an increased starch hydrolysis rate. In addition, the elevated thermal energy effect of HMT enhanced interactions between starch molecular chains. These resulted in a decrease in crystallinity, short-range ordering, and the content of double-helix structure within starch granules. These structural transformations led to higher pasting temperatures, improved hot and cold paste stability, and a decrease in peak viscosity, breakdown, setback, and enthalpy of pasting of the starch granules. CONCLUSION: The combined analysis of microstructure, physicochemical properties, and in vitro digestion characteristics has enabled us to further enhance our understanding of the interaction mechanisms between endogenous proteins, lipids, and starches during HMT. © 2024 Society of Chemical Industry.

The effect of heavy precipitation on the leaching of heavy metals from tropical coastal legacy tailings.

The continued growth in demand for mineral resources has led to a large amount of mining wastes, which is a major challenge in the context of carbon neutrality and climate change. In this study, runoff migration, batch leaching, and column experiments were used to investigate the short-, medium-, and long-term leaching of heavy metals from legacy tailings, respectively; the cumulative metal release kinetic equations were established, and the long-term effects of tailings leaching were verified by HYDRUS-1D. In runoff migration experiments, surface dissolution of tailings and the co-migration of adsorbed soil particles by erosion were the main carriers in the early stages of leachate formation (Mn âˆ¼ 65 mg/L and SO42- up to 2697.2 mg/L). Batch leaching tests showed that the concentration of heavy metals in soil leached by acid rain were 0.1 âˆ¼ 22.0 µg/L for Cr, 0.7 âˆ¼ 26.0 µg/L for Cu, 4.8 âˆ¼ 5646.0 µg/L for Mn, 0.3 âˆ¼ 232.4 µg/L for Ni, and 1.3 âˆ¼ 448.0 µg/L for Zn. The results of column experiments indicated that some soluble components and metals with high mobility showed a significant decreasing trend at cumulative L/S ≤ 2. Additionally, the metals have higher leaching rates under TCLP conditions, as shown by Mn > Co > Zn > Cd > Ni > Cu > Pb > Cr. The fitting results of Langmuir equation were closer to the cumulative release of metals in the real case, and the release amounts of Mn, Zn, Co, and Ni were higher with 55, 5.84, 2.66, and 2.51 mg/kg, respectively. The water flow within tailings affects the spatial distribution of metals, which mainly exist in relatively stable chemical fractions (F3 + F4 + F5 > 90 %) after leaching. Numerical simulation verified that Mn in leachate has reached 8 mg/L at a scale of up to 100 years. The research results are expected to provide technical basis for realizing the resource utilization of tailings in the future.

TO-UGDA: target-oriented unsupervised graph domain adaptation.

Graph domain adaptation (GDA) aims to address the challenge of limited label data in the target graph domain. Existing methods such as UDAGCN, GRADE, DEAL, and COCO for different-level (node-level, graph-level) adaptation tasks exhibit variations in domain feature extraction, and most of them solely rely on representation alignment to transfer label information from a labeled source domain to an unlabeled target domain. However, this approach can be influenced by irrelevant information and usually ignores the conditional shift of the downstream predictor. To effectively address this issue, we introduce a target-oriented unsupervised graph domain adaptive framework for graph adaptation called TO-UGDA. Particularly, domain-invariant feature representations are extracted using graph information bottleneck. The discrepancy between two domains is minimized using an adversarial alignment strategy to obtain a unified feature distribution. Additionally, the meta pseudo-label is introduced to enhance downstream adaptation and improve the model's generalizability. Through extensive experimentation on real-world graph datasets, it is proved that the proposed framework achieves excellent performance across various node-level and graph-level adaptation tasks.

4-Hydroxyphenylpyruvate Dioxygenase Inhibitors: From Molecular Design to Synthesis.

Weed resistance is a critical issue in crop production. Among the known herbicides, 4-hydroxyphenylpyruvate dioxygenase (HPPD) inhibitors are crucial for addressing weed resistance. HPPD inhibitors constitute a pivotal aspect of contemporary crop protection strategies. The advantages of these herbicides are their broad weed spectrum, flexible application, and excellent compatibility with other herbicides. They also exhibit satisfactory crop selectivity and low toxicity and are environmentally friendly. An increasing number of new HPPD inhibitors have been designed by combining computer-aided drug design with conventional design approaches. Herein, the molecular design and structural features of innovative HPPD inhibitors are reviewed to guide the development of new HPPD inhibitors possessing an enhanced biological efficacy.

In situ electrochemical Mn vacancies in CoMnHCF for a high level of zinc storage.

CoMnHCF is utilized in aqueous sodium/zinc mixed ion batteries and exhibits a high reversible capacity with good rate and cycle performances. At 0.05 A g-1 current density, the CoMnHCF can deliver a specific capacity for 180.4 mA h g-1, and have 99.3% capacity retention after 300 cycles at 0.3 A g-1. Such high reversible capacity profits from Mn vacancies that generate in situ during the first cycle, which provides more active sites for Zn storage. The de-intercalation of Na+ further elevates this good electrochemical performance. Co atoms in the framework are not only involved in the redox reactions, but help to support the structure, thus achieving better cycle stabilities.

Evaluation of Wetting Behaviors of Liquid Sodium on Transition Metals: An Experimental and Molecular Dynamics Simulation Study.

In sodium-cooled fast reactors, the wettability of sodium with materials is closely related to sodium-related operations and the detection accuracy of instruments and meters, so how to achieve the selection of materials with different wettability requirements is a key problem in engineering design. To meet these requirements, the wetting behaviors of liquid sodium with nine transition metals were investigated using scanning electron microscopy (SEM), X-ray diffraction (XRD), and molecular dynamics (MD) simulations. The results show that metals such as zinc and gold, which react with sodium to form intermetallic compounds at the interface, exhibit superior wettability. Followed by the metals that have strong interatomic interactions even though they do not react with sodium or dissolve each other, such as cobalt, nickel and copper, while the wettability of these systems tends to be poor at low temperatures. Systems that do not react with each other or have strong interatomic affinities proved to be the most difficult to wet. Notably, metals with the closest-packed crystal structures of fcc and hcp generally have better wettability than those with a bcc structure. They can be a valuable guide for experimental research and technical control.

Enhancing Clinical Predictive Modeling through Model Complexity-Driven Class Proportion Tuning for Class Imbalanced Data: An Empirical Study on Opioid Overdose Prediction.

Class imbalance issues are prevalent in the medical field and significantly impact the performance of clinical predictive models. Traditional techniques to address this challenge aim to rebalance class proportions. They generally assume that the rebalanced proportions are derived from the original data, without considering the intricacies of the model utilized. This study challenges the prevailing assumption and introduces a new method that ties the optimal class proportions to model complexity. This approach allows for individualized tuning of class proportions for each model. Our experiments, centered on the opioid overdose prediction problem, highlight the performance gains achieved by this approach. Furthermore, rigorous regression analysis affirms the merits of the proposed theoretical framework, demonstrating a statistically significant correlation between hyperparameters controlling model complexity and the optimal class proportions.

Enhanced performance and mechanism of adsorption pretreatment for alleviating membrane fouling in AGMBR: Impact of structural variations in carbon adsorbents.

The structural variances of adsorbents play a crucial role in determining the number of effective adsorption sites and pretreatment performance. However, there is still a gap in comprehending the impact of different carbon structural adsorbents on membrane fouling. Therefore, this study aimed to compare the efficacy of granular activated carbon (GAC), powdered activated carbon (PAC), and activated carbon fiber (ACF) in mitigating membrane fouling during municipal sewage reclamation using an aerobic granular sludge membrane bioreactor (AGMBR). The results demonstrated that the utilization of PAC significantly enhanced the normalized flux and reduced fouling resistance in comparison to GAC and ACF systems. PAC effectively adsorbed low and medium-molecular-weight pollutants present in raw sewage, resulting in an increase in average particle size and a decrease in foulant content on the membrane surface. The Hermia model indicated that adsorption pretreatment minimized standard blocking while promoting the formation of a sparse and porous cake layer. Moreover, according to the extended Derjaguin-Landau-Verwey-Overbeek theory, PAC has been demonstrated as the optimal antifouling system owing to its enhanced repulsion between membrane-foulant and foulant-foulant interactions. Correlation analysis revealed that the exceptional antifouling performance of the PAC system was due to its high removal rates of chemical oxygen demand (~78 %) and suspended solids (~97 %). This research offers valuable insights into the mitigation of membrane fouling through the utilization of adsorbents featuring diverse carbon structures.

3'-Deoxyadenosin alleviates methamphetamine-induced aberrant synaptic plasticity and seeking behavior by inhibiting the NLRP3 inflammasome.

JOURNAL/nrgr/04.03/01300535-202410000-00028/figure1/v/2024-02-06T055622Z/r/image-tiff Methamphetamine addiction is a brain disorder characterized by persistent drug-seeking behavior, which has been linked with aberrant synaptic plasticity. An increasing body of evidence suggests that aberrant synaptic plasticity is associated with the activation of the NOD-like receptor family pyrin domain containing-3 (NLRP3) inflammasome. 3'-Deoxyadenosin, an active component of the Chinese fungus Cordyceps militaris, has strong anti-inflammatory effects. However, whether 3'-deoxyadenosin attenuates methamphetamine-induced aberrant synaptic plasticity via an NLRP3-mediated inflammatory mechanism remains unclear. We first observed that 3'-deoxyadenosin attenuated conditioned place preference scores in methamphetamine-treated mice and decreased the expression of c-fos in hippocampal neurons. Furthermore, we found that 3'-deoxyadenosin reduced the aberrant potentiation of glutamatergic transmission and restored the methamphetamine-induced impairment of synaptic plasticity. We also found that 3'-deoxyadenosin decreased the expression of NLRP3 and neuronal injury. Importantly, a direct NLRP3 deficiency reduced methamphetamine-induced seeking behavior, attenuated the impaired synaptic plasticity, and prevented neuronal damage. Finally, NLRP3 activation reversed the effect of 3'-deoxyadenosin on behavior and synaptic plasticity, suggesting that the anti-neuroinflammatory mechanism of 3'-deoxyadenosin on aberrant synaptic plasticity reduces methamphetamine-induced seeking behavior. Taken together, 3'-deoxyadenosin alleviates methamphetamine-induced aberrant synaptic plasticity and seeking behavior by inhibiting the NLRP3 inflammasome.

The role of exogenous hydrogen sulfide in mitigating cadmium toxicity in plants: A comprehensive meta-analysis.

Reducing the accumulation of cadmium (Cd) and mitigating its toxicity are pivotal strategies for addressing Cd pollution's threats to agriculture and human health. Hydrogen sulfide (H2S) serves as a signaling molecule, playing a crucial role in plant stress defense mechanisms. Nevertheless, a comprehensive assessment of the impact of exogenous H2S on plant growth, antioxidant properties, and gene expression under Cd stress remains lacking. In this meta-analysis, we synthesized 575 observations from 27 articles, revealing that exogenous H2S significantly alleviates Cd-induced growth inhibition in plants. Specifically, it enhances root length (by 8.71%), plant height (by 15.67%), fresh weight (by 15.15%), dry weight (by 22.54%), and chlorophyll content (by 27.99%) under Cd stress conditions. H2S boosts antioxidant enzyme activity, particularly catalase (CAT), by 39.51%, thereby reducing Cd-induced reactive oxygen species (ROS) accumulation. Moreover, it impedes Cd translocation from roots to shoots, resulting in a substantial 40.19% reduction in stem Cd content. Additionally, H2S influences gene expression in pathways associated with antioxidant enzymes, metal transport, heavy metal tolerance, H2S biosynthesis, and energy metabolism. However, the efficacy of exogenous H2S in alleviating Cd toxicity varies depending on factors such as plant species, concentration of the H2S donor sodium hydrosulfide (NaHS), application method, and cultivation techniques. Notably, NaHS concentrations exceeding 200 µM may adversely affect plants. Overall, our study underscores the role of exogenous H2S in mitigating Cd toxicity and elucidates its mechanism, providing insights for utilizing H2S to combat Cd pollution in agriculture.

Toward nitrogen recovery: Co-cultivation of microalgae and bacteria enhances the production of high-value nitrogen-rich cyanophycin.

The algal-bacterial wastewater treatment process has been proven to be highly efficient in removing nutrients and recovering nitrogen (N). However, the recovery of the valuable N-rich biopolymer, cyanophycin, remains limited. This research explored the synthesis mechanism and recovery potential of cyanophycin within two algal-bacterial symbiotic reactors. The findings reveal that the synergy between algae and bacteria enhances the removal of N and phosphorus. The crude contents of cyanophycin in the algal-bacterial consortia reached 115 and 124 mg/g of mixed liquor suspended solids (MLSS), respectively, showing an increase of 11.7 %-20.4 % (p < 0.001) compared with conventional activated sludge. Among the 170 metagenome-assembled genomes (MAGs) analyzed, 50 were capable of synthesizing cyanophycin, indicating that cyanophycin producers are common in algal-bacterial systems. The compositions of cyanophycin producers in the two algal-bacterial reactors were affected by different lighting initiation time. The study identified two intracellular synthesis pathways for cyanophycin. Approximately 36 MAGs can synthesize cyanophycin de novo using ammonium and glucose, while the remaining 14 MAGs require exogenous arginine for production. Notably, several MAGs with high abundance are capable of assimilating both nitrate and ammonium into cyanophycin, demonstrating a robust N utilization capability. This research also marks the first identification of potential horizontal gene transfer of the cyanophycin synthase encoding gene (cphA) within the wastewater microbial community. This suggests that the spread of cphA could expand the population of cyanophycin producers. The study offers new insights into recycling the high-value N-rich biopolymer cyanophycin, contributing to the advancement of wastewater resource utilization.

Degradation of Complex Carbon Sources in Organic Fertilizers Facilitates Nitrogen Fixation.

Biological nitrogen fixation is crucial for agriculture and improving fertilizer efficiency, but organic fertilizers in enhancing this process remain debated. Here, we investigate the impact of organic fertilizers on biological nitrogen fixation through experiments and propose a new model where bacterial interactions with complex carbon sources enhance nitrogen fixation. Field experiments showed that adding organic fertilizers increased the nitrogenase activity by 57.85%. Subculture experiments revealed that organic fertilizer addition enriched genes corresponding to complex carbon and energy metabolism, as well as nifJ involved in electron transfer for nitrogenase. It also enhanced bacterial interactions and enhanced connectors associated with complex carbon degradation. Validation experiments demonstrated that combinations increased nitrogenase activity by 2.98 times compared to the single. Our findings suggest that organic fertilizers promoted nitrogen fixation by enhancing microbial cooperation, improved the degradation of complex carbon sources, and thereby provided utilizable carbon sources, energy, and electrons to N-fixers, thus increasing nitrogenase activity and nitrogen fixation.

Overexpression of NtGPX8a Improved Cadmium Accumulation and Tolerance in Tobacco (Nicotiana tabacum L.).

Cadmium (Cd)-induced oxidative stress detrimentally affects hyperaccumulator growth, thereby diminishing the efficacy of phytoremediation technology aimed at Cd pollution abatement. In the domain of plant antioxidant mechanisms, the role of glutathione peroxidase (GPX) in conferring Cd tolerance to tobacco (Nicotiana tabacum) remained unclear. Our investigation employed genome-wide analysis to identify 14 NtGPX genes in tobacco, revealing their organization into seven subgroups characterized by analogous conserved domain patterns. Notably, qPCR analysis highlighted NtGPX8a as markedly responsive to Cd2+ stress. Subsequent exploration through yeast two-hybridization unveiled NtGPX8a's utilization of thioredoxins AtTrxZ and AtTrxm2 as electron donors, and without interaction with AtTrx5. Introduction of NtGPX8a into Escherichia coli significantly ameliorated Cd-induced adverse effects on bacterial growth. Transgenic tobacco overexpressing NtGPX8a demonstrated significantly augmented activities of GPX, SOD, POD, and CAT under Cd2+ stress compared to the wild type (WT). Conversely, these transgenic plants exhibited markedly reduced levels of MDA, H2O2, and proline. Intriguingly, the expression of NtGPX8a in both E. coli and transgenic tobacco led to increased Cd accumulation, confirming its dual role in enhancing Cd tolerance and accumulation. Consequently, NtGPX8a emerges as a promising candidate gene for engineering transgenic hyperaccumulators endowed with robust tolerance for Cd-contaminated phytoremediation.

Overexpression of SQUAMOSA promoter binding protein-like 4a (NtSPL4a) alleviates Cd toxicity in Nicotiana tabacum.

Squamosa Promoter Binding Protein-Like (SPL) plays a crucial role in regulating plant development and combating stress, yet its mechanism in regulating resistance to Cd toxicity remains unclear. In this study, we cloned a nuclear-localized transcription factor, NtSPL4a, from the tobacco cultivar TN90. Transient co-expression results showed that miR156 significantly reduced the expression of NtSPL4a by binding to the 3'-UTR of its transcript. We obtained transgenic tobacco overexpressing NtSPL4a (including the 3'-UTR) and NtSPL4aΔ (lacking the 3'-UTR) through Agrobacterium-mediated genetic transformation. Compared to the wild type (WT), overexpression of NtSPL4a/NtSPL4aΔ shortened the flowering time and exhibited a more developed root system. The transgenic tobacco showed significantly reduced Cd content, being 85.1% (OE-NtSPL4a) and 46.7% (OE-NtSPL4aΔ) of WT, respectively. Moreover, the upregulation of NtSPL4a affected the mineral nutrient homeostasis in transgenic tobacco. Additionally, overexpression of NtSPL4a/NtSPL4aΔ effectively alleviated leaf chlorosis and oxidative stress induced by Cd toxicity. One possible reason is that the overexpression of NtSPL4a/NtSPL4aΔ can effectively promote the accumulation of non-enzymatic antioxidants. A comparative transcriptomic analysis was performed between transgenic tobacco and WT to further unravel the global impacts brought by NtSPL4a. The tobacco overexpressing NtSPL4a had 183 differentially expressed genes (77 upregulated, 106 downregulated), while the tobacco overexpressing NtSPL4aΔ had 594 differentially expressed genes (244 upregulated, 350 downregulated) compared to WT. These differentially expressed genes mainly included transcription factors, metal transport proteins, flavonoid biosynthesis pathway genes, and plant stress-related genes. Our study provides new insights into the role of the transcript factor SPL in regulating Cd tolerance.