Search Results (47)

This study evaluated the effects of different light-emitting diode (LED) spectral qualities on the early growth of kale at the baby-leaf harvest stage in a plant factory with artificial lighting (PFAL) by integrating morphological traits, biomass accumulation, plant quality indices, vegetation indices, and chlorophyll a fluorescence. Two kale (Brassica oleracea L.) cultivars, ‘Jellujon’ and ‘Manchoo Collard’, were grown for four weeks under monochromatic red, green, and blue LEDs, a purple composite LED with far-red wavelengths, and three white LEDs with different correlated color temperatures (3000, 4100, and 6500 K). Blue LED increased shoot height by approximately 14–28%, depending on cultivar and comparison among the white LED treatments, but this elongation did not translate into superior biomass production. In contrast, white LEDs, particularly at 3000–4100 K, increased leaf area to 24.2–24.9 cm² and SPAD units to 47.3–50.2, whereas blue or green LEDs generally resulted in smaller leaves and lower SPAD units. Shoot dry weight under 3000–4100 K white LEDs reached 0.25–0.26 g in ‘Jellujon’ and 0.26–0.29 g in ‘Manchoo Collard’, approximately twofold higher than under blue or green LEDs. Compactness, Dickson quality index, root investment ratio, and leaf efficiency index were also more favorable under white LEDs, indicating improved plant sturdiness and structural stability. Green LED light was associated with lower maximum photochemical efficiency (Φ_Po) and greater energy dissipation (Φ_Do and DI_o/RC), whereas photochemical reflectance index and PI_ABS tended to be more favorable under selected white LED treatments, although these responses were partly cultivar- and treatment-dependent. Taken together, among the LED spectral quality treatments tested, 3000–4100 K white LEDs provided the most consistently favorable conditions for producing structurally robust, high-quality kale at the early growth stage in PFAL systems. The purple LED showed partial advantages in leaf development and selected physiological responses, but these effects were less consistent across cultivars and indices. Full article

Acidithiobacillus caldus is perpetually exposed to multiple extreme environmental stresses. CsrA, functioning as a post-transcriptional regulator of physiological metabolism, acts as a differential modulator, facilitating more economical and efficient adaptation to extreme environments. The csrA expression recombinant strain was constructed in A. caldus MTH-04 by conjugative transfer technology pJD215. Physiological characterization revealed enhanced acid tolerance, significantly elongated flagella, elevated extracellular secretion, and altered biofilm composition. Notably, intracellular concentrations of free glutamate and aspartate increased to 24.18 mg/L and 16.07 mg/L, respectively. The secondary structure of CsrA protein was determined in vitro through circular dichroism spectroscopy and size-exclusion chromatography. Electrophoretic Mobility Shift Assay (EMSA) successfully demonstrated in vitro binding activity of CsrA to the rpoS leader mRNA. CsrA suppresses rpoS mRNA translation by competing with ribosomes for binding sites, thereby negatively regulating rpoS expression. Critical binding sites were further validated through site-directed mutagenesis. Through EMSA, RT-qPCR and the translation reporter system, it was also found that CsrA has a dual regulatory function for nearby flagella- and motility-related gene clusters (flgC, 07035, motD, 15040), which also implies the global regulatory role of CsrA. In summary, a potential overall post-transcriptional regulatory mechanism based on CsrA and rpoS by extremophile A. caldus was proposed. Finally, the efficiency of bioleaching application by csrA overexpression strain was improved by 20.81%. Full article

Amid the intensifying global mandate for sustainable aquafeed strategies, this study investigates the functional efficacy and biochemical implications of black soldier fly larvae oil (BLO), extracted via recently approved patent method depending on cold-aqueous process, as a substitute for conventional fish oil (FO) in zebrafish (Danio rerio) diets. The refined extraction technique, representing an advancement over traditional aqueous methodologies, was engineered to selectively preserve bioactive lipid fractions while minimizing environmental footprint and processing residues. Over a 28-day feeding period, adult zebrafish were allocated into triplicate groups and fed diets comprising 0%, 50%, and 100% substitution of FO with BLO and growth, lipid composition, and dietary fatty acid profiles of both diets and flesh were rigorously evaluated. Zebrafish fed the BLO₁₀₀ diet exhibited the most pronounced somatic growth (2.47 ± 0.01 g), significantly elevated specific growth rates (3.88 ± 0.82% day⁻¹), and the most efficient feed conversion, without compromising survival. Flesh lipid analysis revealed a substantial enrichment in saturated fatty acids—most notably lauric acid (C12:0)—corresponding to increasing dietary BLO levels. Although dietary EPA and DHA levels were reduced, DHA concentrations in fish tissues remained comparable to those of the control group, indicating a compensatory capacity mediated by endogenous elongation and desaturation pathways. These findings substantiate the dual potential of BLO as both a nutritionally viable lipid source and a vector for enhancing aquafeed sustainability. The cold-aqueous extraction method demonstrated here underscores a pivotal advancement in green lipid processing, aligning oil quality with ecological stewardship. This integrative approach not only reinforces BLO’s candidacy as a strategic fish oil substitute but also delineates a pathway toward scalable, species-adapted feed innovation. Future investigations should prioritize the modulation of fatty acid profiles through dietary and extraction optimization to fully realize the translational potential of insect-derived lipids in aquaculture. Full article

Background/Objectives: The selection and validation of species-specific housekeeping genes (HKGs) have become increasingly common in functional genomics, with application of quantitative Polymerase Chain Reaction (qPCR) or cDNA-based qPCR (RT-qPCR). Despite the Macrobrachium amazonicum having RNA-seq studies available, there are still no data on the most stable and consistent HKGs for use in relative gene expression analyses. Therefore, the present study aimed to identify and validate seven HKGs in M. amazonicum: Eukaryotic Translation Initiation Factor (EIF), 18S ribosomal RNA (18S), Ribosomal Protein L18 (RPL18), β-actin, α-tubulin (α-tub), Elongation Factor 1-α (EF-1α), and Glyceraldehyde-3-phosphate Dehydrogenase (GAPDH). Methods: The HKGs were identified in the M. amazonicum transcriptome, characterized for identity confirmation, and compared against public databases. Subsequently, RT-qPCR assays were prepared using muscle, hepatopancreas, gills, testis, androgenic gland, and ovary to assess the stability of the HKG markers, employing the comparative ∆Ct, BestKeeper, NormFinder, and GeNorm methods. Results: All candidate HKGs identified showed high similarity with other decapods. Reactions performed with these markers demonstrated high specificity, PCR efficiency, and elevated coefficients of determination. The comprehensive ranking, indicated that no single HKG was stable across all tissues, with HKGs showing the best stability being tissue-specific. The most stable HKGs were RPL18 and 18S. GAPDH, historically used as an HKG, showed the poorest performance in stability ranking for most tissues tested, whereas β-actin was most suitable only for ovarian. Conclusions: These data reinforce the need for species-specific HKG validation and provide an appropriate panel of reference markers for gene expression studies in the M. amazonicum. Full article

Species of Trichoderma are currently in high demand as eco-friendly and commercial biocontrol agents due to the proliferation of organic farming methods. This study focused on the potential biocontrol agents of Trichoderma against plant-pathogenic fungi. Trichoderma strains were isolated from different sources (soil, grapevine tissues, lemon fruit, and maize seeds), and were characterized morphologically on two culture media, i.e., Potato Dextrose Agar and Malt Extract Agar, and molecularly using two gene regions: translation elongation factor 1 (TEF) and nuclear ribosomal internal transcribed spacer (ITS). Phylogenetic trees were constructed. As a result, two Trichoderma species were identified, i.e., T. afroharzianum and T. longibrachiatum. The biocontrol effects of all isolated strains of Trichoderma on Fusarium plant damping-off and the promotion of plant growth were evaluated. Additionally, the antagonistic efficiency of Trichoderma spp. against F. proliferatum using the dual-culture method was evaluated. Under greenhouse conditions, T. afroharzianum strains AEMCTa3 and AEMCTa6 were used to treat maize plants infected with Fusarium. The application of Trichoderma significantly reduced the disease index to 15.6% and 0%, respectively. Additionally, maize seedlings showed significant improvements in shoot and root lengths and fresh and dry weights and increased photosynthetic pigment contents compared to Fusarium-infected plants and the untreated control. The gas chromatography–mass spectrometry (GC-MS) analysis of T. afroharzianum extracts identified a variety of bioactive compounds. These compounds included antifungal substances like N-ethyl-1,3-dithioisoindoline, as well as plant growth-promoting hormones like 6-pentyl-α-pyrone and gibberellic acid. Interestingly, the analysis also revealed new phenylacetic acid derivatives that may play important roles in both plant health and disease resistance. From a practical perspective, developing diverse application methods for Trichoderma is essential to optimize its role as a biocontrol agent and a plant growth promoter, thereby supporting sustainable agriculture through improved adaptability and effectiveness across different farming systems. Full article

N, as plants’ most essential nutrient, profoundly shapes root system architecture (RSA), with LRs being preferentially regulated. This review synthesizes the intricate molecular mechanisms underpinning N sensing, signaling, and its integration into developmental pathways governing LR initiation, primordium formation, emergence, and elongation. We delve deeply into the roles of specific transporters (NRT1.1, nitrate transporter 2.1 (NRT2.1)), transcription factors (Arabidopsis nitrate regulated 1 (ANR1), NLP7, TGACG motif-binding factor (TGA), squamosa promoter-binding protein-like 9 (SPL9)) and intricate hormone signaling networks (auxin, abscisic acid, cytokinins, ethylene) modulated by varying N availability (deficiency, sufficiency, excess) and chemical forms (NO₃⁻, NH₄⁺, organic N). Emphasis is placed on the systemic signaling pathways, including peptide-mediated long-distance communication (CEP—C-terminally encoded peptide receptor 1 (CEPR1)) and the critical role of the shoot in modulating root responses. Furthermore, we explore the emerging significance of carbon–nitrogen (C/N) balance, post-translational modifications (ubiquitination, phosphorylation), epigenetic regulation, and the complex interplay with other nutrients (phosphorus (P), sulfur (S)) and environmental factors in shaping N-dependent LR plasticity. Recent advances utilizing single-cell transcriptomics and advanced imaging reveal unprecedented cellular heterogeneity in LR responses to N. Understanding this sophisticated regulatory network is paramount for developing strategies to enhance nitrogen use efficiency (NUE) in crops. This synthesis underscores how N acts as a master regulator, dynamically rewiring developmental programs through molecular hubs that synchronize nutrient sensing with root morphogenesis—a key adaptive strategy for resource acquisition in heterogeneous soils. Full article

Translational control is crucial for maintaining cellular homeostasis, yet the distinct features and regulatory requirements governing protein synthesis during erythropoiesis remain unclear. Here, we reveal that erythroid cells exhibit an extraordinarily high demand for protein synthesis, which is required for their differentiation but also implies the need for tight regulation to prevent excessive erythropoiesis. Notably, we identify significant phosphorylation of eukaryotic elongation factor 2 (eEF2) at threonine 56 during erythroid differentiation, which reduces protein synthesis and acts as a molecular brake to limit unchecked erythropoiesis. This is evidenced by elevated red blood cell counts in peripheral blood and increased incidence of blood hyperviscosity and thrombosis in eEF2_T56M mice, which are deficient in eEF2 phosphorylation. Mechanistic studies demonstrate that eEF2 phosphorylation selectively regulates the translation of a subset of proteins, including NFE2, which partially mediates the effects of eEF2 modification. Collectively, our findings highlight a previously unappreciated role for translational control in achieving efficient and balanced erythropoiesis, with eEF2 phosphorylation serving as a critical protective mechanism against hyperactive erythropoiesis and offering a potential therapeutic target for hematologic disorders such as polycythemia vera. Full article

Background: The Arabidopsis FCS-LIKE ZINC FINGER (FLZ) family proteins play crucial roles in responses to various biotic and abiotic stresses, but the functions of many family members remain uncharacterized. Methods: In this study, we investigated the function of FLZ12, a member of the FLZ family, using a reverse genetic approach. Results: We found that overexpression of FLZ12 impaired root hair development, as evidenced by marked reductions in both root hair length and number under normal growth conditions. However, deprivation of phosphate could partially restore root hair formation, although it still impeded root hair elongation. Notably, FLZ12-overexpressing lines exhibited greatly enhanced tolerance to iron deficiency, with seedlings exhibiting more vigorous and robust growth compared to wild-type plants. In contrast, knockout of FLZ12 resulted in slight impact on seedling development. Further analysis revealed that FLZ12 accumulation was increased in vascular tissues of plants subjected to iron starvation, and the protein was predominantly localized within the nucleus. Conclusions: Integrating these findings with existing evidence, we propose that FLZ12 functions as a translational regulator through interacting with other proteins, playing dual roles in root hair development and iron-deficiency responses in Arabidopsis. These findings provide new insights into the FLZ-domain-containing proteins and offer molecular strategies to enhance iron uptake efficiency in crops, highlighting FLZ12 as a promising candidate for future breeding efforts. Full article

Background: Chinese hamster ovary (CHO) cells are extensively used in the pharmaceutical industry for producing complex proteins, primarily because of their ability to perform human-like post-translational modifications. However, the efficiency of high-quality protein production can vary significantly for monoclonal antibody-producing cell lines, within the CHO host cell lines or by extrinsic factors. Methods: To investigate the complex cellular mechanisms underlying this variability, a phosphoproteomics analysis was performed using label-free quantitative liquid chromatography after a phosphopeptide enrichment of recombinant CHO cells producing two different antibodies and a tunicamycin treatment experiment. Using MaxQuant and Perseus for data analysis, we identified 2109 proteins and quantified 4059 phosphosites. Results: Significant phosphorylation dynamics were observed in nuclear proteins of cells producing the difficult-to-produce 2G12 mAb. It suggests that the expression of 2G12 regulates nuclear pathways based on increases and decreases in phosphorylation abundance. Furthermore, a substantial number of changes in the phosphorylation pattern related to tunicamycin treatment have been detected. TM treatment affects, among other phosphoproteins, the eukaryotic elongation factor 2 kinase (Eef2k). Conclusions: The alterations in the phosphorylation landscape of key proteins involved in cellular processes highlight the mechanisms behind stress-induced cellular responses. Full article

The Escherichia coli (E. coli)-based protein synthesis using recombinant elements (PURE) system is a cell-free protein synthesis system reconstituted from purified factors essential for E. coli translation. The PURE system is widely used for basic and synthetic biology applications. One of the major challenges associated with the PURE system is that the protein yield of the system varies depending on the protein. Studies have reported that the efficiency of translation is significantly affected by nucleotide and amino acid sequences, especially in the N-terminal region. Here, we investigated the inherent effect of various N-terminal sequences on protein synthesis using the PURE system. We found that a single amino acid substitution in the N-terminal region significantly altered translation efficiency in the PURE system, especially at low temperatures. This result gives us useful suggestions for the expression of the protein of interest in vitro and in vivo. Full article

Drought and salinity stress reduce root hydraulic conductivity of plant seedlings, and melatonin application positively mitigates stress-induced damage. However, the underlying effect of melatonin priming on root hydraulic conductivity of seedlings under drought–salinity combined remains greatly unclear. In the current report, we investigated the influence of seeds of three wheat lines’ 12 h priming with 100 μM of melatonin on root hydraulic conductivity (Lpr) and relevant physiological indicators of seedlings under PEG, NaCl, and PEG + NaCl combined stress. A previous study found that the combined PEG and NaCl stress remarkably reduced the Lpr of three wheat varieties, and its value could not be detected. Melatonin priming mitigated the adverse effects of combined PEG + NaCl stress on Lpr of H4399, Y1212, and X19 to 0.0071 mL·h⁻¹·MPa⁻¹, 0.2477 mL·h⁻¹·MPa⁻¹, and 0.4444 mL·h⁻¹·MPa⁻¹, respectively, by modulating translation levels of aquaporin genes and contributed root elongation and seedlings growth. The root length of H4399, Y1212, and X19 was increased by 129.07%, 141.64%, and 497.58%, respectively, after seeds pre-treatment with melatonin under PEG + NaCl combined stress. Melatonin -priming appreciably regulated antioxidant enzyme activities, reduced accumulation of osmotic regulators, decreased levels of malondialdehyde (MDA), and increased K⁺ content in stems and root of H4399, Y1212, and X19 under PEG + NaCl stress. The path investigation displayed that seeds primed with melatonin altered the modification of the path relationship between Lpr and leaf area under stress. The present study suggested that melatonin priming was a strategy as regards the enhancement of root hydraulic conductivity under PEG, NaCl, and PEG + NaCl stress, which efficiently enhanced wheat resistant to drought–salinity stress. Full article

Different bacterial species have dramatically different generation times, from 20–30 min in Escherichia coli to about two weeks in Mycobacterium leprae. The translation machinery in a cell needs to synthesize all proteins for a new cell in each generation. The three subprocesses of translation, i.e., initiation, elongation, and termination, are expected to be under stronger selection pressure to optimize in short-generation bacteria (SGB) such as Vibrio natriegens than in the long-generation Mycobacterium leprae. The initiation efficiency depends on the start codon decoded by the initiation tRNA, the optimal Shine–Dalgarno (SD) decoded by the anti-SD (aSD) sequence on small subunit rRNA, and the secondary structure that may embed the initiation signals and prevent them from being decoded. The elongation efficiency depends on the tRNA pool and codon usage. The termination efficiency in bacteria depends mainly on the nature of the stop codon and the nucleotide immediately downstream of the stop codon. By contrasting SGB with long-generation bacteria (LGB), we predict (1) SGB to have more ribosome RNA operons to produce ribosomes, and more tRNA genes for carrying amino acids to ribosomes, (2) SGB to have a higher percentage of genes using AUG as the start codon and UAA as the stop codon than LGB, (3) SGB to exhibit better codon and anticodon adaptation than LGB, and (4) SGB to have a weaker secondary structure near the translation initiation signals than LGB. These differences between SGB and LGB should be more pronounced in highly expressed genes than the rest of the genes. We present empirical evidence in support of these predictions. Full article

As the size of deep learning models continues to expand, the elongation of inference time has gradually evolved into a significant challenge to efficiency and practicality for autoregressive models. This work introduces a hybrid model acceleration strategy based on branch prediction, which accelerates autoregressive model inference without requiring retraining and ensures output consistency with the original model. Specifically, the algorithm employs two models with different parameter sizes aimed at the same task. The smaller model generates a series of potential tokens that are then parallelly validated by the larger model to determine their acceptability. By orchestrating the workflow of the large and small models through a branch-prediction strategy, the algorithm conceals the validation time of the larger model when predictions are successful, thereby accelerating inference. We propose a binomial distribution-based prediction function that blends theoretical principles with empirical evidence, specifically designed for the nuanced requirements of accelerating inference within a hybrid model framework. The entire algorithm was designed and implemented on the llama model for text generation and translation tasks. The experimental results indicate significant improvements. The proposed algorithm achieves a 1.2× to 3.4× increase in inference speed compared to the original model, consistently outperforming the speculative sampling inference acceleration algorithm. Full article

During the emergence of infectious diseases, evaluating the efficacy of newly developed vaccines requires antigen proteins. Available methods enhance antigen protein productivity; however, structural modifications may occur. Therefore, we aimed to construct a novel transient overexpression vector capable of rapidly producing large quantities of antigenic proteins in mammalian cell lines. This involved expanding beyond the exclusive use of the human cytomegalovirus (CMV) promoter, and was achieved by incorporating a transcriptional enhancer (CMV enhancer), a translational enhancer (woodchuck hepatitis virus post-transcriptional regulatory element), and a promoter based on the CMV promoter. Twenty novel transient expression vectors were constructed, with the vector containing the human elongation factor 1-alpha (EF-1a) promoter showing the highest efficiency in expressing foreign proteins. This vector exhibited an approximately 27-fold higher expression of enhanced green fluorescent protein than the control vector containing only the CMV promoter. It also expressed the highest level of severe acute respiratory syndrome coronavirus 2 receptor-binding domain protein. These observations possibly result from the simultaneous enhancement of the transcriptional activity of the CMV promoter and the human EF-1a promoter by the CMV enhancer. Additionally, the synergistic effect between the CMV and human EF-1a promoters likely contributed to the further enhancement of protein expression. Full article

Programmed ribosomal frameshifting (PRF) exists in all branches of life that regulate gene expression at the translational level. The eukaryotic translation initiation factor 5A (eIF5A) is a highly conserved protein essential in all eukaryotes. It is identified initially as an initiation factor and functions broadly in translation elongation and termination. The hypusination of eIF5A is specifically required for +1 PRF at the shifty site derived from the ornithine decarboxylase antizyme 1 (OAZ1) in Saccharomyces cerevisiae. However, whether the regulation of +1 PRF by yeast eIF5A is universal remains unknown. Here, we found that Sc-eIF5A depletion decreased the putrescine/spermidine ratio. The re-introduction of Sc-eIF5A in yeast eIF5A mutants recovered the putrescine/spermidine ratio. In addition, the Sc-eIF5A depletion decreases +1 PRF during the decoding of Ty1 retrotransposon mRNA, but has no effect on −1 PRF during the decoding of L-A virus mRNA. The re-introduction of Sc-eIF5A in yeast eIF5A mutants restored the +1 PRF rate of Ty1. The inhibition of the hypusine modification of yeast eIF5A by GC7 treatment or by mutating the hypusination site Lys to Arg caused decreases of +1 PRF rates in the Ty1 retrotransposon. Furthermore, mutational studies of the Ty1 frameshifting element support a model where the efficient removal of ribosomal subunits at the first Ty1 frame 0 stop codon is required for the frameshifting of trailing ribosomes. This dependency is likely due to the unique position of the frame 0 stop codon distance from the slippery sequence of Ty1. The results showed that eIF5A is a trans-regulator of +1 PRF for Ty1 retrotransposon and could function universally in yeast. Full article