Ammonia stress was detrimental to shrimp, but how water ammonia nitrogen (ammonia-N) influences the shrimp’s health remains unclear. Thus, this study was designed to investigate the effects of water ammonia-N on hemolymph ammonia-N concentration, hepatopancreas structure, and the intestinal microbiota of Litopenaeus vannamei with four experiments. We found that the concentration of ammonia-N in shrimp hemolymph was significantly higher than that in pond water, indicating that water ammonia-N stimulates the accumulation of hemolymph ammonia-N. Results also indicated that the hemolymph ammonia-N accumulation would disrupt the hepatopancreas structure and alter the intestinal microbial composition. The concentration of hemolymph ammonia-N and severity of hepatopancreas damage positively correlated with water ammonia-N concentration. However, though the diversity of intestinal microbiota was varied by ammonia-N, there were no significant differences between groups, suggesting that the variation was relatively minimal. Furthermore, returning shrimp to pristine water after ammonia-N exposure could reduce the hemolymph ammonia-N concentration and the mortality rate. This study provides evidence of temporal variations in hemolymph ammonia-N concentration, hepatopancreatic structure, and intestinal microbiota under different water ammonia-N levels, which might shed insights into ecological cognition on scientific management of shrimp culture and microecological prevention of shrimp health.

SARS-CoV-2 (Severe acute respiratory syndrome coronavirus 2) Variants of Concern (VOCs), such as the Omicron sub-variants, present significant challenges in pandemic control due to their capacity to escape antibodies and breach vaccine protections. Discovering antibodies that can tolerate mutations in VOCs and understanding their underlying mechanisms is crucial for developing therapeutics for COVID-19 patients, particularly those for whom other therapies may be unsuitable. Here, we report the neutralization of the Omicron variant by FD20, a broadly active human monoclonal antibody. In contrast to a clinically approved control antibody, FD20 neutralizes Omicron with comparable IC₅₀ values to those observed for previously circulating VOCs and the original strain reported in Wuhan. Leveraging structural information, we provide insights into its resilience against mutations in Omicron. The results encourage the prospective development of FD20 as a therapeutic option for COVID-19 caused by current and potentially future VOCs.

RNA is an intermediary between DNA and protein, a catalyzer of biochemical reactions, and a regulator of genes and transcripts. RNA structures are essential for complicated functions. Recent years have witnessed rapid advancements in RNA secondary structure probing techniques. These technological strides provided comprehensive insights into RNA structures, which significantly contributed to our understanding of diverse cellular regulatory processes, including gene regulation, epigenetic regulation, and post-transactional regulation. Meanwhile, they have facilitated the creation of therapeutic tools for tackling human diseases. Despite their therapeutic applications, RNA structure probing methods also offer a promising avenue for exploring the mechanisms of human diseases, potentially providing the key to overcoming existing research constraints and obtaining the in-depth information necessary for a deeper understanding of disease mechanisms.

Alternative splicing (AS) significantly enriches the diversity of transcriptomes and proteomes, playing a pivotal role in the physiology and development of eukaryotic organisms. With the continuous advancement of high-throughput sequencing technologies, an increasing number of novel transcript isoforms, along with factors related to splicing and their associated functions, are being unveiled. In this review, we succinctly summarize and compare the different splicing mechanisms across prokaryotes and eukaryotes. Furthermore, we provide an extensive overview of the recent progress in various studies on AS covering different developmental stages in diverse plant species and in response to various abiotic stresses. Additionally, we discuss modern techniques for studying the functions and quantification of AS transcripts, as well as their protein products. By integrating genetic studies, quantitative methods, and high-throughput omics techniques, we can discover novel transcript isoforms and functional splicing factors, thereby enhancing our understanding of the roles of various splicing modes in different plant species.

• High-level secretion expression (~250 mg L-1) of divalent nanobodies in Pichia.

• Detergent washing effectively removes yellow pigment from secreted nanobodies.

• Nanobodies after pigment removal remain biologically active.

Mass spectrometry imaging (MSI) serves as a valuable tool enabling researchers to scrutinize various compounds, peptides, and proteins within a sample, providing detailed insights at both elemental and molecular levels. This innovative technology transforms information obtained from a mass spectrometer— encompassing ionic strength, mass-to-charge ratio, and ionized molecule coordinates—within a defined region into a pixel-based model. Consequently, it reconstructs the spatial distribution of ions, allowing for a comprehensive understanding of molecular landscapes. The significance of MSI lies in its ability to offer multiple advantages, including straightforward sample preparation and remarkable sensitivity, all achieved without the necessity for labeling. Particularly in the realm of plant biology, MSI finds frequent application in examining the distribution of target metabolites and other components within plant tissues. This review delves into the fundamental principles, distinguishing features, merits, and applications of three prominent MSI technologies. Furthermore, we aim to assist readers in navigating the utilization of MSI in their plant biology research by discussing primary challenges, proposing potential solutions, and elucidating future prospects associated with this cutting-edge technology.

Porcine epidemic diarrhea virus (PEDV), Transmissible gastroenteritis virus (TGEV), Porcine deltacoronavirus (PDCoV) and Swine acute diarrhea syndrome coronavirus (SADS-CoV) rank among the most frequently encountered swine enteric coronaviruses (SECoVs), leading to substantial economic losses to the swine industry. The availability of a rapid and highly sensitive detection method proves beneficial for the monitoring and surveillance of SECoVs. Based on the N genes of four distinct SECoVs, a novel detection method was developed in this study by combining recombinant enzyme polymerase isothermal amplification (RPA) with clustered regularly interspaced short palindromic repeats (CRISPR)-associated proteins (Cas) 12a. Results showed that the cut-off value of CRISPR-Cas12a assay for SADS-CoV, PEDV, PDCoV and TGEV was 2.19×10⁴ Relative Fluorescence Units (RFU), 1.57×10⁴ RFU, 3.07×10⁴ RFU and 1.64×10⁴ RFU, respectively. The coefficient of variation (CV) of within and between runs by CRISPR-Cas12a assay for 6 clinical diarrhea samples were both less than 10%. The CRISPR-Cas12a assay demonstrated high specificity for TGEV, PEDV, PDCoV, and SADS-CoV with no cross-reactivity to other common swine viruses. This method also exhibited a low limit of detection of 2 copies for each virus. Additionally, the results demonstrated a perfect agreement (100%) between the CRISPR-Cas12a assay and the RT-qPCR assay. Finally, a total of 494 pig samples from the field tested by CRISPR-Cas12a assay showed that positive rate for SADS-CoV, TGEV, PDCoV and PEDV was 0, 0, 1.2% and 48.6%, respectively. The results suggested the great potential of CRISPR-Cas12a assay to detect SECoVs in the field.

While biotechnologies offer eco-friendly solutions for eliminating air contaminants, there is a scarcity of research examining the impacts of microbial purification of air pollutants on the structure and function of air microbial communities. In this study, we explored a Lactobacillus paracasei B1 (LAB) agent for removing ammoniacal odour. The impacts of LAB on air bacterial community were revealed. by analyzing the air samples before (BT) and after (AT) LAB bioagent treatment. Remarkably, the LAB bioagent significantly reduced the air ammonia concentration by 96.8%. This reduction was associated with a notable decline in bacterial diversity and a significant shift in community composition. The relative abundance of Staphylococcus, a common pathogen, plummeted from 1.91% to 0.03%. Moreover, other potential pathogens decreased by over 87%, signifying the bioagent's impactful role in diminishing health risks. The dominance of OTU-4 (Lactobacillus) highlighted its crucial role not only in competitive interactions but also potentially in shaping the metabolic pathways or community dynamics within the treated air microbial ecosystem. This shift towards deterministic assembly processes post-treatment, as highlighted by the normalized stochasticity ratio (NST), sheds light on the underlying mechanisms dictating the microbial community's response to bioagent interventions. The bioagent-purified air microbial community showed a strong preference for variable selection (88.9%), likely due to the acidity generated by the LAB. In conclusion, our findings emphasized the positive impact of LAB bioagent in enhancing air quality, which associated with the changes in microbial community.

Engineering microbial cell factories have achieved much progress in producing fuels, natural products and bulk chemicals. However, in industrial fermentation, microbial cells often face various predictable and stochastic disturbances resulting from intermediate metabolites or end product toxicity, metabolic burden and harsh environment. These perturbances can potentially decrease productivity and titer. Therefore, strain robustness is essential to ensure reliable and sustainable production efficiency. In this review, the current strategies to improve host robustness were summarized, including knowledge-based engineering approaches, such as transcription factors, membrane/transporters and stress proteins, and the traditional adaptive laboratory evolution based on natural selection. Computation-assisted (e.g. GEMs, deep learning and machine learning) design of robust industrial hosts was also introduced. Furthermore, the challenges and future perspectives on engineering microbial host robustness are proposed to promote the development of green, efficient and sustainable biomanufacturers.

Biomolecular condensates, also referred to as membrane-less organelles, function as fundamental organizational units within cells. These structures primarily form through liquid–liquid phase separation, a process in which proteins and nucleic acids segregate from the surrounding milieu to assemble into micron-scale structures. By concentrating functionally related proteins and nucleic acids, these biomolecular condensates regulate a myriad of essential cellular processes. To study these significant and intricate organelles, a range of technologies have been either adapted or developed. In this review, we provide an overview of the most utilized technologies in this rapidly evolving field. These include methods used to identify new condensates, explore their components, investigate their properties and spatiotemporal regulation, and understand the organizational principles governing these condensates. We also discuss potential challenges and review current advancements in applying the principles of biomolecular condensates to the development of new technologies, such as those in synthetic biology.

Plant lipids are a diverse group of biomolecules that play essential roles in plant architecture, physiology, and signaling. To advance our understanding of plant biology and facilitate innovations in plant-based product development, we must have precise methods for the comprehensive analysis of plant lipids. Here, we present a comprehensive overview of current research investigating plant lipids, including their structures, metabolism, and functions. We explore major lipid classes, i.e. fatty acids, glyceroglycolipids, glycerophospholipids, sphingolipids, and phytosterols, and discuss their subcellular distributions. Furthermore, we emphasize the significance of lipidomics research techniques, particularly chromatography-mass spectrometry, for accurate lipid analysis. Special attention is given to lipids as crucial signal receptors and signaling molecules that influence plant growth and responses to environmental challenges. We address research challenges in lipidomics, such as in identifying and quantifying lipids, separating isomers, and avoiding batch effects and ion suppression. Finally, we delve into the practical applications of lipidomics, including its integration with other omics methodologies, lipid visualization, and innovative analytical approaches. This review thus provides valuable insights into the field of plant lipidomics and its potential contributions to plant biology.