Genome-Wide Identification of Calmodulin-Binding Protein 60 Gene Family and the Function of GhCBP60B in Cotton Growth and Development and Abiotic Stress Response.

The calmodulin-binding protein 60 (CBP60) family is a gene family unique to plants, and its members play a crucial role in plant defense responses to pathogens and growth and development. Considering that cotton is the primary source of natural cotton textile fiber, the functional study of its CBP60 gene family members is critical. In this research, we successfully identified 162 CBP60 members from the genomes of 21 species. Of these, 72 members were found in four cotton species, divided into four clades. To understand the function of GhCBP60B in cotton in depth, we conducted a detailed analysis of its sequence, structure, cis-acting elements, and expression patterns. Research results show that GhCBP60B is located in the nucleus and plays a crucial role in cotton growth and development and response to salt and drought stress. After using VIGS (virus-induced gene silencing) technology to conduct gene silencing experiments, we found that the plants silenced by GhCBP60B showed dwarf plants and shortened stem nodes, and the expression of related immune genes also changed. In further abiotic stress treatment experiments, we found that GhCBP60B-silenced plants were more sensitive to drought and salt stress, and their POD (peroxidase) activity was also significantly reduced. These results imply the vital role of GhCBP60B in cotton, especially in regulating plant responses to drought and salt stress. This study systematically analyzed CBP60 gene family members through bioinformatics methods and explored in depth the biological function of GhCBP60B in cotton. These research results lay a solid foundation for the future use of the GhCBP60B gene to improve cotton plant type and its drought and salt resistance.

GhCKX14 responding to drought stress by modulating antioxi-dative enzyme activity in Gossypium hirsutum compared to CKX family genes.

BACKGROUND: Cytokinin oxidase/dehydrogenase (CKX) plays a vital role in response to abiotic stress through modulating the antioxidant enzyme activities. Nevertheless, the biological function of the CKX gene family has yet to be reported in cotton. RESULT: In this study, a total of 27 GhCKXs were identified by the genome-wide investigation and distributed across 18 chromosomes. Phylogenetic tree analysis revealed that CKX genes were clustered into four clades, and most gene expansions originated from segmental duplications. The CKXs gene structure and motif analysis displayed remarkably well conserved among the four groups. Moreover, the cis-acting elements related to the abiotic stress, hormones, and light response were identified within the promoter regions of GhCKXs. Transcriptome data and RT-qPCR showed that GhCKX genes demonstrated higher expression levels in various tissues and were involved in cotton's abiotic stress and phytohormone response. The protein-protein interaction network indicates that the CKX family probably participated in redox regulation, including oxidoreduction or ATP levels, to mediate plant growth and development. Functionally identified via virus-induced gene silencing (VIGS) found that the GhCKX14 gene improved drought resistance by modulating the antioxidant-related activitie. CONCLUSIONS: In this study, the CKX gene family members were analyzed by bioinformatics, and validates the response of GhCKX gene to various phytohormone treatment and abiotic stresses. Our findings established the foundation of GhCKXs in responding to abiotic stress and GhCKX14 in regulating drought resistance in cotton.

Comprehensive identification and functional characterization of GhpPLA gene family in reproductive organ development.

BACKGROUND: Phospholipases As (PLAs) are acyl hydrolases that catalyze the release of free fatty acids in phospholipids and play multiple functions in plant growth and development. The three families of PLAs are: PLA1, PLA2 (sPLA), and patatin-related PLA (pPLA). The diverse functions that pPLAs play in the growth and development of a broad range of plants have been demonstrated by prior studies. METHODS: Genome-wide analysis of the pPLA gene family and screening of genes for expression verification and gene silencing verification were conducted. Additionally, pollen vitality testing, analysis of the pollen expression pattern, and the detection of POD, SOD, CAT, MDA, and H2O2 were performed. RESULT: In this study, 294 pPLAs were identified from 13 plant species, including 46 GhpPLAs that were divided into three subfamilies (I-III). Expression patterns showed that the majority of GhpPLAs were preferentially expressed in the petal, pistil, anther, and ovule, among other reproductive organs. Particularly, GhpPLA23 and GhpPLA44, were found to be potentially important for the reproductive development of G. hirsutum. Functional validation was demonstrated by VIGS which showed that reduced expression levels of GhpPLA23 and GhpPLA44 in the silenced plants were associated with a decrease in pollen activity. Moreover, a substantial shift in ROS and ROS scavengers and a considerable increase in POD, CAT, SOD, and other physiological parameters was found out in these silenced plants. Our results provide plausibility to the hypothesis that GhpPLA23 and GhpPLA44 had a major developmental impact on cotton reproductive systems. These results also suggest that pPLAs are important for G. hirsutum's reproductive development and suggest that they could be employed as potential genes for haploid induction. CONCLUSIONS: The findings of the present research indicate that pPLA genes are essential for the development of floral organs and sperm cells in cotton. Consequently, this family might be important for the reproductive development of cotton and possibly for inducing the plant develop haploid progeny.

Flexible Recyclable Cellulose Paper Templated Cu-Doped Polydopamine Membranes with Dual Enzyme-Like Activity.

The development of high-efficiency enzyme mimics is of great significance in the field of biocatalysis. However, it remains challenging to design novel enzyme mimics with multiple enzyme-like activities, excellent stability, and good reusability. Herein, a facile molecular assembly strategy to construct dialdehyde cellulose (DAC) templated Cu-doped polydopamine (DAC@PDA/Cu) membrane with dual enzyme-like activities is presented. The Schiff base bonds formed between polydopamine (PDA) and DAC can not only accelerate the adhesion of PDA thin layer but also contribute to Cu-loading and high stability of DAC@PDA/Cu membrane. Importantly, the assembled DAC@PDA/Cu membrane exhibits a remarkable catalytic activity that is superior to the natural laccase along with high stability and excellent reusability. Moreover, the DAC@PDA/Cu membrane also demonstrates peroxidase-like activity, and it is successfully applied in the sensitive detection of ascorbic acid (AA). This work will provide a new paradigm methodology for rational design and practical applications of enzyme mimics based on bioinspired molecular assemblies.

A mini-review on peptide-based self-assemblies and their biological applications.

Peptide-based supramolecular self-assembly from peptide monomers into well-organized nanostructures, has attracted extensive attentions towards biomedical and biotechnological applications in recent decades. This spontaneous and reversible assembly process involving non-covalent bonding interactions can be artificially regulated. In this review, we have elaborated different strategies to modulate the peptide self-assembly through tuning the physicochemical and environmental conditions, includingpH, light, temperature, solvent, and enzyme. Detailed introduction of biological applications and future potential of the peptide-based nano-assemblies will also be given.

A Dipeptide-Based Hierarchical Nanoarchitecture with Enhanced Catalytic Activity.

Achieving synthetic architectures with simple structures and robust biomimetic catalytic activities remains a great challenge. Herein, we explore a facile supramolecular assembly approach to construct a dipeptide-based hierarchical nanoarchitecture with enhanced enzyme-like catalytic activity. In this nanoarchitecture, nanospheres are put in a chain-like arrangement through coordination-driven directional self-assembly. The reversible transformation of anisotropic nanochains to isotropic nanospheres switches biomimetic activity. Notably, the assembled nanoarchitecture exhibits a high enzyme-like activity and remarkable long-term stability to promote hydroquinone oxidation, superior to the natural counterpart. This work will pave the way to develop reversible and reusable supramolecular biocatalysts with ordered hierarchical structures for accelerating chemical transformations.

The Ultrafast Assembly of a Dipeptide Supramolecular Organogel and its Phase Transition from Gel to Crystal.

A gel-to-crystal phase transition of a dipeptide supramolecular assembly mediates active water transportation in oils. The addition of water into ultrafast-assembling dipeptide organogels can induce a lamellar-to-hexagonal structural transformation of dipeptide molecular arrangement. Consequently, a phase transition from gel to crystal occurs and in turn water is transported in the dipeptide crystal via well-defined channels. On a macroscopic scale, water transport in the bulk system exhibits an anisotropic characteristic, which can be tuned by the presence of ions in the Hofmeister series. These favorable features enable the automatic separation of dispersed nanoparticles from dissolved electrolytes in aqueous solution. These findings demonstrate the potential of this assembled system for active filtration without external pressure.

Optically Matched Semiconductor Quantum Dots Improve Photophosphorylation Performed by Chloroplasts.

A natural-artificial hybrid system was constructed to enhance photophosphorylation. The system comprises chloroplasts modified with optically matched quantum dots (chloroplast-QD) with a large Stokes shift. The QDs possess a unique optical property and transform ultraviolet light into available and highly effective red light for use by chloroplasts. This favorable feature enables photosystem II contained within the hybrid system to split more water and produce more protons than chloroplasts would otherwise do on their own. Consequently, a larger proton gradient is generated and photophosphorylation is improved. At optimal efficiency activity increased by up to 2.3 times compared to pristine chloroplasts. Importantly, the degree of overlap between emission of the QDs and absorption of chloroplasts exerts a strong influence on the photophosphorylation efficiency. The chloroplast-QD hybrid presents an efficient solar energy conversion route, which involves a rational combination of a natural system and an artificial light-harvesting nanomaterial.

A Photoinduced Reversible Phase Transition in a Dipeptide Supramolecular Assembly.

Tunable supramolecular assembly has found various applications in biomedicine, molecular catalysis, optoelectronics, and nanofabrication. Unlike traditional covalent conjugation, non-covalent introduction of a photoswitchable moiety enables reversible photomodulation of non-photosensitive dipeptide supramolecular assembly. Under light illumination, a long-lived photoacid generator releases a proton and mediates the dissociation of dipeptide-based organogel, thereby resulting in sol formation. Under darkness, the photoswitchable moiety entraps a proton, resulting in gel regeneration. Furthermore, accompanying the isothermal recycled gel-sol transition in a spatially controlled manner, renewable patterns are spontaneously fabricated. This new concept of light-controlled phase transition of amino acid-based supramolecular assemblies will open up the possibility of wide applications.

Enhanced Photophosphorylation of a Chloroplast-Entrapping Long-Lived Photoacid.

Enhancing solar energy conversion efficiency is very important for developing renewable energy, protecting the environment, and producing agricultural products. Efficient enhancement of photophosphorylation is demonstrated by coupling artificial photoacid generators (PAGs) with chloroplasts. The encapsulation of small molecular long-lived PAGs in the thylakoid lumen is improved greatly by ultrasonication. Under visible-light irradiation, a fast intramolecular photoreaction of the PAG occurs and produces many protons, remarkably enhancing the proton gradient in situ. Consequently, compared to pure chloroplasts, the assembled natural-artificial hybrid demonstrates approximately 3.9 times greater adenosine triphosphate (ATP) production. This work will provide new opportunities for constructing enhanced solar energy conversion systems.

Noncombustible gel polymer electrolyte inspired by bio-radicalchemistry for high voltage and high safety Ni-rich lithium batteries.

For high energy density lithium-ion batteries (LIBs) with nickel-rich ternary cathodes, the chemical degradation of electrolytes caused by free radical reactions and the hazards of thermal runaway have always been significant challenges. Inspired by the free radical scavenging of living organisms and multiphase synergistic flame retardant mechanism, we innovatively designed and prepared a multifunctional flame retardant HCCP-TMP that combines flame retardancy and free radical scavenging by combining hindered amine and cyclophosphazene. Only 1 wt% HCCP-TMP can make the polyacrylate-based gel polymer electrolyte (GPE) incombustible. Moreover, the equipped NCM811//Graphite pouch cells don't exhibit combustion behavior after thermal runaway and can resist mechanical abuse. Based on the above noncombustible GPE, the NCM811//Li battery exhibits capacity retention rate of 82.2 % after 100 cycles at a current density of 2 C and in the voltage range of 3.0-4.7 V, exhibiting excellent cyclability under high voltage. This simple molecular design simultaneously improves the fire safety and high voltage stability, demonstrating enormous application potential in the field of advanced LIBs with high safety and high energy density.

Effects of exercise on muscle fiber conversion, muscle development and meat quality of Sunit sheep.

This study aimed to investigate the effects of exercise on muscle fiber conversion, muscle development and meat quality in the biceps femoris (BF) muscle of Sunit sheep. Twelve Sunit sheep with similar body weight were divided into two groups: control group (C group) and exercise group (E group), E group lambs underwent 6 km of exercise training per day for 90 d. The findings revealed that compared with the C group, exercise training enhanced the expression of MyHC IIa mRNA, decreased the number ratio of type IIB muscle fibers and the expression of MyHC IIb mRNA (P < 0.05). Furthermore, the E group lamb displayed higher creatine kinase (CK) activity, and lactic acid levels (P < 0.05), while glycogen content and lactic dehydrogenase (LDH) activity showed opposite trends (P < 0.05). Exercise significantly up-regulated the mRNA expression of AMP-activated protein kinase α1 (AMPKα1), sirtuin1 (SIRT1), peroxisome proliferator-activated receptor gamma coactivator 1-α (PGC-1α), cytochrome c oxidase IV (COX IV), protein kinase B (Akt), mammalian target of rapamycin (mTOR) and p70 Ribosomal S6 Kinase 1 (p70s6k1) (P < 0.05), suggesting exercise promoted muscle fiber conversion by mediating AMPK/PGC-1α pathway, and improved skeletal muscle development via Akt/mTOR pathway. Besides, backfat thickness and pH45min value in the E group decreased significantly, while the pH24, a*, and shear force value increased significantly (P < 0.05). To conclude, this study suggested that exercise training can be used to alter muscle fiber characteristics and muscle development in lamb production.

Oxyhaemoglobin saturation NIR-IIb imaging for assessing cancer metabolism and predicting the response to immunotherapy.

In vivo quantitative assessment of oxyhaemoglobin saturation (sO2) status in tumour-associated vessels could provide insights into cancer metabolism and behaviour. Here we develop a non-invasive in vivo sO2 imaging technique to visualize the sO2 levels of healthy and tumour tissue based on photoluminescence bioimaging in the near-infrared IIb (NIR-IIb; 1,500-1,700 nm) window. Real-time dynamic sO2 imaging with a high frame rate (33 Hz) reveals the cerebral arteries and veins through intact mouse scalp/skull, and this imaging is consistent with the haemodynamic analysis results. Utilizing our non-invasive sO2 imaging, the tumour-associated-vessel sO2 levels of various cancer models are evaluated. A positive correlation between the tumour-associated-vessel sO2 levels and the basal oxygen consumption rate of corresponding cancer cells at the early stages of tumorigenesis suggests that cancer cells modulate the tumour metabolic microenvironment. We also find that a positive therapeutic response to the checkpoint blockade cancer immunotherapy could lead to a dramatic decrease of the tumour-associated-vessel sO2 levels. Two-plex dynamic NIR-IIb imaging can be used to simultaneously observe tumour-vessel sO2 and PD-L1, allowing a more accurate prediction of immunotherapy response.

Effects of Probiotics Supplementation on the Intestinal Metabolites, Muscle Fiber Properties, and Meat Quality of Sunit Lamb.

The development of animal husbandry is closely related to the meat quality of small ruminants. Intestinal metabolites and the muscle fiber types of lambs are important factors that affect their meat quality, but few studies have examined the regulation of the "intestinal muscle axis" by probiotics. In this study, 12 Sunit lambs were divided into a control group (C) and a probiotics group (P). The gene expressions of the myosin heavy chain, metabolic enzyme activity, and short-chain fatty acids in the intestines were analyzed using gas chromatography-mass spectrometry (GC-MS) and quantitative real-time PCR. The results showed that levels of propionic acid and butyric acid in the intestines of group P were significantly higher than in group C (p < 0.05). In addition, probiotics increased the number and area ratio of type I muscle fibers. They also increased the mRNA expression of MyHC IIA and the activity of malate dehydrogenase (MDH) and succinate dehydrogenase (SDH). Propionic acid was negatively correlated with the number ratio of type IIB muscle fibers. Butyric acid was found to be significantly positively correlated with the number ratio of type IIA muscle fibers. Cooking loss, pH24h, and shear force decreased significantly in group P. In conclusion, intestinal metabolites (SCFAs) altered the activity of oxidative-myofibril-metabolizing enzymes and the expression of myosin heavy-chain type IIA, reduced the meat shear values, and improved meat tenderness. This study provides a new basis for improving the production and meat quality of small ruminants.

Characteristic of GEX1 genes reveals the essential roles for reproduction in cotton.

GEX1 (gamete expressed 1) proteins are critical membrane proteins conserved among flowering plants that are involved in the nuclear fusion and embryonic development. Herein, we identified the 32 GEX1 proteins from representative land plants. In cotton, GEX1 genes expressed in various tissues across all stages of the life cycle, especially in pollen. Subcellular localization indicated the position of GhGEX1 protein was localized in the endoplasmic reticulum. Experimental research has demonstrated that GhGEX1 has the potential to improve the partial abortion phenotype in Arabidopsis. CRISPR/Cas9-mediated knockout of GhGEX1 exhibited the seed abortion. Paraffin section of the ovule revealed that the polar nuclear fusion of ghgex1 plants remains at a standstill when the wild type has developed into a normal embryo. Comparative transcriptome analysis showed that the DEGs of reproductive-related processes and membrane-related processes were repressed in the pollen of knockout lines. The predicted protein interactions showed that GhGEX1 probably functioned through interactions with proteins related to reproduction and membrane. From all these investigations, it was possible to conclude that the GEX1 proteins are evolutionarily conserved in flowering plants and elucidated the pivotal roles during fertilization and early embryonic development in cotton.

Characterization and Functional Analysis of GhNAC82, A NAM Domain Gene, Coordinates the Leaf Senescence in Upland Cotton (Gossypium hirsutum L.).

In the process of growth and development, cotton exhibits premature senescence under various abiotic stresses, impairing yield and fiber quality. NAC (NAM, ATAF1,2, and CUC2) protein widely distributed in land plants, play the critical role in responding to abiotic stress and regulating leaf senescence. We have identified and functional analyzed a NAM domain gene GhNAC82 in upland cotton, it was located on the A11 chromosome 4,921,702 to 4,922,748 bp, only containing one exon. The spatio-temporal expression pattern analysis revealed that it was highly expressed in root, torus, ovule and fiber development stage. The results of qRT-PCR validated that GhNAC82 negatively regulated by salt stress, drought stress, H2O2 stress, IAA treatment, and ethylene treatment, positively regulated by the ABA and MeJA treatment. Moreover, heterologous overexpression of GhNAC82 results in leaf premature senescence and delays root system development in Arabidopsis thaliana. The phenotype of delayed-senescence was performed after silencing GhNAC82 by VIGS in premature cotton. Taken together, GhNAC82 was involved in different abiotic stress pathways and play important roles in negatively regulating leaf premature senescence.

Lactobacillus improves meat quality in Sunit sheep by affecting mitochondrial biogenesis through the AMPK pathway.

Sunit sheep are famous for their high meat quality, but the meat quality of them has declined due to the change in feeding methods. Lactobacillus has a variety of probiotic effects and is widely used in animal diets to optimize meat quality. This study aimed to investigate the effect of dietary supplementation with different levels of Lactobacillus on meat quality. A total of 24 3-month-old Sunit sheep with an average body weight of 19.03 ± 3.67 kg were randomly divided into control (C), 1% (L1), 2% (L2), and 3% Lactobacillus groups (L3), with 6 sheep in each group. Myofiber characteristics, meat quality, and metabolic enzyme activity were detected. Moreover, the regulatory mechanism of Lactobacillus on meat quality was explored by using Western blotting and real-time Quantitative polymerase chain reaction (RT-qPCR). The results showed that dietary addition of Lactobacillus decreased LDH activity in the Biceps femoris of Sunit sheep (P < 0.05). Compared to the other groups, the 1% Lactobacillus group showed the conversion of myofibers from the glycolytic to the oxidative type, and the increasing b* values (P < 0.05), decreasing shear force and cooking loss of meat (P < 0.05) and the relative gene and protein expression levels of AMPK, PGC-1α, NRF1, TFAM, and COX IV (P < 0.05) in the Biceps femoris were also increased in the 1% Lactobacillus group. Therefore, the addition of Lactobacillus to the diet of Sunit sheep could regulate the AMPK signaling pathway to promote myofiber type conversion, which improves meat quality. This study provided a theoretical and data basis for improving the meat quality of sheep and supplied a novel way of applying Lactobacillus.

Monitoring the distribution of internalized silica nanoparticles inside cells via direct stochastic optical reconstruction microscopy.

Understanding the exact localization of nanoparticles within cell is of particular importance for rational design of high-effective nanomedicines. In the present study, direct stochastic optical reconstruction microscopy (dSTORM) is employed to elucidate the precise localization of nanoparticles within cells owing to its superiority of nanometric resolution, multicolour ability and minimal invasiveness. The localization of the Cy5 labelled mesoporous silica nanoparticles (MSNs-Cy5) in MCF-7 cells are monitored by dSTORM and conventional fluorescence microscopy, respectively. The dSTORM images demonstrate much higher spatial resolution for locating MSNs-Cy5 within cells compared to that of the conventional fluorescence images. Moreover, the distribution of MSNs-Cy5 within three cell lines over time are obtained. For the MCF-7 and HeLa cells, MSNs-Cy5 nanoparticles distribute nearly all around the cytoplasm after 5 h incubation. In contrast, MSNs-Cy5 nanoparticles within NIH 3T3 cells are quite different that they are found to be either attached to or embedded into cell membranes, without penetrating into the cytoplasm. Overall, we provide a practical method to reveal the in situ precise imaging of nanoparticles in cells with nanometric resolution precision. This method may open up new opportunities for organelle-specific targeting drug delivery to achieve maximum therapeutic benefit.

The Application Value of Metagenomic and Whole-Genome Capture Next-Generation Sequencing in the Diagnosis and Epidemiological Analysis of Psittacosis.

Background: To evaluate the value of metagenomic next-generation sequencing (mNGS) for the early diagnosis of psittacosis, and to investigate its epidemiology by whole-genome capture. Methods: Twenty-one bronchoalveolar lavage fluid (BALF) and blood samples of 16 psittacosis patients from multiple centers during August 2019 to September 2021 were analyzed retrospectively. mNGS with normal datasets (10 M 75-bp single-end reads after sequencing) and larger datasets (30 M 150-bp paired-end reads after sequencing) as well as quantitative real-time polymerase chain reaction (qPCR) were used to detect the pathogen. Also, whole-genome capture of Chlamydophila psittaci was applied to draw the phylogenetic tree. Results: mNGS successfully detected the pathogen in all 16 cases (100%), while qPCR was positive only in 5 out of 10 cases (50%), indicating a significantly higher sensitivity of mNGS than qPCR (p < 0.01). BALF-mNGS performed better than blood-mNGS (16/16 versus 3/5, p < 0.05). In addition, larger datasets (the read counts have tripled, and the base number was 12-fold larger compared to clinical mNGS with a normal dataset) of mNGS showed significantly increased contents of human DNA (p < 0.05) and decreased reads per million of the pathogen, suggesting no improvement. Whole-genome capture results of five samples (>60% coverage and >1 depth) were used to construct the phylogenetic tree. Conclusion: Significant advantages of mNGS with normal datasets were demonstrated in early diagnosing psittacosis. It is the first study to use whole-genome capture to analyze C. psittaci epidemiological information.

Dopamine-Mediated Biomineralization of Calcium Phosphate as a Strategy to Facilely Synthesize Functionalized Hybrids.

Organic-inorganic hybrid materials have been considered to be promising carriers or immobilization matrixes for biomolecules due to their high efficiency and significantly enhanced activities and stabilities of biomolecules. Here, the well-defined dopamine/calcium phosphate organic-inorganic hybrids (DACaPMFs) are fabricated via one-pot dopamine-mediated biomineralization, and their structure and properties are also characterized. Direct stochastic optical reconstruction microscopy (dSTORM) is first used to probe the distribution of organic components in these hybrids. Combined with spectroscopic data, the direct observation of dopamine in the hybrids helps to understand the formation of a physical chemistry mechanism of the biomineralization. The obtained DACaPMFs with multiple-level pores allow the loading of doxorubicin with a high loading efficiency and a pH-responsive property. Furthermore, thrombin is entrapped by the hybrids to prove the controlled release. It is expected that such organic-inorganic hybrid materials may hold great promise for application in drug delivery as well as scaffold materials in bone tissue engineering and hemostatic material.