Soluble Protein Research Articles

Advances in Research on the Improvement of Low-Salt Meat Product Through Ultrasound Technology: Quality, Myofibrillar Proteins, and Gelation Properties.

The high sodium content in meat products poses health risks to consumers and does not align with modern green and healthy living standards. Current strategies for directly reducing the sodium content in meat products are limited by their negative impact on the sensory or quality attributes of the products. In recent years, there has been great interest in applying ultrasound technology to reduce sodium content. This paper discusses the advantages and disadvantages of current mainstream strategies for reducing the sodium content in meat products, as well as the potential mechanisms by which ultrasound-assisted marination improves the quality of low-salt meat products. The main findings indicate that ultrasound, through its cavitation and mechanical effects, facilitates the transition of proteins from stable insoluble aggregates to stable soluble complexes, exposing more hydrophilic groups and, thus, enhancing protein solubility. At the same time, ultrasound promotes a greater number of proteins to participate in the formation of interfacial layers, thereby increasing emulsifying activity. Furthermore, ultrasound treatment promotes the interaction between proteins and water, leading to partial unfolding of protein chains, which allows polar residues to more readily capture water in the gel, thereby improving the water-holding capacity of the gel. These effects will contribute to the formation of high-quality low-salt meat products. However, variations in the frequency, intensity, and duration of ultrasound treatment can lead to differing effects on the quality improvement of low-salt meat products.

Cardiac Amyloidosis: State-of-the-Art Review in Molecular Pathology.

Amyloidosis refers to a group of diseases caused by extracellular deposits of misfolded proteins, which alter tissue function and structure, potentially affecting any organ. The term "amyloid" was introduced in the 19th century and later associated with pathological protein deposits. Amyloid fibrils, which are insoluble and resistant to degradation, originate from soluble proteins that undergo misfolding. This process can be triggered by several factors, such as aging, elevated protein concentrations, or pathogenic variants. Amyloid deposits damage organs both by disrupting tissue architecture and through direct cytotoxic effects, leading to conditions such as heart failure. Amyloidosis can be classified into acquired or inherited forms and can be systemic or localized. Diagnosing cardiac amyloidosis is complex and often requires tissue biopsies, which are supported by Congo Red dye staining. In some cases, bisphosphonate bone scans may provide a less invasive diagnostic option. In this state-of-the-art review, we focus on the most common forms of cardiac amyloidosis, from epidemiology to therapy, emphasizing the differences in molecular mechanisms and the importance of pathological diagnosis for appropriate treatment using a multidisciplinary approach.

Physiological and Transcriptomic Analyses Reveal the Role of the Antioxidant System and Jasmonic Acid (JA) Signal Transduction in Mulberry (Morus alba L.) Response to Flooding Stress

In recent decades, the frequency of flooding has increased as a result of global climate change. Flooding has become one of the major abiotic stresses that seriously affect the growth and development of plants. Mulberry (Morus alba L.) is an important economic tree in China. Flooding stress is among the most severe abiotic stresses that affect the production of mulberry. However, the physiological and molecular biological mechanisms of mulberry responses to flooding stress are still unclear. In the present study, reactive oxygen species (ROS) metabolism, antioxidant mechanism, and plant hormones in mulberry associated with the response to flooding stress were investigated using physiological and transcriptomic analysis methods. The results showed significant increases in the production rate of superoxide anion (O2•−) and the content of hydrogen peroxide (H2O2) in leaves on the 5th day of flooding stress. This led to membrane lipid peroxidation and elevated malondialdehyde (MDA) levels. Antioxidant enzymes such as catalase (CAT), superoxide dismutase (SOD), and peroxidase (POD) exhibited enhanced activities initially, followed by fluctuations. The ascorbic acid–glutathione (AsA-GSH) cycle played a crucial role in scavenging ROS, promoting the reduction of oxidized glutathione (GSSG) to reduced glutathione (GSH). Transcriptomic analysis revealed the up-regulation of the gene-encoding antioxidant enzymes (APX, MDHAR, GPX, GR, GST) involved in ROS scavenging and stress tolerance mechanisms. Jasmonic acid (JA) levels and the expression of JA synthesis-related genes increased significantly in mulberry leaves under flooding stress. This activation of the JA signaling pathway contributed to the plant’s adaptability to flooding conditions. Proline (Pro) and soluble sugar (SS) contents increased notably in response to flooding stress. Proline helped maintain cell turgor and protected enzymes and membranes from damage, while soluble sugars supported anaerobic respiration and energy supply. However, soluble protein (SP) content decreased, suggesting inhibition of protein synthesis. The study provides insights into mulberry’s flooding tolerance mechanisms, guiding future molecular breeding efforts. This summary captures the key findings and implications of the study on mulberry’s response to flooding stress, focusing on physiological and molecular mechanisms identified in the research.

Biochemical Analysis of Different Wheat Varieties Grown under Various Zones of India

Background: The study aimed to evaluate the flour quality of different wheat varieties (Triticum durum L.) for various end-use products such as chapati, bread, biscuits and pasta. The focus was on identifying varieties with superior nutritional and processing qualities by analyzing various qualitative parameters. Methods: The study assessed multiple wheat varieties using a range of qualitative parameters, including moisture, ash, total soluble sugar, starch, protein, lysine, tryptophan, dry and wet gluten, gluten index, water binding capacity, β-carotene content, sedimentation value, thousand kernel weight and mineral content (Fe and Zn). The wheat varieties evaluated included A-206, GW-1, HI 8498 (Kota), PDW 291 (Hissar) and NIDW 295 (Pune). Result: The study identified A-206 and GW-1 as wheat varieties with superior protein content, gluten quality and water binding capacity, making them ideal for various end-use products. A-206 also had the highest lysine and tryptophan levels, while GW-1 and HI 8498 (Kota) had elevated ash content. PDW 291 (Hissar) was notable for its higher total soluble sugar and NIDW 295 (Pune) excelled in starch content, β-carotene and thousand kernel weight. Overall, NIDW 295 showed the most promise for pasta production due to its comprehensive quality traits.

Growth, physiological and N, P, K accumulation responses of Erythropalum scandens Bl. Seedlings under different substrates

Erythropalum scandens Bl. is a medicinal woody vegetable found in southern China and parts of Southeast Asia. Studies have shown improper substrate hindered E. scandens seedling growth, causing water accumulation and nutrient deficiency. In pursuit of an ideal growth medium for E. scandens seedlings during the early stages, this study conducted a pot experiment to identify a mixed substrate with optimal water permeability and fertility. In this study, pure Alfisols soil treatment as the control (CK), and two soilless substrates (peat soil and perlite) were combined with Alfisols soil into different volume ratios, in order to better use soil resources from understory space and balance the texture of mixed substrates. The growth, physiological characteristics and nutrient status of 24-month-old E. scandens seedlings were determined after planting in different mixed ratios. The results showed that as the proportion of peat soil increased in the mix, most indexes exhibited an initial increase followed by a decline, while soluble protein content decreased consistently. Conversely, an increasing perlite ratio resulted in a general decline in most growth and physiological indexes. Root growth, biomass accumulation and chlorophyll content, peaked in the 66.67% Alfisols soil + 33.33% perlite (T4) treatment. Notably, T3 (66.67% Alfisols soil + 33.33% peat soil) showcased the best above-ground growth, while T1 (50.00% Alfisols soil + 50.00% peat soil) excelled in element content accumulation. In conclusion, the cultivation substrate should primarily consist of Alfisols soil, constituting at least 50%. The addition of peat soil enhances above-ground growth and nutrients accumulation, while perlite contributes to robust root development. One third of peat soil and a small amount of perlite can be added to the substrate during E. scandens seedling cultivation, and proper fertilization should also be used in order to increase nutrient accumulation in aboveground and underground parts. This research provides valuable insights into maximizing the potential of E. scandens seedlings through precise cultivation methods.

Combined effect of molybdenum and nitrogen fertilization on nitrogen metabolism and amino acid content in tobacco leaves

IntroductionThis study investigated the combined effects of molybdenum (Mo) and nitrogen (N) fertilization on N metabolism and amino acid content in the leaves of flue-cured tobacco (Nicotiana tabacum L.) during its mature stage through a pot experiment.MethodsDifferent application levels of Mo (0, 0.15, 0.30 mg/kg soil) and N (0.06, 0.24, 0.42 g/kg soil) were set to observe and analyze changes in leaf quality, N, and amino acid content in the tobacco plants.ResultsThe results revealed that the N fertilizer application level exhibited a primary effect on regulating the total nitrogen, nitrate nitrogen, soluble protein, and amino acid nitrogen concentrations within tobacco leaves, while the effectiveness of Mo fertilization was influenced by the level of N applied. Specifically, under the conditions of 0.24 g/kg soil N and 0.30 mg/kg soil Mo application, the N content, N accumulation, and dry matter mass of the tobacco plants increased significantly by 110%, 204%, and 48%, respectively. Concurrently, nitrate reductase activity increased by 107-fold, and the nitrate nitrogen content was relatively low, contributing to enhanced tobacco yield and safety. Moreover, this treatment led to a notable (170%) increase in free amino acid nitrogen content, with minimal impact on total amino acids and soluble proteins. Notably, it effectively increased the content of free amino acids beneficial to the sensory quality of tobacco (such as histidine, arginine, aspartic acid, isoleucine, and glutamic acid) without reducing the total amino acid content, while simultaneously reducing other amino acids that might affect quality. Therefore, the combined application of 0.30 mg/kg soil Mo and 0.24 g/kg soil N specifically optimized the amino acid composition in tobacco leaves, positively impacting overall quality and market competitiveness.DiscussionThis study provides a theoretical basis for the rational application of Mo fertilizer in Mo-deficient areas to improve the yield and quality of flue-cured tobacco.

Recombinant Plasminogen Activator of the Sandworm (Perinereis aibuhitensis) Expression in Escherichia coli.

As an essential thrombolytic agent, the tissue plasminogen activator receives increasing attention due to its longer half-life, lower immunogenicity, and easier administration, which are superior to other thrombolytic agents. In this study, the isolated and purified plasminogen activator from the sandworm (Perinereis aibuhitensis) was expressed in E. coli (Escherichia coli) to investigate its potential for simplifying the development process. The sandworm plasminogen activator was previously successfully cloned and expressed in E. coli with low yield and activity in the culture supernatant. This low yield and activity prompted us to optimize its DNA sequence. Furthermore, to raise the efficiency in the separation of the target protein, the protein's solubility was enhanced by fusing it with maltose-binding protein (MBP) tags. Eventually, the fibrinolytic activity was successfully restored after digestion with tobacco etch virus (TEV) protease. This study provides an innovative method of efficiently expressing and purifying plasminogen activators from sandworm in E. coli and broadens its applications in therapeutic treatment of cardiovascular diseases, including thrombosis, stroke, and coronary atherosclerotic heart disease.

Mitigation of microplastic toxicity in soybean by synthetic bacterial community and arbuscular mycorrhizal fungi interaction: Altering carbohydrate metabolism, hormonal transduction, and genes associated with lipid and protein metabolism

Microplastics (MPs) have become ubiquitous environmental pollutants, eliciting concerns about their negative impacts on terrestrial and agroecosystems. However, employing sustainable remediation approaches holds the potential to mitigate these negative impacts and bolster MPs tolerance in both the seeds and vegetative structures of plants. The current investigation explores the impact of arbuscular mycorrhizal fungi (AMF) and synthetic bacterial community (SynCom) as potential alleviators of MP-induced stress. Results exhibited that co-applied SynCom and AMF, resulted in the highest plant biomass, leaf area, pod per plant, and 100-seed weight under control (no MPs) and MPs stress. The combined application of SynCom and AMF enhanced colonization, association, and arbuscule abundance in the non-stressed plants (no MPs) as compared to stressed plants. The sole or co-application of SynCom and AMF enhanced the primary metabolites content and mitigated the MPs-induced reduction in soluble sugars, lipids, protein, and oil contents. Soybean inoculation stimulated the genes expression involved in lipid and protein biosynthesis, while a contradictory drift was noted for genes associated with lipid and protein degradation, underpinning the observed increment in protein and lipid content. Soybean inoculated with AMF exhibited the utmost α-amylase and β-amylase activities, demonstrating enhanced osmolyte (soluble sugar) production, mainly under MPs stress. Remarkably, sole or co-application further reinforces the positive effect of MPs stress on the osmoprotectants and antioxidant levels, for instance, phenol, flavonoid, glycine betaine contents, and glutathione-S-transferase (GST) activities. Subsequent to the stress release, a stress hormones known as abscisic acid (ABA) reduced in the seeds of inoculated soybean while gibberellin (GA), trans-zeatin riboside (ZR), and indole acetic acid (IAA) were increased. Therefore, it is concluded that the combined application of SynCom and AMF protected the soybean plants from MPs-induced oxidative damage by upregulating osmoprotectants and antioxidant levels. This study provides insights into the potential of sustainable remediation approaches in alleviating the complex impact of MPs on soybean, contributing to future strategies for environmental sustainability. Future studies could explore the optimization of SynCom and AMF formulations for different crops and environmental conditions, potentially leading to practical applications in sustainable agriculture and pollution management.

Spray Drying of Inca Peanut Meal Protein Hydrolysate to Produce Protein Powder Drink Mix

The optimal conditions (maltodextrin concentration and inlet air temperature) for spray drying Inca peanut meal protein hydrolysate (IMPH) were determined. The maltodextrin concentration was varied at 5, 10 and 15 % by weight, and inlet air temperature was varied at 150, 160 and 170 °C. The IMPH was subjected to spray drying, where the feed solution was atomized into fine droplets and exposed to a hot air stream, leading to rapid drying and the formation of powder. The moisture, protein, yield, water activity and water solubility index (WSI) of IMPH powder were analyzed. The optimal condition for spray drying IMPH powder utilized 10 % maltodextrin and an inlet air temperature of 160 °C. Under these conditions, the moisture, protein, yield and WSI of IMPH powder were 2.93, 36.93, 38.95 and 99.59 %, respectively, while water activity was 0.41. IMPH powder particles were spherical and non-agglomerate. Inca peanut meal protein powder drink mix (IMPD) with cocoa flavor was the most acceptable. The IMPD with cocoa flavor could be promoted as a high protein and high dietary fiber product. It could be kept at 35 ± 2 °C for 12 weeks, whilst retaining good microbiological properties. HIGHLIGHTS The optimal conditions for spray drying Inca peanut meal protein hydrolysate (IMPH) were determined to be 10% maltodextrin and an inlet air temperature of 160 °C. This resulted in high yield, protein content, and water solubility, while keeping moisture and water activity low. IMPH powder had spherical, non-agglomerated particles, and cocoa-flavored Inca peanut protein powder drink mix (IMPD) was the most preferred in sensory evaluations. The IMPD with cocoa flavor was identified as a high-protein, high-dietary-fiber product, suitable for storage at 35 °C for up to 12 weeks while maintaining good microbiological quality. The study highlights the nutritional benefits of the cocoa-flavored IMPD, which includes essential amino acids, omega-3 fatty acids, and antioxidant properties. GRAPHICAL ABSTRACT

Influence of Temperature and Host Plant on the Digestion of Frankliniella intonsa (Trybom) Revealed by Molecular Detection.

Frankliniella intonsa (Trybom) (Thysanoptera: Thripidae) is an important type of thrip and a polyphagous pest, which poses a serious threat to many crops, especially those in tropical regions of China. Its feeding behavior and the damage caused vary among different host plant species and are affected by ambient temperature and plant nutrients as well. The digestion rate is an important index for directly observing the digestion process, but there have been no studies directly measuring the digestion in thrips under the influence of different temperatures and host plants. Here, the digestion rate of F. intonsa was assessed by using a molecular diagnostic tool. We also determined the nutrient content in three host plant (mango, cowpea, and pepper), including soluble proteins, free fatty acids, soluble sugars, and water. The results showed that the high and low temperatures (16 °C and 32 °C) both seemed to accelerate the digestion of F. intonsa compared to the optimal temperature (26 °C) and the protein content of plants played an important role in the digestive response of F. intonsa to temperature changes. The findings can help reveal the feeding damage caused by F. intonsa to different plants and help to better understand its feeding ecology, according to its interaction with the host plant.

Effects of Salt Stress on Seed Germination and Embryo Growth of Oxytropis coerulea

Background: Oxytropis coerulea is considered to be an exceptional forage. The salinization of soil is a prevalent ecological issue worldwide. In order to study the effects of saline solution on seed germination and embryo growth of O. coerulea. Methods: An experiment was carried out using the culture dish filter paper method. Three salt stresses, namely, Na2CO3, NaCl and double salt were set, each of which was set to 0%, 0.3%, 0.6% and 0.9% concentration. Result: The results indicated that as the concentration of salt increased, there was a decrease in the germination rate, germination index and embryo growth. The activities of SOD, POD and CAT, as well as soluble protein content, initially increased and then decreased with increasing salt stress. The toxic effects of the three salts ranged from strong to weak as Na2CO3 greater than double salt greater than NaCl. The salt resistance indexes of 0.3% Na2CO3, NaCl and double salt were 53.87, 57.00 and 38.73, respectively and the salt resistance index of 0.6% NaCl was 50.21, indicating a high salt-tolerant degree. It showed certain resistance to 0.3% Na2CO3, 0.6% NaCl and 0.3% double salt. In short, the various soluble mechanisms of O. coerulea in different stages of salt stress and different salt stress levels have different reaction mechanisms, which reflect the salt stress of O. coerulea seeds. O. coerulea seeds can be planted in a suitable concentration of saline-alkali land, which can utilize the feed value and medicinal value of O. coerulea to improve economic benefits and utilize saline soil to improve the ecological environment.

Establishment, Multiplication, and Biochemical Analysis of Embryogenic Lines of the Amazonian Palm Euterpe precatoria Mart. under Suspension Culture

The palm Euterpe precatoria holds great social, cultural, and environmental importance. The heart of palm and the fruit are the main products used for industrialization due to their energetic properties. Thus, the aim of this study was to establish a suspension cultivation protocol for the species using different explant sources. For this, eight lineages of E. precatoria embryogenic calluses were tested, with five in liquid medium Murashige and Skoog (MS) with 5 μM Picloram and three for comparison in semisolid medium MS with 20 μM Picloram and 5 μM 2iP. The growth curve was obtained by weighing the calli from 60 to 180 days of cultivation. The Gompertz model was applied, and growth kinetics were evaluated. At 100 days, the contents of total soluble sugars (TSSs) and total soluble proteins (TSPs) were determined. Principal components (PCA) were measured. According to the analysis of the data, the cultivation of E. precatoria lineages in liquid medium was successfully carried out, and the establishment was achieved. The model can be considered adequate since the R2 values found describe more than 90% of the growth kinetics of the lineages. In the liquid system, lineages L1 (from leaf explants and multiplied in semisolid medium—SM), L2 (from leaf explants and multiplied in SM), and L6 (from zygotic embryo explants and multiplied in liquid medium—LM) showed the shortest time to double the biomass accumulation. Multivariate analysis reveals a significant increase in masses in liquid cultures, represented by lineages L6 and L2. There was statistical difference in the amount of TSSs extracted; the highest TSS levels were observed in lineages cultivated in LM. The protein content found was very low, showing statistical differences among the lineages. In this work, the establishment and multiplication of embryogenic calli of E. precatoria are described for the first time, and they emerge as viable alternatives for the vegetative propagation of the species.

Understanding the role of soluble proteins and exosomes in non-invasive urine-based diagnosis of preeclampsia

Preeclampsia is a hypertensive disorder of pregnancy that can lead to stillbirth and preterm birth if not treated promptly. Currently, the diagnosis of preeclampsia relies on clinical symptoms such as hypertension and proteinuria, along with invasive blood tests. Here, we investigate the role of soluble proteins and exosomes in noninvasive diagnosing preeclampsia non-invasively using maternal urine and urine-derived exosomes. We quantified the levels of particles and the presence of TSG101 and CD63 in urine and urinary exosomes via the biologically intact exosome separation technology (BEST) platform. Then, we obtained higher levels of soluble proteins such as fms-like tyrosine kinase-1 (sFlt-1) and placental growth factor (PlGF) from urine as it was than urinary exosomes. Compared to commercial blood tests, the sensitivity of the sFlt-1/PlGF ratio was found to be 4.0 times higher in urine tests and 1.5 times higher in tests utilizing urine-derived exosomes. Our findings offer promising possibilities for the early and non-invasive identification of high-risk individuals at risk of preeclampsia, allowing for comprehensive preventive management.

Pre-fusion AAA+ remodeling of target-SNARE protein complexes enables synaptic transmission.

Membrane fusion is driven by SNARE complex formation across cellular contexts, including vesicle fusion during synaptic transmission. Multiple proteins organize trans-SNARE complex assembly and priming, leading to fusion. One target membrane SNARE, syntaxin, forms nanodomains at the active zone, and another, SNAP-25, enters non-fusogenic complexes with it. Here, we show that the AAA+ protein NSF (N-ethylmaleimide sensitive factor) and SNAP (soluble NSF attachment protein) must act prior to fusion. We show that syntaxin clusters are conserved, that NSF colocalizes with them, and characterize SNARE populations within and near these clusters using cryo-EM. Supercomplexes of NSF, α-SNAP, and either a syntaxin tetramer or two binary complexes of syntaxin-SNAP-25 reveal atomic details of SNARE processing and show how sequential ATP hydrolysis drives disassembly. These results suggest a functional role for syntaxin clusters as reservoirs and a corresponding role for NSF in syntaxin liberation and SNARE protein quality control preceding fusion.

Exogenous γ-aminobutyric acid (GABA) enhances rye (Secale cereale) seedling resistance to combined freeze-thaw and cadmium stress.

Freeze-thaw is a common stress at high altitudes in northern China. There is a risk of cadmium (Cd) contamination in the region. γ-aminobutyric acid (GABA) is a natural product that regulates plant growth. Rye (Secale cereale ) was used as research material to investigate the physiological effects of exogenous GABA on rye seedlings under the single and combined stresses of freeze-thaw and cadmium. The results showed that the combined stress severely inhibited shoot length, root length, fresh weight, and dry weight, increased malondialdehyde and hydrogen peroxide contents, and significantly decreased superoxide dismutase (SOD) activity. Foliar application of 5mM GABA alleviated the negative effects of stress on seedling growth, increased soluble protein content, and reduced malondialdehyde and hydrogen peroxide contents. Exogenous GABA application also enhanced the activities of SOD and peroxidase (POD). Additionally, the presence of exogenous GABA activated the GABA metabolic process and encouraged the accumulation of phytochelatins, glutathione, and non-protein thiol. These results indicate that exogenous GABA can effectively improve the resistance of rye seedlings to freeze-thaw and Cd by regulating the antioxidant enzyme system and enhancing its own detoxification mechanism, and they provide a basis for future applications of exogenous GABA, which is beneficial for ecological protection.

Productive, Morphological and Nutritional Indicators of Cactus Pear in a Semiarid Region

This study aimed to evaluate the effect of irrigation depths (IDs) with brackish water and levels of organic fertilizer (OF) on the morphological aspects, production, chemical composition and in vitro digestibility of cactus pear grown in a semiarid region. This experiment was conducted in an area already established for 2 years with cactus pear [Opuntia stricta (Haw.)] and started 18 months after the standardization cut. This was a 5 × 4 factorial design of five irrigation depths (0, 12.5, 25, 37.5 and 50% evapotranspiration—ETo) and four levels of organic fertilizer (0, 15, 30, and 45 megagram per hectare—Mg/ha), with four replications. The water used in the experiment was classified with high salinity and low sodium content (C3S1), and presented an electrical conductivity of 1.73 dS/m. There was no effect of the ID × OF interaction on the morphological and productive characteristics of cactus pear (p > 0.05); however, the ID × OF interaction promoted effects on ether extract, crude protein, neutral detergent insoluble protein, neutral detergent insoluble ash, total carbohydrates and non-fiber carbohydrates (p < 0.05). The ID and OF levels separately influenced the productive, morphological and nutritional characteristics of cactus pear. Under experimental conditions, we recommend the use of organic fertilizer at a level up to 45 Mg/ha, which is associated with the use of lower brackish water levels for the cultivation of cactus pear. This study’s findings provide new insights into reducing the use of potable water in crop irrigation for dryland regions and other regions.

In silico and in vitro evaluation of a PE38 and Nb-based recombinant immunotoxin targeting the GRP78 receptor in cancer cells.

Cancer is a global health problem despite the most developed therapeutic modalities. The delivery of specific therapeutic agents to a target increases the effectiveness of cancer treatment by reducing side effects and post-treatment issues. Our aim in this study was to design a recombinant protein consisting of nanobody molecules and exotoxin that targets the surface GRP78 receptor on tumor cells. Bioinformatics methods make drug design and recombinant protein evaluation much easier before the laboratory steps. Two constructs were designed from a single-variable domain on heavy chain nanobody domains and PE toxin domains II, Ib, and III. The physicochemical properties, secondary structure, and solubility of the chimeric protein were analyzed using different software. Prostate cancer DU-145 and breast cancer MDA-MB-468 cell lines were used as GRP78-positive and negative controls, respectively. Accordingly, the cytotoxicity, binding affinity, cell internalization, and apoptosis were evaluated using MTT, enzyme-linked immunosorbent assay, and western blot. The results showed that in the DU-145 cell line, the cytotoxicity of two recombinant immunotoxins is dose and time-dependent. In MDA-MB-468 and HEK-293 cells, such an event does not occur. It is possible that two constructs designed for immunotoxins can attach to GRP78-positive cancer cells and then eradicate cancer cells by internalization and apoptosis. As our in vitro results were in line with in silico data confirming the Bioinformatics predictions, it can be concluded that the designed recombinant immunotoxins may exhibit therapeutic potential against GRP78-positive tumor cells.

Exploring Plant Growth-Promoting Traits of Endophytic Fungi Isolated from Ligusticum chuanxiong Hort and Their Interaction in Plant Growth and Development.

Endophytic fungi inhabit various plant tissues and organs without inducing evident disease symptoms. They can contribute positively to the growth of plants, bolster plants resilience to environmental and biological stresses, and facilitate the accumulation of secondary metabolites. These microbial resources possess significant developmental and utilization value in various applications. Hence, this study focused on exploring the plant growth-promoting (PGP) traits of 14 endophytic fungi from Ligusticum chuanxiong Hort (CX) and elucidating the effects and mechanisms that facilitate plant growth. According to PGP activity evaluation, the majority of strains demonstrated the capacity to produce IAA (78.57%), siderophores (50.00%), ammonia (35.71%), potassium solubilization (21.43%), nitrogen fixation (57.14%), and phosphate solubilization (42.86%). Further investigations indicated that the levels of IAA ranged from 13.05 to 301.43 μg/mL, whereas the soluble phosphorus levels ranged from 47.32 to 125.95 μg/mL. In cocultivation assays, it was indicated that Fusarium sp. YMY5, Colletotrichum sp. YMY6, Alternaria sp. ZZ10 and Fusarium sp. ZZ13 had a certain promoting effect on lateral root number and fresh weight of tobacco. Furthermore, ZZ10 and ZZ13 significantly enhanced the germination potential, germination index, and vigor index of tobacco seeds. The subsequent potted trials demonstrated that the four endophytic fungi exhibited an enhancement to growth parameters of tobacco to a certain extent. ZZ10 and ZZ13 treatment had the best promotion effect. Inoculation with YMY5 increased the chlorophyll a and total chlorophyll content. ZZ10 and ZZ13 treatment remarkably increased the net photosynthetic rate, soluble sugars and soluble protein content, catalase and peroxidase activities, and lowered malondialdehyde content in tobacco leaves. In addition, YMY5 remarkably elevated superoxide dismutase activities. ZZ13 upregulated the expression of growth-related gene. Among them, ZZ13 had a better growth-promoting effect. In conclusion, these endophytic fungi possessing multi-trait characteristics and the capacity to enhance plant growth exhibit promising potential as biofertilizers or plant growth regulators.

Genome-wide analysis of glutathione S-transferase genes in four Prunus species and the function of PmGSTF2, activated by PmMYBa1, in regulating anthocyanin accumulation in Prunus mume

Glutathione S-transferases (GSTs) are proteases with multiple physiological functions and play an important role in plant responses to abiotic stresses. Nevertheless, there is a paucity of systematic research on GST genes in Prunus genus. Here, 330 GST genes in four Prunus species were identified for the first time and classified into eight subgroups based on protein sequence and conserved structure, among which Tau subfamily genes had the largest number. The amino acid lengths of GST-encoded proteins in the four species ranged from 66 to 1152 aa, most of which were soluble proteins and located in the cytoplasm and chloroplasts. The GST family was propelled by tandem duplications, yet robust purifying selection constrained its divergence. Conserved motif and domain analysis revealed that the majority of PmGSTs exhibited a highly conserved GST-N structure. The expression pattern of PmGSTs exhibited tissue specificity and spatiotemporal specificity. qRT-PCR validated the transcriptome results and 11 genes were differentially expressed in varieties with different flower and stem colors. In addition, we discovered an anthocyanin-related gene PmGSTF2, which can effectively restore the anthocyanin and proanthocyanidin deficiency-related phenotypes of the Arabidopsis tt19 mutant. Recombinant PmGSTF2 enhanced the water solubility of cyanidin and cyanidin-3-O-glucoside in vitro. Moreover, PmMYBa1 could directly bind to the promoter of PmGSTF2 and activate its expression. The findings revealed that GSTs were preserved in Prunus species and that PmGSTF2 was critical in regulating anthocyanin accumulation.

Effects of Combined Biochar and Chemical Fertilizer Application on Soil Fertility and Properties: A Two-Year Pot Experiment

A two-year pot experiment was conducted to investigate the effects of the combined application of biochar and chemical fertilizer on soil quality and vegetable growth by adding different proportions of chemical fertilizer and biochar to the soil in 2022 and no fertilizer in 2023. It was concluded that the combined treatment improved the soil’s properties. After two consecutive years of planting vegetables, the improvement of soil properties was the most significant with the 1.5 g biochar + 80% chemical fertilizer optimal fertilizer application (BCF6) treatment. In comparison to the control (CK), soil pH, electrical conductivity, and dissolved organic carbon increased by 0.59 units, 166.6%, and 282.6%, respectively. Soil fertility also improved significantly, indicating that the combined treatments resulted in the slow release of nutrients to enhance the effectiveness of the fertilizers. Co-application significantly increased the yield of the edible parts of Chinese cabbage and improved its quality. The most significant effects of vitamin C content and soluble protein were observed in Chinese cabbage under BCF6 treatment, which were 3.33 and 1.42 times more than the CK, respectively. Utilizing biochar as a partial substitute for chemical fertilizers can improve soil structure and fertility over the long term while reducing the reliance on chemical fertilizers, ultimately providing sustained economic and ecological benefits for agricultural production.