Soluble Protein Research Articles

Effects of host plants on aphid feeding behavior, fitness, and Buchnera aphidicola titer.

Aphids are sap-feeding plant pests that depend on their symbiotic relationships with the primary endosymbiont Buchnera aphidicola to adapt to impoverished diets. However, how the host plant affects the aphid primary symbiont and aphid adaptation to host plant transfer are poorly known. In this study, aphid symbiont screening and genotype identification were used to establish 2 aphid strains (Rhopalosiphum maidis [Rm] and Rhopalosiphum padi [Rp] strains) containing only Buchnera without any secondary symbionts for both wheat aphid species (R. maidis and R. padi). Aphid fitness and Buchnera titers were unstable on some of these host plants after transferring to novel host plants (G1-G5), which were influenced by host plant species and generations; however, they stabilized after prolonged feeding on the same plants for 10 generations. The electropenetrography (EPG) records showed that the allocation of aphid feeding time was significantly distinct in the 6 host plants; aphids had more intracellular punctures and spent more nonprobing time on green bristlegrass which was not conducive to its growth compared with other plants. The content of soluble sugar, soluble protein, and amino acid in the leaves of the 6 host plants were also clearly separated. The correlation coefficient analysis showed that the nutrient contents of host plants had significant correlations with aphid feeding behaviors, fitness, and Buchnera titers. In the meantime, aphid fitness, and Buchnera titers were also affected by aphid feeding behaviors. Also, Buchnera titers of aphid natural populations on 6 host plants showed a visible difference. Our study deepened our understanding of the interaction among aphids, endosymbionts, and host plants, indicating that the host plant nutrient content is a predominant factor affecting aphid adaptation to their diet, initially affecting aphid feeding behaviors, and further affecting aphid fitness and Buchnera titers, which would further contribute to exploiting new available strategies for aphid control.

Comparison of plasma-activated saline prepared with plasma gases with different N2/O2 ratios activated by gliding arc discharge

Methicillin-resistant Staphylococcus aureus (MRSA) presents a significant threat due to the multiple resistance to antibiotics, leading to severe and challenging-to-treat infections. Plasma-activated saline (PAS) prepared by plasma gases, could efficiently inactivate various pathogenic bacteria including both sensitive and antibiotic-resistant bacteria. In this study, the PAS was prepared by plasma gases with different ratios of N2 and O2 activated by gliding arc discharge. First, the gaseous reactive species in the plasma gases were compared, revealing that the highest levels of NO x including NO2 and N2O5 were generated in the gases with the N2/O2 ratios of 4:6, 5:5, and 6:4. Subsequently, the PAS prepared by the two plasma-activated gases at the N2/O2 ratios of 5:5 and 6:4 exhibited the strongest inactivation effects on both planktic MRSA and biofilms. Furthermore, the aqueous reactive species in the PAS exhibited varied change trends with the increasing N2/O2 ratios. Additionally, ultraviolet spectroscopy combined with the probe of N, N-diethyl-p-phenylenediamine was applied for the detection of O2NOO− in the PAS, and the levels of O2NOO− in the PAS were positively correlated with the inactivation effects. Moreover, the PAS induced varying levels of nitration modification on the soluble proteins in MRSA cells, which were related to the intensities of O2NOO− in the PAS. This study regulated the reactive species in the PAS through gas composition and explored the inactivation mechanism of the PAS, providing a new strategy to promote the preparation efficiency of plasma-activated solutions for biomedical applications.

The effects of Marrubium alysson and Torilis arvensis natural and nano extracts on priming of wheat seeds in response to drought

AbstractAgriculture and climate change are inextricably linked in various aspects. Droughts have become more frequent as a result of climate change, having a significant impact on crop productivity. As a result, the current study investigated the effect of seed priming with natural plant extract and biosynthesized nano plant extract as an environmentally friendly tool for mitigating the drought effect on wheat as an economic crop. The study investigates the biosynthesis of Ag-nano particles from extracts of Marrubium alysson and Torilis arvensis. The UV–Vis spectrophotometer was used to characterize the biosynthesized AgNPs. Wheat grains were primed with Marrubium alysson and Torilis arvensis, along with their nanoextracts, and grown in different water regimes (100%, 60% and 40% field capacity), as well as hydropriming. Leaves were collected to determine the photosynthetic pigments, phenolics, flavonoids, CAT, GPX, H2O2, MDA, soluble sugars, and soluble proteins. In comparison with hydropriming seeds, the study discovered that natural and nano extracts significantly increased the CAT and GPX, as well as soluble proteins. Phenolics, flavonoids, soluble sugars, H2O2, and MDA content all decreased significantly, but pigment content remained unchanged. The study believed that priming wheat with natural and nano extracts, improved drought tolerance through the use of their metabolites, which included soluble sugars, phenolics, and flavonoids, accumulating in other metabolites like lignin, starch, and flavolignan to increase plant tolerance and reduce oxidative damage. Furthermore, nano extracts of Torilis arvensis and Marrubium alysson may be more effective than plant extracts since they separate from each other in PCA analysis.

Architecting Multicompartmentalized, Giant Vesicles with Recombinant Fusion Proteins.

We present a straightforward strategy for constructing giant, multicompartmentalized vesicles using recombinant fusion proteins. Our method leverages the self-assembly of globule-zipper-elastin-like polypeptide fusion protein complexes in aqueous conditions, eliminating the need for organic solvents and chemical conjugation. By employing the thin-film rehydration method, we have successfully encapsulated a diverse range of bioactive macromolecules and engineered organelle-like compartments─ranging from soluble proteins and coacervate droplets to vesicles─within these protein-assembled giant vesicles. This approach also facilitates the integration of water-soluble block copolymers, enhancing the structural stability and functional versatility of the vesicles. Our results suggest that these multicompartment giant protein vesicles not only mimic the complex architecture of living cells but also support biochemically distinct reactions regulated by functionally folded proteins, providing a robust model for studying cellular processes and designing microreactor systems. This work highlights the transformative potential of self-assembling recombinant fusion proteins in artificial cell design.

Phosphorus and selenium compounding mitigates Cr stress in peanut seedlings by enhancing growth homeostasis and antioxidant properties.

Peanut is an economically important crop, but it is susceptible to Cr contamination. In this study, we used peanut as experimental material to investigate the effects of exogenous P, Se interacting with Cr on the nutrient growth and antioxidant system of peanut seedlings by simulating Cr (0μM, 50μM, and 100μM) stress environment. The results showed that exogenous P, Se supply could mitigate irreversible damage to peanut seedlings by altering the distribution of Cr in roots and aboveground, changing root conformation, and repairing damaged cells to promote growth. When the Cr concentration is 100μM, it exhibits the highest toxicity. Compared to the control group P and Se (0 MM), the treatment with simultaneous addition of P + Se (0.5 + 6.0) resulted in a significant increase in root length and root tip number by 248.7% and 127.4%, respectively. Additionally, there was a 46.9% increase in chlorophyll content, a 190.2% increase in total surface area of the seedlings, and a respective increase of 149.1% and 180.3% in soluble protein content in the shoot and roots. In addition, by restricting the absorption of Cr and reducing the synthesis of superoxide dismutase SOD (Superoxide dismutase), CAT (Catalase), POD (Peroxidase), and MDA (Malonaldehyde), it effectively alleviates the oxidative stress on the antioxidant system. Therefore, the exogenous addition of P (0.5 MM) and Se (6.0 MM) prevented the optimal concentration of chromium toxicity to peanuts. Our research provides strong evidence that the exogenous combination of P and Se reduces the risk of peanut poisoning by Cr, while also exploring the optimal concentration of exogenous P and Se under laboratory conditions, providing a basis for further field experiments.

Validation of Molecular Markers for Low Kunitz Trypsin Inhibitor Content in European Soybean (Glycine max L. Merr.) Germplasm

Trypsin inhibitors (TI) in raw soybean grain, mainly represented by the Kunitz trypsin inhibitor protein (KTI), prevent the normal activity of the digestive enzymes trypsin and chymotrypsin in humans and monogastric livestock. The inactivation of TI is achieved through costly and time-consuming heat treatment. Thermal processing also impairs the solubility and availability of the soybean grain protein. Therefore, the genetic elimination of KTI has been proposed as a suitable alternative to heat treatment. The aim of this study was to screen the collection of European soybean cultivars with six genetic markers (one SSR marker and five SNP markers) previously proposed as tightly linked to the KTI3 gene encoding the major Kunitz trypsin inhibitor seed protein of soybean and validate their usability for marker-assisted selection (MAS). The six markers were validated on a subset of 38 cultivars with wide variability in KTI content and in the F2 and F3:5 progenies of two crosses between the known high- and low-KTI cultivars. Three genetic markers (SSR Satt228 and two SNP markers, Gm08_45317135_T/G and Gm08_45541906_A/C) were significantly associated with KTI content in a subset of 38 cultivars. Low-KTI alleles were detected in both low- and high-KTI genotypes and vice versa, high-KTI alleles were found in both high- and low-KTI genotypes, indicating a tight but not perfect association of these markers with the KTI3 gene. The genetic marker SSR Satt228 showed a significant association with KTI content in the F2 progeny, while the SNP markers Gm08_45317135 and Gm08_45541906 allowed significant discrimination between progeny with high- vs. low-KTI progenies in the F3:5 generation. These three markers could be applied in MAS for low-KTI content but not without the additional phenotyping step to extract the desired low-KTI genotypes.

The acute effects of variations in the flour composition of crackers on the glycemic index and glycemic responses in healthy adults.

This study assessed the glycemic index (GI) and glycemic load (GL) of three crackers formulated with different flours: a control cracker (CC) made with conventional flour, one with 30% whole wheat flour substitution (WWC), and another with 30% sunflower seed flour substitution (SFC). This study aimed to explore the impact of these substitutions, which vary in protein and fiber content, on the glycemic responses compared to a reference glucose drink. In a randomized controlled, crossover design, 11 healthy participants (mean age 23.5 ± 1 years; 7 women; BMI 23 ± 1 kg/m2), consumed cracker meals (CC, WWC, and SFC) each providing 50 g of available carbohydrates, and a 50 g glucose reference in separate sessions. The SFC crackers provided low GI and GL values (GI: 53 on the glucose scale, GL: 6 per serving), whereas the WWC and CC crackers provided high GI (GI: 77 and 90 on the glucose scale, respectively) and medium GL values (11 and 12 per serving, respectively). Compared with the glucose reference and CC crackers, only SFC induced lower postprandial glucose concentrations, lower glucose excursions, and lower peak glucose values. All crackers were rated as enjoyable and associated with increased satiety. SFC moderated postprandial glycemic responses compared to CC and the reference (D-glucose), but not WWC. These effects may be attributed to the soluble fibers and protein content of the SFC. These findings suggest potential benefits for body weight management and glycemic control, warranting further investigation of the role of flour substitutions in healthy snack options. This trial has been registered at Clinicaltrials.gov (NCT05702372).

Calcium oxide enhances the anaerobic co-digestion of excess sludge and plant waste: performance and mechanism

ABSTRACT The study investigates the effect of the oxidant calcium oxide (CaO) on the codigestion of excess sludge (ES) and plant waste (PW) under mesophilic anaerobic conditions to enhance methane production. The findings indicate that CaO significantly elevated methane yield in the codigestion system, with an optimum CaO addition of 6% resulting in a maximum methane production of 461 mL/g volatile solids, which is approximately 1.3 times that of the control group. Mechanistic exploration revealed that CaO facilitated the disintegration of organic matter, enhanced the release of soluble chemical oxygen demand, and increased the concentrations of soluble proteins and polysaccharides within the codigestion substrate. The presence of CaO was conducive to the generation and biological transformation of volatile fatty acids, with a notable accumulation of acetic acid, a smaller carboxylic acid within the VFAs. The proportion of acetate in the CaO-amended group increased to 32.6–36.9%. Enzymatic analysis disclosed that CaO enhanced the activity of hydrolytic and acidogenic enzymes associated with the ES and PW codigestion process but suppressed the activity of coenzyme F420. Moreover, CaO augmented the nutrient load in the fermentation liquid. The study provides an alternative scheme for the efficient resource utilization of ES and PW.

Effects of germination on the physicochemical, thermal, in vitro protein digestibility, Antinutrients, and antioxidant properties of sorghum flour

SummaryThe present study investigates the impact of germination (72 h) on physicochemical, protein solubility, in vitro protein digestibility, antinutrient, antioxidant, and structural properties of sorghum flour. Germination resulted in a reduction of crude protein, fat, fibre, and ash. However, it significantly improved the DPPH (58.42%–69.44%), total phenolic content (1.31–3.58 mg GAE g−1), and total flavonoid content (57.66–80.16 mg QE g−1). Germination also increased in vitro protein digestibility (50.12%–75.52%) and Fe2+ chelating activity (11.18%–48.06%), while reducing antinutrients such as phytic acid (176 to 86 mg/100 g) and tannin (3.53–2.16 mg/100 g). Structural analyses (XRD and FTIR) revealed modifications in crystalline and molecular structures, while scanning electron microscopy showed disrupted starch granules in germinated flour. These findings suggest that germination enhances the functional and nutritional properties of sorghum flour, making it more suitable for various applications in food products.

Salt–Alkali Tolerance Evaluation for Bermudagrass and Critical Indicator Screening at the Seedling Stage

The adaptability of bermudagrass genotypes to high-pH saline–alkali conditions was investigated through a comprehensive evaluation of 38 genotypes during the seedling stage. For this purpose, two distinct treatments were established: exposure to saline–alkali solution composed of 45% NaCl, 5% Na2SO4, 5% NaHCO3, and 45% Na2CO3 (pH 10.0), and exposure to distilled water as control. On 6th day of treatment, eight physiological indicators were measured. Compared with the control, the net photosynthetic rates, leaf water content, and chlorophyll content of the test genotypes decreased under stress. In contrast, the soluble protein content, proline levels, malondialdehyde concentration, and conductivity exhibited an increase. The salt–alkali tolerance coefficients of each indicator ranged from 0.24 to 8.54, and the variable coefficient was from 9.77% to 62.82%. Based on the salt–alkali tolerance coefficients, the comprehensive evaluation value (D) and resistance coefficient (CSAC) for each genotype were calculated. Subsequently, 38 genotypes were classified into three salt–alkali tolerance clusters by hierarchical clustering analysis, with Cluster I consisting of 10 genotypes with the most salt–alkali tolerance, and Cluster II with intermediate tolerance. Cluster III was comprised of 18 genotypes showing the lowest tolerance. The predictive model for assessing salt–alkali tolerance in bermudagrass is (D) = −0.238 + 0.106 × SACChlb + 0.209 × SACRWC + 0.015 × SACPro + 0.284 × SACProtein + 0.051 × SACPn. Notably, Cluster I genotypes were more vigorous and showed lower damage under saline stress compared to Cluster III. Moreover, stepwise regression analysis pinpointed Chlb, RWC, and Pro as crucial indicators for evaluating salt–alkali tolerance in bermudagrass genotypes.

Effect of ultrasound and marination on functional and sensory properties, protein profiles and bioaccessibility of amino acids in turkey meat

SummaryThis study investigates the impact of ultrasound treatment and marination on turkey meat proteins, focusing on functional and sensory properties and protein hydrolysis. Three different treatments, marination (M), ultrasound (US) and a combination of ultrasound and marination (USM), were applied to turkey meat. The USM caused the highest water‐holding capacity, increased tenderness and improved the general acceptability of turkey meat. SEM images of raw samples showed that US and M changed the structure of muscle fibres, exhibiting more cavities and irregular, broken and separated fibres. Cooking changed the muscle fibres of the sample, forming several cavities and separate fibres. SDS‐PAGE profiles showed that different treatments affected protein solubility during gastric digestion. Ultrasound treatment caused the loss of some essential amino acids in the supernatant of intestinal digests, including lysine, leucine and tyrosine. In vitro bioaccessibility of leucine, threonine, lysine, valine and isoleucine was the highest in the untreated sample.

Unveiling the Catalytic Roles of DsBBS1 and DsBBS2 in the Bibenzyl Biosynthesis of Dendrobium sinense

Dendrobium sinense, an endemic medicinal herb in Hainan Island, is rich in bibenzyl compounds. However, few studies have explored the molecular mechanisms of bibenzyl biosynthesis. This study presents a comprehensive analysis of DsBBS1 and DsBBS2 function in D. sinense. A molecular docking simulation revealed high-resolution three-dimensional structural models with minor domain orientation differences. Expression analyses of DsBBS1 and DsBBS2 across various tissues indicated a consistent pattern, with the highest expression being found in the roots, implying that they play a pivotal role in bibenzyl biosynthesis. Protein expression studies identified optimal conditions for DsBBS2-HisTag expression and purification, resulting in a soluble protein with a molecular weight of approximately 45 kDa. Enzyme activity assays confirmed DsBBS2’s capacity to synthesize resveratrol, exhibiting higher Vmax and lower Km values than DsBBS1. Functional analyses in transgenic Arabidopsis demonstrated that both DsBBS1 and DsBBS2 could complement the Atchs mutant phenotype. The total flavonoid content in the DsBBS1 and DsBBS2 transgenic lines was restored to wild-type levels, while the total bibenzyl content increased. DsBBS1 and DsBBS2 are capable of catalyzing both bibenzyl and flavonoid biosynthesis in Arabidopsis. This study provides valuable insights into the molecular mechanisms underlying the biosynthesis of bibenzyl compounds in D. sinense.

Antioxidative and Metabolic Responses in Canola: Strategies with Wood Distillate and Sugarcane Bagasse Ash for Improved Growth under Abiotic Stress

In the context of increasing agricultural challenges posed by soil salinity and drought stress, the main importance of the present study was to evaluate some novel treatments for improving canola productivity and resilience by applying wood distillate (WD) in combination with bagasse ash (SBA). A two-year field experiment using a split plot design was conducted and evaluated several physiological and biochemical parameters under different irrigation regimes conducted at 80% and 50% field capacity. While there were considerable moderation effects of SBA and WD on soil salinity, expressed as exchangeable sodium percentage (ESP), under both well-irrigated and drought conditions, more importantly, the ESP was reduced to 31% under drought stress with combined WD and SBA applications over any single factor. WD and SBA treatments of canola leaves showed reduced Na content with increased K levels, and the plants maintained physiological attributes—chlorophyll content, stomatal conductance, and relative water content—to the level of controls of well-irrigation. Besides, they significantly alleviated oxidative stress by decreasing the hydrogen peroxide (H2O2), malondialdehyde (MDA), and electrolyte leakage (EL) levels and increasing the activities of antioxidant enzymes like superoxide dismutase (SOD) and ascorbate peroxidase (APX). Nonenzymatic antioxidants such as total soluble sugars (TSS), total soluble proteins (TSP), total phenolic content (TPC), and total flavonoid content (TFC) were significantly increased under stress conditions with a special accent on combined treatment, whereas the levels of proline and GB that increased in alignment with drought reduced under the combined application. Various growth parameters of plants like plant height, number of branches, and siliques per plant were significantly improved with WD and SBA under drought stress. Principal component analysis (PCA) and Pearson correlation further confirmed the relationships among these parameters and thus underpinned that WD and SBA can evoke a synergistic effect to enhance growth promotion and stress tolerance in canola. This, therefore, infers that the combined application of WD and SBA can be key, offering very high potential as viable options to better canola productivity under adverse environmental conditions.

Effect of lipoxygenase-catalyzed linoleic acid oxidation and 4-hydroxy-2-nonenal on digestibility and gel properties of myofibrillar protein

The oxidative modification of myofibrillar proteins (MPs) has been identified as a crucial factor affecting meat quality during processing. We compared the effects of 4-hydroxy-2-nonenal (HNE) and lipoxygenase (LOX)-catalyzed linoleic acid (LA) oxidation products on the digestibility and gel properties in MPs from bighead carp. Both treatments resulted in decreased free amino acid content, reduced digestibility, and loss of amino acids in MPs. The HNE treatment enhanced gel strength by increasing hydrophobic interactions within the gel matrix. Conversely, the LOX-catalyzed LA oxidation enhanced disulfide bonds, leading to an agglomerated microstructure. Reduced myofibrillar protein (MP) solubility, primarily due to protein aggregation, was observed in the LA group but not in the HNE group. This distinction suggests that the impact on MP functionality is predominantly influenced by the protein aggregation induced by LOX-catalyzed LA oxidation rather than by HNE alone.

Coating seeds with purified-hydrocolloids extracted from conventional activated sludge affects the growth physiology of wheat under gradient salt stress

With continuous innovations in wastewater treatment technology, the use of excess sludge has become a research focus. Purified-hydrocolloids obtained from aerobic granular sludge (AGS) and conventional activated sludge (CAS) have many practical applications, including waterproofing, adsorption, flame retardance, slow-release properties, and seed coating. In this study, purified-hydrocolloids extracted from CAS was used as a new seed coating, which was uniformly sprayed onto the surface of wheat seeds. Purified-hydrocolloids was most effective in resisting salt stress in wheat seedlings at a NaCl concentration of 125mmol/L (soil salinity 2.74g/Kg). The germination percentage, plant height, K+/Na+ ratio, soluble sugar and soluble protein contents of purified-hydrocolloids-coated seeds increased by 13.33%, 46.32%, 37.25%, 8.89%, and 36.22%, respectively, compared with wheat seeds without purified-hydrocolloids coating. Inhibition of O2− production activity and malondialdehyde content was reduced by 38.62% and 48.00%, respectively. Therefore, the purified-hydrocolloids coating had a mitigating effect on wheat seed germination and seedling growth in salt-stressed environments, and this application would be beneficial to the use of saline soil resources and the improvement of ecosystems in coastal areas.

Low-cost, immediate, general-purpose, and high-throughput (LIGHt) smartphone colorimetric screening assay for water-soluble protein

An efficient and rapid method for the detection of total soluble protein in tobacco leaves, utilizing a smartphone-based colorimetric approach has been developed. The proposed low-cost, immediate, general-purpose, and high-throughput (LIGHt) smartphone colorimetric screening assay integrates commercially available microplates, enabling on-site, high-throughput screening of tobacco leaf quality. The study involves preparing protein standard solutions and constructing standard curves using both spectrophotometric and smartphone-based methods. The LIGHt smartphone colorimetry yielded an average relative standard deviation of 10.6 %, a limit of detection of 2 μg/mL, and an average recovery of 93 %. The results demonstrated a comparable performance between intensities from the blue channel and the absorbance values in reflecting protein concentrations, validating the feasibility of utilizing smartphone colorimetry for protein concentration determination. Our approach demonstrates the potential for practical implementation in the field, providing a cost-effective and user-friendly solution for rapid quality assessment in the tobacco industry. The LIGHt smartphone colorimetry enhances quality control practices in the tobacco sector and offers a promising tool for on-site production quality testing in various industries, such as fruits and vegetables.

Biopurification using non-growing microorganisms to improve plant protein ingredients

Securing a sustainable global food supply for a growing population requires a shift toward a more plant-based diet. The application of plant-based proteins is therefore increasing, but unpleasant off-flavors complicate their use. Here, we screened 97 microorganisms for their potential to remove off-flavors in a process with limiting amounts of fermentable sugar. This allowed the production of a more neutral-tasting, purified food ingredient while limiting microbial growth and the production of typical fermentation end products. We demonstrate that various lactic acid bacteria (LAB) and yeasts remove “green” aldehydes and ketones. This conversion can be carried out in less than one hour in almond, pea, potato, and oat proteins. Heterofermentative LAB was best at aldehyde and ketone neutralization with minimum de novo formation of microbial volatiles such as ethylacetate (sweet, fruity) or alpha-diketones (butter- and cheese-like). While sensory properties were improved, changes in protein solubility, emulsification, foaming, and in vitro digestibility were limited.

Morphological and physiological investigations reveal the regulatory effect of exogenous paclobutrazol on flowering promotion by winter warming in Chaenomeles speciosa ‘Changshouguan’

The application of exogenous paclobutrazol (PP333) can improve the ability of winter warming to promote flowering in Chaenomeles speciosa, but the underlying mechanism is unclear. In this study, the cultivar ‘Changshouguan’ was sprayed with different concentrations of PP333 during flower bud differentiation, and the changes in the anatomical structures and physiological characteristics of the flower buds during the differentiation process, as well as the growth state of the flower buds and the effect on flowering promotion after winter warming treatment, were comprehensively investigated. The results showed that different concentrations of PP333 could advance the flowering time of ‘Changshouguan’ by 15–24 d under the warming treatment and increase the flowering duration to 17 d compared with those under the warming treatment alone (CK), and 1000 mg/L was the best treatment. Compared with the CK treatment, the PP333 treatment decreased the contents of indole acetic acid (IAA) and gibberellic acid (GAs) and increased the contents of zeatin ribosides (ZRs) and abscisic acid (ABA), thus changing the balance of hormones during flower bud differentiation. The inflection point (low point) of the curve shapes of the ZRs/GAs and ZRs/IAA ratios appeared significantly earlier, which showed a pattern consistent with soluble sugar and protein content and antioxidant activity. Interestingly, the above changes also corresponded to earlier flowering times during the warming process. Taken together, these results indicate that spraying an appropriate concentration of PP333 in the early stage of ‘Changshouguan’ flower bud differentiation promotes the early differentiation of flower buds and early flowering under winter warming treatment by altering their endogenous hormone content and homeostasis and changing their physiological state. The key to maintaining a relatively long flowering period in plants in the PP333 treatment group after flowering promotion was the increased accumulation of sugars and proteins.

Leveraging Immunofocusing and Virus-like Particle Display to Enhance Antibody Responses to the Malaria Blood-Stage Invasion Complex Antigen PfCyRPA

A vaccine protecting against malaria caused by Plasmodium falciparum is urgently needed. The blood-stage invasion complex PCRCR consists of the five malarial proteins PfPTRAMP, PfCSS, PfRipr, PfCyRPA, and PfRH5. As each subcomponent represents an essential and highly conserved antigen, PCRCR is considered a promising vaccine target. Furthermore, antibodies targeting the complex can block red blood cell invasion by the malaria parasite. However, extremely high titers of neutralizing antibodies are needed for this invasion-blocking effect, and a vaccine based on soluble PfRH5 protein has proven insufficient in inducing a protective response in a clinical trial. Here, we present the results of two approaches to increase the neutralizing antibody titers: (A) immunofocusing and (B) increasing the immunogenicity of the antigen via multivalent display on capsid virus-like particles (cVLPs). The immunofocusing strategies included vaccinating with peptides capable of binding the invasion-blocking anti-PfCyRPA monoclonal antibody CyP1.9, as well as removing non-neutralizing epitopes of PfCyRPA through truncation. Vaccination with PfCyRPA coupled to the AP205 cVLP induced nearly two-fold higher IgG responses compared to vaccinating with soluble PfCyRPA protein. Immunofocusing using a linear peptide greatly increased the neutralizing capacity of the anti-PfCyRPA antibodies. However, significantly lower total anti-PfCyRPA titers were achieved using this strategy. Our results underline the potential of a cVLP-based malaria vaccine including full-length PfCyRPA, which could be combined with other leading malaria vaccine antigens presented on cVLPs.

Multiple aspects of amyloid dynamics in vivo integrate to establish prion variant dominance in yeast

Prion variants are self-perpetuating conformers of a single protein that assemble into amyloid fibers and confer unique phenotypic states. Multiple prion variants can arise, particularly in response to changing environments, and interact within an organism. These interactions are often competitive, with one variant establishing phenotypic dominance over the others. This dominance has been linked to the competition for non-prion state protein, which must be converted to the prion state via a nucleated polymerization mechanism. However, the intrinsic rates of conversion, determined by the conformation of the variant, cannot explain prion variant dominance, suggesting a more complex interaction. Using the yeast prion system [PSI+], we have determined the mechanism of dominance of the [PSI+]Strong variant over the [PSI+]Weak variant in vivo. When mixed by mating, phenotypic dominance is established in zygotes, but the two variants persist and co-exist in the lineage descended from this cell. [PSI+]Strong propagons, the heritable unit, are amplified at the expense of [PSI+]Weak propagons, through the efficient conversion of soluble Sup35 protein, as revealed by fluorescence photobleaching experiments employing variant-specific mutants of Sup35. This competition, however, is highly sensitive to the fragmentation of [PSI+]Strong amyloid fibers, with even transient inhibition of the fragmentation catalyst Hsp104 promoting amplification of [PSI+]Weak propagons. Reducing the number of [PSI+]Strong propagons prior to mating, similarly promotes [PSI+]Weak amplification and conversion of soluble Sup35, indicating that template number and conversion efficiency combine to determine dominance. Thus, prion variant dominance is not an absolute hierarchy but rather an outcome arising from the dynamic interplay between unique protein conformations and their interactions with distinct cellular proteostatic niches.