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Abstract Banana is sensitive to cold stress and often suffers from chilling injury with browning peel and failure to normal ripening. We have previously reported that banana chilling injury is accompanied by a reduction of miR528 accumulation, alleviating the degradation of its target gene MaPPO and raising ROS levels that cause peel browning. Here, we further revealed that the miR528-MaPPO cold-responsive module was regulated by miR156-targeted SPL transcription factors, and the miR156c-MaSPL4 module was also responsive to cold stress in banana. Transient overexpression of miR156c resulted in a more severe chilling phenotype by decreasing the expression of MaSPL4 and miR528. Conversely, the browning was alleviated in STTM-miR156c silencing and OE-MaSPL4 samples. Furthermore, DNA affinity purification sequencing and MaSPL4-overexpressing transcriptome jointly revealed that MaSPL4 may mediate the transcription of genes related to lipid metabolism and antioxidation, in addition to the miR528-MaPPO module, demonstrating MaSPL4 as a master regulator in the fruit cold response network. In summary, our results suggest that the miR156c-MaSPL4 module can mediate the chilling response in banana by regulating the miR528-MaPPO module and multiple other pathways, which provides evidence for the crosstalk between TFs and miRNAs that can be used for the molecular breeding of fruit cold tolerance.

Background: Fumonisins (FUMs) are among the most common mycotoxins in plant-derived food products. FUMs contamination has considerably impacted human and animal health, while causing significant economic losses. Hence, management of FUMs contamination in food production and supply chains is needed. The toxicities of FUMs have been widely investigated. FUMs management has been reported and several available strategies have been developed successfully to mitigate FUMs contamination present in foods. However, currently available management of FUMs contamination from different phases of food chains and the mechanisms of some major strategies are not comprehensively summarized. Aim of review: This review comprehensively characterize the occurrence, impacts, and management of FUMs contamination across food production and supply chains. Pre- and post-harvest strategies to prevent FUMs contamination also are reviewed, with an emphasis on the potential applications and the mechanisms of major mitigation strategies. The presence of modified FUMs products and their potential toxic effects are also considered. Importantly, the potential application of biotechnological approaches and emerging technologies are enunciated. Key scientific concepts of review: Currently available pre- and post-harvest management of FUMs contamination primarily involves prevention and decontamination. Prevention strategies are mainly based on limiting fungal growth and FUMs biosynthesis. Decontamination strategies are implemented through alkalization, hydrolysis, thermal or chemical transformation, and enzymatic or chemical degradation of FUMs. Concerns have been raised about toxicities of modified FUMs derivatives, which presents challenges for reducing FUMs contamination in foods with conventional methodologies. Integrated prevention and decontamination protocols are recommended to control FUMs contamination across entire value chains in developed countries. In developing countries, several other approaches, including cultivating, introducing Bt maize, simple sorting/cleaning, and dehulling, are suggested. Future studies should focus on biotechnological approaches, emerging technologies, and metagenomic/genomic identification of new degradation enzymes that could allow better opportunities to manage FUMs contamination in the entire food system.

The degradation of cell wall polysaccharides in pineapple fruit during softening was investigated in the present study. Two pectin fractions and two hemicellulose fractions were extracted from the cell wall materials of ‘Comte de Paris’ pineapple fruit at five softening stages, and their compositional changes were subsequently analyzed. The process of softening of the fruit corresponded to an increase in the water-soluble pectin (WSP) and 1 M KOH-soluble hemicellulose (HC1) fractions, and a decrease in the acid-soluble pectin (ASP) fraction, which suggested the solubilization and conversion of cellular wall components. However, the content of 4 M KOH-soluble hemicellulose (HC2) decreased and then returned to the initial level. Furthermore, WSP, ASP, and HC1 showed an increment in the content of low molecular weight polymers while a decline in the high molecular weight polymers throughout softening, and not significant change in the contents of different molecular polymers of HC2 was observed. Moreover, the galacturonic acid (GalA) content in the main chain of WSP was maintained at a relatively constant level, but the major branch monosaccharide galactose (Gal) in WSP decreased. Different from WSP, the molar percentages of Gal and GalA in ASP decreased. The Gal or Arabinose (Ara) in HC1 exhibited a gradual decline while the molar percentages of xylose (Xyl) and glucose (Glu) in the main chain increased. These suggested that the main chain of ASP degraded while the branched chains of ASP, WSP and HC1 depolymerized during pineapple softening. Overall, fruit softening of ‘Comte de Paris’ pineapple was found to be the result of differential modification of pectin and hemicellulose.

Soymilk is a natural nanocarrier. However, the performance of flavonoid-soymilk nano-complex remains unclear. In this work, icariin-soymilk nano-complexes (ISNCs) were successfully fabricated and characterized. The effects of high-pressure homogenization (HPH) treatment on structure and physicochemical properties of soymilk and nano-complexes were investigated. HPH treatment could significantly improve the surface hydrophobicity and interfacial activity of soymilk. The soymilk with HPH treatment could significantly improve the water solubility (20 folds), thermal stability and bioavailability of icariin. The highest encapsulation efficiency (93.28%), loading capacity (39.09 μg/mg), ζ-potentia (absolute value, 31.20 mV) and bioavailability (72.14%) were observed in HSI-200 (200 bar of homogenization pressure). While HSI-500 (500 bar of homogenization pressure) showed the smallest particle size (183.73 nm). ISNCs showed a rougher surface and an irregular lamellar structure with large amount of fi ne particles by using Cryo-SEM, suggesting that icariin was encapsulated in soymilk. These data supplied a novel strategy to improve the performance of icariin in functional foods.

Abstract Longan (Dimocarpus longan) is an important subtropical evergreen fruit tree (family Sapindaceae) in southern China and exhibits a high commercial and nutritional value. Endocarp browning of longan fruit once harvested considerably reduces the commercial value. The excessive accumulation of reactive oxygen species (ROS) can induce the rapid endocarp browning of longan fruit during storage, but the ROS‐mediated mechanism remains relatively unclear. In this investigation, a hydrogen peroxide scavenger, 1,3‐dimethylthiourea (DMTU), was used to treat longan fruit to elucidate the physiological and molecular bases of the ROS‐mediated endocarp browning of longan fruit. Application of 5 mM DMTU significantly delayed endocarp browning, inhibited respiration rate, suppressed the accumulations of H2O2 and malondialdehyde (MDA), and activated the antioxidant system of longan fruit during storage, such as 3.8 of the DMTU‐treated fruit versus 1.6 of non‐DMTU‐treated fruit of the browning index, 398 versus 346 μg kg−1 S−2 of the respiration rate, and 47.5 versus 37.8 nmol g−1 of the MDA content after 6 days of storage. Furthermore, the DMTU treatment significantly affected the expression levels of the H2O2 receptors and hydrogen peroxide‐induced Ca2+ increases (HPCA)‐LIKEs (HPCALs). Additionally, the regulatory network comprising transcription factors controlling the HPCAL expression in longan was constructed. The major transcription factors, including GATA1, GATA28, TCP20, and AGL20, may contribute to the ROS‐mediated endocarp browning of longan fruit during storage by regulating the HPCAL expression levels. This study could provide new insights into the involvement of the HPCALs in the ROS‐mediated endocarp browning of longan fruit during storage.

In this study, the structural characters, antioxidant activities and bile acid-binding ability of sea buckthorn polysaccharides (HRPs) obtained by the commonly used hot water (HRP-W), pressurized hot water (HRP-H), ultrasonic (HRP-U), acid (HRP-C) and alkali (HRP-A) assisted extraction methods were investigated. The results demonstrated that extraction methods had significant effects on extraction yield, monosaccharide composition, molecular weight, particle size, triple-helical structure, and surface morphology of HRPs except for the major linkage bands. Thermogravimetric analysis showed that HRP-U with filamentous reticular microstructure exhibited better thermal stability. The HRP-A with the lowest molecular weight and highest arabinose content possessed the best antioxidant activities. Moreover, the rheological analysis indicated that HRPs with higher galacturonic acid content and molecular weight showed higher viscosity and stronger crosslinking network (HRP-C, HRP-W and HRP-U), which exhibited stronger bile acid binding capacity. The present findings provide scientific evidence in the preparation technology of sea buckthorn polysaccharides with good antioxidant and bile acid binding capacity which are related to the structure affected by the extraction methods.

The wheel hub is an important component of the wheel, and a good hub design can significantly improve vehicle handling, stability, and braking performance, ensuring safe driving. This article optimized the hub structure through morphological aspects, where reducing the hub weight contributed to enhanced fuel efficiency and overall vehicle performance. By referencing honeycombed structures, a bionic hub design is numerically simulated using finite element analysis and response surface optimization. The results showed that under the optimization of the response surface analytical model, the maximum stress of the optimized bionic hub was 109.34 MPa, compared to 119.77 MPa for the standard hub, representing an 8.7% reduction in maximum stress. The standard hub weighs 34.02 kg, while the optimized hub weight was reduced to 29.89 kg, a decrease of 12.13%. A fatigue analysis on the optimized hub indicated that at a stress of 109.34 MPa, the minimum load cycles were 4.217 × 105 at the connection point with the half-shaft, meeting the fatigue life requirements for commercial vehicle hubs outlined in the national standard GB/T 5334-2021.

Abstract The traditional identification methods of direct current‐to‐direct current (DC–DC) converters usually consider the common situation where the training and testing sets have the same input voltage. In real‐world applications, the actual input voltage may fluctuate beyond its original value, thus negatively affecting the parameter identification accuracy. Therefore, this paper presents a parameter identification method under fluctuating input voltages. Firstly, to increase the identification accuracy, the paper proposes the coefficient of variation method to extract key features from all time‐ and frequency‐domain features as training data. Then, for fluctuating input voltages, the paper adds the input voltages into the training data to build the parameter identification network based on Dendrite Net. When using the established network to identify component parameters in practical applications, the actual input voltage may change to an unknown value because of irregular fluctuation. Hence, the actual input voltage needs to be identified before identifying the component parameters of a DC–DC converter. Finally, the paper builds the input voltage identification network. By combining the identification results of the actual input voltage, the parameter identification under fluctuating input voltages can be completed. The simulation and hardware experiments results validate the practicability and effectiveness of the proposed method.

Abstract Fusarium proliferatum is one of the pathogenic fungi causing crown rot that results in quality deterioration and commercial value loss of banana fruit during postharvest storage. Secreted proteins from pathogenic fungi can be delivered into hosts to suppress plant immunity and establish infection. We reported here a secreted aspartic proteinase OPSB (FpOPSB) that was active during F. proliferatum infection on banana peel. FpOPSB exhibited protease activity based on in vitro assay. Immunohistochemistry assay along with subcellular localization analysis demonstrated that FpOPSB can be secreted into banana peel and localized in cytoplasm. FpOPSB deletion mutant (Δopsb) showed reduced virulence on banana fruit but no obvious effect on fungal growth compared with wild‐type. Subsequently, scanning electron microscopy results suggested that FpOPSB was important for conidial production and infectious growth as well as spreading of F. proliferatum in banana peel. Heterologous expression of FpOPSB in tobacco significantly enhanced plant susceptibility to F. proliferatum. Further, seven proteins were identified as potential targets of FpOPSB in banana peel using immunoprecipitation–mass spectrometry (MS)/MS, including pathogenesis‐related 10, thioredoxin H1, nonsymbiotic hemoglobin 2, fructokinase‐1, patellin‐1, leucine aminopeptidase 2, and l‐lactate dehydrogenase A. Taken together, our results confirmed that the secreted FpOPSB is a critical virulence factor of F. proliferatum when infecting banana fruit. Additionally, we postulated that FpOPSB might interfere the host immune system or neutralize the host defenses, thereby contributing to the pathogenicity of F. proliferatum. We propose aspartic proteinase as a new potential target for controlling fungal disease of postharvest fruit.

Abstract Fresh fruits have good flavor and high nutritional value, but in the postharvest stage they will age and decay rapidly, so we need to find green postharvest fruit preservatives. And natural organic acids are promising natural fruit preservatives due to their safety and effectiveness. The present work reviews recent applications of natural organic acids in postharvest fruit quality management and discussed its potential biochemical mechanisms. This work indicates that numerous natural organic acids are effective postharvest fruit preservatives, such as oxalic acid, salicylic acid, citric acid, ascorbic acid, phenolic acid and terpenic acid. Natural organic acid treatments can improve postharvest fruit quality, as reflected by delaying senescence, alleviating chilling injury, controlling disease, and inhibiting browning. And natural organic acid can have multiple benefits on postharvest fruit. Natural organic acids also play a variety of roles in postharvest fruit disease control. In addition, chemical modification and cotreatment of natural organic acids can be performed to improve the efficiency of postharvest fruit preservation applications. The work provides an important reference for postharvest fruit quality management and the development of natural OAs fruit preservatives.

【Objective】This study aims to understand the roles of MIR164, NAC gene families and miR164-NAC regulatory modules in banana ripening and response to low temperature stress, so as to provide a theoretical basis for banana variety improvement and molecular breeding.【Method】'Brazil banana' was used as test material. Through high-throughput sequencing and bioinformatics analysis using miRBase, NCBI database and Clustal, TBtools, MCScanX and iTOL softwares, miR164 and NAC family members in banana were characterized, including their chromosomal location, structure, physical/ chemical properties, phylogenetic relationships, etc. Multiple miR164-NAC regulatory modules in bananas were identified through degradome sequencing and experimental validation combining transcriptome data. Next, the expression patterns of miR164-NAC regulatory modules during ripening and under cold stress were analyzed by small RNA northern blot and qRT-PCR.【Result】A total of six miR164 family members were identified in banana, of which four were located within the coding genes and two in the intergenic region. Phylogenetic analysis showed that several banana MIR164 precursors with high abundance were clustered together with papaya, suggesting that the origin of banana MIR164 gene family was closer to dicotyledonous plants. The banana genome encodes a total of 222 NAC members, unevenly distributed across all 11 chromosomes. A total of 134 homologous gene pairs were identified in these banana NACs, including 4 tandem repeats and 130 segment-replicating repeats, indicating that the main driving force of banana NAC genes evolution came from segmentreplicating events. Comparative phylogenetic analysis of all NAC proteins in banana, Arabidopsis thaliana and Oryza sativa divided this family into 23 subgroups, and transcriptome data revealed extensive redundancy and expression specificity of banana NAC genes. Physicochemical analysis showed that almost all banana NAC proteins were hydrophilic, and less than 15% were stable proteins. The miR164-NAC176/165 regulatory module in banana was verified, and the accumulation of miR164 in banana was induced by ethylene and gradually increased with fruit ripening, while the expression of MaNAC176/165 negatively regulated by miR164 in this module was gradually decreased during fruit ripening. Under the cold stress, miR164 was also obviously induced, resulting in the downregulation of its targets MaNAC176 and MaNAC165.【Conclusion】This study suggested that MaNAC176 and MaNAC165 may be transcriptional repressors of banana fruit ripening, while miR164 promotes ripening by negatively regulating MANAC176/165. This module may also be a key regulatory pathway of banana chilling injury. This study identified key miR164-NAC candidate modules in banana fruit ripening and cold stress response, which laid a foundation for subsequent gene cloning and functional analysis.

Abstract Macroinvertebrates play a vital role in coastal ecosystems and are an important indicator of ecosystem quality. Both anthropogenic activity and environmental changes may lead to significant changes in the marine macroinvertebrate community. However, the assembly process of benthic biodiversity and its mechanism driven by environmental factors at large scales remains unclear. Here, using the benthic field survey data of 15 years at large spatial and temporal scales from the Yellow Sea Large Marine Ecosystem, we investigated the relative importance of environmental selection, dispersal processes, random‐deterministic processes of macroinvertebrates community diversity assembly, and the responses of this relative importance driven by temperature and nutrients. Results showed that the macroinvertebrates community diversity is mainly affected by dispersal. Nitrogen and phosphorus are the most important negative factors among environmental variables, while geographical distance is the main limiting factor of β diversity. Within the range of 0.35–0.70 mg/L of nutrients, increasing nutrient concentration can significantly facilitate the contribution of the decay effect to β diversity. Within the temperature range studied (15.0–18.0°C), both warming and cooling can lead to a greater tendency for species diversity assembly processes to be dominated by deterministic processes. The analysis contributes to a better understanding of the assembly process of the diversity of coastal marine macroinvertebrates communities and how they adapt to global biogeochemical processes.

BackgroundOsteoarthritis (OA) is featured as cartilage loss, joint pain and loss of labor, which the inflammatory reaction may play critical roles. Ononin is an isoflavone isolating from medicinal plants and has anti-inflammatory effects. Our study investigated the anti-inflammation response of ononin on OA.MethodsAnterior cruciate ligament transection (ACLT)-induced OA operation was used to establish research model, then treated with ononin for 8 weeks. The condition of joint injury was assessed using pathological staining. The concentration of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6) in serum were measured by Elisa kit. The expression of collagen II and matrix metalloproteinase 13 (MMP-13) proteins to assess cartilage metabolism level by immunohistochemistry and Western blot. We detected the expression of proteins involved in the MAPK and NF-κB signaling pathways. Finally, we used molecular docking to assess the affinity of ononin for the target proteins ERK1/2, JNK1/2, p38 and p65.ResultsOur results confirmed that ononin ameliorated cartilage impairment through histopathological analysis by improving the morphological structures and cartilage tidal lines and decreasing Osteoarthritis Research Society International (OARSI) scores in OA rats. Moreover, ononin inhibited the secretion of above factors in OA rats. Furthermore, ononin has been shown to improve cartilage content levels in OA rats. In addition, ononin inhibited the reactivity of MAPK and NF-κB pathways in OA rats. And molecular docking indicated the ligand molecules could stably bind to the proteins of above receptors.ConclusionOur results demonstrated that ononin may ameliorate cartilage damage and inflammatory response in OA rats by downgrading MAPK and NF-κB pathways, thus identifying ononin as a potential novel drug to treat OA.

Abstract Postharvest handling is an important approach to reduce loss and ensure the global supply of fresh fruits and vegetables (FVs). Recently, nanomaterials are increasingly recognized for their great potentials in extending shelf life and maintaining the quality of postharvest FVs owing to their unique properties and outstanding functions with extremely small sizes. This study reviews critically applications of nanotechnology in shelf life extension and quality maintenance of postharvest FVs via nano‐packaging, nano‐coatings, and micro/nano‐bubbles. The advantages of these nanotechnologies to effectively compensate for the shortcomings of the traditional methods are highlighted alongside future research directions at a large low‐cost scale. Generally, nano‐packaging, nano‐coatings, and micro/nano‐bubbles have the functions to modify gas or solvent permeability, inhibit pathogens infection, enhance antimicrobial capacity, and regulate the physiological activity of postharvest FVs. With the development of nanotechnology and quality control measures, it can be expected that green nano‐preservation technology will be an important and novel alternative for the storage and transportation of postharvest FVs.

The cytochrome b6f (Cyt b6f) complex is a multisubunit protein complex in chloroplast thylakoid membranes required for photosynthetic electron transport. Here we report the isolation and characterization of the new tiny albino 1 (nta1) mutant in Arabidopsis, which has severe defects in Cyt b6f accumulation and chloroplast development. Gene cloning revealed that the nta1 phenotype was caused by disruption of a single nuclear gene, NTA1, which encodes an integral thylakoid membrane protein conserved across green algae and plants. Overexpression of NTA1 completely rescued the nta1 phenotype, and knockout of NTA1 in wild-type plants recapitulated the mutant phenotype. Loss of NTA1 function severely impaired the accumulation of multiprotein complexes related to photosynthesis in thylakoid membranes, particularly the components of Cyt b6f. NTA1 was shown to directly interact with four subunits (Cyt b6/PetB, PetD, PetG, and PetN) of Cyt b6f through the DUF1279 domain and C-terminal sequence to mediate their assembly. Taken together, our results identify NTA1 as a new and key regulator of chloroplast development that plays essential roles in assembly of the Cyt b6f complex by interacting with multiple Cyt b6f subunits.

Resveratrol is regarded as neutraceuticals with multiple health benefits. The introduction of prenyl can enhance the bioactivity. In this work, the cancer preventive activities and mechanisms of 18 prenylated reseveratrol and derivatives were investigated. The results showed that prenyl increased the antiproliferative activities of resveratrol, oxyresveratrol and piceatannol against cancer cells, and their antiproliferative activities were time- and dose-dependent. 4-C-prenylation was important for the antiproliferative activity of stilbenoids. The 4-C-prenyl stilbenoids showed better antiproliferative activities than other prenylated stilbenoids. 4-C-prenyl piceatannol showed the best antiproliferative activity. Human hepatoellular carcinomas (HepG2) cell was more sensitive to prenylated stilbenoids than human MCF-7 breast carcinoma cell. 4-C-prenyl piceatannol had high affinities to Caspase-3, Caspase-9, CDK2 and Cyclin A2. The possible amino acids involved in binding 4-C-prenyl piceatannol were revealed. The expression of Caspase-3 and Caspase-9 were upregulated by 4-C-prenyl piceatannol and the expression of CDK2 and Cyclin A2 in HepG2 cells were downregulated, which contributed to apoptosis. The above results eludicated the possible antiproliferative mechanisms of prenylated stilbenoids.

Ultrasound is effective to fabricate nanocomplex. Soymilk is a natural nanocarrier with good compatibility. However, information about soymilk-nutraceuticals nanocomplex is limited. In this work, soymilk was used to encapsulate icariin, a well known nutraceutical with poor bioavailability. The effect of ultrasound on the quality of icariin-soymilk nanocomplexes (ISNCs) was investigated. Ultrasound could reduce the particle size, improve the surface hydrophobicity and change the microstructure of soymilk. With increasing ultrasound treatment time, an increased surface hydrophobicity was observed. The highest encapsulation efficiency (89.67 %) and loading capacity (28.92 µg/mg) were found for USI-20, whereas the smallest particle size (132.47 nm) was observed for USI-120. USI-60 showed the lowest ζ-potential (-31.33 mV) and the highest bioaccessibility (76.08 %). Ultrasound could enhance the storage stability of ISNCs. The data of NMR and fluorescence indicated that ISNCs were mainly stabilized by hydrophobic interaction.

The α-glucosidase inhibitor is of interest to researchers due to its association with type-II diabetes treatment by suppressing postprandial hyperglycemia. Hesperidin is a major flavonoid in orange fruit with diverse biological properties. This paper evaluates the effects of hesperidin on α-glucosidase through inhibitory kinetics, fluorescence quenching, and molecular docking methods for the first time. The inhibition kinetic analysis shows that hesperidin reversibly inhibited the α-glucosidase activity with an IC50 value of 18.52 μM and the inhibition was performed in an uncompetitive type. The fluorescence quenching studies indicate that the intrinsic fluorescence of α-glucosidase was quenched via a static quenching process and only one binding site was present between the hesperidin and α-glucosidase. The interaction between them was spontaneous and mainly driven by hydrogen bonds, as well as hydrophobic forces. Furthermore, the molecular docking results suggest that hesperidin might bond to the entrance or outlet part of the active site of α-glucosidase through a network of five hydrogen bonds formed between hesperidin and the four amino acid residues (Trp709, Arg422, Asn424, and Arg467) of α-glucosidase and the hydrophobic effects. These results provide new insight into the inhibitory mechanisms of hesperidin on α-glucosidase, supporting the potential application of a hesperidin-rich orange product as a hypoglycemic functional food.

Titanium dioxide nanoparticles (TiO2NPs) are widely produced and used nanoparticles. Yet, TiO2NP exposure may possess toxic effects to different cells and tissues, including the brain. Recent studies significantly expanded the understanding of the molecular mechanisms underlying TiO2NP neurotoxicity implicating a number of both direct and indirect mechanisms. In view of the significant recent progress in research on TiO2NP neurotoxicity, the objective of the present study is to provide a narrative review on the molecular mechanisms involved in its neurotoxicity, with a special focus on the studies published in the last decade. The existing data demosntrate that although TiO2NP may cross blood-brain barrier and accumulate in brain, its neurotoxic effects may be mediated by systemic toxicity. In addition to neuronal damage and impaired neurogenesis, TiO2NP exposure also results in reduced neurite outgrowth and impaired neurotransmitter metabolism, especially dopamine and glutamate. TiO2NP exposure was also shown to promote α-synuclein and β-amyloid aggregation, thus increasing its toxicity. Recent findings also suggest that epigenetic effects and alterations in gut microbiota biodiversity contribute to TiO2NP neurotoxicity. Correspondingly, in vivo studies demosntrated that TiO2NPs induce a wide spectrum of adverse neurobehavioral effects, while epidemiological data are lacking. In addition, TiO2NPs were shown to promote neurotoxic effects of other toxic compounds. Here we show the contribution of a wide spectrum of molecular mechanisms to TiO2NP-induced neurotoxicity; yet, the role of TiO2NP exposure in adverse neurological outcomes in humans has yet to be fully appreciated.

Cowpea aphid (Aphis craccivora Koch) is a plant pest that causes serious damage to vegetable crops. Extensive use of synthetic chemical pesticides causes deleterious effects on consumers as well as the environment. Hence, the search for environmentally friendly insecticides in the management of cowpea aphids is required. The present work aims to investigate the aphicidal activity of pomelo seed oil (PSO) on cowpea aphids, the possible insecticidal mechanisms, its chemical constituent profile, as well as the toxicity of its primary compounds. The results of the toxicity assay showed that PSO had significant insecticidal activity against aphids with a 72-hour LC50 value of 0.09 μg/aphid and 3.96 mg/mL in the contact and residual toxicity assay, respectively. The enzymatic activity of both glutathione S-transferase (GST) and acetyl cholinesterase (AChE) significantly decreased, as well as the total protein content, after PSO treatment, which suggested that the reduction of AChE, GST, and the total protein content in aphids treated with PSO might be responsible for the mortality of A. craccivora. The GC-MS analysis revealed that PSO contained limonene (22.86%), (9Z,12Z)-9,12-octadecadienoic acid (20.21%), n-hexadecanoic acid (15.79%), (2E,4E)-2,4-decadienal (12.40%), and (2E,4Z)-2,4-decadienal (7.77%) as its five major compounds. Furthermore, (9Z,12Z)-9,12-octadecadienoic acid showed higher toxicity to aphids than both PSO and thiamethoxam (positive control). This study emphasized the potential of PSO as a natural plant-derived insecticide in controlling cowpea aphids and also provided a novel approach for the value-added utilization of pomelo seed.

Peptides are good food stabilizers and thickeners. They also present good bioactivities. The structure characteristics of peptides are related to the health benefits. In this study, immunoregulatory peptides were produced by Alcalase® and neutrase hydrolysis of soybean proteins. UHPLC-MS/MS was applied for structure identification. The immumoregulatory activities and mechanisms were investigated. Eighty five peptides were identified from the hydrolysates and 84 peptides might be immunoregulatory peptides. Most of them were novel immunoregulatory peptides. These peptides had good immunoregulatory activities, including increasing the pinocytotic capacity, and inducing the production and genes expression of NO, IL-6 and TNF-α in a dose-dependent manner. Specific inhibitors of NF-κB and JNK could significantly suppress NO production stimulated by soybean peptides, implying that NF-κB and JNK pathways were involved in their immunoregulatory mechanisms. Our results suggested that the soybean peptides prepared in this study were promising immunomodulatory neutraceuticals.

Objectives To explore the main factors affecting the knowledge, attitude and practice about influenza and influenza vaccine as well as the intention to receive influenza vaccination among the same group of medical students before (2019) and after (2021) the COVID-19 outbreak.Design A population-based prospective cohort study.Setting A longitudinal cohort study of a selected medical school in Chongqing, China, which ran from 2019 to 2021.Participants A total of 803 medical students participated in the study in 2019 and only 484 students responded in 2021. The response rate for our survey was only 60.27% due to graduation, emails being abandoned, etc.Results The influenza vaccination rate of students at this medical school was 6.7% in 2019, compared with 25.8% in 2021. The awareness rate of medical students about influenza and influenza vaccine was 82.8% in 2019 and 86% in 2021, and there was no significantly statistical difference between the 2 years (p=0.134); the number of medical students with supportive attitude towards influenza vaccine was 95.1% in 2019 and 97.1% in 2021, and there was no statistically significant difference between the 2 years (p=0.078); the number of students who actively learnt about knowledge related to influenza vaccine rose from 183 (22.8%) in 2019 to 195 (40.3%) in 2021.Conclusions The COVID-19 outbreak prompted an increase in influenza vaccination rates among medical students in Chongqing, with almost all students (96.0%) believing that the spread of COVID-19 promoted their knowledge about influenza and influenza vaccine, and the vast majority (74.8%) believing that the spread of COVID-19 promoted their willingness to receive influenza vaccine.

Blue and green molds, the common phenotypes of post-harvest diseases in fruits, are mainly caused by Penicillium fungal species, including P. italicum, P. digitatum, and P. expansum. We sequenced and assembled the genome of a P. italicum strain, which contains 31,034,623 bp with 361 scaffolds and 627 contigs. The mechanisms underlying the evolution of host specificity among the analyzed Penicillium species were associated with the expansion of protein families, genome restructuring, horizontal gene transfer, and positive selection pressure. A dual-transcriptome analysis following the infection of Valencia orange (Citrus sinensis) by P. italicum resulted in the annotation of 9,307 P. italicum genes and 24,591 Valencia orange genes. The pathogenicity of P. italicum may be due to the activation of effectors, including 51 small secreted cysteine-rich proteins, 110 carbohydrate-active enzymes, and 12 G protein-coupled receptors. Additionally, 211 metabolites related to the interactions between P. italicum and Valencia orange were identified by gas chromatography-time of flight mass spectrography, three of which were further confirmed by ultra-high performance liquid chromatography triple quadrupole mass spectrometry. A metabolomics analysis indicated that P. italicum pathogenicity is associated with the sphingolipid and salicylic acid signaling pathways. Moreover, a correlation analysis between the metabolite contents and gene expression levels suggested that P. italicum induces carbohydrate metabolism in Valencia orange fruits as part of its infection strategy. This study provides useful information regarding the genomic determinants that drive the evolution of host specificity in Penicillium species and clarifies the host-plant specificity during the infection of Valencia orange by P. italicum. IMPORTANCE P. italicum GL_Gan1, a local strain in Guangzhou, China, was sequenced. Comparison of the genome of P. italicum GL_Gan1 with other pathogenic Penicillium species, P. digitatum and P. expansum, revealed that the expansion of protein families, genome restructuring, HGT, and positive selection pressure were related to the host range expansion of the analyzed Penicillium species. Moreover, gene gains or losses might be associated with the speciation of these Penicillium species. In addition, the molecular basis of host-plant specificity during the infection of Valencia orange (Citrus sinensis) by P. italicum was also elucidated by transcriptomic and metabolomics analysis. The data presented herein may be useful for further elucidating the molecular basis of the evolution of host specificity of Penicillium species and for illustrating the host-plant specificity during the infection of Valencia orange by P. italicum.

Abstract Background The activation of NOD-like receptor protein 3 (NLRP3) inflammasome-dependent pyroptosis has been shown to play a vital role in the pathology of manganese (Mn)-induced neurotoxicity. Sodium para-aminosalicylic acid (PAS-Na) has a positive effect on the treatment of manganism. However, the mechanism is still unclear. We hypothesized that PAS-Na might act through NLRP3. Methods The microglial cell line BV2 and male Sprague-Dawley rats were used to investigate the impacts of PAS-Na on Mn-induced NLRP3 inflammasome-dependent pyroptosis. The related protein of the NF-κB pathway and NLRP3-inflammasome-dependent pyroptosis was detected by western blot. The reactive oxygen species and mitochondrial membrane potential were detected by immunofluorescence staining and flow cytometry. The activation of microglia and the gasdermin D (GSDMD) were detected by immunofluorescence staining. Results Our results showed that Mn treatment induced oxidative stress and activated the NF-κB pathway by increasing the phosphorylation of p65 and IkB-α in BV2 cells and in the basal ganglia of rats. PAS-Na could alleviate Mn-induced oxidative stress damage by inhibiting ROS generation, increasing mitochondrial membrane potential and ATP levels, thereby reducing the phosphorylation of p65 and IkB-α. Besides, Mn treatment could activate the NLRP3 pathway and promote the secretion of IL-18 and IL-1β, mediating pyroptosis in BV2 cells and in the basal ganglia and hippocampus of rats. But an inhibitor of NF-κb (JSH-23) treatment could significantly reduce LDH release, the expression of NLRP3 and Cleaved CASP1 protein and IL-1β and IL-18 mRNA level in BV2 cells. Interestingly, the effect of PAS-Na treatment in Mn-treated BV2 cells is similar to those of JSH-23. Besides, immunofluorescence results showed that PAS-Na reduced the increase number of activated microglia, which stained positively for GSDMD. Conclusion PAS-Na antagonized Mn-induced NLRP3 inflammasome dependent pyroptosis through inhibiting NF-κB pathway activation and oxidative stress.

BackgroundInfluenza is associated with a large number of disease burdens, and it is generally recommended that all healthcare workers (HCWs) get an influenza vaccination. However, the vaccination rates among HCWs are still low. This study aimed to assess HCWs' knowledge, attitude, and practice (KAP) about the influenza vaccine, and by establishing a structural equation model (SEM) to explore the influencing factors of medical personnel's influenza vaccination in Chongqing, China.MethodsFrom September to November 2019, we conducted a cross-sectional survey in four sentinel hospitals and four non-sentinel hospitals in Chongqing, China. We calculated knowledge, attitude, and behavior scores for each study participant and assessed the level of knowledge, attitude, and behavior of the medical staff. An SEM was used to evaluate the relationship between latent variables, and the path graph between knowledge, attitude, and behavior was established.ResultsA total of 1,412 valid questionnaires were collected in this survey, including four sentinel hospitals (N = 606, 42.92%) and four non-sentinel hospitals (N = 806, 57.08%). Women (N = 1,102, 78.05%) were more than men (N = 310, 21.95%), with an average age of 32.36 ± 7.78 years old and under 30 years old (N = 737, 52.20%), respectively. Nurses (741, 52.48%) were the main subjects, followed by physicians (457, 32.37%). The final SEM model was obtained after the model was modified and adjusted. A bootstrap analysis of path coefficients was carried out on the final model. Knowledge has a direct influence on behavior. The normalized path coefficient is 0.071 (95% CI: 0.002–0.161), and the value of P of the hypothesis test result of the path coefficient is 0.042. The direct influence of knowledge on attitude standardization was 0.175 (95% CI: 0.095–0.281). The direct influence of attitude on practice standardization was 0.818 (95% CI: 0.770–0.862). The indirect effect of knowledge on the standardization of practice through attitude was 0.144 (95% CI: 0.076–0.235).ConclusionsAccording to the SEM, there is a direct positive correlation between KAP and the influenza vaccine. The indirect influence of knowledge on the standard of behavior through attitude is about two times as much as the direct influence on behavior, indicating that attitude plays a strong mediating role between knowledge and practice.

The last 30 years have witnessed the degradation and loss of function in the ecosystem of the Bohai Sea, China. Great efforts, such as artificial reefs, have been used to restore the ecosystem of the Bohai Sea in recent years. In this study, we examined the seasonal variation of food webs in artificial reefs of Juehua Island by the Ecopath model. Results showed that the number of species, range of trophic level, energy cycle proportion, energy complexity, nutrition transfer efficiency, and ecosystem maturity have increased from summer to autumn. However, the food web stability of artificial reefs has no obvious change from summer to autumn. Loop analysis revealed that the heaviest feedback loops are the interactions among benthos, zooplankton, and phytoplankton, and the predator-prey negative interaction strength between zooplankton and phytoplankton dominated the stability of the artificial reef, whether in summer or autumn. This study suggests that stability is a more robust indicator for ecosystem assessment and could provide useful information in future restoration plans of the marine ecosystem.

Abstract Background Banana anthracnose, caused by Colletotrichum musae, is one of the most severe postharvest diseases in banana. Melatonin is widely known for its role in enhancing plant stress tolerance. However, little is known about the control of melatonin on anthracnose in postharvest banana fruit. Results In this study, exogenous melatonin treatment could significantly reduce the incidence of anthracnose in ripe yellow banana fruit and delay fruit senescence. However, melatonin treatment did not affect the growth of Colletotrichum musae in vitro. Transcriptomic analysis of banana peel showed that 339 genes were up-regulated and 241 were down-regulated in the peel after melatonin treatment, compared with the control. Based on GO terms and KEGG pathway, these up-regulated genes were mainly categorized into signal transduction, cell wall formation, secondary metabolism, volatile compounds synthesis and response to stress, which might be related to the anti-anthracnose of banana fruit induced by melatonin treatment. This view was also supported by the increase of volatile compounds, cell wall components and IAA content in the melatonin-treated fruit peel via the metabolomic analysis. After melatonin treatment, auxin, ethylene and mitogen-activated protein kinase (MAPK) signaling pathways were enhanced, which might be involved in the enhanced fruit resistance by regulating physiological characteristics, disease-resistant proteins and metabolites. Conclusions Our results provide a better understanding of the molecular processes in melatonin treatment delaying banana fruit senescence and anthracnose incidence.

Gannan navel orange (Citrus sinensis Osbeck cv. Newhall) is an economically important fruit, but postharvest loss occurs easily during storage. In this study, the effects of different temperatures, light illuminations, and low-temperature plasma treatments on the water loss and quality of the Gannan navel orange were investigated. The fruit began to rot after 90 d of storage at 5 °C and 20–45 d at 26 °C. Navel oranges stored at 26 °C had 7.2-fold and 3.1-fold higher rates of water loss at the early and late storage stages, respectively, as compared with those stored at 5 °C. Storage at 5 °C decreased the contents of total soluble solids at the early storage stage and the contents of titratable acids at the late storage stage, whereas storage at 26 °C decreased the contents of total soluble solids at the late storage stage and the contents of titratable acids at the early storage stage, respectively. Application of low-temperature plasma produced by air ionization for 6 min, or continuous blue or red light illumination significantly inhibited water loss within 7 and 21 d of storage at 22 °C, respectively, but exhibited no significant effect on fruit quality. Furthermore, the low-temperature plasma treatment protected against fruit rot. Thus, treatment with low-temperature plasma followed by storage at a low temperature under continuous red or blue light illumination was of potential value as a green technology for preserving Gannan navel orange during storage.

The effects of five different drying methods, namely, freeze drying (FD), shade drying (SD), hot-air oven drying at 50 °C (OD50), 70 °C (OD70), and microwave drying (MD) on the bioactive phytochemicals and antioxidant capacity of navel orange peel were assessed and comprehensively discussed in detail. Compared with other drying methods, MD-treated peel contained the lowest total phenolic content (TPC) and total flavonoid content (TFC). The peel subjected to OD70 treatment was superior in TPC relative to other treatments and the highest TFC was found in the peels treated with FD. HPLC analysis identified thirteen flavonoids involving three flavanone glycosides (FGs) and ten polymethoxyflavones (PMFs) in navel orange peel and revealed that PMFs in peel were stable under all these drying methods, whereas the three major FGs (narirutin, hesperidin, and didymin) in peel significantly degraded in response to MD treatment. The peels subjected to OD50/OD70 treatments had the most potent antioxidant capacity when compared to other drying methods. Furthermore, Pearson’s correlation analysis was performed. The results revealed here allow us to recommend the use of OD50 or OD70 for the drying of orange peel, both of which help the maintenance of bioactive compounds in the peel and improve its antioxidant capacity.

Abstract Background Temperature stress is a major environmental factor affecting not only plant growth and development, but also fruit postharvest life and quality. MicroRNAs (miRNAs) are a class of non-coding small RNAs that play important roles in various biological processes. Harvested banana fruit can exhibit distinct symptoms in response to different temperature stresses, but the underlying miRNA-mediated regulatory mechanisms remained unknown. Results Here, we profiled temperature-responsive miRNAs in banana, using deep sequencing and computational and molecular analyses. In total 113 known miRNAs and 26 novel banana-specific miRNAs were identified. Of these miRNAs, 42 miRNAs were expressed differentially under cold and heat stresses. Degradome sequencing identified 60 target genes regulated by known miRNAs and half of these targets were regulated by 15 temperature-responsive miRNAs. The correlative expression patterns between several miRNAs and their target genes were further validated via qRT-PCR. Our data showed that miR535 and miR156 families may derive from a common ancestor during evolution and jointly play a role in fine-tuning SPL gene expression in banana. We also identified the miRNA-triggered phased secondary siRNAs in banana and found miR393-TIR1/AFB phasiRNA production displaying cold stress-specific enrichment. Conclusions Our results provide a foundation for understanding the miRNA-dependent temperature stress response in banana. The characterized correlations between miRNAs and their response to temperature stress could serve as markers in the breeding programs or tools for improving temperature tolerance of banana.

Abstract Banana as a typical climacteric fruit soften rapidly, resulting in a very short shelf life after harvest. Sodium dichloroisocyanurate (NaDCC) is reported to be an effectively antibacterial compound. Here, we investigated the effects of NaDCC on ripening and senescence of harvested banana fruit at physiological and molecular levels. Application of 200 mg L−1 NaDCC solution effectively inhibited the ripening and senescence of banana fruit after harvest. NaDCC treatment reduced greatly ethylene production rate and expressions of genes encoding 1-aminocyclopropane-1-carboxylate synthetase, 1-aminocyclopropane-1-carboxylate oxidase, ethylene-responsive transcription factor and EIN3-binding F-box protein. Meanwhile, NaDCC treatment down-regulated markedly the expressions of xyloglucan endotransglucosylase/hydrolase and pectinesterase genes. Furthermore, NaDCC treatment affected significantly the accumulation of ripening-related primary metabolites such as sugars and organic acids. Additionally, NaDCC treatment decreased the production of hydroxyl radical and increased 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging activity, reducing power and hydroxyl radical scavenging activity. In conclusion, NaDCC delayed effectively the ripening and senescence of harvested banana fruit via the reduced ethylene effect and enhanced antioxidant activity.

Reactive oxygen species (ROS) play an active role in plant defense. Polyphenol oxidase (PPO) and peroxidase (POD) participate in the synthesis of phytoalexins. The comparative activities of ROS, including hydrogen peroxide (H2O2), superoxide anions (O2−), and hydroxyl radicals (·OH), against the fungal pathogen Colletotrichum gloeosporioides from papaya fruit were evaluated. The effects of ROS on PPO and POD activities in papaya fruit inoculated with C. gloeosporioides and the development of natural decay in intact fruit were also investigated. ·OH was the most effective in inhibiting conidial germination and mycelial growth of C. gloeosporioides in vitro. However, 20 or 30 mM H2O2 exhibited the best control of the three ROS treatments at ameliorating the disease symptoms associated with the highest levels of PPO and POD activities in papaya fruit. Furthermore, the activities of PPO and POD negatively correlated with the disease index. Overall, H2O2 treatments can induce the resistance of papaya fruit against C. gloeosporioides owing to the enhanced activities of PPO and POD. Treatment with 20 mM H2O2 also significantly reduced the incidence of decay in intact papaya fruit in semi-commercial experiment, which could make it a potential alternative manner to control postharvest disease in papaya fruit.

Abstract There has been continued interest in bacteriocins research from an applied perspective as bacteriocins have potential to be used as natural preservative. Four bacteriocinogenic lactic acid bacteria (LAB) strains of Lactobacillus curvatus (Arla-10), Enterococcus faecium (JFR-1), Lactobacillus paracasei subsp. paracasei (JFR-5) and Streptococcus thermophilus (TSB-8) were previously isolated and identified in our lab. The objective of this study was to determine the optimal growth conditions for both LAB growth and bacteriocins production. In this study, various growth conditions including culture media (MRS and BHI), initial pH of culture media (4.5, 5.5, 6.2, 7.4 and 8.5), and incubation temperatures (20, 37 and 44 °C) were investigated for LAB growth measured as optical density (OD), bacteriocin activity determined as arbitrary unit and viability of LAB expressed as log CFU ml−1. Growth curves of the bacteriocinogenic LAB were generated using a Bioscreen C. Our results indicated that Arla-10, JFR-1, and JFR-5 strains grew well on both MRS and BHI media at growth temperature tested whereas TSB-8 strain, unable to grow at 20 °C. LAB growth was significantly affected by the initial pH of culture media (p

A novel method of generating multi-frequency test stimuli for incipient faults is presented to improve the fault detection accuracy of analog circuits. This paper analyzes the primary cause of low incipient fault detection accuracy and indicates that the high aliasing between the incipient fault response and the normal response seriously impairs the fault recognition ability of fault classifiers. Therefore, this paper builds an aliasing measuring model (AMM) to generate the multi-frequency test stimuli set for incipient faults. The principle part of the AMM is the aliasing measuring algorithm (AMA), which uses the response aliasing as the pivotal index to evaluate the test frequency. The test frequencies with smaller response aliasing will be selected. The other part of the AMM contains the genetic algorithm and the greedy algorithm, which can advance the AMA to quickly generate the multi-frequency test stimuli set for the incipient fault and can ensure that the obtained test set covers the entire circuit and contains fewer test frequencies. The simulation experimental results validate two conclusions: the test frequencies obtained by the AMM remarkably increase the incipient fault detection accuracy and reduce the time for test frequency generation and the multi-frequency test stimuli set contains fewer elements. The hardware experimental results demonstrate that the proposed method is practicable and effective.

Astragalus membranaceus is a valuable medicinal plant species widely distributed in Asia. Its root is the main medicinal tissue rich in methoxylated flavonoids. Origin can highly influence the chemical composition and bioactivity. To characterize the principal chemicals influenced by origin and provide more information about their antioxidant profile, the extracts of A. membranaceus roots from four origins were analysed by UHPLC-MS/MS. Thirty-four flavonoids, including thirteen methoxylated flavonoids, fifteen flavonoid glycosides and six flavonols, were identified. By principal component analysis, eighteen identified compounds were considered to be principal compounds. They could be used to differentiate A. membranaceus from Shanxi, Inner Mongolia, Heilongjiang and Gansu. The antioxidant activity was analysed by ORAC assay, DPPH radical scavenging activity assay and cell antioxidant activity assay. ‘Inner Mongolia’ extract showed the highest antioxidant activity. These results were helpful to understand how origin influenced the quality of A. membranaceus.

In the actual fault diagnosis process of an analog circuit, there is often a problem due to the lack of fault samples, leading to the low-accuracy of diagnostic models. Therefore, using positive samples that are easy to obtain to establish diagnostic models became a research hotspot in the field of analog circuit fault diagnosis. This paper proposes a method based on Support Vector Data Description (SVDD) and Dempster–Shafer evidence theory (D–S evidence theory) for fault diagnosis of modular analog circuit. Firstly, the principle of circuit module partition is proposed to divide the analog circuit under test, and the output port of each module is selected as test point. Secondly, the paper extracts the feature of the time-domain and frequency-domain output signals of the circuit module through Principal Component Analysis (PCA). Thirdly, four state detection models based on SVDD are established to judge the working state of each circuit module, including TSG, TSP, FSG, and FSP state detection model. Finally, the D–S theory is introduced to integrate the test results of each model for locating fault circuit module. To verify the effectiveness of the proposed method, the dual bandpass filter circuit is selected for simulation and hardware experiment. The results show that the proposed method can locate the analog fault effectively and has a higher diagnosis accuracy.

The fruit of Averrhoa carambola L. (Oxalidaceae) is a commercially important fruit. Our study on the chemical constituents present in the fresh sweet fruit led to the isolation of eleven non-flavonoid phenolic compounds. Their structures were identified by analyses of spectroscopic data. They were two new alkyl phenol diglucosides, carambolasides K (1) and L (2), four phenylpropanoids, (+)-isolariciresinol 9-O-β-d-glucoside (3), (+)-lyoniresinol 9-O-β-d-glucoside (4), (−)-lyoniresinol 9-O-β-d-glucoside (5), and 1-O-feruloyl-β-d-glucose (6), three benzoic acids, protocatechuic acid (7), 1-O-vanilloyl-β-d-glucose (8), and tecomin (9), a simple phenol, koaburaside (10), and a naphthoquinone, (+)-cryptosporin (11). All of them were reported from A. carambola fruit for the first time. Compounds 1–8 and 10 showed 2,2′-azinobis-(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) radical cation scavenging activity comparable to l-ascorbic acid, while compounds 3, 5, and 7 exhibited potent ferric reducing antioxidant power (FRAP). In addition, compounds 6 and 11 demonstrated weak porcine pancreatic lipase inhibitory activity.

Ascorbate peroxidase (APX) is a key antioxidant enzyme that is involved in diverse developmental and physiological process and stress responses by scavenging H2O2 in plants. APX itself is also subjected to multiple posttranslational modifications (PTMs). However, redox-mediated PTM of APX in plants remains poorly understood. Here, we identified and confirmed that MaAPX1 interacts with methionine sulfoxide reductase B2 (MsrB2) in bananas. Ectopic overexpression of MaAPX1 delays the detached leaf senescence induced by darkness in Arabidopsis. Sulfoxidation of MaAPX1, i.e., methionine oxidation, leads to loss of the activity, which is repaired partially by MaMsrB2. Moreover, mimicking sulfoxidation by mutating Met36 to Gln also decreases its activity in vitro and in vivo, whereas substitution of Met36 with Val36 to mimic the blocking of sulfoxidation has little effect on APX activity. Spectral analysis showed that mimicking sulfoxidation of Met36 hinders the formation of compound I, the first intermediate between APX and H2O2. Our findings demonstrate that the redox state of methionine in MaAPX1 is critical to its activity, and MaMsrB2 can regulate the redox state and activity of MaAPX1. Our results revealed a novel post-translational redox modification of APX.

Chilling injury is especially prominent in postharvest bananas stored at low temperature below 13 °C. To elucidate better the relationship between cell membrane lipids and chilling injury, an untargeted lipidomics approach using ultra-performance liquid chromatography–mass spectrometry was conducted. Banana fruit were stored at 6 °C for 0 (control) and 4 days and then sampled for lipid analysis. After 4 days of storage, banana peel exhibited a marked chilling injury symptom. Furthermore, 45 lipid compounds, including glycerophospholipids, saccharolipids, and glycerolipids, were identified with significant changes in peel tissues of bananas stored for 4 days compared with the control fruit. In addition, higher ratio of digalactosyldiacylglycerol (DGDG) to monogalactosyldiacylglycerol (MGDG) and higher levels of phosphatidic acid (PA) and saturated fatty acids but lower levels of phosphatidylcholine (PC), phosphatidylethanolamine (PE), and unsaturated fatty acids were observed in banana fruit with chilling injury in contrast to the control fruit. Meanwhile, higher activities of phospholipase D (PLD) and lipoxygenase (LOX) were associated with significantly upregulated gene expressions of MaPLD1 and MaLOX2 and higher malondialdehyde (MDA) content in chilling injury-related bananas. In conclusion, our study indicated that membrane lipid degradation resulted from reduced PC and PE, but accumulated PA, while membrane lipid peroxidation resulted from the elevated saturation of fatty acids, resulting in membrane damage which subsequently accelerated the chilling injury occurrence of banana fruit during storage at low temperature.

Melatonin (MLT) is a vital signaling molecule that regulates multiple physiological processes in higher plants. In the current study, the role of MLT in regulating chilling tolerance and its possible mechanisms in litchi fruit during storage at ambient temperatures after its removal from refrigeration was investigated. The results show that the application of MLT (400 μM, dipping for 20 min) to ‘Baitangying’ litchi fruit effectively delayed the development of chilling injury (CI) while inhibiting pericarp discoloration, as indicated by higher chromacity values (L*, a*, b*) and anthocyanin levels. MLT treatment suppressed the enhancements of the relative electrical conductivity (REC) and malondialdehyde (MDA) content, which might contribute to the maintenance of membrane integrity in litchi fruit. MLT treatment slowed the decline in cellular energy level, as evidenced by higher adenosine triphosphate (ATP) content and a higher energy charge (EC), which might be ascribed to the increased activities of enzymes associated with energy metabolism including H+-ATPase, Ca2+-ATPase, succinate dehydrogenase (SDH), and cytochrome C oxidase (CCO). In addition, MLT treatment resulted in enhanced proline accumulation, which was likely a consequence of the increased activities of ornithine-δ-aminotransferase (OAT) and Δ1-pyrroline-5-carboxylate synthase (P5CS) and the suppressed activity of proline dehydrogenase (PDH). These results suggest that the enhanced chilling tolerance of litchi fruit after MLT treatment might involve the regulation of energy and proline metabolism.

Red light irradiation can effectively prolong the shelf-life of many fruit. However, little is known about red light-induced metabolite and enzyme activities. In this study, pitaya fruit was treated with 100 Lux red light for 24 h. Red light irradiation significantly attenuated the variation trend of senescence traits, such as the decrease of total soluble solid (TSS) and TSS/acidity (titratable acidity, TA) ratio, the increase of TA, and respiratory rate. In addition, the reactive oxygen species (ROS) related characters, primary metabolites profiling, and volatile compounds profiling were determined. A total of 71 primary metabolites and 67 volatile compounds were detected and successfully identified by using gas chromatography mass spectrometry (GC-MS). Red light irradiation enhanced glycolysis, tricarboxylic acid (TCA) cycle, aldehydes metabolism, and antioxidant enzymes activities at early stage of postharvest storage, leading to the reduction of H2O2, soluble sugars, organic acids, and C-6 and C-7 aldehydes. At a later stage of postharvest storage, a larger number of resistance-related metabolites and enzyme activities were induced in red light-treated pitaya peel, such as superoxide dismutase (SOD), ascorbate peroxidase (APX), 1,1-diphenyl-2-picryl-hydrazyl (DPPH) radical-scavenging, reducing power, fatty acids, and volatile aroma.

Fusarium proliferatum is an important pathogen and causes a great economic loss to fruit industry. Environmental pH-value plays a regulatory role in fungi pathogenicity, however, the mechanism needs further exploration. In this study, F. proliferatum was cultured under two initial pH conditions of 5 and 10. No obvious difference was observed in the growth rate of F. proliferatum between two pH-values. F. proliferatum cultured under both pH conditions infected banana fruit successfully, and smaller lesion diameter was presented on banana fruit inoculated with pH 10-cultured fungi. Proteomic approach based on two-dimensional electrophoresis (2-DE) was used to investigate the changes in secretome of this fungus between pH 5 and 10. A total of 39 differential spots were identified using matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry (MALDI-TOF/TOF-MS) and liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS). Compared to pH 5 condition, proteins related to cell wall degrading enzymes (CWDEs) and proteolysis were significantly down-regulated at pH 10, while proteins related to oxidation-reduction process and transport were significantly up-regulated under pH 10 condition. Our results suggested that the downregulation of CWDEs and other virulence proteins in the pH 10-cultured F. proliferatum severely decreased its pathogenicity, compared to pH 5-cultured fungi. However, the alkaline environment did not cause a complete loss of the pathogenic ability of F. proliferatum, probably due to the upregulation of the oxidation-reduction related proteins at pH 10, which may partially compensate its pathogenic ability.

The Dicer protein is one of the most important components of RNAi machinery because it regulates the production of small RNAs (sRNAs) in eukaryotes. Here, Dicer1-like gene (Pit-DCL1) and Dicer2-like gene (Pit-DCL2) RNAi transformants were generated via pSilent-1 in Penicillium italicum (Pit), which is the causal agent of citrus blue mold. Neither transformant showed a change in mycelial growth or sporulation ability, but the pathogenicity of the Pit-DCL2 RNAi transformant to citrus fruits was severely impaired, compared to that of the Pit-DCL1 RNAi transformant and the wild type. We further developed a citrus wound-mediated RNAi approach with a double-stranded fragment of Pit-DCL2 generated in vitro, which achieved an efficiency in reducing Pi-Dcl2 expression and virulence that was similar to that of protoplast-mediated RNAi in P. italicum, suggesting that this approach is promising in the exogenous application of dsRNA to control pathogens on the surface of citrus fruits. In addition, sRNA sequencing revealed a total of 69.88 million potential sRNAs and 12 novel microRNA-like small RNAs (milRNAs), four of which have been predicated on target innate immunity or biotic stress-related genes in Valencia orange. These data suggest that both the Pit-DCL1 and Pit-DCL2 RNAi transformants severely disrupted the biogenesis of the potential milRNAs, which was further confirmed for some milRNAs by qRT-PCR or Northern blot analysis. These data suggest the sRNAs in P. italicum that may be involved in a molecular virulence mechanism termed cross-kingdom RNAi (ck-RNAi) by trafficking sRNA from P. italicum to citrus fruits.

The pericarp of Litchi chinensis was reported to be a rich source of antioxidants. This study aimed to identify the chemical structures present in litchi pericarp with antioxidant activity. Four lignans, (+)-isolariciresinol 9-O-α-l-arabinopyranoside (1), burselignan 9-O-α-l-arabinopyranoside (2), (−)-secoisolariciresinol 9-O-α-l-arabinopyranoside (3), and sisymbrifolin (4), a chromane, 2α-methoxychroman-3α,5,7-triol (5), a benzophenone, garcimangosone D (6), and two sesquiterpenes, β-d-glucopyranosyldihydrophaseate (7) and citroside A (8) were isolated. Their structures were elucidated by spectroscopic methods. Compounds 1–3, 5, and 7 were new metabolites and the others were reported from this species for the first time. Among them, compounds 1–5 showed moderate antioxidant activity in ferric reducing antioxidant power (FRAP) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assays.

Litchi (Litchi chinensis Sonn.) is a well-known traditional medicine since ancient times. The crude extract of litchi leaf exhibited good bioactivity against cancer cells. In this work, three sesquilignans, ehletianol C (1), sesquipinsapol B (2) and sesquimarocanol B (3), and one lignan glycoside schizandriside (4) were isolated from the ethyl acetate-soluble extract of litchi leaf by column chromatography. Their structures were further elucidated by ESI-MS and NMR spectral studies. Ehletianol C, sesquipinsapol B and sesquimarocanol B were detected from this species for the first time. Sesquipinsapol B (EC50 values less than 2.0 μg/mL) and sesquimarocanol B (EC50 values less than 2.4 μg/mL) showed very high cytotoxicities against nasopharynx carcinoma cells CNE1 and CNE2 cells, respectively. Thus, they could be utilized as good drug candidates against nasopharynx cancer. Moreover, three sesquilignans showed effective in vitro cytotoxicities against human cancer cells HepG2 and Hela. Four lignans showed better antioxidant activities than butylated hydroxytoluene (BHT).

Biotransformation with Aspergillus awamori could degrade the bound bioactive compounds of plant tissues into free form and induce the biosynthesis of novel chemicals. In this work, two bioactive compounds with high levels were produced from litchi pericarp via A. awamori fermentation. They were purified and identified to be nigragillin and dihydrophaseic acid by nuclear magnetic resonance spectroscopy and mass spectrometry. The anticancer activity, DNA protection effect and antioxidant activity of these two compounds were evaluated. Dihydrophaseic acid had significantly inhibitory effect against HepG2 and HeLa cells with IC50 values of 41.92 ± 6.15 and 79.37 ± 9.78 μg/mL, respectively. Nigragillin showed a better DPPH radical scavenging activity than dihydrophaseic acid. Dihydrophaseic acid possessed good lipid peroxidation inhibition effect, hydroxyl radical scavenging activity, and DNA protection effect even at a low dosage. The results indicated that biotransformation with A. awamori was a good way to produce bioactive compounds.

The banana is one of the most important fruits in the world. Bananas undergo a rapid ripening process after harvest, resulting in a short shelf. In this study, the mechanism underlying pulp ripening of harvested bananas was investigated using integrated transcriptomic, proteomic, and metabolomic analysis. Ribonucleic acid sequencing (RNA-Seq) revealed that a great number of genes related to transcriptional regulation, signal transduction, cell wall modification, and secondary metabolism were up-regulated during pulp ripening. At the protein level, 84 proteins were differentially expressed during pulp ripening, most of which were associated with energy metabolism, oxidation-reduction, cell wall metabolism, and starch degradation. According to partial least squares discriminant analysis, 33 proteins were identified as potential markers for separating different ripening stages of the fruit. In addition to ethylene’s central role, auxin signal transduction might be involved in regulating pulp ripening. Moreover, secondary metabolism, energy metabolism, and the protein metabolic process also played an important role in pulp ripening. In all, this study provided a better understanding of pulp ripening of harvested bananas.

β-aminobutyric acid (BABA) is a new environmentally friendly agent to induce disease resistance by priming of defense in plants. However, molecular mechanisms underlying BABA-induced priming defense are not fully understood. Here, comprehensive analysis of priming mechanism of BABA-induced resistance was investigated based on mango-Colletotrichum gloeosporioides interaction system using iTRAQ-based proteome approach. Results showed that BABA treatments effectively inhibited the expansion of anthracnose caused by C. gleosporioides in mango fruit. Proteomic results revealed that stronger response to pathogen in BABA-primed mango fruit after C. gleosporioides inoculation might be attributed to differentially accumulated proteins involved in secondary metabolism, defense signaling and response, transcriptional regulation, protein post-translational modification, etc. Additionally, we testified the involvement of non-specific lipid-transfer protein (nsLTP) in the priming acquisition at early priming stage and memory in BABA-primed mango fruit. Meanwhile, spring effect was found in the primed mango fruit, indicated by inhibition of defense-related proteins at priming phase but stronger activation of defense response when exposure to pathogen compared with non-primed fruit. As an energy-saving strategy, BABA-induced priming might also alter sugar metabolism to provide more backbone for secondary metabolites biosynthesis. In sum, this study provided new clues to elucidate the mechanism of BABA-induced priming defense in harvested fruit.

The juice sacs of pummelo fruit is susceptible to softening during storage at 25 °C, which causes quality deterioration and flavor loss during postharvest pummelo storage. This study investigated the changes in metabolisms of antioxidant and cell wall in juice sacs of three pummelo cultivars—Hongroumiyou (HR), Bairoumiyou (BR) and Huangroumiyou (HuR)—during postharvest storage. The results revealed that, with the extension of storage, the juice sacs of three pummelo cultivars exhibited a decrease in total antioxidant capacity (TAC), DPPH and ABTS radical scavenging activity; a decline in total phenols (TP) content and an increase firstly then a decrease in total ascorbic acid (TAA) content; and a decrease in lipoxygenase (LOX) activity and a rise initially, but a decline in activities of ascorbate peroxidase (APX) and glutathione peroxidase (GPX). Additionally, increased water-soluble pectin (WSP), but declined propectin, ionic-soluble pectin (ISP) and chelator-soluble pectin (CSP); as well as an increase from 0 d to 60 d then followed by a decline in activities of pectinesterase (PE), polygalacturonase (PG) and pectate lyase (PL) were observed. These results suggested that the metabolisms of antioxidant and cell wall could result in softening and senescence of pummelo fruit.