Development of antifogging double-layer film using cellulose nanofibers and carboxymethyl chitosan for white Hypsizygus marmoreus preservation.

Films with simultaneously excellent mechanical and anti-fog properties are of great importance for food packaging. A novel strategy is described here to prepare long-lasting anti-fog film with antibacterial and antioxidant capabilities via a simple, green approach. The CMC (carboxymethyl chitosan) gel was integrated with CNF/TA (cellulose nanofibers/tannic acid) composite solution based on layer-by-layer assembly to form a membrane with a bilayer structure. The anti-fog performance of the bilayer film could be adjusted by regulating the CNF/TA layer thickness. On the whole, the developed anti-fog film had high mechanical strength and excellent UV shielding properties, as well as good antibacterial and antioxidant properties, and could be non-fogging for a long time under water vapor (40 °C). The effect of double layer anti-fog film (3%CmFT-3) on the fresh-keeping effect of white Hypsizygus marmoreus was compared at room temperature (28 °C) with commercially available anti-fog PVC film. The results showed that the bilayer anti-fog film could effectively prevent the generation of fog, delay the Browning, inhibit mildew, improve the overall acceptability, and effectively extend the shelf life of white Hypsizygus marmoreus. This biomass-based anti-fog film offers great potential for the development of multifunctional green food packaging.

Lowering glycemic levels via gastrointestinal tract factors: the roles of dietary fiber, polyphenols, and their combination.

Dietary fiber (DF) and polyphenols (DP) are typical blood sugar-lowering components, and both play distinct yet interconnected roles in exerting their blood sugar-lowering effects. We comprehensively summarized the single and combined effects of DF and DP on blood glucose homeostasis through regulating the relevant factors in the upper gastrointestinal tract (UGT) and lower gastrointestinal tract (LGT). In the UGT, DF slowed down glucose metabolism by enhancing digesta viscosity and hindering enzyme-substrate interaction. DP primarily targeted enzymes and substrates. When combined, DP enhanced the adsorption capacity of DF for glucose. DF weakened DP's inhibitory effect on enzymes. Both DF and DP disrupted glucose intestinal uptake via physical or genomic modulation, but the co-consumption of DF and DP demonstrated a lower inhibitory effect on glucose uptake than DP alone. In the LGT, DF and DP showed synergistic or antagonistic effects on gut microbiota. Remarkably, whole foods exhibited potent prebiotic effects due to their compound-rich matrix, potentially enhancing glucose homeostasis and expanding dietary options for glucose regulation research.

Effect of L-cysteine treatment to induce postharvest disease resistance of Monilinia fructicola in plum fruits and the possible mechanisms involved.

Plum is an important stone fruit in China, but the fruit is easily perishable and susceptible to infection by pathogens. Traditionally, synthetic fungicides are used to control diseases. However, the side effects of fungicides should not be ignored. Cysteine, generally recognized as safe (GRAS) amino acid, has been reported to play roles in the plant abiotic stress response, but little is known about the role of cysteine to control postharvest diseases in fruits. Therefore, this study was designed to investigate the effect of L-cysteine treatment on control of postharvest brown rot in artificially inoculated plum fruits and the possible biocontrol mechanisms involved. Postharvest plum fruits were inoculated with 1, 10, 100 and 1000 mg L-1 L-cysteine. 100 mg L-1 L-cysteine treatment effectively controlled brown rot in artificially inoculated plum fruits by inducing resistance. Furthermore, 100 mg L-1 L-cysteine treatment increased the activities of glucose-6-phosphate dehydrogenase (G6PDH) and 6-phosphogluconate dehydrogenase (6PGDH), enhanced the content of NADPH of the pentose phosphate pathway, as well as improved the contents of H2O2 and some amino acids in the artificially inoculated plum fruits. 100 mg L-1 L-cysteine treatment also elevated the antioxidant content (AsA, GSH) and the antioxidant enzymes activities (APX, GR, MDAR, DHAR) of the ascorbate-glutathione (AsA-GSH) pathway. The protective effects of L-cysteine treatment on postharvest plum fruits likely be due to activating some defense-related responses of the fruit against infection. L-cysteine treatment is a safe promising method for controlling postharvest brown rot in plum fruits.

Dietary fiber of Tartary buckwheat bran modified by steam explosion alleviates hyperglycemia and modulates gut microbiota in db/db mice.

Type 2 diabetes is a serious threat to human health. Tartary buckwheat bran dietary fiber has good hypoglycemic activity, with its modification widely studied. However, the hypoglycemic activity of steam explosion modified Tartary buckwheat bran soluble dietary fiber (SE-SDF) has not been reported. This research aimed at investigating the hypoglycemic effect with its underlying mechanism of SE-SDF on type 2 diabetic db/db mice. Results found SE-SDF decreased the levels of fasting blood glucose and glycosylated hemoglobin while improved oral glucose tolerance, insulin resistance, and injuries of liver, pancreas, and colon in diabetic db/db mice. Additionally, SE-SDF up-regulated the protein expression levels of hepatic phosphatidylinositol 3 kinase (PI3K), G protein-coupled receptor43 (GPR43), and phospho-adenosine monophosphate activated protein kinase (p-AMPK), whereas inhibited the protein expression levels of hepatic fork-head transcription factor O1 (FoxO1), phosphoenolpyruvate carboxy kinase (PEPCK) and glucose-6-phosphatase (G-6-Pase). Moreover, SE-SDF increased the production of fecal short chain fatty acids (SCFAs) and the expression of colon GPR43 and the concentration of serum glucagon like peptide-1 (GLP-1), leading to reduced ratio of Firmicutes/Bacteroidetes but increased relative abundance of Parabacteroides, norank_f_Muribaculaceae, Alloprevotella, Ruminiclostridium_9, unclassified_f_Ruminococcaceae, and Lachnospiraceae_NK4A136_group. These findings suggested that SE-SDF ameliorated type 2 diabetes via activating the liver PI3K/Akt/FoxO1 and GPR43/AMPK signaling pathways and modulating the gut microbiota-SCFAs-GPR43/GLP-1 signaling axis.

Anti-aging effects of the fermented anthocyanin extracts of purple sweet potato on Caenorhabditis elegans.

Anthocyanins have anti-inflammatory, anticarcinogenic and antioxidant properties and anti-aging effects as well as potential application as pigments. The metabolism of anthocyanins in fermented food has attracted increasing attention. However, the effect of lactic acid bacteria (LAB) fermentation on its anti-aging activity remains mostly unknown. The current study aimed to investigate the compositions, antioxidant activities and anti-aging effect of fermented purple sweet potato anthocyanins (FSPA) on aging Caenorhabditis elegans compared to raw purple sweet potato anthocyanins (PSPA). Results showed that anthocyanins were degraded into more bioavailable phenolic acids by Weissella confusa fermentation. PSPA and FSPA can extend the lifespan of C. elegans by 26.7% and 37.5%, respectively, through improving the activity of antioxidant enzymes as well as decreasing MDA content, ROS levels and lipofuscin accumulation. Pretreatment of the worms with PSPA and FSPA induced their potential to resist to thermal tolerance and oxidative stress, and FSPA exerted a higher anti-stress effect than PSPA. Moreover, FSPA supplementation upregulated the mRNA expressions of genes daf-16, hsp-16.2, sir-2.1, skn-1 and sod-3 and downregulated the expression of daf-2 in the nematodes, whereas PSPA only induced the increase in the expressions of sir-2.1, skn-1 and sod-3. Overall, FSPA can improve stress resistance and extend the lifespan of C. elegans by both insulin/IGF-1 signaling pathway and dietary restriction pathway, providing a theoretical basis for the application of PSPA in fermented food as functional pigments.

Metschnikowia citriensis FL01 antagonize Geotrichum citri-aurantii in citrus fruit through key action of iron depletion.

Metschnikowia citriensis FL01 has great potential for biocontrol applications for its excellent biocontrol efficacy on postharvest diseases of citrus fruit, and the iron depletion by pulcherriminic acid (PA) and then formation of insoluble pigment pulcherrimin had been speculated as an important action mechanism. To identify the genes involved in pulcherrimin synthesis and reutilization in M. citriensis FL01, we de novo assembled the genome of M. citriensis FL01 based on long-read PacBio sequencing. The final assembled genome consisted of 12 contigs with a genome size of 25.74 Mb, G + C content of 49.16% and 9310 protein-coding genes. The genome-wide BLAST of the PUL genes of M. pulcherrima APC 1.2 showed that the four PUL genes were clustered and located on Contig 4 of M. citriensis FL01. In order to further clarify the role of pulcherrimin pigment on biocontrol of M. citriensis FL01, CRISPR/cas9 technology was used to knock out PUL2 gene that was responsible for PA synthesis and the pigmentless mutants with stable phenotype were obtained. The mutant strains of M. citriensis FL01 lost the ability to produce pulcherrimin pigment, and simultaneously lost the ability to inhibit the growth of Geotrichum citri-aurantii in vitro. Moreover, the biocontrol efficacy of pigmentless mutant strains against sour rot was about 80% lower than that of wild-type M. citriensis FL01. These results directly proved that the iron depletion was an important mechanism of M. citriensis FL01.

Promotion of the anticancer activity of curcumin based on a metal-polyphenol networks delivery system.

Curcumin (Cur), a hydrophobic active pharmaceutical ingredient with high anticancer activity, has poor water solubility and low bioavailability. Although many delivery systems have been developed to improve their bioavailability, some limitation such as low drug loading efficiency and poor stability are still remained. The metal-polyphenol networks (MPNs) delivery system designed in this subject solved above problems and effectively improved the anticancer activity of Cur. The synthesized Cur@EGCG-Fe(III) is consisting of epigallocatechin gallate (EGCG), iron chloride (FeCl3) and Cur, and the well-designed structure endow Cur@EGCG-Fe(III) high loading efficiency, good water solubility and stability. After the Cur@EGCG-Fe(III) nanoparticles were internalized by MCF-7 cells, the Cur could be released in endo/lysosomal microenvironment (pH = 5.0), and the Cur delivery in the deep tumor could be realized. The distribution of Cur@EGCG-Fe(III) in MCF-7 cells was analyzed by laser confocal, and Cur@EGCG-Fe(III) could effectively deliver more Cur into MCF-7 cells in comparison with free Cur. In addition, the results of flow cytometry and western blot further indicated that Cur@EGCG-Fe(III) had a stronger ability to induce apoptosis than free Cur. Transwell cell migration and invasion experiments showed that Cur and EGCG-Fe(III) had a synergistic effect in inhibiting MCF-7 cell migration and invasion. In vitro hemolysis and in vivo experiments showed that the Cur@EGCG-Fe(III) had negligible effect on the blood environment and a great tumor-inhibition efficacy, indicating that the MPNs delivery system had a good blood compatibility and antitumor activity. Our results indicated that MPNs-coated Cur nanoparticle could be a new form of Cur delivery system for anticancer application.

Transcription Factor CsWRKY65 Participates in the Establishment of Disease Resistance of Citrus Fruits to Penicillium digitatum.

Penicillium digitatum is the primary pathogen that causes serious yield losses worldwide. In our previous study, CsWRKY transcription factors (TFs) and some genes associated with immunity were identified in citrus fruits after P. digitatum infection, but little information is available in the literature on the mechanisms of TFs in citrus disease resistance. In this study, the possible mechanisms of CsWRKY65 participating in the establishment of disease resistance were investigated. Results show that CsWRKY65 was a transcriptional activator in the nucleus. The dual-luciferase transient assays and electrophoretic mobility shift assays showed that CsWRKY65 bound with CsRbohB, CsRbohD, CsCDPK33, and CsPR10 promoters to activate gene transcription. Besides, the transient overexpression of CsWRKY65 induced reactive oxygen species accumulation and increased PR gene expression in Nicotiana benthamiana leaves. The transient overexpression of CsWRKY65 in the citrus peel enhanced the disease resistance against P. digitatum. In conclusion, CsWRKY65 is likely to be involved in regulating the disease resistance to P. digitatum of citrus fruits by directly activating the expressions of CsRbohB, CsRbohD, CsCDPK33, and CsPR10. This study provides new information for the mechanism of citrus WRKY TFs participating in the establishment of disease resistance.

Pichia galeiformis Induces Resistance in Postharvest Citrus by Activating the Phenylpropanoid Biosynthesis Pathway.

This study aimed to investigate the effect of Pichia galeiformis on disease resistance and elucidate the changes in phenylpropane biosynthesis treated by P. galeiformis in postharvest citrus. The results showed that P. galeiformis reduced the disease incidence and lesion diameters without direct contact with the pathogen Penicillium digitatum. Transcriptome analysis revealed that phenylpropanoid biosynthesis was triggered by P. galeiformis. Genes encoding phenylpropanoid biosynthesis were upregulated, including phenylalanine ammonia-lyase (PAL), 4-coumaroyl-CoA ligase (4CL), cinnamate-4-hydroxylase (C4H), peroxidase (POD), cinnamyl alcohol dehydrogenase (CAD), O-methyltransferase, and hydroxyl cinnamoyl transferase. Moreover, P. galeiformis increased the activity of PAL, 4CL, C4H, POD, polyphenol oxidase, and CAD in citrus pericarp. In addition, P. galeiformis treated citrus displayed higher levels of total phenolic compounds, flavonoid, and lignin and higher amounts of ferulic and sinapic acid. In conclusion, these results suggested that P. galeiformis could induce resistance through modulating the pathway of phenylpropanoid biosynthesis in postharvest citrus.

Variations in chlorophyll and carotenoid contents and expression of genes involved in pigment metabolism response to oleocellosis in citrus fruits.

Yellow or green spots related to pigment changes found at the early stage of oleocellosis can cause severe economic damage. However, little information exists on pigment changes during oleocellosis development, so this study investigated the main changes in chlorophyll and carotenoid metabolites and related gene expression. Among the variations, the increased contents of chlorophyll a and b, and decreased concentrations of lutein, ß-cryptoxanthin, zeaxanthin, violaxanthin, α-carotene and ß-carotene were responsible for chlorophyll and carotenoid changes, respectively. Regarding gene expression, the up-regulated genes, magnesium chelatase subunit H (MgCh), magnesium-protoporphyrin IX monomethyl ester (oxidative) cyclase 1/2 (MPEC1/2), protochlorophyllide reductase a, chloroplastic 1/2 (PORA1/2) and chlorophyllide a oxygenase (CAO), regarding chlorophyll synthesis as well as the down-regulated genes, phytoene synthase (PSY), phytoene dehydrogenase (PDS), lycopene ß-cyclase (LCYb), and zeaxanthin epoxidase 1/2 (ZEP 1/2) and the up-regulated genes (+)-abscisic acid 8'-hydroxylase 1/2 (ABA-HX 1/2), regarding carotenoid metabolism, constituted the major variations in oleocellosis peels.

Analysis of changes in volatile constituents and expression of genes involved in terpenoid metabolism in oleocellosis peel.

Oleocellosis is a serious physiological disorder in citrus fruit that mainly results in appearance and quality deterioration. It has been well established that the occurrence of oleocellosis is highly correlated with the release of peel oil from citrus fruit, while there is little information on the dynamic changes in the content of the volatile constituents and the expression of genes involved in terpenoid metabolism during oleocellosis development. In the present research, large changes in the volatile profiles and gene expression in terpenoid metabolism were observed in oleocellosis peels compared to healthy ones. Among volatiles, the decreased contents of α-pinene, d-limonene, ß-myrcene, linalool, ß-caryophyllene, α-terpineol, nonanal, neryl acetate and (-)-carvone played a major role in these changes. For gene expressions in terpenoid metabolism, the up-regulated genes aldehyde dehydrogenase (NAD+) (ALDH) and the down-regulated genes ß-caryophyllene synthase 1 (BCS1), α-terpineol synthase 2 (TES2) and myrcene synthase (MS) were the main differences in oleocellosis peels.

Exogenous nitric oxide-induced postharvest disease resistance in citrus fruit to Colletotrichum gloeosporioides.

BACKGROUND: Nitric oxide (NO) is an important signaling molecule involved in numerous plant responses to biotic and abiotic stresses. To investigate the effects of NO on the control of postharvest anthracnose caused by Colletotrichum gloeosporioides in citrus fruit and its possible mechanisms, citrus fruit were treated with an NO donor. RESULTS: The results showed that exogenous NO released from 50 µmol L(-1) sodium nitroprusside aqueous solution could effectively reduce the disease incidence and lesion diameter of citrus fruit inoculated with C. gloeosporioides during storage at 20 °C. Exogenous NO could regulate hydrogen peroxide levels, stimulate the synthesis of phenolic compounds, and induce phenylalanine ammonia-lyase, peroxidase, polyphenol oxidase, catalase activities, and the ascorbate-glutathione cycle. Furthermore, exogenous NO could inhibit weight loss, improve the ascorbic acid and titratable acidity content, and delay the increase in total soluble solids content in citrus fruit during storage at 20 °C. CONCLUSIONS: The results suggest that the use of exogenous NO is a potential method for inducing the disease resistance of fruit to fungal pathogens and for extending the postharvest life of citrus fruit.