L-malic Acid Research Articles

The effects and cell phenotype changes in Escherichia coli O157:H7 induced into VBNC state by l-malic acid at low temperature

Under environmental stress conditions, microorganisms can form a viable but non-culturable (VBNC) state, which poses a potential risk to food safety and human health. l-Malic acid (L-MA) is widely found in low-temperature fruit and vegetable juices as a common natural organic acid as well as a food additive. However, there is no report on whether L-MA in food processing can promote the formation of VBNC states. In this study, we investigated the pattern of the L-MA-induced VBNC state of Escherichia coli O157:H7 as a model organism, as well as the characteristics at the cellular and molecular levels. We discovered that L-MA can induce the VBNC state in a significant number of E. coli cells, a process accelerated by low temperatures (4 °C). Our analysis revealed that VBNC cells induced by both L-MA and 4 °C resuscitated earlier than those subjected to either condition alone, indicating increased food safety risks. For survival, L-MA-induced VBNC E. coli cells became substantially smaller and exhibited a significant decrease in respiratory activity while maintaining the basic function of the cell membrane. These cellular changes were supported by molecular-level evidence, reinforcing the implications for food safety under these conditions. This study underscores the importance of recognizing the potential risks posed by L-MA and low temperatures in food preservation, particularly in terms of underestimating the presence of harmful bacteria in food products. The insights gained from this research could contribute to the development of more effective food safety measures, particularly in the handling and processing of fruit and vegetable juices.

Metabolic engineering of Aspergillus niger for accelerated malic acid biosynthesis by improving NADPH availability.

Microbial production of L-malic acid from renewable carbon sources has attracted extensive attention. The reduced cofactor NADPH plays a key role in biotransformation because it participates in both biosynthetic reactions and cellular stress responses. In this study, NADPH or its precursors nicotinamide and nicotinic acid were added to the fermentation medium of Aspergillus niger RG0095, which significantly increased the yield of malic acid by 11%. To further improve the titer and productivity of L-malic acid, we increased the cytoplasmic NADPH levels of A. niger by upregulating the NAD kinases Utr1p and Yef1p. Biochemical analyses demonstrated that overexpression of Utr1p and Yef1p reduced oxidative stress, while also providing more NADPH to catalyze the conversion of glucose into malic acid. Notably, the strain overexpressing Utr1p reached a malate titer of 110.72±1.91gL-1 after 108h, corresponding to a productivity of 1.03±0.02gL-1h-1. Thus, the titer and productivity of malate were increased by 24.5% and 44.7%, respectively. The strategies developed in this study may also be useful for the metabolic engineering of fungi to produce other industrially relevant bulk chemicals.

Effects of verjus acidification on muscat canelli grape juice and wines

Our study aimed to investigate the use of verjus (green grape juice) as an acidifying agentfor improving the overall acid profile and balance of wine, lowering the pH and alcohol content and creating a sustainable winemaking practice that can be utilised in warm grape-growing regions, such as Texas, where pH levels range from 3.5 to over 4.0 in white juice and wine. Muscat Canelli wines were produced using two different treatments for pH and acidity balance. Verjus was produced by pressing under-ripe grapes resulting from crop thinning practices and added at two different rates of 2 % and 11 % to juice obtained from ripe grapes prior to fermentation. For all the wines produced, pH, titratable acidity (expressed as g/L Tartaric Acid), alcohol, glucose-fructose, l-malic acid, tartaric acid, potassium, polyphenols were analysed, and preference testing was performed via consumer sensory panels. Results obtained indicate that verjus can be used to balance pH and acidity as well as lower alcohol content. Consumer preference ranking indicated no significant differences between any of the treatments, while acidity rankings correctly distinguished the wines according to their acid levels. This work shows that the use of verjus as a pre-fermentation acidification agent could be a potential tool for winemaking to help mitigate the negative effects associated with climate change.

Broiler Spaghetti Meat Abnormalities: Muscle Characteristics and Metabolomic Profiles.

Spaghetti meat (SM) is a newly identified muscle abnormality that significantly affects modern broiler chickens, consequently exerting a substantial economic impact on the poultry industry worldwide. However, investigations into the meat quality and the underlying causative factors of SM in broilers remain limited. Therefore, this study was undertaken to systematically evaluate meat quality and muscle fiber characteristics of SM-affected meat. To elucidate the disparities between SM-affected and normal (NO) muscles in broiler chickens reared under identical conditions, we selected 18 SM-affected breast tissues and 18 NO breast tissues from 200 broiler chickens raised according to commercial standards under the same conditions for our study. The results showed that compared with the NO group, the muscle surface of the SM group lost integrity, similar to strip and paste. The brightness and yellowness values were significantly higher than those of the NO group. On the contrary, the shear force and protein were significantly lower in the SM group. Microscopic examination revealed that the muscle fibers in the SM group were lysed, necrotic, and separated from each other, with a large number of neutrophils diffusely distributed on the sarcolemma and endometrium. Thirty-five significantly different metabolites were observed in the breast muscles between both groups. Among them, the top differential metabolites-14,15-DiHETrE, isotretinoin, L-malic acid, and acetylcysteine-were mainly enriched in lipid metabolism and inflammatory pathways, including linoleic acid, arachidonic acid, phenylalanine, and histidine metabolism. Overall, these findings not only offer new insights into the meat quality and fiber traits of SM but also contribute to the understanding of potential mechanisms and nutritional regulators for SM myopathy.

Characterization of malolactic fermentation by Lactiplantibacillus plantarum in red grape must

Malolactic fermentation (MLF) is a desirable biotransformation in red and some high-acidity white wines. This process involves the decarboxylation of l-malic acid into l-lactic acid by lactic acid bacteria (LAB). Among them, Oenococcus oeni and Lactiplantibacillus plantarum are the most common, being the last usually used as starter to perform MLF in wine. Compared with O. oeni, L. plantarum is less tolerant to wine conditions such as pH and ethanol; thus, it is preferable to inoculate it in must. The current literature has not yet addressed the impact of some parameters on the inoculation of this bacterium in grape must. Therefore, the aim of this work was to characterize the effect of the inoculum concentration, pH and ethanol tolerance, as well as the inoculation strategy, on the MLF performance of L. plantarum in grape must. We observed that a population < 108 CFU/mL could compromise MLF performance. Additionally, L. plantarum was able to survive at low pH values, such as 2.50, but in the presence of ethanol or a fermenting yeast, it could tolerate pH values in the range of 3.00–3.25. Finally, coinoculation or sequential inoculation with S. cerevisiae allows for the completion of MLF before the end of alcoholic fermentation.

Insights into tolerance mechanisms of earthworms (Eisenia fetida) in copper-contaminated soils by integrating multi-omics analyses

Earthworms can resist high levels of soil copper (Cu) contamination and play an essential role in absorbing them effectively. However, the molecular mechanisms underlying Cu tolerance in earthworms are poorly understood. To address this research gap, we studied alterations of Eisenia fetida in antioxidant enzymes, gut microbiota, metabolites, and genes under varying levels of Cu exposure soils (0, 67.58, 168.96, 337.92 mg/kg). Our results revealed a reduction in antioxidant enzyme activities across all treatment groups, indicating an adaptive response to alleviate Cu-induced oxidative stress. Analysis of gut microbiota revealed a significant increase in the abundance of bacteria associated with nutrient uptake and Cu2+ excretion under Cu stress. Furthermore, metabolomic analysis discovered an increase in certain metabolites associated with energy metabolism, such as pyruvic acid, L-malic acid, and fumaric acid, as Cu concentration escalated. These results suggested that enhanced energy supply contributes to the elevated tolerance of E. fetida towards Cu. Additionally, transcriptome analysis not only identified crucial detoxification genes (Hsp70, CTSL, GST, CHAC, and GCLC), but also confirmed the critical role of glutathione metabolism as a key pathway in E. fetida Cu detoxification processes. These findings provide a new perspective on the molecular mechanisms of Cu tolerance in earthworms.

Distinct Changes in Metabolic Profile and Sensory Quality with Different Varieties of Chrysanthemum (Juhua) Tea Measured by LC-MS-Based Untargeted Metabolomics and Electronic Tongue.

Chrysanthemum tea, a typical health tea with the same origin as medicine and food, is famous for its unique health benefits and flavor. The taste and sensory quality of chrysanthemum (Juhua) tea are mainly determined by secondary metabolites. Therefore, the present research adopted untargeted metabolomics combined with an electronic tongue system to analyze the correlation between the metabolite profiles and taste characteristics of different varieties of chrysanthemum tea. The results of sensory evaluation showed that there were significant differences in the sensory qualities of five different varieties of chrysanthemum tea, especially bitterness and astringency. The results of principal component analysis (PCA) indicated that there were significant metabolic differences among the five chrysanthemum teas. A total of 1775 metabolites were identified by using untargeted metabolomics based on UPLC-Q-TOF/MS analysis. According to the variable importance in projection (VIP) values of the orthogonal projections to latent structures discriminant analysis (OPLS-DA), 143 VIP metabolites were found to be responsible for metabolic changes between Huangju and Jinsi Huangju tea; among them, 13 metabolites were identified as the key metabolites of the differences in sensory quality between them. Kaempferol, luteolin, genistein, and some quinic acid derivatives were correlated with the "astringency" attributes. In contrast, l-(-)-3 phenyllactic acid and L-malic acid were found to be responsible for the "bitterness" and "umami" attributes in chrysanthemum tea. Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis showed that the flavonoid and flavonol biosynthesis pathways had important effects on the sensory quality of chrysanthemum tea. These findings provide the theoretical basis for understanding the characteristic metabolites that contribute to the distinctive sensory qualities of chrysanthemum tea.

Analysis of blood biochemistry and non-targeted metabolomics of endometritis in dairy cows

The incidence of bovine endometritis, which has a negative impact on the reproduction of dairy cows, has been recently increasing. In this study, the differential markers and metabolites of healthy cows and cows with endometritis were analyzed by measuring blood biochemical indicators and immune factors using biochemical and enzyme-linked immunosorbent assay kits combined with nontargeted metabolomics. The LC-QTOF platform was used to evaluate the serum metabolomics of healthy cows and cows with endometritis after 21–27 days of calving. The results showed that glucose, free fatty acid, calcium, sodium, albumin, and alanine aminotransferase levels were significantly lower in the serum of cows with endometritis than in healthy cows (P < 0.05). However, the serum potassium, interleukin-1, interleukin-6, and tumor necrosis factor levels were significantly higher in cows with endometritis (P < 0.05). In addition, the serum metabolome data analysis of the two groups showed that the expression of 468 metabolites was significantly different (P < 0.05), of which 291 were upregulated and 177 were downregulated. These metabolites were involved in 78 metabolic pathways, including amino acid, nucleotide, carbohydrate, lipid, and vitamin metabolism pathways; signal transduction pathways, and other biological pathways. Taken together, negative energy balance and immune activation, which are related to local abnormalities in amino acid, lipid, and carbohydrate metabolism, were the important causes of endometritis in dairy cows. Metabolites such as glucose, carnosine, dehydroascorbic acid, L-malic acid, tetrahydrofolic acid, and UDP-glucose may be used as key indicators in the hematological diagnosis and treatment of endometritis in dairy cows.

Aspergillus oryzae as a Cell Factory: Research and Applications in Industrial Production.

Aspergillus oryzae, a biosafe strain widely utilized in bioproduction and fermentation technology, exhibits a robust hydrolytic enzyme secretion system. Therefore, it is frequently employed as a cell factory for industrial enzyme production. Moreover, A. oryzae has the ability to synthesize various secondary metabolites, such as kojic acid and L-malic acid. Nevertheless, the complex secretion system and protein expression regulation mechanism of A. oryzae pose challenges for expressing numerous heterologous products. By leveraging synthetic biology and novel genetic engineering techniques, A. oryzae has emerged as an ideal candidate for constructing cell factories. In this review, we provide an overview of the latest advancements in the application of A. oryzae-based cell factories in industrial production. These studies suggest that metabolic engineering and optimization of protein expression regulation are key elements in realizing the widespread industrial application of A. oryzae cell factories. It is anticipated that this review will pave the way for more effective approaches and research avenues in the future implementation of A. oryzae cell factories in industrial production.

Functional carbon dots-hydrogel complex for selective antibacterial and detection applications

The carbon dots (CDs) with excellent optical properties and their hydrogel complex are of great significance in biomedicine, healthcare and biochemical detection fields. This paper reports the preparation of green-emitting CDs (MA-CDs) through one-step hydrothermal route with citric acid as reducing agent, L-malic acid as carbon source and N-(2-hydroxyethyl)ethylenediamine as nitrogen source. To expand its application in biology, MA-CDs were coupled with vancomycin to obtain multifunctional CDs (VMA-CDs). The prepared VMA-CDs exhibit selective antibacterial behavior to Gram-positive bacteria, and it could be used as a fluorescent probe to selectively label Staphylococcus aureus (S. aureus). Moreover, thanks to the excellent optical properties of VMA-CDs, it has been used as a fluorescent sensor to detect Au3+ with detection range of 6.50 nM–21.93 μM and detection limit 3.98 nM. By introducing the fluorescence of CDs as the reference signal, and VMA-CDs as a response signal, the hydrogel (V-SP) was prepared and realized the detection of Au3+ in microfluidics with assistance of a smartphone to collect and analyze data.

Conditional expression of FumA in Aspergillus niger enhances synthesis of L-malic acid.

Currently, the L-malic acid titer achieved through Aspergillus niger fermentation reaches 201 g/L, meeting industrial demands satisfactorily. However, the co-presence of structurally similar fumaric acid and succinic acid in fermentation products suggests a theoretical potential for further improvement in L-malic acid production. In the tricarboxylic acid cycle, fumarate reductase mediates the conversion of succinic acid to fumaric acid. Subsequently, fumarase catalyzes the conversion of fumaric acid to L-malic acid. Notably, both enzymatic reactions are reversible. Our investigation revealed that A. niger contains only one mitochondria-located fumarase FumA. Employing CRISPR-Cas9 technology, we performed a replacement of the fumA promoter with a doxycycline-induced promoter Tet. Under non-inducing condition, the conditional strain exhibited increased levels of fumaric acid and succinic acid. It strongly suggests that FumA mainly promotes the flow of fumaric acid to L-malic acid. Furthermore, a promoter PmfsA that is exclusively activated in a fermentation medium by calcium carbonate was identified through RNA-sequencing screening. Utilizing PmfsA to regulate fumA expression led to a 9.0% increase in L-malic acid titer, an 8.75% increase in yield (glucose to L-malic acid), and an 8.86% enhancement in productivity. This research serves as a significant step toward expediting the industrialization of L-malic acid synthesis via biological fermentation. Additionally, it offers valuable insights for the biosynthesis of other organic acids.IMPORTANCEThis study focuses on enhancing L-malic acid synthesis by modifying the tricarboxylic acid cycle within the mitochondria of Aspergillus niger. We emphasize the significant role of fumarase in converting fumaric acid into L-malic acid, enhancing our understanding of metabolic pathways in A. niger. The precise regulation of fumA is highlighted as a key factor in enhancing L-malic acid production. Furthermore, this research introduces a stringent conditional promoter (PmfsA), exclusively activated by CaCO3. The utilization of PmfsA for fumA expression resulted in heightened L-malic acid titers. The progress in metabolic engineering and bioprocess optimization holds promise for expediting industrial L-malic acid synthesis via biological fermentation. Moreover, it carries implications for the biosynthesis of various other organic acids.

A metabolomics study on carcinogenesis of ground-glass nodules.

This study aimed to identify differential metabolites and key metabolic pathways between lung adenocarcinoma (LUAD) tissues and normal lung (NL) tissues using metabolomics techniques, to discover potential biomarkers for the early diagnosis of lung cancer. Forty-five patients with primary ground-glass nodules (GGN) identified on computed tomography imaging and who were willing to undergo surgery at Shanghai General Hospital from December 2021 to December 2022 were recruited to the study. All participants underwent video thoracoscopy surgery with segmental or wedge resection of the lung. Tissue samples for pathological examination were collected from the site of ground-glass nodules (GGN) lesion and 3 cm away from the lesion (NL). The pathology results were 35 lung adenocarcinoma (LUAD) cases (13 invasive adenocarcinoma, 14 minimally invasive adenocarcinoma, and eight adenocarcinoma in situ), 10 benign samples, and 45 NL tissues. For the untargeted metabolomics technique, 25 LUAD samples were assigned as the case group and 30 NL tissues as the control group. For the targeted metabolomics technique, ten LUAD samples were assigned as the case group and 15 NL tissues as the control group. Samples were analyzed by untargeted and targeted metabolomics, with liquid chromatography-tandem mass spectrometry detection used as part of the experimental procedure. Untargeted metabolomics revealed 164 differential metabolites between the case and control groups, comprising 110 up regulations and 54 down regulations. The main metabolic differences found by the untargeted method were organic acids and their derivatives. Targeted metabolomics revealed 77 differential metabolites between the case and control groups, comprising 69 up regulations and eight down regulations. The main metabolic changes found by the targeted method were fatty acids, amino acids, and organic acids. The levels of organic acids such as lactic acid, fumaric acid, and malic acid were significantly increased in LUAD tissue compared to NL. Specifically, an increased level of L-lactic acid was found by both untargeted (variable importance in projection [VIP] = 1.332, fold-change [FC] = 1.678, q = 0.000) and targeted metabolomics (VIP = 1.240, FC = 1.451, q = 0.043). Targeted metabolomics also revealed increased levels of fumaric acid (VIP = 1.481, FC = 1.764, q = 0.106) and L-malic acid (VIP = 1.376, FC = 1.562, q = 0.012). Most of the 20 differential fatty acids identified were downregulated, including dodecanoic acid (VIP = 1.416, FC = 0.378, q = 0.043) and tridecane acid (VIP = 0.880, FC = 0.780, q = 0.106). Furthermore, increased levels of differential amino acids were found in LUAD samples. Lung cancer is a complex and heterogeneous disease with diverse genetic alterations. The study of metabolic profiles is a promising research field in this cancer type. Targeted and untargeted metabolomics revealed significant differences in metabolites between LUAD and NL tissues, including elevated levels of organic acids, decreased levels of fatty acids, and increased levels of amino acids. These metabolic features provide valuable insights into LUAD pathogenesis and can potentially serve as biomarkers for prognosis and therapy response.

The forced activation of asexual conidiation in Aspergillus niger simplifies bioproduction

Aspergillus niger is an efficient cell factory for organic acids production, particularly l-malic acid, through genetic manipulation. However, the traditional method of collecting A. niger spores for inoculation is labor-intensive and resource-consuming. In our study, we used the CRISPR-Cas9 system to replace the promoter of brlA, a key gene in Aspergillus conidiation, with a xylose-inducible promoter xylP in l-malic acid-producing A. niger strain RG0095, generating strain brlAxylP. When induced with xylose in submerged liquid culture, brlAxylP exhibited significant upregulation of conidiation-related genes. This induction allowed us to easily collect an abundance of brlAxylP spores (>7.1 × 106/mL) in liquid xylose medium. Significantly, the submerged conidiation approach preserves the substantial potential of A. niger as a foundational cellular platform for the biosynthesis of organic acids, including but not limited to l-malic acid. In summary, our study offers a simplified submerged conidiation strategy to streamline the preparation stage and reduce labor and material costs for industrial organic acid production using Aspergillus species.

Enhancing l-Malic Acid Production in Aspergillus niger via Natural Activation of sthA Gene Expression.

The efficient production of l-malic acid using Aspergillus niger requires overcoming challenges in synthesis efficiency and excessive byproduct buildup. This study addresses these hurdles, improving the activity of NADH-dependent malate dehydrogenase (Mdh) in the early stages of the fermentation process. By employing a constitutive promoter to express the Escherichia coli sthA responsible for the transfer of reducing equivalents between NAD(H) and NADP(H) in A. niger, the l-malic acid production was significantly elevated. However, this resulted in conidiation defects of A. niger, limiting industrial viability. To mitigate this, we discovered and utilized the PmfsA promoter, enabling the specific expression of sthA during the fermentation stage. This conditional expression strain showed similar phenotypes to its parent strain while exhibiting exceptional performance in a 5 L fermenter. Notably, it achieved a 65.5% increase in productivity, reduced fermentation cycle by 1.5 days, and lowered succinic acid by 76.2%. This work marks a promising advancement in industrial l-malic acid synthesis via biological fermentation, showcasing the potential of synthetic biology in A. niger for broader applications.

Eco-Friendly and Efficient Extraction of Polysaccharides from Acanthopanax senticosus by Ultrasound-Assisted Deep Eutectic Solvent.

A green extraction method was developed using deep eutectic solvent extraction for the polysaccharide from Acanthopanax senticosus (A. senticosus). Among the eight types of DES prepared, the DES with a ratio of 1:4 L-malic acid to L-proline was found to be a suitable extraction solvent based on the extraction efficiency. The extraction parameters were optimized by Plackett-Burman and response surface methodology (RSM). The best extraction conditions were found for L-malic acid. Under the conditions of an L-malic acid/L-proline ratio of 1:4, ultrasonic power of 240 W, material-liquid ratio of 31.068 g/mL, water content of 32.364%, extraction time of 129.119 min, and extraction temperature of 60 °C, the extraction rate of A. senticosus polysaccharides was 35.452 ± 0.388 mg-g-1. This rate was higher than that of polysaccharides obtained by hot water extraction (13.652 ± 0.09 mg-g-1). The experimental results were best fitted by the quasi-secondary kinetic model when compared to two other kinetic models. Electron microscopic observations showed that DESs were more destructive to plant cells. The polysaccharide extracted from DESs had more monosaccharide components, a lower molecular weight, a higher antioxidant capacity, and superior anti-glycation activity compared to polysaccharides extracted from water (ASPS-PW). This study demonstrates the effectiveness of DESs in obtaining polysaccharides from A. senticosus.

Maternal Malic Acid May Ameliorate Oxidative Stress and Inflammation in Sows through Modulating Gut Microbiota and Host Metabolic Profiles during Late Pregnancy.

Sows suffer oxidative stress and inflammation induced by metabolic burden during late pregnancy, which negatively regulates reproductive and lactating performances. We previously found that L-malic acid (MA) alleviated oxidative stress and inflammation and improved reproductive performances in sows. However, the mechanism underlying the MA's positive effects remains unexplored. Here, twenty Large White × Landrace sows with similar parity were randomly divided into two groups and fed with a basal diet or a diet supplemented with 2% L-malic acid complex from day 85 of gestation to delivery. The gut microbiome, fecal short-chain fatty acids, and untargeted serum metabolome were determined. Results showed that Firmicutes, Bacteroidota, and Spirochaetota were the top abundant phyla identified in late pregnancy for sows. Maternal MA supplementation modulated the composition but not the richness and diversity of gut microbiota during late pregnancy. Correlation analysis between gut microbiota and antioxidant capacity (or inflammation indicators) revealed that unclassified_f_Ruminococcaceae, unclassified_f_Lachnospiraceae, UCG-002, norank_f_norank_o_RF3, and Lactobacillus might play a role in anti-oxidation, and Lachnospiraceae_XPB1014_group, Lachnospiraceae_NK4A136_group, UCG-002, unclassified_f_Ruminococcaceae, Candidatus_Soleaferrea, norank_f_UCG-010, norank_f_norank_o_RF39, and unclassified_f_Lachnospiraceae might be involved in the anti-inflammatory effect. The improved antioxidant and inflammation status induced by MA might be independent of short chain fatty acid changes. In addition, untargeted metabolomics analysis exhibited different metabolic landscapes of sows in the MA group from in the control group and revealed the contribution of modified amino acid and lipid metabolism to the improved antioxidant capacity and inflammation status. Notably, correlation results of gut microbiota and serum metabolites, as well as serum metabolites and antioxidant capacity (or inflammation indicators), demonstrated that differential metabolism was highly related to the fecal microorganisms and antioxidant or inflammation indicators. Collectively, these data demonstrated that a maternal dietary supply of MA can ameliorate oxidative stress and inflammation in sows through modulating gut microbiota and host metabolic profiles during late pregnancy.

Study of Shanxi aged vinegar by non-targeted metabolomics techniques and antioxidant activity characteristics

The quality and flavor of Shanxi Aged Vinegar (SAV) are closely related to its aging time. In this study, for the first time, UPLC-Q-TOF/MS combined with non-targeted metabolomics techniques were employed to analyze SAV samples from 5 different years. The antioxidant activities were evaluated using ABTS and DPPH assays. The non-targeted metabolomics results identified a total of 99 metabolites, among which 24 were significantly associated with the aging time of SAV and were identified as potential markers of vintage, including key components such as organic acids, amino acids, and polyphenols. They contribute significantly to the unique flavor and nutritional value of SAV. Specifically, in SAV aged for 10 years, the contents of citric acid, L-malic acid, L-glutamic acid, protocatechuic acid, and hydroferulic acid were significantly higher than in other vintages. The results of ABTS and DPPH antioxidant activities indicated a positive correlation between the antioxidant activity of SAV and its aging time. Among the 12 identified polyphenols, chlorogenic acid may be the primary contributor to the antioxidant results of SAV. This study provides important references for further exploration of the health value and antioxidant mechanisms of SAV.

Differences of Pine Wood Nematode (Bursaphelenchus xylophilus) Developmental Stages under High-Osmotic-Pressure Stress.

Under ion imbalance, water deficiency, and salt stress, the osmotic pressure of the tree sap increases, and pine wood nematodes (Bursaphelenchus xylophilus, PWN) parasitizing in the trees may be subjected to high-osmotic-pressure stress. KCl, L-malic acid, sucrose, and glycerol solutions were used as osmolytes to explore the highest osmotic concentration that PWN can tolerate. Survival analysis showed that when the treatment concentration exceeded 90%, only a few nematodes in the glycerol group survived under 6 h treatment, and most of the survivors were third-stage dispersal juveniles (DJ3). Further examination revealed that under different concentrations of glycerol-induced high osmotic pressure, the survival rate and body length change rate were the highest in the DJ3 and the lowest in the second-stage propagative juveniles. In order to explore the molecular mechanism of resistance of DJ3 to high osmotic stress, transcriptome sequencing was performed at each developmental stage of PWN and differentially expressed genes that were up-regulated or down-regulated only in DJ3 were screened. The expression of genes related to CoA in DJ3, a key enzyme in metabolism, was significantly higher than the other developmental stages. In addition, the expression of the anti-reversal signal pathway-related gene AKT-1 in DJ3 was significantly lower than in the other development stages. Therefore, the specific expression of genes in DJ3 under high osmotic pressure may help them rapidly produce and accumulate energy-related compounds and activate the adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK) pathway to respond to damage caused by high-osmotic-pressure stress in time, thus promoting survival.

Effects of microwave radiation on the physicochemical properties, structure, and digestibility of the synthesized different crystal forms of malic acid starch ester

The effects of microwave combined with L-malic acid treatment on the degree of substitution (DS), structure, physicochemical properties, and digestibility of sweet potato starch (A-type), potato starch (B-type), and pea starch (C-type) were evaluated. The order of DS obtained was: DSM-POS > DSM-SPS > DSM-PES. Fourier transform-infrared spectroscopy (FT-IR) showed that the obtained modified starch produced a new absorption band at 1735 cm−1. Scanning electron microscopy (SEM) and polarized light microscopy indicated that different types of native starches exhibited different granular morphologies and appeared to have different degrees of damage, but still had polarized crosses after modification. Sweet potato starch had the smallest particle size, while potato starch had the largest. X-ray diffractometry (XRD) showed that the modified starches still retained the same crystal structure as the native starches, but the relative crystallinity decreased. The apparent viscosity and swelling power of modified starches dropped, but their water/oil holding capacity, amylose content, and resistant starch content all increased. The results demonstrate that the degree of influence on the structure, physicochemical properties, and digestibility of different starches varies under the same modification conditions.

Cultivar-dependent grape berry dehydration in later ripening phases may be associated with energy regulation and ionic homeostasis

Climate warming and drought may alter the progression of sugar accumulation by the grape berry, and contribute to pre-harvest berry shrivel and cell vitality loss which were hypothesised to be associated with solute accumulation cessation and energy status in the berries. To explore the relationship between energy and cultivar-dependent berry cell death, we characterised the variations of pericarp constituents and energy status across five time points, five ripening phases, and three cultivars with various levels of berry shrivel and cell death. Grape berries of Shiraz, Chardonnay and Flame Seedless at the Eichorn-Lorenz stages 33 to 39 were sampled at 5:00, 9:00, 13:00, 17:00 and 21:00 on each sampling day, which were used in the analysis of water percentage, adenosine triphosphate (ATP), ethanol, sugar, L-malic acid, as well as elements with vascular mobility, including potassium (K) and sodium (Na). Shiraz and Chardonnay ceased accumulating sugar and phloem mobile elements during the later ripening phase, while Flame Seedless did not undergo water loss or cease to accumulate solutes in pericarps. Shiraz berry ripening was distinguished by greater pericarp water loss, ATP reduction, increased ion disorder susceptibility as indicated by a lower K/Na ratio, and higher ethanol concentration in the later ripening phase. The Chardonnay pericarp contained the highest ATP concentration and K/Na ratio among the three cultivars. The Flame Seedless pericarp maintained a low but stable K/Na ratio throughout berry ripening. Berry ripening phase impacted the diurnal variations of pericarp water percentage and ATP concentration. The results indicated that energy regulation and ionic homeostasis may be associated with cultivar-dependent grape pericarp hydration, solute accumulation and cell vitality during berry ripening.