Putrescine Content Research Articles

Competitive network of polyamines metabolic and ethylene biosynthesis pathways during gibberellin-induced parthenocarpic grape fruit setting

Polyamines (PAs) and ethylene are involved in the modulation of plant growth and development. However, their roles in fruit-set, especially in exogenous gibberellin (GA3)-induced grape parthenocarpic berries, and the related competitive action mode are poorly understood. For this, we, here performed their content determination, bioinformatics and expression pattern analysis of genes to identify the key ones in the competitive network of polyamines metabolic and ethylene biosynthesis (PMEB) pathways. The content of putrescine (Put) significantly increased; while 1-aminocyclopropanecarboxylic acid (ACC) sharply decreased during the fruit-set process of GA3-induced grape parthenocarpic seedless berries. Totally, twenty-five genes in PMEB pathways, including 20 polyamines metabolic (PM) genes and 5 ethylene biosynthesis (EB) ones were identified in grape, of which 8 PM and 2 EB genes possessed the motifs responsive to phytohormone GA. The expression levels of most PMEB genes kept changing during grape fruit-set generating a competitive action mode of GA3-mediated two metabolic fluxes toward PAs and ethylene synthesis. Exogenous GA3 might enhance grape fruit-set of parthenocarpic berries via up-regulation of VvSAMS4, VvSAMDC1/2, VvODC1, VvSPDS1, and VvPAO1 to promote PAs accumulation, whereby repressing the ethylene synthesis by down-regulation of VvACS1 and VvACO2. Our findings provide novel insights into GA3-mediated competitive inhibition of ethylene by PAs to promote the fruit-set of parthenocarpic berries in grape, which has important implications for molecular breeding of seedless grape with high fruit-setting rate.

Preinoculation with Bradyrhizobium japonicum enhances the salt tolerance of Glycine max seedlings by regulating polyamine metabolism in roots

Rhizobia are common symbiotic microorganisms in the root system of leguminous plants that can usually provide nitrogen to the host through nitrogen fixation. Studies have shown that rhizobium-preinoculated soybean plants usually exhibit improved salt tolerance, but the underlying mechanism is not fully understood. In this paper, transcriptome sequencing (RNA-seq) revealed that preinoculation with rhizobia affected polyamine (PA) metabolism in soybean roots. The assay of PA contents showed that preinoculation with rhizobia significantly increased the putrescine (Put) content in roots and leaves during short-term salt treatment (0-5d). Long-term salt treatment (5-7d) resulted in a high Put content and significantly increased Spm and Spd contents, resulting in a rapid increase in the Put/(Spd+Spm) ratio (0-5d) and subsequent decrease. Moreover, rhizobium preinoculation of soybean plants resulted in increased contents of conjugated and bound PAs under salt stress. Further transcriptome sequencing, PA contents, PA synthase expression and activity analysis revealed that GmADC may be a key gene related to salt tolerance in rhizobium-preinoculated soybean plants, and the GmADC-overexpressing soybean hairy-root composite plants exhibited less ROS damage, lower Cl-/NO3- ratios and Na+/K+ ratios, and stabilized ion homeostasis. Taken together, preinoculation with rhizobia increased the expression level and enzyme activity of arginine decarboxylase (ADC) in soybean roots, increased the content of Put in roots and leaves, and increased the content of conjugated and bound PAs in soybean plants, thereby alleviating the oxidative and ionic injuries of soybean plants and enhancing the salt tolerance.

Polyamine impact on physiology of early stages of reef-building corals-insights from rearing experiments and RNA-Seq analysis.

Polyamines are involved in various functions related to the cellular-level responses. To assess effects of polyamines on marine organisms, rearing experiments and comprehensive gene expression analyses were conducted on Acropora digitifera and Acropora sp.1, representative reef-building corals along the west-central coast of Okinawa, Japan, to evaluate effects of putrescine. Concentrations of putrescine ≥ 1mM dissolved tissues of juvenile polyps and increased mortality of planula larvae. RNA-Seq analysis of juvenile polyps exposed to putrescine at the stage before effects became visible revealed dynamic fluctuations in gene expression in the putrescine-treated samples, with increased expression of stress-responsive genes (e.g. NAD-dependent protein deacylase sirtuin-6) and the polyamine transporter Slc18b1-like protein. These results also suggest that putrescine affects expression of genes related to ribosomes and translation. This study provides important insights into roles of polyamines and future directions regarding physiological responses of corals.

The role and the determination of the LuxI protein binding targets in the formation of biogenic amines in Hafnia alvei H4

Hafnia alvei is a spoilage microorganism that possesses the LuxI/LuxR-type quorum sensing (QS) system. Biogenic amines (BAs) are important in food spoilage and safety, yet the role of QS in BA formation remains poorly understood. This study investigated the ability of H. alvei H4 to produce BAs in fish flesh and decarboxylase culture media. The findings showed that H. alvei H4 produced substantial amounts of putrescine and cadaverine in turbot flesh, with its enhanced amine-producing capacity potentially leading to the eventual deterioration of the fish. Furthermore, the deletion of the QS element—AHL synthase gene luxI—affected the concentrations of both BAs. Based on these observations, the present study conducted multifaceted experiments, including phenotypic assessments and analyses of gene expression, to explore the role of luxI and to identify its specific binding targets. The results indicated that putrescine formation in H. alvei H4 primarily occurred via the arginine deiminase (ADI) pathway, with luxI playing a positive role in the conversion of arginine to ornithine and subsequently to putrescine. The reduction in putrescine content observed in a luxI mutant (ΔluxI) was attributed to the direct binding of the LuxI protein to the promoters of the argF and speC genes, which code for ornithine carbamoyltransferase and ornithine decarboxylase, respectively. The findings of this study provided the basis to understand the influence of QS on BA production in H. alvei, by specifically demonstrating the involvement of the luxI gene on putrescine and cadaverine production.

Chitosan coatings containing thyme essential oil enhance the quality of snakehead (Channa striata) during chilled storage verified by metabolomics approaches.

In the present study, we investigated the preservative effects of chitosan (CS) coatings, with and without thyme essential oil (TEO), combined with vacuum impregnation (VI) on maintaining the quality of snakehead fillets during chilled storage. The results showed that the VI treatment significantly inhibited drip loss, discoloration, microbial proliferation, and the accumulation of biogenic amines (BAs) in the sneakhead fillets. Compared to the control, the fillets treated with VI of 1% (w/w) CS and 1.5% (w/w) TEO (i.e., CSTEO) showed significant reductions in both psychrophiles and mesophiles, with a 2.66 log CFU/g decrease in total viable count (TVC) on day 3 and a 1.89 log CFU/g decline in TVC on day 9, respectively. In addition, the content of histamine and putrescine in the CSTEO groups was maintained at ∼1.14 and 3.23mg/kg during the 12-day chilled storage, respectively. A total of 100 chemical compounds were tentatively identified using untargeted metabolomics approaches. The multivariate analysis further revealed that the combination of VI and CSTEO maintained fish quality mainly through preventing lipid oxidation and protein degradation. Overall, the VI-CSTEO treatment effectively maintained the fish quality during storage at 4°C, with minimum microbial proliferation and accumulation of BAs. PRACTICAL APPLICATION: The preservative effect of chitosan coatings containing thyme essential oil combined with vacuum impregnation on snakehead quality during the 12-day chilled storage was verified, and the underlying mechanisms were deciphered through integrated metabolomics approaches. Our study could provide a promising strategy for the preservation of aquatic products.

Association of Polyamine Intake, Other Dietary Components, and Fecal Content of N-acetyl Putrescine and Cadaverine with Patients' Colorectal Lesions.

Colorectal cancer (CRC) is the second leading cause of cancer death worldwide. Early detection and the modification of risk factors, such as diet, can reduce its incidence. Among food components, polyamines are important for maintaining gastrointestinal health and are metabolites of gut microbiota. Their disruption is linked to CRC, making polyamines a potential marker of the disease. This study analyzed the relationship between dietary components, including polyamines, and the presence of polyamines in feces to determine whether their presence could contribute to predicting the occurrence of colorectal lesions in patients. In total, 59 participants of both sexes (aged 50 to 70 years) who had undergone colonoscopy screening for CRC (18 without and 41 with colorectal lesions) participated in the study. A nutritional survey and determination of fecal polyamine content were performed. Specific dietary components and putrescine levels were higher in patients with colorectal lesions. The diet ratio of putrescine-spermidine and the fecal content of N-acetyl putrescine and cadaverine were elevated in patients with precancerous lesions and adenocarcinomas, showing a potential predictive value for the presence of colorectal lesions. These findings suggest that N-acetyl putrescine and cadaverine could be complementary markers for the diagnosis of suspected colorectal lesions.

Fermentation Parameters, Amino Acids Profile, Biogenic Amines Formation, and Bacterial Community of Ensiled Stylo Treated with Formic Acid or Sugar.

Substantial proteolysis occurs and free amino acids can be degraded to biogenic amines by decarboxylation during stylo (Stylosanthes guianensis) ensiling. High biogenic amine concentrations in silage are harmful to the health of ruminant animals. The purposes of this work were to (1) analyze the biogenic amines and amino acids concentrations, bacterial composition, and fermentation profile of spontaneously fermented stylo silage, (2) explore the effect of formic acid or sugar additive on these silage parameters, and (3) further reveal the correlations between silage amines and fermentation parameters, amino acids, and bacteria. Freshly chopped stylo was treated with distilled water (control), formic acid (4 mL/kg), and sugar (20 g/kg) and fermented for 28 days. The results indicated that putrescine (321 mg/kg dry matter), cadaverine (384 mg/kg dry matter), and tyramine (127 mg/kg dry matter) rapidly increased in concentration and become predominant in the control silage after 28 days of fermentation. Applying formic acid and sugar at ensiling, especially the acidifier, significantly decreased putrescine, cadaverine, tyramine, and total biogenic amine concentrations compared with the control treatment (p < 0.0001). Clostridium pabulibutyricum, Weissella cibaria and W. paramesenteroides were the predominant bacteria in the control silage, and the application of both additives remarkably lowered their relative abundance in comparison with the control treatment (p < 0.001). Correlation analysis showed that putrescine, cadaverine, and tyramine were positively related to pH, butyric acid, non-protein nitrogen, and ammonia nitrogen (p < 0.01). These amines also had significant correlations with C. pabulibutyricum, W. cibaria and W. paramesenteroides (p < 0.001). Putrescine, cadaverine, and tyramine were the main biogenic amines and C. pabulibutyricum was the predominant undesirable bacterium in naturally fermented stylo silage. C. pabulibutyricum, W. cibaria and W. paramesenteroides were positively related to putrescine, cadaverine, and tyramine formation. The application of formic acid or sugar significantly reduced the undesirable bacterial population and improved the fermentation and hygienic quality of the stylo silage. These findings lay the foundation for further elucidating the microbial mechanism underlying the main biogenic amine formation during fermentation of stylo silage.

Effects of Lactic Acid Bacteria on Reducing the Formation of Biogenic Amines and Improving the Formation of Antioxidant Compounds in Traditional African Sourdough Flatbread Fermentation.

This study investigated the safety and functionality of traditional African sourdough flatbread (kisra), based on the content of biogenic amines (BAs) and antioxidant compounds and their improvement using lactic acid bacteria (LAB) species. The primary BAs detected in naturally fermented kisra were tyramine, histamine, putrescine, and cadaverine, with putrescine being the most abundant after baking. In vitro BA production of microorganisms isolated from kisra sourdough revealed that the Enterococcus genus contributed to tyramine accumulation, whereas presumptive yeasts may contribute to putrescine and cadaverine accumulation. The use of LAB species, including Lactiplantibacillus plantarum, Limosilactobacillus fermentum, Levilactobacillus brevis, and Weissella cibaria, significantly reduced putrescine content to less than about 23% of that of naturally fermented kisra, and eliminated tyramine, histamine, and cadaverine formation. Meanwhile, DPPH scavenging activity, total polyphenolic content, and tannin content in naturally fermented kisra were 85.16%, 1386.50 µg/g, and 33.16 µg/g, respectively. The use of LAB species did not affect the DPPH scavenging activity or tannin content but significantly increased the total phenolic content by up to 20% compared to naturally fermented kisra. Therefore, fermentation with LAB starter cultures might be a promising approach to improve the safety related to BAs as well as the functionality of kisra bread.

Identification and Characterization of Polyamine Metabolism in Citrus in Response to 'Candidatus Liberibacter asiaticus' Infection.

Citrus Huanglongbing, one of the most devastating citrus diseases, is caused by 'Candidatus Liberibacter asiaticus' (CLas). Polyamines are aliphatic nitrogen-containing compounds that play important roles in disease resistance and are synthesized primarily by two pathways: an arginine decarboxylation pathway and an ornithine decarboxylation pathway. However, it is unclear whether polyamines play a role in the tolerance of citrus to infection by CLas and, if so, whether one or both of the core polyamine metabolic pathways are important. We used high-performance liquid chromatography and ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry to detect the contents of nine polyamine metabolism-related compounds in six citrus cultivars with varying levels of tolerance to CLas. We also systematically detected the changes in polyamine metabolism-related compounds and H2O2 contents and compared the gene expression levels and the activities of enzymes involved in the polyamine metabolic pathway among healthy, asymptomatic, and symptomatic leaves of Newhall navel oranges infected with CLas. The tolerant and moderately tolerant varieties showed higher polyamine metabolism-related compound levels than those of susceptible varieties. Compared with the healthy group, the symptomatic group showed significantly increased contents of arginine, ornithine, γ-aminobutyric acid, and putrescine by approximately 180, 19, 1.5, and 0.2 times, respectively, and upregulated expression of biosynthetic genes. Arginase and ornithine decarboxylase enzyme activities were the highest in the symptomatic group, whereas arginine decarboxylase and agmatine deiminase enzyme activities were the highest in the asymptomatic group. The two polyamine biosynthetic pathways showed different trends with the increase of the CLas titer, indicating that polyamines were mainly synthesized through the arginine decarboxylase pathway in the asymptomatic leaves and were synthesized via the ornithine decarboxylase pathway in symptomatic leaves. These findings provide new insight into the changes in polyamine metabolism in citrus infected with CLas.

DETERMINATION OF BIOGENIC AMINES IN CULTIVATED AND PROCESSED MUSHROOMS INTENDED FOR THE SLOVAK MARKET

Cultivated and processed white button mushrooms are widely used in the menu of the Slovak consumers. However, these food items can potentially contain biogenic amines, attributed to the presence of precursor compounds and their susceptibility to microbial spoilage. Biogenic amines significantly influence food quality and can pose health risks to consumers. This study is focused on determination of spermidine, putrescine, tyramine, cadaverine, histamine, spermine, and 2-phenylethylamine in processed mushroom products available in the Slovak market. The findings revealed considerable variability in biogenic amine content across individual products, influenced by the producer. Spermidine, putrescine and spermine emerged as the predominant biogenic amines with concentration ranges 2349.8 -7412.8, 37.2 - 607.5 and 74.2 - 267.6 mg/kg DW, respectively. Tyramine and histamine were detected in 22.2% of processed mushroom samples with one product exceeding concentration 200 mg/kg DW. Cadaverine was identified in 44.4% samples. The presence of histamine has to be concerned regarding potential health risks for consumers. Comparison of the mushroom products showed significant differences between the groups of products with whole mushrooms vs. sliced mushrooms in brine in case of the content of spermine (p&lt;0.001), water content (p&lt;0.01), putrescine, histamine and tyramine (p&lt;0.05 for all these BAs). No significant differences were found between the groups for the cadaverine, spermidine and total content of biogenic amines. In house rapid validation was performed in term of determination of limits of detection and limits of quantification and intraday precision of retention time for individual biogenic amines in this study. Limits of detection ranged from 0.013 to 0.029 µg/mL and limits for quantification were in the range from 0.039 to 0.087 µg/mL. The RSD for the retention time in standard solutions did not exceed 0.12%.

Luminal Polyamines Disrupt Intestinal Barrier and Undergo Complex Regulation by Host and Commensal Microbiota

Background: Polyamines produced by host cells and intestinal microbiota and are critical to overall health. Studies have shown putrescine, a polyamine, can strengthen, or in some instances disrupt, gastrointestinal (GI) barrier integrity, yet the regulation of polyamine concentrations in the GI tract is unknown. Host colonocytes and microbiota both metabolize polyamines, and in GI diseases like irritable bowel syndrome (IBS) where intestinal microbiota is significantly altered, polyamine metabolism may be affected. Aim: Determine regulation of luminal polyamine concentration and its effects on intestinal barrier integrity. Hypothesis: Luminal polyamine concentration is suppressed by commensal intestinal microbiota and increasing concentration of putrescine results in intestinal barrier disruption. Methods: Microbiota effects on luminal polyamine concentrations were assessed by humanizing germ-free (GF) mice with a single oral gavage of commensal human microbiota. Polyamine effect on barrier integrity was assessed in vitro using Caco-2 cells and ex vivo on Ussing using mouse colonic tissue with 300Da fluorescein and 4kDa FITC-Dextran tracers. Changes in transepithelial resistance (TER) and rate of tracer flux was recorded. Untargeted metabolomics with shotgun metagenomics was completed on human fecal samples and sigmoid colonic biopsies were collected for bulk RNAseq. Bray-Curtis distance was determined at the species level using the PERMANOVA method and used to test relationships between beta-diversity and fecal polyamine concentration. Results: Humanization with human commensal microbiota (1.2% C. aerofaciens) lead to a 20-fold increase in cecal putrescine and a 20-fold decrease in fecal putrescine compared to GF controls. In vitro addition of putrescine (100mM) caused increased fluorescein flux (3.50±0.24 vs 1.44±0.04 mg/mL) and a rapid loss of transepithelial resistance compared to controls (% of Baseline TER, 36.6±8.2 vs 102.8±5.4, 3h postexposure). Ex vivo addition of putrescine caused a loss of TER (57% vs 89% of baseline by 3h) and an increased rate of 4kDa FITC-Dextran flux compared to untreated controls (1236±131 vs 562±70 ng/hr/cm2). Transcriptomic profiles show decreased expression of ornithine decarboxylase antienzyme (OAZ1, FDR=0.018) in IBS which inhibits the rate limiting enzyme for putrescine production. Increased lactulose excretion (2-24h), suggestive of increased distal colonic permeability, correlated with N-acetylputrescine concentration (p=0.07). N-acetylputrescine also correlated positively with increased stool frequency (p=0.009) and looser consistency (p=0.013). C. aerofaciens, a polyamine consumer, negatively correlated with n-acetylputrescine (FDR=0.02) suggesting a loss of specific taxa may lead to insuffcient polyamine metabolism. Conclusion: Polyamines, namely putrescine, disrupts intestinal barrier integrity, correlate with in vivo permeability, and are decreased in the colon by commensal microbiota. These observations identify putrescine as a negative effector of barrier function and suggest a novel role for the commensal microbiome in regulating barrier integrity through polyamine metabolism. Further work investigating host and microbiota mechanisms of polyamine metabolism, signaling and the specific taxa involved is needed to better understand the effects polyamines have on GI physiology. NIH DK 127998 (MG), support from Mayo Clinic Center for Individualized Medicine (CIM), and Mayo Clinic Center for Cell Signalling in Gastroenterology (C-Sig) P30DK084567. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Arginine and its metabolites stimulate proliferation, differentiation, and physiological function of porcine trophoblast cells through β-catenin and mTOR pathways.

Arginine, which is metabolized into ornithine, proline, and nitric oxide, plays an important role in embryonic development. The present study was conducted to investigate the molecular mechanism of arginine in proliferation, differentiation, and physiological function of porcine trophoblast cells (pTr2) through metabolic pathways. The results showed that arginine significantly increased cell viability (P < 0.05). The addition of arginine had a quadratic tendency to increase the content of progesterone (P = 0.06) and protein synthesis rate (P = 0.03), in which the maximum protein synthesis rate was observed at 0.4 mM arginine. Arginine quadratically increased (P < 0.05) the intracellular contents of spermine, spermidine and putrescine, as well as linearly increased (P < 0.05) the intracellular content of NO in a dose-dependent manner. Arginine showed a quadratic tendency to increase the content of putrescine (P = 0.07) and a linear tendency to increase NO content (P = 0.09) in cell supernatant. Moreover, increasing arginine activated (P < 0.05) the mRNA expressions for ARG, ODC, iNOS and PCNA. Furthermore, inhibitors of arginine metabolism (L-NMMA and DFMO) both inhibited cell proliferation, while addition of its metabolites (NO and putrescine) promoted the cell proliferation and cell cycle, the mRNA expressions of PCNA, EGF and IGF-1, and increased (P < 0.05) cellular protein synthesis rate, as well as estradiol and hCG secretion (P < 0.05). In conclusion, our results suggested that arginine could promote cell proliferation and physiological function by regulating the metabolic pathway. Further studies showed that arginine and its metabolites modulate cell function mainly through β-catenin and mTOR pathways.

The Influence of Different Levels of Sodium Chloride, Sodium Nitrite, and Glucose on Biogenic Amines and Microbial Communities in Fermented Goat Meat Sausage.

The influence of different levels of sodium chloride, sodium nitrite, and glucose on the quality characteristics of spontaneously fermented goat meat sausage was investigated. The amounts of total biogenic amines in all the sausages ranged from 324.70 to 388.77 mg/kg; among them, spermine was the most abundant, with amounts ranging from 230.96 to 275.78 mg/kg. Increasing sodium chloride from 15 to 35 g/kg, the content of cadaverine, putrescine, tyramine, phenylethylamine, tryptamine, and total amines decreased, and Enterobacteriaceae counts decreased at the same time. Increasing sodium nitrite from 150 to 250 mg/kg, the content of cadaverine, histamine, and total amines decreased, while Enterobacteriaceae counts decreased simultaneously. Increasing glucose from 10 to 40 g/kg, the content of cadaverine, spermidine, and total amines decreased. Enterococcus was the most abundant genus across all the samples, and the relative abundance of Enterococcus was reduced obviously by increasing sodium nitrite and glucose levels. The top 10 differential bacterial taxa for each additive group were respectively obtained, and microbial biomarkers for each level of additive within its group were acquired, respectively. Through Pearson correlation, Lactobacillus was positively correlated with phenylethylamine, tryptamine, tyramine, and cadaverine, Bacteroides and Sediminibacterium were positively correlated with phenylethylamine and putrescine, respectively, suggesting they have the potential to produce biogenic amines. The results provided references for controlling the accumulation of biogenic amines in fermented goat meat sausage via the addition of auxiliary additives during the processing.

Effect of New Methods for Inhibiting Malolactic Fermentation on the Analytical and Sensory Parameters of Wines

This study focuses on the impact of new methods for inhibiting malolactic fermentation in white wines on their analytical and sensory properties. Enological preparations with different mechanisms of effect were tested: fumaric acid, chitosan, Estaan (a preparation based on tannin inhibition), medium-chain fatty acids (MCFAs), sulphur dioxide and a control variant. Malolactic fermentation (MLF) was also performed. The samples underwent analysis through HPLC (high-performance liquid chromatography) to determine the concentrations of malic and lactic acid, as well as biogenic amines. GC (gas chromatography) analysis was used to monitor volatile substances, alongside sensory evaluation. This study demonstrated a significant influence of individual enological preparations on the aromatic profile of the examined wines. The SO2 and MCFA variants exhibited the highest concentrations of volatile substances within the esters group, specifically isoamyl acetate, 1-hexyl acetate and phenylethyl acetate. Conversely, the fumaric acid and Estaan variants displayed the lowest concentrations of these esters. The most notable disparities were observed in acetoin concentration, with the MCFA variant exhibiting the lowest values. Additionally, the chitosan variant showed higher concentrations of putrescine and spermidine compared to the MCFA and fumaric acid variants, which presented the lowest levels.

Functional divergence of two arginine decarboxylase genes in tropane alkaloid biosynthesis and root growth in Atropa belladonna

Putrescine, produced via the arginine decarboxylase (ADC)/ornithine decarboxylase (ODC)-mediated pathway, is an initial precursor for polyamines metabolism and the root-specific biosynthesis of medicinal tropane alkaloids (TAs). These alkaloids are widely used as muscarinic acetylcholine antagonists in clinics. Although the functions of ODC in biosynthesis of polyamines and TAs have been well investigated, the role of ADC is still poorly understood. In this study, enzyme inhibitor treatment showed that ADC was involved in the biosynthesis of putrescine-derived metabolites and root growth in Atropa belladonna. Further analysis found that there were six ADC unigenes in the A. belladonna transcriptome, with two of them, AbADC1 and AbADC2, exhibiting high expression in the roots. To investigate their roles in TAs/polyamines metabolism and root growth, RNA interference (RNAi) was used to suppress either AbADC1 or AbADC2 expression in A. belladonna hairy roots. Suppression of the AbADC1 expression resulted in a significant reduction in the putrescine content and hairy root biomass. However, it had no noticeable effect on the levels of N-methylputrescine and the TAs hyoscyamine, anisodamine, and scopolamine. On the other hand, suppression of AbADC2 expression markedly reduced the levels of putrescine, N-methylputrescine, and TAs, but had no significant effect on hairy root biomass. According to β-glucuronidase (GUS) staining assays, AbADC1 was mainly expressed in the root elongation and division region while AbADC2 was mainly expressed in the cylinder of the root maturation region. These differences in expression led to functional divergence, with AbADC1 primarily regulating root growth and AbADC2 contributing to TA biosynthesis.

Molecular Insights into the Synergistic Effects of Putrescine and Ammonium on Dinoflagellates.

Ammonium and polyamines are essential nitrogen metabolites in all living organisms. Crosstalk between ammonium and polyamines through their metabolic pathways has been demonstrated in plants and animals, while no research has been directed to explore this relationship in algae or to investigate the underlying molecular mechanisms. Previous research demonstrated that high concentrations of ammonium and putrescine were among the active substances in bacteria-derived algicide targeting dinoflagellates, suggesting that the biochemical inter-connection and/or interaction of these nitrogen compounds play an essential role in controlling these ecologically important algal species. In this research, putrescine, ammonium, or a combination of putrescine and ammonium was added to cultures of three dinoflagellate species to explore their effects. The results demonstrated the dose-dependent and species-specific synergistic effects of putrescine and ammonium on these species. To further explore the molecular mechanisms behind the synergistic effects, transcriptome analysis was conducted on dinoflagellate Karlodinium veneficum treated with putrescine or ammonium vs. a combination of putrescine and ammonium. The results suggested that the synergistic effects of putrescine and ammonium disrupted polyamine homeostasis and reduced ammonium tolerance, which may have contributed to the cell death of K. veneficum. There was also transcriptomic evidence of damage to chloroplasts and impaired photosynthesis of K. veneficum. This research illustrates the molecular mechanisms underlying the synergistic effects of the major nitrogen metabolites, ammonium and putrescine, in dinoflagellates and provides direction for future studies on polyamine biology in algal species.

Mechanisms involved in plant growth promotion by Enterobacter cloacae DJ under salinity-alkalinity stress

BackgroundPlant-promoting bacteria are safer alternatives to pesticides and fertilizers, reduce environmental pollution, and increase crop yields. We isolated an Enterobacter cloacae strain DJ with plant-promoting effects from the rhizosphere soil of a plant (Leymus chinensis (Trin.) Tzvel) in the western region of Jilin Province of China and investigated the mechanisms underlying the adaptation of the DJ bacteria to salinity-alkalinity environments and the molecular mechanisms of the cross-talk between DJ bacteria and cucumber seedlings.ResultsThe average diameter of the colonies on the salinity-alkalinity medium after incubation for 24 h was 3.3 cm, and this was significantly higher than the 1.9 cm diameter in ADF medium (p < 0.01). Comparative proteomic analysis revealed that 188 differentially expressed proteins, comprising 116 upregulated and 72 downregulated proteins, significantly changed in salinity-alkalinity groups compared to the control groups. The top one upregulated pathway of KEGG enrichment was bacterial chemotaxis, DJ bacteria adapted to salinity-alkalinity environments by upregulating the genes associated with bacterial chemotaxis. The contents of putrescine in salinity-alkalinity and control groups were 4.73 μg/mL and 3.02 μg/mL, whereas the contents of spermidine were 46.18 ng/mL and 0.34 ng/mL, respectively. Comparing to the control cohorts, the concentrations of both polyamines in the experimental cohorts exhibited statistically significant increases (p < 0.01). The expression of Pt gene encoding polyamine transporter protein was sharply up-regulated in cucumber roots after treatment with DJ bacteria under salinity-alkalinity stress; the expression was more than tenfold higher than that in the control groups. The enzyme activities of POD, SOD, and CAT in cucumber seedlings were higher compared to those in the control groups (p < 0.01). The stem height, number of lateral roots, and fresh weight of cucumber seedlings in the DJ treatment groups were 6.0 cm, 17 roots, and 0.42 g, respectively, whereas those of the control groups were 3.8 cm, 14 roots, and 0.28 g, respectively, with a notable difference between two cohorts (p < 0.01).ConclusionsDJ bacteria can live in salinity-alkalinity conditions by upregulating the expression of genes associated with chemotaxis. The resistance of cucumber seedlings under salinity-alkalinity conditions through the antioxidant pathway was increased by polyamines produced by DJ bacteria.Graphical

Selenium treatment alters the accumulation of osmolytes in arsenic-stressed rice (Oryza sativa L.).

Arsenic (As), one of the major pollutants in the soil, is an important environmental concern as its consumption can cause adverse health symptoms in living organisms. Its contamination of rice grown over As-contaminated areas is a serious concern in South Asian countries. Selenium (Se) has been reported to influence various osmolytes under metal stress in plants. The present study reports the role of Se in mitigating As stress in rice by modulating osmolyte metabolism. Rice plants grown in As-amended soil (2.5-10mgkg-1) in pots were treated with sodium selenate (0.5-1.0mg Se kg-1 soil) in glass house conditions and leaf samples were collected at 60 and 90days after sowing (DAS). As-treated rice leaves displayed a reduction in relative water content (RWC) and dry weight than control with a maximum reduction of 1.68- and 2.47-fold in RWC and 1.95- and 1.69-fold in dry weight in As10 treatment at 60 and 90 DAS, respectively. Free amino acids (1.38-2.26-fold), proline (3.88-3.93-fold), glycine betaine (GB) (1.27-1.72-fold), choline (1.67-3.1-fold), total soluble sugars (1.29-1.61-fold), and reducing sugars (1.67-2.19-fold) increased in As-treated rice leaves as compared to control at both stages. As stress increased the γ-aminobutyric acid (GABA), putrescine content, and glutamate decarboxylase activity whereas diamine oxidase and polyamine oxidase activities declined by 1.69-1.88-fold and 1.52-1.86-fold, respectively. Se alone or in combination with As improved plant growth, RWC, GB, choline, putrescine, and sugars; lowered proline and GABA; and showed a reverse trend of enzyme activities related to their metabolism than respective As treatments. As stress resulted in a higher accumulation of osmolytes to combat its stress which was further modulated by the Se application. Hence, the current investigation suggested the role of osmoprotectants in Se-induced amelioration of As toxicity in rice plants.

Physiological, metabolic and hormonal responses of two Pinus spp. with contrasting susceptibility to brown-spot needle blight disease.

Needle blights are serious fungal diseases affecting European natural and planted pine forests. Brown-spot needle blight (BSNB) disease, caused by the fungus Lecanosticta acicola, causes canopy defoliation and severe productivity losses, with consequences depending on host susceptibility. To gain new insights into BSNB plant-pathogen interactions, constitutive and pathogen-induced traits were assessed in two host species with differential disease susceptibility. Six-month-old Pinus radiata D. Don (susceptible) and Pinus pinea L. (more resistant) seedlings were needle inoculated with L. acicola under controlled conditions. Eighty days after inoculation, healthy-looking needles from symptomatic plants were assessed for physiological parameters and sampled for biochemical analysis. Disease progression, plant growth, leaf gas-exchanges and biochemical parameters were complemented with hormonal and untargeted primary metabolism analysis and integrated for a holistic analysis. Constitutive differences between pine species were observed. Pinus pinea presented higher stomatal conductance and transpiration rate and higher amino and organic acids, abscisic acid as well as putrescine content than P. radiata. Symptoms from BSNB disease were observed in 54.54% of P. radiata and 45.45% of P. pinea seedlings, being more pronounced and generalized in P. radiata. For both species, plant height, sub-stomatal CO2 concentration and water-use efficiency were impacted by infection. In P. radiata, total soluble sugars, starch and total flavonoids content increased after infection. No differences in hormone content after infection were observed. However, secondary metabolism was induced in P. pinea visible through total phenolics, flavonoids and putrescine accumulation. Overall, the observed results suggest that P. pinea constitutive and induced traits may function as two layers of a defence strategy which contributed to an increased BSNB resistance in comparison with P. radiata. This is the first integrative study linking plant physiological and molecular traits in Pinus-Lecanosticta acicola pathosystem, contributing to a better understanding of the underlying resistance mechanisms to BSNB disease in pines.

Putrescine promotes MMP9-induced angiogenesis in skeletal muscle through hydrogen peroxide/METTL3 pathway

Blood vessels play a crucial role in the development of skeletal muscle, ensuring the supply of nutrients and oxygen. Putrescine, an essential polyamine for eukaryotic cells, has an unclear impact on skeletal muscle angiogenesis. In this study, we observed lower vessel density and reduced putrescine level in the muscle of low-birth-weight piglet models, and identified a positive correlation between putrescine content and muscle vessel density. Furthermore, putrescine was found to promote angiogenesis in skeletal muscle both in vitro and in vivo by targeting matrix metalloproteinase 9 (MMP9). On a mechanistic level, putrescine augmented the expression of methyltransferase like 3 (METTL3) by attenuating hydrogen peroxide production, thereby increasing the level of N6-methyladenosine (m6A)-modified MMP9 mRNA. This m6A-modified MMP9 mRNA was subsequently recognized and bound by the YTH N6-methyladenosine RNA binding protein 1 (YTHDF1), enhancing the stability of MMP9 mRNA and its protein expression, consequently accelerating angiogenesis in skeletal muscle. In summary, our findings suggest that putrescine enhances MMP9-mediated angiogenesis in skeletal muscle via the hydrogen peroxide/METTL3 pathway.