Accumulation Of Putrescine Research Articles

Arbuscular mycorrhiza induced putrescine degradation into γ-aminobutyric acid, malic acid accumulation, and improvement of nitrogen assimilation in roots of water-stressed maize plants

Water shortage limits plant growth and development by inducing physiological and metabolic disorders, while arbuscular mycorrhizal (AM) symbiosis can improve plant adaptation to drought stress by altering some metabolic and signaling pathways. In this study, root growth and levels of some metabolites (polyamines, amino acids, and malic acid [MA]) and key enzymes were examined in AM-inoculated and non-inoculated (NM) maize seedlings under different water conditions. The results showed that AM symbiosis stimulated root growth and the accumulation of putrescine (Put) during initial plant growth. Root Put concentration significantly decreased in AM compared with NM plants under water stress; correspondingly, Put degradation via diamine oxidase into γ-aminobutyric acid (GABA) occurred. Moreover, glutamine concentration and the activity of N assimilation enzymes (nitrate reductase and glutamine synthetase) were higher in roots of AM than NM plants under moderate water stress. The activity of GABA transaminase and malic enzyme, and MA concentration were also higher in roots of AM than NM plants under moderate water stress. Our results indicated that Put catabolism along with improved N assimilation and the accumulation of GABA and MA were the key metabolic processes in roots of AM maize plants in response to water stress.

Effect of grape seed extract on quality and microbiota community of container-cultured snakehead (Channa argus) fillets during chilled storage

Herein, the effects of grape seed extract (GSE) on the microflora and biochemical changes of container cultured snakehead (Channa argus) fillets during 11 days of chilled storage were investigated. The sensory analysis, the total number of viable colonies, the total amount of volatile basic nitrogen, and k-value analysis revealed that GSE retarded the deterioration of snakehead fillets. The degradation of inosine 5′-monophosphate and the accumulation of inosine and hypoxanthine in the GSE group were slower than these in the control group. Moreover, GSE treatment effectively decreased the accumulation of putrescine, cadaverine, and histamine. Illumina-MiSeq high throughput sequencing results showed that GSE inhibited the growth of Aeromonas on snakehead fillets. Based on the microbial enumeration, sensory analysis, and k-value, GSE prolonged the shelf life of fillets for 3 days, suggesting its potential for snakehead fillets preservation.

Role of genes and metabolites involved in polyamines synthesis pathways and nitric oxide synthase in stomatal closure on Rosa damascena Mill. under drought stress

In order to evaluate the genes involved in polyamines synthesis pathway and the role of nitric oxide synthase (NOS) and H2O2 in stomatal closure under drought stress, a research conducted with three irrigation levels (100, 50 and 25% field capacity) at 1, 3, 6 and 12 days on Rosa damascena Mill. HPLC and qPCR results showed that putrescine (Put) accumulation occurred at first day in both 50 and 25% of field capacity and then decreased the other days. Furthermore, Put accumulation in the indirect pathway (ADC, AIH and CPA) was more effective related to the direct pathway (ODC) under severe stress. Increased expression of genes involved in production of spermidine (Spd) and spermine (Spm) i.e., SAMDC, SPDS and SPMS correlated with the highest accumulation of Spd and Spm under 50% FC at 6 d and 25% FC at 12 d, respectively. Moreover, results showed that Put reduction simultaneously accumulated H2O2, which subsequently increased NOS expression suggesting a key signal for stomatal closure.

Change in taste-altering non-volatile components of blood and common orange fruit during cold storage.

Cold storage may cause changes in the volatile and non-volatile components of orange fruit, in association with the decrement of the characteristic fruit flavour and sensory acceptability. The aim of this work was to evaluate the changes of some non-volatile taste-altering components (total and individual sugars, acids, anthocyanins, putrescine and limonin) that may affect the organoleptic perception of cold-stored orange fruit. Three blood orange varieties ('Tarocco TDV', 'Tarocco Gallo', and 'Moro') and a common variety ('Washington navel') were stored at 6±1°C and 90-95% Relative Humidity (RH) for 60 d. Chemical and sensory assessments were performed during fruit storage at 15 d intervals. During storage, no dramatic change of the physicochemical parameters was recorded and the ascorbic acid content remained almost unchanged in all varieties. As expected, total anthocyanins significantly increased during storage. Limonin significantly decreased in all varieties. A consistent and significant increase in putrescine occurred during storage in the fruit of the pigmented varieties, not recorded in the common orange variety. Putrescine behaviour showed direct correlation with the accumulation of off-flavour in cold-stored 'Moro' and 'T. TDV' fruit, showing a clear influence of its relative concentration on the sensory perception of fruit. Finally, principal component analysis showed that the complete quality profile of the four investigated varieties represented clear differentiation without overlapping clusters. Our results suggest that the arise of a negative sensory perception in cold stored blood orange fruit might be linked to their accumulation of putrescine.

Effect of taxifolin on physicochemical and microbiological parameters of dry-cured pork sausage

The effect of taxifolin (TXF) with starter cultures (SC), such as Leuconostoc carnosum, or a mixture of strains Pediococcus pentosaceus and Staphylococcus xylosus, on the TXF stability was evaluated. UPLC analysis demonstrated that after 181 days of storage total TXF content was the highest in samples with TXF and L. carnosum (60%), compared to the 1st day of storage. The sausages with TXF and the mixture of P. pentosaceus and S. xylosus (56%) followed next. The samples treated only with TXF retained 40% of TXF, compared to the 1st day of storage. TXF had no significant effect on the growth of lactic acid bacteria. The accumulation of biogenic amines (BA), including histamine and putrescine, was more effectively reduced in sausages inoculated with the TXF plus P. pentosaceus and S. xylosus mixture. Using this mixture, the rate of lipolysis and processes of lipid oxidation were effectively slowed down. Fatty acid (FA) composition was stable in all cases.

Enhanced accumulation of gibberellins rendered rice seedlings sensitive to ammonium toxicity.

Ammonium (NH4+) phytotoxicity is a worldwide phenomenon, but the primary toxic mechanisms are still controversial. In the present study, we investigated the physiological function of gibberellins (GAs) in the response of rice plants to NH4+ toxicity and polyamine accumulation using GA biosynthesis-related rice mutants. Exposure to NH4+ significantly decreased GA4 production in shoots of wild-type (WT) plants. Both exogenous GA application to the WT and increases in endogenous GA levels in eui1 mutants rendered them more sensitive to NH4+ toxicity. In contrast, growth of sd1 GA-deficient mutants was more tolerant to NH4+ toxicity than that of their WT counterparts. The role of polyamines in GA-mediated NH4+ toxicity was evaluated using WT rice plants and their GA-related mutants. The eui1 mutants with GA overproduction displayed a higher endogenous putrescine (Put) accumulation than WT plants, leading to an enhanced Put/[spermidine (Spd)+spermine (Spm)] ratio in their shoots. In contrast, mutation of the SD1 gene encoding a defective enzyme in GA biosynthesis resulted in a significant increase in Spd and Spm production, and reduction in the Put/(Spd+Spm) ratio when exposed to a high NH4+ medium. Exogenous application of Put exacerbated symptoms associated with NH4+ toxicity in rice shoots, while the symptoms were alleviated by an inhibitor of Put biosynthesis. These findings highlight the involvement of GAs in NH4+ toxicity, and that GA-induced Put accumulation is responsible for the increased sensitivity to NH4+ toxicity in rice plants.

Metabolomic Profiling of the Host Response of Tomato (Solanum lycopersicum) Following Infection by Ralstonia solanacearum.

Tomato (Solanum lycopersicum) is an important dietary source of bioactive phytochemicals and active breeding programs constantly produce new cultivars possessing superior and desirable traits. The phytopathogenic Ralstonia solanacearum, the causal agent of bacterial wilt, is a highly destructive bacterial disease with a high economic impact on tomato production. This study followed an untargeted metabolomic approach involving four tomato cultivars and aimed at the identification of secondary metabolites involved in plant defense after infection with R. solanacearum. Liquid chromatography coupled to mass spectrometry (LC-MS) in combination with multivariate data analysis and chemometric modelling were utilized for the identification of discriminant secondary metabolites. The total of 81 statistically selected features were annotated belonging to the metabolite classes of amino acids, organic acids, fatty acids, various derivatives of cinnamic acid and benzoic acids, flavonoids and steroidal glycoalkaloids. The results indicate that the phenylpropanoid pathway, represented by flavonoids and hydroxycinnamic acids, is of prime importance in the tomato defense response. The hydroxycinnamic acids esters of quinic acid, hexoses and glucaric acids were identified as signatory biomarkers, as well as the hydroxycinnamic acid amides to polyamines and tyramine. Interestingly, the rapid and differential accumulation of putrescine, dopamine, and tyramine derivatives, along with the presence of a newly documented metabolite, feruloyl serotonin, were documented in the infected plants. Metabolite concentration variability in the different cultivar tissues point to cultivar-specific variation in the speed and manner of resource redistribution between the host tissues. These metabolic phenotypes provide insights into the differential metabolic signatures underlying the defense metabolism of the four cultivars, defining their defensive capabilities to R. solanacearum.

Biochemical features of primary cells from a pediatric patient with a gain-of-function ODC1 genetic mutation.

We recently described a new autosomal dominant genetic disorder in a pediatric patient caused by a heterozygous de novo mutation in the ornithine decarboxylase 1 (ODC1) gene. The new genetic disorder is characterized by global developmental delay, alopecia, overgrowth, and dysmorphic features. We hypothesized that this new mutation (c.1342 A>T) leads to a C-terminal truncation variant of the ODC protein that is resistant to normal proteasomal degradation, leading to putrescine accumulation in cells. ODC (E.C. 4.1.1.17) is a rate-limiting enzyme in the biosynthesis of polyamines (putrescine, spermidine, and spermine) that plays a crucial role during embryogenesis, organogenesis, and tumorigenesis. In this study, we show that primary dermal fibroblasts derived from a skin biopsy of a 3-year-old patient contain large amounts of ODC protein and putrescine compared with primary dermal (neonatal and adult) fibroblast control cells. Importantly, the accumulated ODC protein variant remained functionally active as we detected exceptionally high ODC enzyme activity in both primary dermal fibroblasts (12-17-fold of controls) and red blood cells (RBCs) (125-137-fold of controls), using a specific 14C radioactive ODC activity assay. Exposure of primary dermal fibroblasts to ODC inhibitor α-difluoromethylornithine (DFMO) reduced the ODC activity and putrescine to levels observed in controls without adversely affecting cell morphology or inducing cell death. In conclusion, our patient and potentially other patients that carry a similar ODC1 gain-of-function mutation might benefit from treatment with DFMO, a drug with a good safety profile, to suppress the exceptionally high ODC activity and putrescine levels in the body.

Increased bound putrescine accumulation contributes to the maintenance of antioxidant enzymes and higher aluminum tolerance in wheat

The accumulation of bound and conjugated polyamines (PAs) is an important protective trait in plants under adverse environmental conditions. However, their role in plant responses to aluminum (Al) stress remains largely unknown. In this study, we showed that Al treatment increased bound putrescine (Put) levels in the wheat root tips of Al-tolerant Xi Aimai-1, with little effect on its bound spermidine and conjugated PAs or that of Al-sensitive Yangmai-5. Terminating bound Put increments with a synthesis inhibitor (Phenanthroline, o-phen) exacerbated Al-induced root inhibition and callose production. However, it had no significant effect on Al uptake or distribution under Al stress. Instead, Al-induced reactive oxygen species (ROS) production and thus, oxidative damage, was greatly exacerbated by o-phen in the roots of Xi Aimai-1. Application of o-phen barely affected the two ROS generating enzymes (plasma membrane NADPH oxidase and cell wall-bound polyamine oxidase) in wheat roots. However, exogenous o-phen significantly reduced antioxidant enzyme (superoxide dismutase, ascorbate peroxidase, and catalase) activity, which positively correlated with the level of bound Put in Xi Aimai-1. These results clearly suggest that bound Put accumulation works to protect against Al-induced oxidative damage, possibly by maintaining antioxidant capacity in wheat.

Formation of Biogenic Amines in Pa (Green Onion) Kimchi and Gat (Mustard Leaf) Kimchi.

In this study, biogenic amine content in Pa (green onion) kimchi and Gat (mustard leaf) kimchi, Korean specialty kimchi types, was determined by high-performance liquid chromatography (HPLC). Many kimchi samples contained low levels of biogenic amines, but some samples had histamine and tyramine content over the safe levels. Based on the comparative analysis between the ingredient information on food labels and biogenic amine content of kimchi samples, Myeolchi-aekjeot appeared to be an important source of biogenic amines in both kimchi. Besides, through the 16s rRNA sequence analysis, Lactobacillus brevis appeared to be responsible for the formation of biogenic amines (tyramine, β-phenylethylamine, putrescine, and cadaverine) in both kimchi, in a strain-dependent manner. During fermentation, a higher accumulation of tyramine, β-phenylethylamine, and putrescine was observed in both or one (for putrescine) of kimchi types when L. brevis strains served as inocula. The addition of Myeolchi-aekjeot affected the initial concentrations of most biogenic amines (except for spermidine in Gat kimchi) in both kimchi. Therefore, this study suggests that using appropriately salted and fermented seafood products for kimchi preparation and using biogenic amine-negative and/or biogenic amine-degrading starter cultures would be effective in reducing biogenic amine content in Pa kimchi and Gat kimchi.

Effect of Biofermentation with Taxifolin on Physicochemical and Microbiological Properties of Cold-Smoked Pork Sausages.

SUMMARYThe aim of this work is to evaluate the effect of taxifolin on the physicochemical and microbiological properties of cold-smoked pork sausages produced using different commercial starter cultures with Leuconostoc carnosum and with a mixture of Pediococcus pentosaceus and Staphylococcus xylosus. Ultra performance liquid chromatography analysis demonstrated that after 181 days of storage total taxifolin content was the highest in samples with taxifolin and L. carnosum (60%), compared to the first day of storage. The sausages with taxifolin and the mixture of P. pentosaceus and S. xylosus (56%) followed next. Taxifolin improved the hygienic quality of sausages without significant effect on the growth of lactic acid bacteria. The accumulation of biogenic amines, including histamine and putrescine, was more effectively reduced in sausages inoculated with the taxifolin and P. pentosaceus and S. xylosus mixture. Using this mixture, the rate of lipolysis and lipid oxidation were effectively slowed down. Samples with taxifolin and L. carnosum showed the highest free radical scavenging activity on the first day of the study ((77.4±1.3) %) (p<0.05 in all samples). Mixtures containing taxifolin and starter cultures bound free radicals better than taxifolin alone. The colour parameters (L*, a* and b*) of preparations and final products were significantly influenced by taxifolin and starter cultures and storage time (p<0.05 in all samples).

Iron Regulation in Clostridioides difficile

The response to iron limitation of several bacteria is regulated by the ferric uptake regulator (Fur). The Fur-regulated transcriptional, translational and metabolic networks of the Gram-positive, pathogen Clostridioides difficile were investigated by a combined RNA sequencing, proteomic, metabolomic and electron microscopy approach. At high iron conditions (15 μM) the C. difficile fur mutant displayed a growth deficiency compared to wild type C. difficile cells. Several iron and siderophore transporter genes were induced by Fur during low iron (0.2 μM) conditions. The major adaptation to low iron conditions was observed for the central energy metabolism. Most ferredoxin-dependent amino acid fermentations were significantly down regulated (had, etf, acd, grd, trx, bdc, hbd). The substrates of these pathways phenylalanine, leucine, glycine and some intermediates (phenylpyruvate, 2-oxo-isocaproate, 3-hydroxy-butyryl-CoA, crotonyl-CoA) accumulated, while end products like isocaproate and butyrate were found reduced. Flavodoxin (fldX) formation and riboflavin biosynthesis (rib) were enhanced, most likely to replace the missing ferredoxins. Proline reductase (prd), the corresponding ion pumping RNF complex (rnf) and the reaction product 5-aminovalerate were significantly enhanced. An ATP forming ATPase (atpCDGAHFEB) of the F0F1-type was induced while the formation of a ATP-consuming, proton-pumping V-type ATPase (atpDBAFCEKI) was decreased. The [Fe-S] enzyme-dependent pyruvate formate lyase (pfl), formate dehydrogenase (fdh) and hydrogenase (hyd) branch of glucose utilization and glycogen biosynthesis (glg) were significantly reduced, leading to an accumulation of glucose and pyruvate. The formation of [Fe-S] enzyme carbon monoxide dehydrogenase (coo) was inhibited. The fur mutant showed an increased sensitivity to vancomycin and polymyxin B. An intensive remodeling of the cell wall was observed, Polyamine biosynthesis (spe) was induced leading to an accumulation of spermine, spermidine, and putrescine. The fur mutant lost most of its flagella and motility. Finally, the CRISPR/Cas and a prophage encoding operon were downregulated. Fur binding sites were found upstream of around 20 of the regulated genes. Overall, adaptation to low iron conditions in C. difficile focused on an increase of iron import, a significant replacement of iron requiring metabolic pathways and the restructuring of the cell surface for protection during the complex adaptation phase and was only partly directly regulated by Fur.

The arginine decarboxylase gene ADC1, associated to the putrescine pathway, plays an important role in potato cold-acclimated freezing tolerance as revealed by transcriptome and metabolome analyses.

Low temperature severely influences potato production as the cultivated potato (Solanum tuberosum) is frost sensitive, however the mechanism underlying the freezing tolerance of the potato is largely unknown. In the present research, we studied the transcriptome and metabolome of the freezing-tolerant wild species Solanum acaule (Aca) and freezing-sensitive cultivated S.tuberosum (Tub) to identify the main pathways and important factors related to freezing tolerance. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway annotation indicated that polyamine and amino acid metabolic pathways were specifically upregulated in Aca under cold treatment. The transcriptome changes detected in Aca were accompanied by the specific accumulation of putrescine, saccharides, amino acids and other metabolites. The combination of transcriptome and metabolome analyses revealed that putrescine exhibited an accumulative pattern in accordance with the expression of the arginine decarboxylase gene ADC1. The primary role of putrescine was further confirmed by analyzing all three polyamines (putrescine, spermidine, and spermine) and the genes encoding the corresponding enzymes in two sets of potato genotypes with distinct freezing tolerance, implying that only putrescine and ADC1 were uniquely enhanced by cold in the freezing-tolerant genotypes. The function of putrescine was further analyzed by its exogenous application and the overexpression of SaADC1 in S.tuberosum cv. E3, indicating its important role(s) in cold-acclimated freezing tolerance, which was accompanied with the activation of C-repeat binding factor genes (CBFs). The present research has identified that the ADC1-associated putrescine pathway plays an important role in cold-acclimated freezing tolerance of potato, probably by enhancing the expression of CBF genes.

Evaluation of psychrophilic, mesophilic, histamine forming bacteria and biogenic amine content in the muscle of mud spiny lobster, Panulirus polyphagus (HERBST, 1793) during ice storage

AbstractThe aim of the present study was to assess the evolution of psychrophilic, mesophilic and histamine forming bacteria along with the accumulation of cadaverine, histamine, putrescine, spermidine, spermine, tryptamine and tyramine during ice storage of mud spiny lobster (Panulirus polyphagus). Total mesophilic, psychrophilic and amine forming bacteria were initially enumerated at 2.62, 2.14 and 2.10 log cfu/g muscle, respectively and reached 7.07, 7.11, and 6.80 log cfu/g muscle, respectively, after 15 days of ice storage. Tryptamine, putrescine and histamine were detected from the 5th day, tyramine, spermidine, and spermine from the 10th day and cadaverine only at the 15th day of storage. Positive correlation of mesophilic and psychrophilic bacteria with tryptamine, putrescine and tyramine was detected. Species belonging to Enterobacter spp., Proteus spp., Klebsiella spp., Citrobacter spp., Photobacterium spp., Serratia spp., and Morganella morganii able to produce histamine, putrescine, cadaverine and tyramine were isolated during storage. The results obtained suggest that P. polyphagus muscle can be stored in ice for short period of time, which may reach 10 days. After the 10th day of ice storage, accumulation of biogenic amines may constitute a potential health hazard.Practical applicationsTo improve our understanding of freshness and bacterial species contribution to biogenic amines production in lobster during ice storage, knowledge of the microecosystem dynamics is necessary. Therefore, we investigated the dynamics of mesophilic, psychrophilic and histamine forming bacteria as well as the evolution of cadaverine, histamine, putrescine, spermidine, spermine, tryptamine and tyramine during ice storage of mud spiny lobster (Panulirus polyphagus). The results of the present study could be useful to assure the safety of lobster by controlling the biogenic amines content.

Modulation of plant and bacterial polyamine metabolism during the compatible interaction between tomato and Pseudomonas syringae

The polyamines putrescine, spermidine and spermine participate in a variety of cellular processes in all organisms. Many studies have shown that these polycations are important for plant immunity, as well as for the virulence of diverse fungal phytopathogens. However, the polyamines’ roles in the pathogenesis of phytopathogenic bacteria have not been thoroughly elucidated to date. To obtain more information on this topic, we assessed the changes in polyamine homeostasis during the infection of tomato plants by Pseudomonas syringae. Our results showed that polyamine biosynthesis and catabolism are activated in both tomato and bacteria during the pathogenic interaction. This activation results in the accumulation of putrescine in whole leaf tissues, as well as in the apoplastic fluids, which is explained by the induction of its synthesis in plant cells and also on the basis of its excretion by bacteria. We showed that the excretion of this polyamine by P. syringae is stimulated under virulence-inducing conditions, suggesting that it plays a role in plant colonization. However, no activation of bacterial virulence traits or induction of plant invasion was observed after the exogenous addition of putrescine. In addition, no connection was found between this polyamine and plant defence responses. Although further research is warranted to unravel the biological functions of these molecules during plant-bacterial interactions, this study contributes to a better understanding of the changes associated with the homeostasis of polyamines during plant pathogenesis.

Ornithine decarboxylase is involved in methyl jasmonate-regulated postharvest quality retention in button mushrooms (Agaricus bisporus).

In the present study, we investigated the role of ornithine decarboxylase (ODC) in the methyl jasmonate (MeJA)-regulated postharvest quality maintenance of Agaricus bisporus (J. E. Kange) Imbach button mushrooms by pretreating mushrooms with a specific irreversible inhibitor called α-difluoromethylornithine (DFMO) before exposure to MeJA vapor. Mushrooms were treated with 0 or 100 µmol L-1 MeJA or a combination of 120 µmol L-1 DFMO and 100 µmol L-1 MeJA, respectively, before storage at 4 °C for 21 days. Treatment with MeJA alone induced the increase in ODC activity whereas this effect was greatly suppressed by pretreatment with DFMO. α-Difluoromethylornithine strongly attenuated the effect of MeJA on decreasing cap opening, slowing the decline rate of soluble protein and total sugar, and accumulating total phenolics and flavonoids. α-Difluoromethylornithine pretreatment also counteracted the ability of MeJA to inhibit polyphenol oxidase and lipoxygenase activities, and malondialdehyde production, and to stimulate superoxide dismutase and catalase activities. It also largely downregulated MeJA-induced accumulation of free putrescine (Put). These results reveal that ODC is involved in MeJA-regulated postharvest quality retention of button mushrooms, and this involvement is likely to be associated with Put levels. © 2018 Society of Chemical Industry.

Effects of inoculating autochthonous starter cultures on biogenic amines accumulation of Chinese traditional fermented fish

The effects of three autochthonous starter cultures (Lactobacillus plantarum 120, Saccharomyces cerevisiae 2018 and Staphylococcus xylosus 135) on biogenic amines (BAs) accumulation in Chinese traditional fermented freshwater fish products (CTFPs) were studied. Some quality parameters of CTFPs, namely microbiological characteristics, α-amino nitrogen, trichloroacetic acid (TCA)-soluble peptides, protease activity, and free amino acids (FFAs) were also detected. No histamine accumulation was observed in each fermentation group. Putrescine and tyramine were the dominating BAs in CTFPs. Putrescine accumulation was significantly (p < 0.05) suppressed by inoculated with L. plantarum 120 and S. xylosus 135, and lowest tyramine content (363.15 mg/kg) was observed in sample inoculated with mixed starter cultures. Low levels of cadaverine (8.22–20.03 mg/kg), spermidine (6.60–8.11 mg/kg) and spermine (7.94–9.62 mg/kg) were observed in all groups at the end of the fermentation. Additionally, total BAs contents showed a significant (p < 0.05) correlation with protease activities and α-amino nitrogen contents. Practical applications The present study demonstrated the changes in some quality parameters and BAs contents of CTFPs influenced by different starter cultures. Results showed that lactic acid bacteria (LAB) and yeast were gradually increased, severe hydrolysis of proteins was occurred during CTFPs fermentation process, histamine accumulation was inhibited completely in all fermentation groups, putrescine was significantly (p < 0.05) suppressed by inoculated with L. plantarum 120 and S. xylosus 135, and significant (p < 0.05) inhibition of tyramine was observed in sample inoculated with mixed starter culture. To date, CTFPs were usually produced by spontaneous fermentation, and the quality was unsteady. Therefore, to improve the quality and avoid the unacceptable levels of BAs accumulation, inoculation of proper autochthonous starter cultures is advisable.

Modulation of polyamine biosynthesis in Arabidopsis thaliana by a drought mitigating Pseudomonas putida strain

Plant growth promoting rhizobacteria (PGPR) are a diverse group of beneficial soil bacteria that help plants in myriad ways. They are implicated in the processes of general growth and development, as well as stress mitigation. Although the physiology of plant-PGPR interaction for abiotic stress tolerance has been well reported, the underlying molecular mechanisms in this phenomenon are not clearly understood. Among the many endogenous molecules that have been reported to impart abiotic stress tolerance in plants are a group of aliphatic amines called polyamines. Here, we report the impact of a free living, drought-mitigating rhizobacterial strain, Pseudomonas putida GAP-P45 on the expression of key genes in the polyamine metabolic pathway and the accumulation of the three major polyamines, putrescine, spermidine and spermine in water-stressed Arabidopsis thaliana. We observed that, inoculation of A. thaliana with P. putida GAP-P45 with or without water-stress, caused significant fluctuations in the expression of most polyamine biosynthetic genes (ADC, AIH, CPA, SPDS, SPMS and SAMDC) and cellular polyamine levels at different days of analysis post treatments. The enhanced accumulation of free cellular putrescine and spermidine observed in this study correlated positively with the water stress tolerant phenotype of A. thaliana in response to P. putida GAP-P45 inoculation reported in our previous study (Ghosh et al., 2017). Our data point towards (a) transcriptional regulation of polyamine biosynthetic genes and (b) complex post transcriptional regulation and/or interconversion/canalization of polyamines, by P. putida GAP-P45 under normal and water-stressed conditions.

Putrescine as indicator of manganese neurotoxicity: Dose-response study in human SH-SY5Y cells

Disrupted polyamine metabolism with elevated putrescine is associated with neuronal dysfunction. Manganese (Mn) is an essential nutrient that causes neurotoxicity in excess, but methods to evaluate biochemical responses to high Mn are limited. No information is available on dose-response effects of Mn on putrescine abundance and related polyamine metabolism.The present research was to test the hypothesis that Mn causes putrescine accumulation over a physiologically adequate to toxic concentration range in a neuronal cell line. We used human SH-SY5Y neuroblastoma cells treated with MnCl2 under conditions that resulted in cell death or no cell death after 48 h. Putrescine and other metabolites were analyzed by liquid chromatography-ultra high-resolution mass spectrometry. Putrescine-related pathway changes were identified with metabolome-wide association study (MWAS). Results show that Mn caused a dose-dependent increase in putrescine over a non-toxic to toxic concentration range. MWAS of putrescine showed positive correlations with the polyamine metabolite N8-acetylspermidine, methionine-related precursors, and arginine-associated urea cycle metabolites, while putrescine was negatively correlated with γ-aminobutyric acid (GABA)-related and succinate-related metabolites (P < 0.001, FDR < 0.01). These data suggest that measurement of putrescine and correlated metabolites may be useful to study effects of Mn intake in the high adequate to UL range.

Polyamine- and Amino Acid-Related Metabolism: The Roles of Arginine and Ornithine are Associated with the Embryogenic Potential.

The mechanisms that control polyamine (PA) metabolism in plant cell lines with different embryogenic potential are not well understood. This study involved the use of two Araucaria angustifolia cell lines, one of which was defined as being blocked, in that the cells were incapable of developing somatic embryos, and the other as being responsive, as the cells could generate somatic embryos. Cellular PA metabolism was modulated by using 5 mM arginine (Arg) or ornithine (Orn) at two time points during cell growth. Two days after subculturing with Arg, an increase in citrulline (Cit) content was observed, followed by a higher expression of genes related to PA catabolism in the responsive cell line; whereas, in the blocked cell line, we only observed an accumulation of PAs. After 14 d, metabolism was directed towards putrescine accumulation in both cell lines. Exogenous Arg and Orn not only caused a change in cellular contents of PAs, but also altered the abundance of a broader spectrum of amino acids. Specifically, Cit was the predominant amino acid. We also noted changes in the expression of genes related to PA biosynthesis and catabolism. These results indicate that Arg and Orn act as regulators of both biosynthetic and catabolic PA metabolites; however, we suggest that they have distinct roles associated with embryogenic potential of the cells.