Increase In Putrescine Content Research Articles

Ethanol induces heat tolerance in plants by stimulating unfolded protein response.

Ethanol priming induces heat stress tolerance by the stimulation of unfolded protein response. Global warming increases the risk of heat stress-related yield losses in agricultural crops. Chemical priming, using safe agents, that can flexibly activate adaptive regulatory responses to adverse conditions, is a complementary approach to genetic improvement for stress adaptation. In the present study, we demonstrated that pretreatment of Arabidopsis with a low concentration of ethanol enhances heat tolerance without suppressing plant growth. We also demonstrated that ethanol pretreatment improved leaf growth in lettuce (Lactuca sativa L.) plants grown in the field conditions under high temperatures. Transcriptome analysis revealed a set of genes that were up-regulated in ethanol-pretreated plants, relative to water-pretreated controls. Binding Protein 3 (BIP3), an endoplasmic reticulum (ER)-stress marker chaperone gene, was among the identified up-regulated genes. The expression levels of BIP3 were confirmed by RT-qPCR. Root-uptake of ethanol was metabolized to organic acids, nucleic acids, amines and other molecules, followed by an increase in putrescine content, which substantially promoted unfolded protein response (UPR) signaling and high-temperature acclimation. We also showed that inhibition of polyamine production and UPR signaling negated the heat stress tolerance induced by ethanol pretreatment. These findings collectively indicate that ethanol priming activates UPR signaling via putrescine accumulation, leading to enhanced heat stress tolerance. The information gained from this study will be useful for establishing ethanol-mediated chemical priming strategies that can be used to help maintain crop production under heat stress conditions.

Effect of silver nitrate and putrescine on in vitro shoot organogenesis of Polygonum multiflorum

Polygonum multiflorum is a flowering plant that belongs to the family Polygonaceae and it is commonly used for medicinal and ornamental purposes. Few studies have been studied about the regeneration of this species. Therefore, we aimed to develop a suitable protocol for regeneration and subsequent growth of shoots by comparing the silver nitrate (AgNO3) (ethylene inhibitor) and the putrescine (polyamine). Internode explants were cultured on shoot regeneration media (Murashige and Skoog (MS) media containing 2 mg L-1 of 6-benzylaminopurine). To analysis, the effect of AgNO3 and putrescine on shoot regeneration and length, different concentrations (mg L-1) of AgNO3 (0, 1, 5, 7, 10, and 20) and putrescine (0, 10, 30, 50, 100, and 200) were added to the MS media. The result showed that at highest concentration (20 mg L-1) of AgNO3 treatment decrease number of shoots (NOS) (1.4 ± 0.2 mm) and shoot length (9.7 ± 1.6 mm) was observed. Putrescine considerably increased the regeneration efficiency, NOS per explant, and shoot length in all the concentrations when compared to AgNO3 treatment. Among the different concentrations, the highest NOS (2.52 ± 0.2 mm) was obtained in cultures supplemented with 30 mg L-1 putrescine, whereas the further increase in putrescine concentration reduced shoot regeneration. The longest shoots (20.5 ± 1.7 mm) were achieved in cultures supplemented with 200 mg L-1 putrescine. The findings of this study indicate that the addition of putrescine to the media could be suitable for P. multiflorum micropropagation and plant transformation.

Drought tolerant maize cultivar accumulates putrescine in roots

We compared putrescine metabolism in roots of tolerant maize cultivar (Karoon) to a susceptible cultivar (260) under drought stress. The experiment was conducted in growth chamber under controlled condition in 80 pots. Drought stress led to significant increase in root growth indices, H2O2 content and activity of antioxidant enzymes (superoxide dismutase, catalase, and peroxidase). The qPCR analysis revealed that expression of maize polyamine oxidase (ZmPAO) genes including ZmPAO1, ZmPAO2, ZmPAO4 and ZmPAO6 as well as ornithine decarboxylase (ZmODC) significantly increased in root of both cultivars in response to drought. However, up-regulation of ZmPAO3 and ZmPAO5 was only observed in root of tolerant cultivar. Consistently, drought caused increased polyamine oxidase activity in the tolerant cultivar. Transcription of spermidine synthase (ZmSPDS) was down-regulated by drought stress. Transcript levels of ZmPOA, ZmODC and ZmSPDS were more affected by drought in tolerant cultivar compared to susceptible cultivar. Increase in putrescine content under drought stress was more prominent in roots of tolerant cultivar, compared to susceptible cultivar. We attribute the accumulation of putrescine to increased rate of putrescine synthesis, due to higher expression of ZmPAOs and ZmODC and decreased rate of its depletion for biosynthesis of higher polyamines due to down-regulation of ZmSPDS.

Manganese Stimulates Putrescine Accumulation and Influences Associated Polyamine, Methionine and Neurotransmitter Metabolism in Human SH-SY5Y Neuroblastoma Cells

Manganese (Mn) is an essential nutrient. However, a longitudinal study showed that exposure to Mn containing welding fumes under federal safety limits may cause a dose-dependent progression of Parkinsonism. Excess Mn causes neurotoxicity, however the underlying molecular mechanism remains unclear. Disrupted polyamine metabolism with increased putrescine levels was observed in suicidal cohorts and also occurs in multiple neurologic disorders, but no information is available on the possible effects of Mn on polyamine metabolism. To test the hypothesis that increased Mn causes increased putrescine and altered polyamine metabolism, human SH-SY5Y neuroblastoma cells were treated with MnCl2 (0,1,5,10,50,100μM for 5 hr; n=9/condition). Results showed that cellular Mn measured was representative of normal to pathological range (6 to 49 ng/mg protein) in the human brain. Putrescine and other untargeted metabolites were analyzed by liquid chromatography-ultra high-resolution mass spectrometry followed by metabolome-wide association study (MWAS) of putrescine. Results showed that cellular Mn caused a dose dependent increase in putrescine content (3.2 nmol/mg protein to 4.5 nmol/mg protein; p

Effect of polyamines on somatic embryogenesis via mature embryo in wheat

Plant tissue culture via somatic embryogenesis plays a key role in wheat genetic transformation studies. Therefore, the production of embryogenic calli with a high regeneration capacity is a prerequisite for efficient plant regeneration. Although immature embryos are the best type of explants for plant regeneration via somatic embryogenesis, the use of mature embryos has remarkable advantages compared to immature embryos as explants. However, the regeneration capacity of mature embryos is still lower than that of immature embryos. Plant regeneration from somatic embryos can only occur when they are mature enough. Moreover, the maturation of somatic embryos and their conversion into plantlets is closely associated with the used plant growth regulators. Polyamines are major components in the formation of somatic embryogenesis and plant regeneration. In this study, the effect of three types of polyamines on somatic embryogenesis and plant regeneration in wheat was evaluated. Although the effect of polyamines on embryogenic callus formation was not significant, their effect on responded embryogenic callus rate and plant regeneration efficiency was very significant. The highest responded embryogenic callus rate and highest regeneration efficiency were obtained on MS medium containing putrescine. Unlike other polyamines, an increase in putrescine concentration promoted a higher number of regenerated plants. The highest values for responded embryogenic callus and regeneration efficiency were determined in 1 mM concentration of putrescine. Plantlets derived from somatic embryos were successfully established in soil, producing fertile seeds.

Influence of Putrescine on Enzymes of Ammonium Assimilation in Maize Seedling

The effect of different concentrations of putrescine on biochemical changes in root and shoot of six days old maize seedlings in terms of enzymes of ammonium assimilation were examined. The results revealed that glutamate dehydrogenase (GDH) activity was enhanced at lower concentration of putrescine but at higher concentration, the activity of this enzyme was declined. Glutamine synthetase (GS) activity decreased with increase in concentration of putrescine and it was highest at 1000 μm concentration. Howe ver, glutamate synthase (GOGAT) activity increased with increase in concentration of putrescine upto 100 μm in root and upto 50 μm in shoot and further increase in concentration resulted in decline of enzymatic activity. Protein and total nitrogen content increased upto 10 μm concentration of putrescine and it decreased further with increase in concentration both in root and shoot of maize seedling.

Endotoxin and biogenic amine levels in Atlantic mackerel (Scomber scombrus), sardine (Sardina pilchardus) and Mediterranean hake (Merluccius merluccius) stored at 22 °C

Whole Atlantic mackerel (Scomber scombrus), sardine (Sardina pilchardus) and Mediterranean hake (Merluccius merluccius) from the Croatian Adriatic were stored at 22 °C and changes in histamine, putrescine, tyramine and cadaverine levels were monitored in relation to bacterial endotoxin. After 12 h, histamine levels in sardine were above the legal limit of 50 mg kg−1, set by the US Food and Drug Administration, and an increase in putrescine content preceded the increase in histamine. After 24 h, histamine contents in mackerel and sardine reached 1090 ± 101 and 577 ± 275 mg kg−1, respectively, which exceeded the toxic threshold of 500 mg kg−1. At the same time, the putrescine content was also high in both fish (353–420 mg kg−1). The time-course of endotoxin production was similar in all fish species stored at 22 °C. A high correlation was found between endotoxin and histamine, and between endotoxin and putrescine in mackerel and sardine. On the other hand, high endotoxin levels in hake, after 24 h, were associated with the low histamine and putrescine content (40–60 mg kg−1).

Calorimetry studies of a chemical oscillatory system

The effect of putrescine (PUT) on KSCN-H2O2-CuSO4-NaOH oscillating system was investigated by calorimetric method. The oscillating reaction was monitored in a closed reactor with stirring, and the result showed that the oscillating period was linearly related with putrescine concentration and the numbers of oscillation decreased with increase in putrescine concentration. When [PUT]=2.83·10−4 M, no oscillation was observed. A possible mechanism is proposed that putrescine is a scavenger of the active-oxygen species. The result of numerical simulation by a simplified mechanism consisting of 18 kinetic steps and 16 variables is consistent with the experimental findings.

Exogenous salicylic acid increases polyamine content but may decrease drought tolerance in maize

It was shown in a previous work that 0.5 mM salicylic acid (SA) added in the hydroponic solution of maize increased its tolerance to low temperature stress [Planta 208 (1999) 175]. The effect of SA and cold treatments on polyamine content in the leaves was investigated using the HPLC technique in this work. Both 0.5 mM SA and cold treatment caused a significant increase in putrescine content. Spermidine increased only when the addition of SA was followed by low temperature stress. The spermine content decreased after both SA and low temperature treatments. The parallel use of 0.5 mM SA and 15% PEG caused a dramatic increase in the electrolyte leakage and a decrease in certain photosynthetic parameters in maize and wheat. It is concluded that the 0.5 mM SA pre-treatment, which increased the chilling tolerance, caused an increased sensitivity to drought.

Effects of Dietary 1, 4-Diaminobutane (Putrescine) on Eggshell Quality and Laying Performance of Older Hens

We conducted an experiment to evaluate the potential for dietary 1,4-diaminobutane (putrescine) to promote eggshell quality and overall laying hen performance. A total of 128, 60-wk-old Barred Rock hens were fed a corn and soybean meal-based layer diet supplemented with 0.0 (control), 0.2, 0.4, and 0.6% free base 1,4-diaminobutane for 4 wk. The feeding of supplemental putrescine decreased feed consumption (P ≤ 0.05) and egg mass (P ≤ 0.05) and tended to decrease egg production (P < 0.08). Albumen quality was not significantly affected (P < 0.09) by the end of the experiment, as determined by Haugh units. Eggshell thickness was not significantly improved with lower levels of dietary putrescine (P < 0.08). Although dietary putrescine did not have any effect on the relative weights of duodenum, jejunum + ileum, or pancreas, there was a linear increase in putrescine concentrations in tissues (P ≤ 0.05). Supplementation of dietary putrescine also resulted in increased putrescine and spermidine concentrations in egg (P ≤ 0.05). Egg weight and eggshell deformation increased over time; however, eggshell weight, eggshell weight as percentage of egg weight, and eggshell thickness decreased (P ≤ 0.05). It appeared that eggshell quality declined regardless of diet over the 4-wk experimental period. It was concluded that the lack of effect of dietary putrescine on egg parameters, with the exception of albumen quality and eggshell thickness, was due to putrescine toxicity. Hens transferred excess dietary putrescine and metabolites to eggs.

Role of polyamines in the cytokinin-dependent physiological processes II. Modulation of polyamine levels during cytokinin-stimulated expansion of cucumber cotyledons

The cucumber cotyledon expansion test was used as a model system to study a possible relationship between cytokinin and polyamines. When kinetin was applied to excised cotyledons incubated in the dark it caused a marked increase in the activity of arginine decarboxylase. As a result of ADC action, putrescine content also rose markedly, whereas the level of spermidine and spermine decreased. However, inhibition of putrescine biosynthesis with D-arginine did not affect cytokinin promotion growth. Applied alone, putrescine had no significant effect on growth. These results indicate that the large increase in putrescine content that derives from cytokinin treatment cotyledons is not essential for cytokinin-induced expansion of cotyledons. Addition of K+ and Ca2+ ions to the cotyledons incubated with cytokinin caused a marked reduction in the putrescine level and ADC activity. The higher level of putrescine (35 %) and spermine (62 %) bound to chromatin and the large increase (174 %) in spermidine content bound to ribosomes which derive from cytokinintreated cotyledons in relation to literature data can indicate that these polyamines may play an important role in gene expression during cytokinin-stimulated expansion of cucumber cotyledons. The inhibition of cytokinin effect, viz. enlargement of the cotyledons by inhibitors of spermidine biosynthesis, additionally suggessted a possible involvement of polyamines in cytokinin action.

Prolactin and polyamine catabolism: specific effect on polyamine oxidase activity in rat thymus

Much evidence suggests that prolactin has an immunoregulatory function and that its effects on cells of the immune system depend on the level and specific forms of the receptors present on the target cells. The effect of administration of prolactin on polyamine catabolism was investigated in thymus of male intact rats by measuring the activities of spermidine/spermine N 1-acetyltransferase and polyamine oxidase, because of the relationships between polyamines (especially putrescine) and the immune system. The administration of prolactin to rats resulted in the rapid induction of spermidine/spermine N 1-acetyltransferase activity in the thymus (1.6-times the level of control rats, within 4 h), and in a marked decrease in polyamine oxidase activity at 24 h. The changes in enzyme activities were accompanied by an increase in putrescine concentration and a decrease in spermidine and spermine concentrations. In the spleen, prolactin increased SAT activity only 24 h after administration and was ineffective on PAO activity.

Effects of foliar potassium concentration on morphology, ultrastructure and polyamine concentrations of Scots pine needles.

We examined the effects of three foliar potassium concentrations (high, intermediate and low) on the morphology, ultrastructure and polyamine concentrations of current-year and 1- and 2-year-old needles of 30-year-old Scots pine (Pinus sylvestris L.) trees. Foliar K concentration had only a slight effect on needle morphology. The sclerenchyma cell walls were thinner, the xylem area was larger, and the resin ducts were smaller in needles with a low K concentration than in needles with a high or intermediate K concentration. In addition, the bundle sheath cells were collapsed in needles having a low K concentration. The secondary growth of phloem tissue and the mesophyll area were greater in needles with a high or intermediate K concentration than in needles with a low K concentration, possibly indicating greater production of photoassimilates in these trees. At the ultrastructural level, mesophyll cells with enlarged central vacuoles and small vacuoles containing electron-dense material were common in needles having a low K concentration. Enlargement of the central vacuole coincided with an exponential increase in putrescine concentration in needles with a low K concentration, suggesting that the central vacuole may function as a storage site for putrescine.

Ozone stress modulates amine oxidase and lipoxygenase expression in lentil ( Lens culinaris) seedlings

The effect of ozone stress on polyamine metabolism and membrane lipid peroxidation in lentil seedlings through the amine oxidase and lipoxygenase activity and expression has been investigated. Ozone is shown to control the expression of these enzymes at the transcriptional level, down-regulating the amine oxidase gene and up-regulating the lipoxygenase gene. The decrease of amine oxidase activity correlated with the increase of putrescine concentration in the ozone-treated plantlets, whereas the increase of lipoxygenase activity was paralleled by enhanced membrane lipid peroxidation. Finally, polyamines are shown to inhibit lipoxygenase activity in lentils.

Loss of Intracellular Putrescine Pool-Size Regulation Induces Apoptosis

Synthesis and uptake are two important regulated mechanisms by which eukaryotic cells maintain polyamine levels. The role that loss of synthesis and/or uptake regulation plays in mediating putrescine toxicity was investigated by comparing toxicity in an ornithine decarboxylase (ODC)-deficient Chinese hamster ovary cell line (C55.7) with a functional putrescine transport system and an ODC-overproducing rat hepatoma cell line (DH23b), which are transport regulation deficient. When C55.7 cells were transfected with either mouse ODC (M) or trypanosome ODC (Tb), intracellular putrescine content increased slightly in C55.7(Tb-ODC), compared to C55.7(M-ODC), due to the lack of response of Tb-ODC to polyamine regulation. The increase in putrescine content resulting from loss of ODC regulation had no impact on cell growth and viability. When the feedback repression of polyamine uptake was blocked with cycloheximide, C55.7 cells transfected with either ODC construct accumulated very high levels of putrescine from the medium, and underwent apoptosis in a putrescine dose-dependent manner. A similar correlation of deregulated putrescine uptake and increased apoptotic cells was observed in DH23b cells. These data demonstrate that loss of feedback regulation on the polyamine transport system, but not ODC activity, is sufficient to induce apoptosis. Thus, downregulation of the transport system is necessary to prevent accumulation of cytotoxic putrescine levels in rodent cells.

In vivo trypanocidal activities of new S-adenosylmethionine decarboxylase inhibitors.

A series of novel aromatic derivatives based on the structure of methylglyoxal bis(guanylhydrazone) (MGBG) was examined for trypanocidal activities in human and veterinary trypanosomes of African origin. One agent, CGP 40215A, a bicyclic analog of MGBG which also resembles the diamidines diminazene (Berenil) and pentamidine, was curative of infections by 19 isolates of Trypanosoma brucei subspecies as well as a Trypanosoma congolense isolate. Several of these isolates were resistant to standard trypanocides. Curative doses were < or = 25 mg/kg of body weight/day for 3 days in these acute laboratory model infections. In addition, CGP 40215A also cured a model central nervous system infection in combination with the ornithine decarboxylase inhibitor DL-alpha-difluoromethylornithine (DFMO; Ornidyl, eflornithine). Curative combinations were 14 days of oral 2% DFMO (approximately 5 g/kg/day) plus 5, 10, or 25 mg/kg/day for 3 or 7 days given by intraperitoneal injection or with a miniosmotic pump. Combinations were most effective if CGP 40215A was given in the second half or at the end of the DFMO regimen. MGBG has modest activity as an inhibitor of trypanosome S-adenosylmethionine decarboxylase (50% inhibitory concentration [IC50]. 130 microM), while CGP 40215A was a more active inhibitor (IC50, 20 microM). Preincubation of trypanosomes with CGP 40215A for 1 h caused a reduction in spermidine content (36%) and an increase in putrescine content (20%), indicating that one possible mechanism of its action may be inhibition of polyamine biosynthesis.

Increased polyamine levels and changes in the sensitivity to convulsions during chronic treatment with cocaine in mice

Polyamines have been demonstrated to modulate seizure activity in animals. Repeated administration of a subthreshold dose of cocaine resulted in the development of sensitization to cocaine-induced seizures during an initial 3 or 4 days, followed by the development of tolerance to seizures on days 5 and 6. In the present study, polyamines, such as putrescine, spermidine and spermine, were measured in regions of the brain obtained from mice that showed differential sensitivity in seizure activity during repeated cocaine injections. Animals were sacrificed for polyamine measurements 24 h after the second and the fifth injections of either cocaine or saline (on day 3 and day 6, respectively), and 3 days after the last injection. On day 3, there were significant increases in putrescine in the striatum, hippocampus and cerebellum, and in spermine in the cerebellum of cocaine-treated mice, as compared to saline-treated mice. On day 6, treatment with cocaine significantly increased putrescine in all regions, and spermidine in striatum and hippocampus, as compared to saline treatment. Cocaine treatment had no effect on any polyamine levels measured 3 days after the last injection, except for spermidine in the cortex. Because putrescine has been shown to be an antagonist of the polyamine-binding site on the N-methyl- d-aspartate receptor and to retard the development of amygdala-kindling, the present results suggest that the increases in putrescine content may be associated with the development of tolerance to convulsant effects observed during the later period of repeated administration of cocaine.

Methyl jasmonate reduces chilling injury in Cucurbita pepo through its regulation of abscisic acid and polyamine levels

Zucchini squash fruit ( Cucurbita pepo L.) were pressure infiltrated (82.7 kPa for 3 min)_with methyl jasmonate (MJ) in aqueous suspension and then stored at a chilling temperature of 5°C. Control fruit were treated with distilled water and handled in a similar manner. Control fruit started to show chilling injury after 4 days of storage, whereas treated fruit had a 2–4 day delay in the onset of chilling injury symptoms. The abscisic acid (ABA) level in exocarp tissue of the control fruit increased after chilling treatment. An even larger increase in ABA level was found in the MJ-treated tissue upon exposure to 5°C, indicating that MJ may stimulate ABA synthesis at chilling temperatures. The treated and control fruit had similar increases in putrescine content when exposed to chilling temperatures. Spermidine and spermine contents decreased during storage at 5°C in treated and control squash. However, the treated fruit maintained higher levels of spermidine and spermine than the control fruit throughout storage at 5°C. These results indicate that MJ may prevent chilling injury by a mechanism which involves regulation of ABA and polyamine levels.

Fungicidal activity of the synthetic putrescine analogue, (E)‐l,4‐diaminobut‐2‐ene, and derivatives

AbstractThe putrescine analogue. (E)‐1,4‐diaminobut‐2‐ene (E‐BED), synthesized as the dihydrochloride salt, controlled five economically important crop pathogens, Erysiphe qraminis DC f.sp. hordei Marchal. Uromyces viciae‐fabae (Pers.) Schroet, Botrytis fabae Sardina, Podosphaera leucotricha (Ell. &amp; Ev.) Salm. and Phytophthora infestans (Mont) De Bary. The Z‐isomer. Z‐BED. was also fungicidal, although less so than E‐BED. Post‐inoculation treatment with E‐BED gave greater control of powdery mildew infection on barley and rust and chocolate spot on broad bean than did pre‐inoculation application. It was also effective in vitro against Botrytis cinerea Pers. ex Fr., Pyricularia oxyzae Br. &amp; Cav. and Pyrenophora avenae Ito &amp; Karibay. When P. avenae was grown in the presence of E‐BED dihydrochloride at 81·5 mg litre−1, growth was reduced by 58% and there were significant reductions in soluble ornithine decarboxylase (ODC) and S‐adenosylmethionine decarboxylase (AdoMetDC) activity. These changes were accompanied by a sevenfold increase in putrescine concentration, a 60% increase in spermine concentration and a 32% reduction in spermidine concentration within the fungal tissue.

Characterization of a COS cell line deficient in polyamine transport

In the present study, we describe the isolation and characterization of a COS cell line deficient in polyamine uptake that may provide an important tool for the molecular cloning of polymaine transporter(s). The cells were selected by isolation for resistance against the cytotoxic agent, methylglyoxal bis(guanylhydrazone) (MGBG), which is entering the cells using the same transport system as the polyamines. The isolated cell line was capable of growing in the presence of 100 μM MGBG, which totally inhibited the growth of the wild-type cells. The transport of putrescine and spermidine was markedly decreased in the COS-MGBG r cells. The decrease in putrescine transport was mainly a result of a 14-fold decrease in V max , wheras the reduced spermidine uptake was due to a 3–4-fold decrease in V max as well a 12-fold increase in K m , indicating the existence of at least two separate transport systems. No major difference in polyamine content was seen between the parental and the COS-MGBG r cells when grown without MGBG. In the presence of MGBG, both cell lines exhibited an increase in putrescine content. Treatment with MGBG also resulted in a decrease in spermidine and spermine contents in the wild-type cells. In the COS-MGBG r cells, on the other hand, there were no statistically significant effects on the spermidine and spermine contents by MGBG treatment. In the wild-type cells, depletion of polyamines, e.g., by treatment with the ornithine decarboxylase inhibitor 2-difluoromethylornithine (DFMO), stimulated the uptake of polyamines (3–7-fold), whereas in the COS-MGBG r cells the effect of DFMO treatment on polyamine transport was only minor. In contrast to the growth-medium of the wild-type cells, large amounts of polyamines accumulated in the medium of the COS-MGBG r cells, presumably indicating that COS cells normally excrete polyamines and then salvage them using the polyamine transport system.