Free Polyamines Research Articles

Transcriptome and Metabolome Analyses Reveal Molecular Responses of Two Pepper (Capsicum annuum L.) Cultivars to Cold Stress.

Low temperature is a significant factor affecting field-grown pepper. The molecular mechanisms behind peppers’ response to cold stress remain unknown. Transcriptomic and metabolomic analyses were used to investigate the responses of two pepper cultivars, XS (cold-sensitive) and GZ (cold-resistant), to cold stress; these were screened from 45 pepper materials. In this study, compared with the control group (0 h), we identified 10,931 differentially expressed genes (DEGs) in XS and GZ, 657 differentially expressed metabolites (DEMs) in the positive ion mode, and 390 DEMs in the negative ion mode. Most DEGs were involved in amino acid biosynthesis, plant hormone signal transduction, and the mitogen-activated protein kinase (MAPK) signaling pathway. Furthermore, metabolomic analysis revealed that the content of free polyamines (PAs), plant hormones, and osmolytes, mainly contained increased putrescine, spermine, spermidine, abscisic acid (ABA), jasmonic acid (JA), raffinose, and proline, in response to cold stress. Importantly, the regulation of the ICE (inducer of CBF expression)-CBF (C repeat binding factors)-COR (cold regulated) pathway by Ca2+ signaling, MAPK signaling, and reactive oxygen species (ROS) signaling plays a key role in regulating responses of peppers to cold stress. Above all, the results of the present study provide important insights into the response of peppers to cold stress, which will reveal the potential molecular mechanisms and contribute to pepper screening and breeding in the future.

Conjugated Polyamines in Root Plasma Membrane Enhanced the Tolerance of Plum Seedling to Osmotic Stress by Stabilizing Membrane Structure and Therefore Elevating H+-ATPase Activity.

Polyamines are small positively charged molecules in plants and play important functions in many biological processes under various environmental stresses. One of the most confounding problems relating to polyamines (PAs) in stresses is the lack of understanding of the mechanisms underlying their function(s). Furthermore, a limited number of studies have addressed this issue at the sub-cellular level, especially in tree plants under drought stress. Therefore, in this research, by simulating natural drought stress with polyethylene glycol (PEG) osmotic stress, the relationship between the levels of conjugated polyamines and the activity of H+-ATPase in the plasma membrane was elucidated with the roots of two plum (Prunus salicina L.) cultivars, which were different in drought tolerance, as experimental materials. Furthermore, free PA levels and the activities of S-adenosylmethionine decarboxylase (SAMDC) and transglutaminase (TGase), which were closely associated with the levels of free and conjugated PAs, were also detected. Results showed that under osmotic stress, the increases of the levels of non-covalently conjugated (non-CC) spermidine (Spd) and spermine (Spm), covalently conjugated (CC) putrescine (Put) and Spd in the plasma membrane of drought-tolerant Ganli No. 5 were more significant than those of drought-sensitive Suli No. 3, indicating that these conjugated PAs might be involved in the tolerance of plum seedlings to stress. Furthermore, the conjugated PAs were closely correlated with plum seedling growth, water retention capacity, plasma membrane damage degree, and hydrogen (H+)-ATPase activity in the plasma membrane. To get more complementary pieces of evidence, we subjected plum seedlings to combined treatments of PEG and exogenous PA (Spd and Spm), and an inhibitor of SAMDC [methylglyoxal-bis (guanylhydrazone), (MGBG)] or TGase (o-phenanthroline). These results collectively suggested that non-CC Spd and Spm, CC Put and Spd in plasma membrane might function in enhancing the tolerance of plum seedlings to osmotic stress by stabilizing membrane structure and therefore elevating H+-ATPase activity.

Orbitides and free polyamines have similarly limited fungicidal activity against three common pathogens of flax in vitro

Fusarium oxysporum f. sp. lini and Septoria linicola are causes of fusarium wilt and pasmo in flax ( Linum usitatissimum). Members of a third fungal genus, Alternaria spp., have also been found in fiber and linseed varieties of flax, and are a source of post-harvest spoilage and mycotoxins in a wide range of crops. We performed a microdilution assay and calculated the median effective concentration (EC50) to compare the potency of cyclolinopeptides (CLPs), two polyamines (spermidine and spermine), and the fungicide carbendazimin in the control of three fungi that have potential pathogenic activity ( F. oxysporum, S. linicola, and Alternaria spp), of which the first two are particulary significant causes of disease in flax. For carbendazim, all EC50 values were <0.6 μg/mL. The observed EC50 ranged from 111 to 340 μg/mL for a mixture of six unique CLPs, 109 to 778 μg/mL for spermine, and 21 to 272 μg/mL for spermidine. Spermidine was most effective against Alternaria sp., with an EC50 of 21 μg/mL. The results presented here showed that polyamines and CLPs possess limited antifungal activities against several fungi, with spermidines the most effective naturally occurring compound tested. Our findings do not support the hypothesis that CLPs act as potent antifungals against the three species of pathogens tested.

Polyamines conjugated to plasma membrane functioned in enhancing the tolerance of cucumber seedlings to osmotic stress via elevating H+-ATPase activity

Polyamine (PA), one of the important plant growth regulators, is closely associated with drought stress. However, the function of conjugated PA is not still clear in the roots of cucumber seedlings under polyethylene glycol (PEG) osmotic stress. Therefore, in this study the relationship between the levels of conjugated polyamines and the activity of H+-ATPase in plasma membrane was elucidated with the roots of two cucumber (Cucumis sativus L.) cultivars, which were different in drought tolerance, as experimental materials. Furthermore, the contents of free PAs and the activities of S-adenosylmethionine decarboxylase (SAMDC) and transglutaminase (TGase), which were closely related to the levels of conjugated polyamines, were also determined. Results showed that under osmotic stress, the increases of the levels of non-covalently conjugated (non-CC) spermidine (Spd) and spermine (Spm), covalently conjugated (CC) putrescine (Put) and Spd in plasma membrane of drought-tolerant Tangshan 5 were more obvious than those of drought-sensitive Jinyou 1. Furthermore, the conjugated PAs mentioned above were closely correlated with increase rate of seedling dry weight, plasma membrane permeability, water content and H+-ATPase activity in plasma membrane. Results of the additional tests, in which exogenous Spd, Spm and two inhibitors, MGBG and phenanthrolin were used, were complementary to the results above. From these results, it could be concluded that non-CC Spd and Spm, CC Put and Spd in plasma membrane functioned in enhancing the tolerance of cucumber seedlings to osmotic stress via elevating H+-ATPase activity.

Dissolved free amino acids and polyamines are two major dissolved organic nitrogen sources for marine bacterioplankton in the northern slope of the South China Sea

Dissolved free amino acids and polyamines are two major dissolved organic nitrogen sources for marine bacterioplankton in the northern slope of the South China Sea

Gibberellin and spermidine synergistically regulate polyamine metabolism during the development of Rhododendron flowers

Polyamines (PAs) are involved in various developmental processes, especially plant flowering. Their significant influences have been established; however, the exact mechanism by which PAs regulate flowering remains unclear. To explore PA metabolism in plant flowering, gibberellic acid (GA3, 0~2400 mg L−1) and spermidine (Spd, 0~1 mM) were applied alone or in combination during the early stage of flower bud formation in Rhododendron simsii. The application of GA3 alone advanced initial flowering, while that of Spd alone delayed initial flowering. Interestingly, GA3 and Spd applied in combination advanced initial flowering by 2 days. Furthermore, from stage 1 to 2, endogenous PA levels and the soluble conjugated and insoluble bound fractions of PAs and key enzymes (e.g., diamine oxidase, arginine decarboxylase, ornithine decarboxylase and S-adenosylmethionine decarboxylase) increased, and the level of PA oxidase decreased. These findings revealed that exogenous GA3 and Spd delay flower senescence by improving PA biosynthesis and preventing PA degradation. Moreover, exogenous GA3 and Spd enhanced the levels of endogenous PA and GA3, while the conversion of free PAs to soluble conjugated and insoluble bound forms delayed Rhododendron senescence. Overall, our findings reveal a potential positive feedback mechanism by which higher endogenous PA contents and the combined effects of exogenous GA3 and Spd synergistically delay Rhododendron senescence by enhancing PA biosynthesis and converting free PA to soluble conjugated and insoluble bound forms, thus reducing PA degradation during flower senescence.

Spermidine Pretreatments Mitigate the Effects of Saline Stress by Improving Growth and Saline Excretion in Frankenia pulverulenta

Climatic change, ecosystem imbalance, and soil salinization and desertification are serious obstacles to the restoration of degraded landscapes. Halophyte cultivation could constitute a way of mitigating these problems. Frankenia pulverulenta is used in the remediation and revegetation of areas affected by salinity and as an ornamental plant on saline soils since it can tolerate salt concentrations up to 200 mM NaCl. To increase saline tolerance, different plant growth regulators (auxins, cytokinins, gibberellins, spermidine, and salicylic acid) were tested in hydroponic conditions before the addition of NaCl (300 mM). At 52 days, growth, proline, saline excretion, free polyamines, and ethylene were determined under both saline and non-saline conditions. All growth regulators improved growth in the salt-free pretreatment; however, under conditions of salinity, pretreatment with spermidine (0.5 mM) and then salicylic acid (0.5 mM) were efficient at reversing the inhibitory effect of salt and improved saline excretion in F. pulverulenta. A strong positive correlation of polyamines and proline, and a negative correlation with ethylene, indicate that pretreatments that increase endogenous polyamine content and osmolytes are the most effective in improving salt tolerance of F. pulverulenta and could be used in the technical guidance of the cultivation of this halophyte.

Stem canker caused by Phomopsis spp. Induces changes in polyamine levels and chlorophyll fluorescence parameters in pecan leaves

Pecan plants are attacked by the fungus Phomopsis spp. that causes stem canker, a serious and emerging disease in commercial orchards. Stem canker, which has been reported in several countries, negatively affects tree canopy health, eventually leading to production losses. The purpose of this study was to inquire into the physiology of pecan plants under stem canker attack by Phomopsis spp. To this end, pecan plants were inoculated with an isolate of Phomopsis spp. and several parameters, such as polyamines, proline, sugars, starch, chlorophyll fluorescence and canopy temperature were analysed. Under artificial inoculation, a high disease incidence was observed with symptoms similar to those in plants showing stem canker under field conditions. Furthermore, the infected stem showed dead tissue with brown necrotic discolouration in the xylem tissue. The free polyamines putrescine, spermidine, and spermine were detected and their levels decreased as leaves aged in the infected plants with respect to the controls. Chlorophyll fluorescence parameters, such as Sm, ψEO, and QbRC decreased under plant infection and therefore the K-band increased. Canopy temperature and proline content increased in the infected plants with respect to the controls while sugar content decreased. These data suggest that stem canker caused by Phomopsis spp. induces physiological changes that are similar to those observed in plants under drought stress. To our knowledge, this is the first study that documents the physiological and biochemical effects derived from pecan-Phomopsis interaction.

Exogenous melatonin alleviated chilling injury in harvested plum fruit via affecting the levels of polyamines conjugated to plasma membrane

Polyamines (PAs) are related closely with many abiotic stresses. However, conjugated PA functions are not clear in alleviating chilling injury in fruit. The contents of free PAs in plum flesh cell and conjugated PAs in plasma membrane (PM), the activities of S-adenosylmethionine decarboxylase (SAMDC) and transglutaminase (TGase), and the degree of chilling injury were investigated in harvested plum (Prunus salicina L.) fruit pre-treated with exogenous melatonin. Chilling treatment resulted in the injury of plasma membrane (PM), as judged by the increased PM permeability, chilling injury index and malondialdehyde (MDA) content, and the decreased sulfhydryl group contents. Exogenous melatonin alleviated chilling injury, in parallel with the increased levels of free spermidine (Spd) and spermine (Spm) in cell, conjugated non-covalently (CNC) Spd and Spm, and conjugated covalently (CC) Put and Spd in PM, resulting from the increased activities of SAMDC and TGase. The results of the additional experiments with two inhibitors, methylglyoxyl-bis (guanylhydrazone) (MGBG) and phenanthrolin, which inhibit the activities of SAMDC and TGase, respectively, were complementary evidences for our research. MGBG and TGase treatment decreased the level of CNC PAs (Spd and Spm) and CC PAs (Put and Spd), respectively, coupled with aggravating the chilling injury. From these above mentioned-results, it could be suggested that exogenous melatonin alleviated chilling injury by elevating the levels of CNC Spd, CNC Spm, CC Put and CC Spd in PM.

Polyamine oxidase (PAO)–mediated polyamine catabolism plays potential roles in peach (Prunus persica L.) fruit development and ripening

Recently, there is an increased interest in the function of polyamine (PA) catabolism during fruit ripening, but little is known about its role during the ripening of peach (Prunus persica L.). In this study, the contents of both free and conjugated PAs markedly decreased as the peach fruit ripened. However, RNA-seq analysis showed that the transcript levels of PA synthesis–related genes increased, suggesting that the decreasing PA content during peach ripening was associated with PA catabolism. To better understanding the function of PA catabolism in peach ripening, genes encoding potential polyamine oxidases (PAOs), which are involved in PA catabolism, were identified across the peach genome. Four putative PAO genes (PpePAO1-PpePAO4) were identified in peach using the recently released genome database. The mRNA level of PpePAO1 was significantly increased during peach fruit ripening. Detection of free PAs in tobacco leaves transiently over-expressing PpePAO1 suggested that this gene is probably involved in terminal catabolism of PA. Treatment with the PAO inhibitor guazatine significantly reduced ethylene production and flesh softening of peach fruit, decreased the expression levels of fruit ripening-related genes, and significantly decreased the expression level of PpePAO1 and PAO activity, while PA contents were dramatically higher compared to control treatments. Our results suggests that PpePAO1-mediated PA catabolism is associated with decreased PA content during peach fruit ripening. These data provide valuable knowledge for better understanding the roles of PA catabolism in peach development and ripening.

Involvement of Polyamine Metabolism in the Response of Medicago truncatula Genotypes to Salt Stress.

Salinity constitutes one of the most important causes leading to severe reduction in plant yield. Several reports correlate the accumulation of polyamines in plants with tolerance to abiotic stress cues. The present study examined three Medicago truncatula genotypes with differing sensitivities to salinity (TN1.11, tolerant; Jemalong A17, moderately sensitive; TN6.18, sensitive), with the aim of examining the genotype-specific involvement of the polyamine metabolic pathway in plant response to salinity. The study was carried out with leaves harvested 48 h after watering plants with 200 mM NaCl. A comprehensive profile of free polyamines was determined using high performance liquid chromatography. All genotypes showed spermidine and spermine as the most abundant polyamines under control conditions. In salinity conditions, spermine levels increased at the expense of putrescine and spermidine, indicating a drift of polyamine metabolism towards the synthesis of increasing polycationic forms as a stress response. The increasing balance between high and low polycationic forms was clearly diminished in the salt-sensitive genotype TN6.18, showing a clear correlation with its sensitive phenotype. The polyamine metabolic profile was then supported by molecular evidence through the examination of polyamine metabolism transcript levels by RT-qPCR. General suppression of genes that are involved upstream in the PA biosynthetic pathway was determined. Contrarily, an induction in the expression of genes involved in the biosynthesis of spermine and spermidine was observed, in agreement with the metabolic analysis. A significant induction in diamino oxidase expression, involved in the catabolism of putrescine, was specifically found in the sensitive genotype ΤΝ6.18, indicating a distinct metabolic response to stress. Present findings highlight the involvement of polyamines in the defense response of Medicago genotypes showing sensitivity to salt stress.

Molecular Mechanisms of the 1-Aminocyclopropane-1-Carboxylic Acid (ACC) Deaminase Producing Trichoderma asperellum MAP1 in Enhancing Wheat Tolerance to Waterlogging Stress.

Waterlogging stress (WS) induces ethylene (ET) and polyamine (spermine, putrescine, and spermidine) production in plants, but their reprogramming is a decisive element for determining the fate of the plant upon waterlogging-induced stress. WS can be challenged by exploring symbiotic microbes that improve the plant’s ability to grow better and resist WS. The present study deals with identification and application of 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase-producing fungal endophyte Trichoderma asperellum (strain MAP1), isolated from the roots of Canna indica L., on wheat growth under WS. MAP1 positively affected wheat growth by secreting phytohormones/secondary metabolites, strengthening the plant’s antioxidant system and influencing the physiology through polyamine production and modulating gene expression. MAP1 inoculation promoted yield in comparison to non-endophyte inoculated waterlogged seedlings. Exogenously applied ethephon (ET synthesis inducer) and 1-aminocyclopropane carboxylic acid (ACC; ET precursor) showed a reduction in growth, compared to MAP1-inoculated waterlogged seedlings, while amino-oxyacetic acid (AOA; ET inhibitor) application reversed the negative effect imposed by ET and ACC, upon waterlogging treatment. A significant reduction in plant growth rate, chlorophyll content, and stomatal conductance was noticed, while H2O2, MDA production, and electrolyte leakage were increased in non-inoculated waterlogged seedlings. Moreover, in comparison to non-inoculated waterlogged wheat seedlings, MAP1-inoculated waterlogged wheat exhibited antioxidant–enzyme activities. In agreement with the physiological results, genes associated with the free polyamine (PA) biosynthesis were highly induced and PA content was abundant in MAP1-inoculated seedlings. Furthermore, ET biosynthesis/signaling gene expression was reduced upon MAP1 inoculation under WS. Briefly, MAP1 mitigated the adverse effect of WS in wheat, by reprogramming the PAs and ET biosynthesis, which leads to optimal stomatal conductance, increased photosynthesis, and membrane stability as well as reduced ET-induced leaf senescence.

Shoot Girdling Improves Rooting Performance of Kalamata Olive Cuttings by Upregulating Carbohydrates, Polyamines and Phenolic Compounds

Girdling (a ring of bark approximately 5 mm wide) was applied on olive mother plants to investigate its effect on the rooting ability of cuttings. Treatment was applied in autumn and in spring. The cuttings were then immersed for 5 s into 2000 mg L−1 indole-3-butyric acid (IBA) in a 45% v/v ethanolic solution. Thirty days after girdling, cuttings were taken from girdled and control shoots from the part just above the girdle zone (basal), as well as from the part right above (middle). The base of the cuttings was collected, and the concentration of polyamines, phenolic compounds and soluble carbohydrates was determined. Girdling enhanced the rooting performance of the cuttings, while autumn proved to be a better season for rooting compared to spring. Girdling positively affected the concentration of all the measured parameters. The main characteristics of autumn cuttings were the high levels of glucose, mannitol, free and total polyamines, hydroxytyrosol, tyrosol, verbascoside, oleuropein, quercetin and luteolin. The cuttings taken from the middle part of the girdled shoot exhibited high sucrose, glucose, mannitol, free polyamines, hydroxytyrosol, luteolin-7-glucoside, total phenols and flavanol concentrations. Nonetheless, further research is needed in order to draw conclusions on the overall efficiency of girdling on inducing rooting of olive cuttings.

Effects of alkali stress on dry matter accumulation, root morphology, ion balance, free polyamines, and organic acids of sugar beet

Sugar beet is strongly resistant to salt-alkalinity. Understanding the physiological alkali stress resistance mechanism of sugar beet is important for fully utilizing saline–alkaline soil. Sugar beet seedlings from cultivars KWS0143 (alkali-tolerant) and Beta464 (alkali-sensitive) were treated with five concentrations of mixed alkaline solutions (NaHCO3: Na2CO3, 2:1), namely, 0 (control), 25, 50, 75, and 100 mM (mole concentration was calculated in Na+). A sharper decrease in dry weight per plant (87.1%) and total root length (91.7%) of Beta464 were observed compared to the 61.5% and 85.0% decrease in those of KWS0143 under 100 mM alkali treatment. With increasing alkaline stress, Na+ accumulation hindered K+ and Ca2+ absorption by roots. Free polyamines contents and phosphoenolpyruvate carboxylase (PEPC) activity in roots of both cultivars were all significantly enhanced by 50 and 75 mM alkali treatments. KWS0143 exhibited higher dissociated putrescine (Put), spermine (Spm), as well as spermidine (Spd) levels within the roots compared to Beta464 under alkali conditions. Root free Spd contents of KWS0143 and Beta464 increased by 154.2 and 64.5% treated with 50 mM alkali in comparison with the control. After treated with the dose of 25 mM, root succinic acid (SA) contents of KWS0143 and Beta464 increased by 90.4 and 14.3%, respectively, compared to the plants subjected to the control. Our results imply that polyamines and PEPC contribute to the tolerance of sugar beet to alkali stress. Those results could be useful for enriching the theory of plant stress response.

Ethylene is a local modulator of jasmonate-dependent phenolamide accumulation during Manduca sexta herbivory in Nicotiana attenuata.

Rapid reconfigurations of interconnected phytohormone signalling networks allow plants to tune their physiology to constantly varying ecological conditions. During insect herbivory, most of the induced changes in defence-related leaf metabolites are controlled by jasmonate (JA) signalling, which, in the wild tobacco Nicotiana attenuata, recruits MYB8, a transcription factor controlling the accumulation of phenolic-polyamine conjugates (phenolamides). In this and other plant species, herbivory also locally triggers ethylene (ET) production but the outcome of the JA-ET cross-talk at the level of secondary metabolism regulation has remained only superficially investigated. Here, we analysed local and systemic herbivory-induced changes by mass spectrometry-based metabolomics in leaves of transgenic plants impaired in JA, ET and MYB8 signalling. Parsing deregulations in this factorial data-set identified a network of JA/MYB8-dependent phenolamides for which impairment of ET signalling attenuated their accumulation only in locally damaged leaves. Further experiments revealed that ET, albeit biochemically interrelated to polyamine metabolism via the intermediate S-adenosylmethionine, does not alter the free polyamine levels, but instead significantly modulates phenolamide levels with marginal modulations of transcript levels. The work identifies ET as a local modulator of phenolamide accumulations and provides a metabolomics data-platform with which to mine associations among herbivory-induced signalling and specialized metabolites in N. attenuata.

Enhancement of γ-aminobutyric acid (GABA) and other health-promoting metabolites in germinated broccoli by mannose treatment

The contents of anthocyanins, total glucosinolates, sulforaphane, and the metabolism of γ-aminobutyric acid (GABA) in broccoli sprouts grown under 1.0 mmol L−1 of mannose were investigated. The results showed that mannose treatment improved glutamate (Glu) content and the activity of diamine oxidase (DAO), and reduced γ-aminobutyric acid transaminase (GABA-T) activity, which enriched GABA biosynthesis. Mannose treatment enhanced the activities of diamine oxidase (DAO), polyamine oxidase (PAO), and 4-aminobutyraldehyde dehydrogenase (ABALDH), and increased the conversion rate of free polyamine. At the same time, it is also beneficial for the enrichment of anthocyanins, total glucosinolates and sulforaphane. These results indicated that mannose treatment has a significant enhancement effect on GABA and other health-promoting metabolites in germinated broccoli.

Changes in polyamine pattern mediates sex differentiation and unisexual flower development in monoecious cucumber (Cucumis sativus L.)

Changes in the levels of polyamines are associated with fundamental physiological processes such as embryogenesis, induction of flowering, fruit development and ripening, senescence, and responses to environmental stresses, but the role of polyamines in sex differentiation and unisexual flower development has not been deeply studied. To extend the knowledge on the regulatory mechanisms of flowering in monoecious plant (producing unisexual flowers), we investigated the morphogenesis and free polyamine levels in Cucumis sativus during sex differentiation and unisexual flower development in vitro using histocytological and biochemical methods. As shown in our study, floral development in vitro was undisturbed and flowers of both sexes were produced. Sex differentiation relied on preventing the development of generative organs of the opposite sex, as we observed carpel repression in male flowers and stamen repression in female flowers. Pollen viability was negatively correlated with female flower development on the same node. Biochemical analysis revealed increased accumulation of aliphatic amines (tri, tetra-amines) in generative (flower buds and flowers) compare to vegetative (axillary buds and leaves) organs. Undifferentiated floral buds contained elevated levels of agmatine, cadaverine, spermidine and spermine. Sex differentiation was associated with significantly decreased levels of agmatine and cadaverine. Our results showed that female flowers contained higher levels of total polyamine than male flowers. The increased level of cadaverine was associated with macrogametogenesis and female flower maturation. Putrescine was important for male flower development. Such results support the hypothesis that aliphatic amines are involved in unisexual flower development.

Oxidative stress due to high solar radiation increases free polyamines content in apple skin

Oxidative stress due to high solar radiation increases free polyamines content in apple skin

TGase regulates salt stress tolerance through enhancing bound polyamines-mediated antioxidant enzymes activity in tomato

The accumulation of free polyamines (PAs) plays critical role in plants under abiotic stress conditions, while protective traits of bound PAs receiving much less attention, our understanding of the regulation of free PAs converted to bound PAs is limited. In this study, we demonstrated that the PA mediator transglutaminase (TGase) play an important role in improving tomato tolerance to salinity stress, in part because its critical role in inducing accumulation of bound PAs under salt stress. TGase expression was induced upon salt stress. Exogenous phenanthroline (O-phen, a TGase and bound PAs synthesis inhibitor) abolished the TGase-mediated salt tolerance with terminating bound PAs accumulation in TGaseOE plants. O-phen also had a negatively altered on Na+/K+ uptake or homeostasis under salt stress in leaves of TGaseOE plants. Instead, the overexpression of TGase effectively alleviated salt-induced oxidative damage and reduced reactive oxygen species (ROS) over-production, but it was significantly aggravated by O-phen in the TGaseOE plants. Although the ROS generating enzymes (PA oxidase, diamine oxidase and NADPH oxidase) was not affected by application of O-phen, the antioxidant enzyme (superoxide dismutase, catalase, ascorbate peroxidase and monodehydroascorbate reductase) activity was significantly decreased by exogenous O-phen in plants. Furthermore, further analysis showed that the antioxidant enzyme activity positively and closely correlated with the level of bound PAs in TGaseOE plant. Together, these results uncovered a probably function mechanism of TGase-mediated antioxidant enzyme activity to reduced salt-induced oxidative damage, which is partially depended on bound PAs accumulation.

CsICE1 Functions in Cold Tolerance by Regulating Polyamine levels May through Interacting with Arginine Decarboxylase in the Tea Tree

Background: The identification of C-repeat binding factor (CBF), and the characterization as an inducer of CBF Expression 1 (ICE1), and a major activator for C-repeat binding factor, were important breakthroughs in the cold signaling network. Methods: In the present study, the full length cDNA of ICE1 was isolated from the tea tree (Camellia sinensis). CsICE1 protein was located in the cell nucleus as revealed by subcellular localization analysis. To investigate the biological functions of CsICE1, a transgenic line fused with the CsICE1 gene in Arabidopsis thaliana (arabidopsis) was generated by the floral dip method. Results: The CsICE1 was expressed differentially in various tea tree tissues, mostly in buds and leaves, and the transcript level of CsICE1 was increased after 1 h and peaked at 2 h under cold treatment. Transcription activity assay indicated that the spermine synthase (SPMS) and arginine decarboxylase (ADC) genes were possible targets of CsICE1. In addition, the values of net photosynthetic rate, transpiration rate, stomatal conductance in transgenic lines declined by less extent than wild-type plants under low temperatures. Furthermore, transcript levels of ADC genes in the transgenic lines had no apparent alteration under normal growth conditions but substantially increased under cold conditions, consistent of changes in free polyamine levels. Taken together, these results demonstrated that CsICE1 plays a positive role in cold tolerance, which may be due to the modulation of polyamine levels through interacting with CsADC.