Effect of drought and combined drought and heat stress on polyamine metabolism in proline-over-producing tobacco plants.

The roles of proline and polyamines (PAs) in the drought stress responses of tobacco plants were investigated by comparing the responses to drought alone and drought in combination with heat in the upper and lower leaves and roots of wild-type tobacco plants and transformants that constitutively over-express a modified gene for the proline biosynthetic enzyme Δ1-pyrroline-5-carboxylate synthetase (P5CSF129A; EC 2.7.2.11/1.2.1.41). In both genotypes, drought stress coincided with a decrease in relative water content (RWC) that was much less severe in the upper leaves than elsewhere in the plant. The drought also increased proline levels in both genotypes. A brief period of heat stress (2 h at 40 °C) at the end of the drought period did not significantly influence the proline levels in the upper leaves and roots but caused a further increase in the lower leaves of both genotypes. The rate at which these elevated proline levels returned to normal during the post-stress recovery period was slower in the transformants and plants that had been subjected to the combined stress. In both genotypes, drought stress significantly reduced the levels of spermidine (Spd) and putrescine (Put) in the leaves and roots relative to those for controls, and increased the levels of spermine (Spm) and diaminopropane (Dap, formed by the oxidative deamination of Spd and Spm). Spd levels may have declined due to its consumption in Spm biosynthesis and/or oxidation by polyamine oxidase (PAO; EC 1.5.3.11) to form Dap, which became more abundant during drought stress. During the rewatering period, the plants' Put and Spd levels recovered quickly and the activity of the PA biosynthesis enzymes in their leaves and roots increased substantially; this increase was more pronounced in transformants than WT plants. The high levels of Spm observed in drought stressed plants persisted even after the 24 h recovery and rewatering phase. The malondialdehyde (MDA) contents of the lower leaves of WTs increased substantially during the drought stress period; a less pronounced increase occurred in the transformants and after the application of the combined stress. After the post-stress recovery period, the MDA contents in the leaves of both genotypes were higher than those in the corresponding controls. The MDA contents of the upper leaves in plants of both genotypes remained relatively constant throughout, indicating that these leaves are preferentially protected against the adverse effects of oxidative stress and demonstrating the efficiency of the plants' induced antioxidative defense mechanisms.

Cvikrová M ，Gemperlová L ，Martincová O ，Vanková R ... -  《-》

Effect of heat stress on polyamine metabolism in proline-over-producing tobacco plants.

The effect of heat stress on the accumulation of proline and on the level of polyamines (PAs) in tobacco plants was investigated. Responses to heat stress were compared in the upper and lower leaves and roots of tobacco plants that constitutively over-express a modified gene for the proline biosynthetic enzyme Δ1-pyrroline-5-carboxylate synthetase (P5CSF129A) and in the corresponding wild-type. In the initial phases of heat stress (after 2h at 40°C), the accumulation of proline increased in the wild type but slightly decreased in the transformants. The response to heat stress in proline-over-producing tobacco plants involved a transient increase in the levels of free and conjugated putrescine (Put) and in the levels of free spermidine (Spd), norspermidine (N-Spd) and spermine (Spm) after a 2-h lag phase, which correlated with stimulation of the activity of the corresponding biosynthetic enzymes. Diamine oxidase (DAO) activity increased in both plant genotypes, most significantly in the leaves of WT plants. Polyamine oxidase (PAO) activity increased in the roots of WT plants and decreased in the leaves and roots of the transformants. After 6h of heat stress, proline accumulation was observed in the transformants, especially in the lower leaves; much more modest increase was observed in the WT plants. A decrease in the levels of free and conjugated Put coincided with down-regulation of the activity of ornithine decarboxylase and marked stimulation of DAO activity in the leaves and roots of the transformants. PAO activity increased in the roots of the transformants but decreased in the leaves. Conversely, in WT tobacco subjected to 6h of heat stress, slight increases in free and conjugated PA levels were observed and the activity of DAO only increased in the roots; PAO activity did not change from the value observed during the initial phase of heat stress. 6 Hours' heat stress had no effect on the level of malondialdehyde (MDA; a product of lipid peroxidation), in the upper leaves of either genotype. After a recovery period (2h at 25°C), most of the studied parameters exhibited values comparable to those observed in untreated plants. The coordination of the proline and polyamine biosynthetic pathways during heat stress conditions is discussed.

Cvikrová M ，Gemperlová L ，Dobrá J ，Martincová O ，Prásil IT ，Gubis J ，Vanková R ... -  《-》

Enriched endogenous free Spd and Spm in alfalfa (Medicago sativa L.) under drought stress enhance drought tolerance by inhibiting H(2)O(2) production to increase antioxidant enzyme activity.

Drought stress is a major factor limiting agricultural development, and exogenous polyamines (PAs) can increase plant drought resistance by enhancing antioxidant activity, but few studies have examined whether endogenous PAs enhance the plant antioxidant system. Here, to investigate the effects of endogenous PAs on the antioxidant system of alfalfa under drought stress and the underlying mechanisms, two alfalfa cultivars, Longzhong (drought resistant) and Gannong No. 3 (drought sensitive), were used as test materials, and their seedlings were treated with polyethylene glycol (PEG-6000) for 8 days at -1.2 MPa to simulate drought stress. The levels of free PAs [putrescine (Put), spermidine (Spd) and spermine (Spm)], hydrogen peroxide (H2O2), malondialdehyde (MDA), key PA metabolism enzyme [arginine decarboxylase (ADC), ornithine decarboxylase (ODC), S-adenosylmethionine decarboxylase (SAMDC), polyamine oxidase (PAO), and diamine oxidase (DAO)] activities, and antioxidant enzyme [superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD)] activities were measured. These physiological indicators were used for correlation analysis to investigate the relationship between PA metabolism and the antioxidant enzyme system. The results showed that PA synthesis in alfalfa under drought stress was dominated by the ADC pathway. Spd and Spm played an important role in improving drought tolerance. The high levels of ADC and SAMDC activities were facilitated by the conversion of Put to Spd and Spm. H2O2 generation by oxidative decomposition of PAs was mainly dependent on the oxidative decomposition of DAO but not PAO. Low DAO activity favored low H2O2 production. Spd, Spm, ADC, ODC and SAMDC were positively correlated with the antioxidant enzymes SOD, CAT and POD in both cultivars under drought. Therefore, we concluded that high ADC and SAMDC activities in alfalfa promoted the conversion of Put to Spd and Spm, leading to high accumulation of Spd and Spm and low Put accumulation. Low Put levels led to low H2O2 production through low DAO activity, and low H2O2 levels induced the expression of antioxidant enzyme-encoding genes to improve antioxidant enzyme activity and reduce MDA accumulation and thereby enhanced drought resistance in alfalfa.

Wang W ，Shi S ，Kang W ，He L ... -  《-》

Application of γ-aminobutyric acid demonstrates a protective role of polyamine and GABA metabolism in muskmelon seedlings under Ca(NO3)2 stress.

The effects of exogenous γ-aminobutyric acid (GABA) application on growth, polyamine and endogenous GABA metabolism in muskmelon leaves and roots were measured. Plants were treated under control or 80 mM Ca(NO3)2 stress conditions with or without foliar spraying 50 mM GABA. Ca(NO3)2 stress significantly suppressed seedling growth and GABA transaminase activity, and enhanced glutamate decarboxylase (GAD) activity and endogenous GABA levels. Polyamine (PA) biosynthesis and degradation capacity increased in parallel with increasing GAD activity. Exogenous GABA application effectively alleviated the growth inhibition caused by Ca(NO3)2 stress, and significantly enhanced the activities of arginine decarboxylase (ADC), ornithine decarboxylase (ODC), S-adenosylmethionine decarboxylase (SAMDC), polyamine oxidase (PAO), and diamine oxidase (DAO). Exogenous GABA also significantly reduced the accumulation of free putrescine (Put) and increased the levels of free spermidine (Spd) and spermine (Spm) in leaves, which improved the capacity for polyamine biosynthesis. Application of exogenous GABA under Ca(NO3)2 stress enables the plants to maintain a higher ratio of free Spd and free Spm with respect to free Put. Our data suggest that exogenous GABA has an important role in improving muskmelon seedling tolerance to Ca(NO3)2 stress by improving biosynthesis of PAs and GABA, and by preventing PA degradation. There is a potential positive feedback mechanism that results from higher endogenous GABA content and the combined effects of Ca(NO3)2 stress and exogenous GABA, which coordinately alleviate Ca(NO3)2 stress injury by enhancing PA biosynthesis and converting free Put to an insoluble bound PA form, and reduce PA degradation in muskmelon seedlings.

Hu X ，Xu Z ，Xu W ，Li J ，Zhao N ，Zhou Y ... -  《-》

Spermidine alleviates the growth of saline-stressed ginseng seedlings through antioxidative defense system.

Protective effects of exogenous spermidine (Spd), activity of antioxygenic enzymes, and levels of free radicals in a well-known medicinal plant, Panax ginseng was examined. Seedlings grown in salinized nutrient solution (150 mM NaCl) for 7d exhibited reduced relative water content, plant growth, increased free radicals, and showing elevated lipid peroxidation. Application of Spd (0.01, 0.1, and 1mM) to the salinized nutrient solution showed increased plant growth by preventing chlorophyll degradation and increasing PA levels, as well as antioxidant enzymes such as CAT, APX, and GPX activity in the seedlings of ginseng. During salinity stress, Spd was effective for lowering the accumulation of putrescine (Put), with a significant increase in the spermidine (Spd) and spermine (Spm) levels in the ginseng seedlings. A decline in the Put level ran parallel to the higher accumulation of proline (Pro), and exogenous Spd also resulted in the alleviation of Pro content under salinity. Hydrogen peroxide (H2O2) and superoxide (O2(-)) production rates were also reduced in stressed plants after Spd treatment. Furthermore, the combined effect of Spd and salt led to a significant increase in diamine oxidase (DAO), and subsequent decline in polyamine oxidase (PAO). These positive effects were observed in 0.1 and 1mM Spd concentrations, but a lower concentration (0.01 mM) had a very limited effect. In summary, application of exogenous Spd could enhance salt tolerance of P. ginseng by enhancing the activities of enzyme scavenging system, which influence the intensity of oxidative stress.

Parvin S ，Lee OR ，Sathiyaraj G ，Khorolragchaa A ，Kim YJ ，Yang DC ... -  《-》