Short-term Low Temperature Stress Research Articles

Hepatic Metabolomic Responses to Low-Temperature Stress in the Invasive Turtle, Trachemys scripta elegans.

Investigating the physiological and biochemical changes of ectothermic species before entering hibernation would contribute to the understanding of how they adapt to low-temperature environments. Here, red-eared slider turtle (Trachemys scripta elegans) hatchlings were maintained under different thermal treatments (24 °C, slowly decreasing temperatures from 24 °C to 14 °C, and to 4 °C). Hepatic metabolite alterations were measured to assess the metabolic impacts of low-temperature stress in this species. Of these differentially changed metabolites, some (e.g., raffinose, spermidine, allocholic acid, taurohyocholate, 2-ketobutyric acid, acetylcysteine) were shown to decrease, while others (e.g., stearolic acid, D-mannose) increased in low-temperature treatments. Our results indicated that short-term low-temperature stress might have limited impacts on lipid and energy metabolism in this species. The changes in other metabolites (e.g., allocholic acid, taurohyocholate, spermine, acetylcysteine) might be associated with a low food intake (and thus reduced digestive performance) and weakened immune ability of low-temperature-exposed animals.

Resilience mechanisms of Trichopria drosophilae (Hymenoptera: Diapriinae) under global extreme cooling: insights into parasitic response and physiological adaptation.

Global climate warming and frequent extreme low-temperature events have made it essential to investigate the impact of low temperatures on parasitic wasps to protect and strengthen farmland biodiversity, which in turn enhances the biological control potential of natural enemies such as parasitic wasps. We systematically examined how low-temperature stress affects the parasitic functional response of Trichopria drosophilae to Drosophila suzukii (Diptera:Drosophilidae)pupae. Our findings indicate that the parasitic behavior of T. drosophilae towards D. suzukii pupae aligns with the Holling II functional response model following exposure to different temperatures. Within the temperature range of 8 °C to -8 °C, lower temperatures correlated decreased instantaneous attack rate of T. drosophilae and an increase in processing time. The search constant Q initially increased and then decreased with declining temperatures. Short-term low-temperature stress negatively impacted the parasitic and searching abilities of T. drosophilae but did not alter its parasitic functional response model. Notably, short-term low-temperature stress had minimal effects on the water content, protein content, and total sugar content of male and female T. drosophilae adults. However, as temperatures decreased, the activities of key enzymes, including GAPDH, SOD, T-AOC, and malondialdehyde (MDA), exhibited an initial increase followed by a decrease. Conversely, the activities of LDH and HOAD decreased, while the activities of CAT and POD increased. Further study on the effect of short-term low temperature on T. drosophilae can provide a research basis for the large-scale production and low-temperature refrigeration technology of T. drosophilae, and provide a scientific basis for its efficient use in the field.

Rapid cold hardening response of the phytoseiid mite Neoseiulus striatus: increased cold tolerance but not reduced predation

The predatory mite Neoseiulus striatus (Wu) (Acari: Phytoseiidae), which has been found on maize plants in Inner Mongolia, is regarded as a promising biological control agent of small sucking pests, especially Tetranychid mites. Temperature is an important abiotic factor, and a sudden drop in temperature may affect its performance when released in areas with big circadian temperature differences. Rapid cold hardening is a type of phenotypic plasticity that allows ectotherms to quickly enhance their cold tolerance in response to a brief chilling exposure. However, it is not clear whether N. striatus possesses such plasticity. To understand how this species of phytoseiid mite copes with short-term low-temperature stress, its rapid cold hardening response was studied in the laboratory by first exploring its critical temperature. Then, the effects of exposure to a series of temperatures (0, 5, 10, 15, and 20°C) for different durations on the survival of N. striatus were investigated to determine the optimal temperature and duration for cold hardening. Moreover, the effect of cold hardening on the consumption of Tetranychus urticae by N. striatus was also investigated, as was the response of immature stages. The critical temperature for N. striatus was - 15°C, at which its survival dropped below 10%. An exposure to 5°C for 2h was optimal for rapid cold hardening, efficiently increasing survival in N. striatus. Rapid cold hardening was found in all developmental stages of N. striatus, and there was no significant effect of exposure on the consumption of spider mites. Our study shows that exposure to low temperatures during a limited period can enhance the cold hardiness but not reduce predation by N. striatus, which will be advantageous to its field applications during a period of large diurnal temperature fluctuations.

Ultrastructural changes in developmental stages of anther and pollen grains as affected by short-term exposure to low temperatures in strawberry

Short-term exposure to low temperatures can lower pollen viability and germination in strawberry. Fruit malformation was found to be negatively correlated with pollen germination. Ultrastructural analysis was performed at various developmental stages of anthers and pollen grains of the strawberry upon exposure to short-term low temperature (STLT). ‘Maehyang’ strawberry was exposed to 2 oC from 6 pm to 9 am for 3 days during different flower developments before anthesis, whereas the plants in control remained in the greenhouse. The epidermis did not develop well and was deformed after STLT. The epidermis showed a significantly lower cell expansion index in the short-term low-temperature treatment than in control at the vacuolated microspore, bicellular pollen, and mature pollen stages, indicating significant deformation of the epidermal cells by low temperature. The projected area of starch was significantly greater in STLT treatment than in control at the mature pollen stage. The relative fluorescence intensity of anthers was significantly higher in STLT treatment than in control at the sporogenous cell and microspore mother cell stages, indicating that low temperature substantially inhibited anther development at early stages. The results showed that STLT stress caused abnormal development of the ultrastructure of the anther wall and pollen grains, along with starch over-accumulation during pollen development. These abnormal developments may lead to poor pollen germination and thus cause fruit malformation in strawberry.

Simulating the effects of low-temperature stress on wheat biomass growth and yield

Low-temperature stress in late spring poses a serious threat to winter wheat production. In three-year environment-controlled phytotron experiments at both elongation and booting stages, we observed that short-term low-temperature stress decreased leaf area and stem, limited leaf photosynthetic system, and severely reduced grain yield, especially in the low-temperature sensitive cultivar. To better quantify the effects of low-temperature stress on leaf photosynthesis rate (Pn), leaf area index (LAI), biomass partitioning, and grain yield, the lethal temperature at 50% mortality (LT50), low-temperature damage index (LDI), and process-based algorithms were proposed and incorporated into the WheatGrow model. The algorithms take into account the effects of low-temperature with different durations and reflected a low-temperature sensitivity of cultivars at different growth periods by using LT50. The improved WheatGrow model could reproduce well the observed dynamics of the decrease of Pn during the treatment period, recovery of Pn after cold treatments, and the damage to LAI and biomass accumulation under the low-temperature stress treatments. Compared to the original model, RMSE of Pn, LAI, total aboveground biomass, and grain yield in the improved WheatGrow model were reduced by 63 to 77% under low-temperature stress. Although the newly developed algorithms expand crop modeling capacity into quantifying the impacts of low-temperature stress on wheat productivity, further assessments with observed data under field conditions are still required, especially for obtaining suitable parameters at the regional scale.

Optimized Phosphorus Application Alleviated Adverse Effects of Short-Term Low-Temperature Stress in Winter Wheat by Enhancing Photosynthesis and Improved Accumulation and Partitioning of Dry Matter

Low-temperature stress has become an important abiotic factor affecting high and stable wheat production. Therefore, it is necessary to take appropriate measures to enhance low-temperature tolerance in wheat. A pot experiment was carried out using Yannong19 (YN19, a cold-tolerant cultivar) and Xinmai26 (XM26, a cold-sensitive cultivar). We employed traditional phosphorus application (TPA, i.e., R1) and optimized phosphorus application (OPA, i.e., R2) methods. Plants undertook chilling (T1 at 4 °C) and freezing treatment (T2 at −4 °C) as well as ambient temperature (CK at 11 °C) during the anther differentiation period to investigate the effects of OPA and TPA on photosynthetic parameters and the accumulation and distribution of dry matter. The net photosynthetic rate (Pn), stomatal conductance (Gs) and transpiration rate (Tr) of flag leaves decreased in low-temperature treatments, whereas intercellular carbon dioxide concentration (Ci) increased. Compared with R1CK, Pn in R1T1 and R1T2 treatments was reduced by 26.8% and 42.2% in YN19 and 34.2% and 54.7% in XM26, respectively. In contrast, it increased by 6.5%, 8.9% and 12.7% in YN19 and 7.7%, 15.6% and 22.6% in XM26 for R2CK, R2T1 and R2T2 treatments, respectively, under OPA compared with TPA at the same temperature treatments. Moreover, low-temperature stress reduced dry matter accumulation at the reproductive growth stage. OPA increased dry matter accumulation of vegetative organs after the flowering stage and promoted the transportation of assimilates to grains. Hence, the grain number per spike (GNPS), 1000-grain weight (TGW) and yield per plant (YPP) increased. The low-temperature treatments of T1 and T2 caused yield losses of 24.1~64.1%, and the yield increased by 8.6~20.5% under OPA treatments among the two wheat cultivars. In brief, OPA enhances low-temperature tolerance in wheat, effectively improves wheat architecture and photosynthesis, increases GNPS and TGW and ultimately lessens yield losses.

Comparative Transcriptome and Interaction Protein Analysis Reveals the Mechanism of IbMPK3-Overexpressing Transgenic Sweet Potato Response to Low-Temperature Stress.

The sweet potato is very sensitive to low temperature. Our previous study revealed that IbMPK3-overexpressing transgenic sweet potato (M3) plants showed stronger low-temperature stress tolerance than wild-type plants (WT). However, the mechanism of M3 plants in response to low-temperature stress is unclear. To further analyze how IbMPK3 mediates low-temperature stress in sweet potato, WT and M3 plants were exposed to low-temperature stress for 2 h and 12 h for RNA-seq analysis, whereas normal conditions were used as a control (CK). In total, 3436 and 8718 differentially expressed genes (DEGs) were identified in WT at 2 h (vs. CK) and 12 h (vs. CK) under low-temperature stress, respectively, whereas 1450 and 9291 DEGs were detected in M3 plants, respectively. Many common and unique DEGs were analyzed in WT and M3 plants. DEGs related to low temperature were involved in Ca2+ signaling, MAPK cascades, the reactive oxygen species (ROS) signaling pathway, hormone transduction pathway, encoding transcription factor families (bHLH, NAC, and WRKY), and downstream stress-related genes. Additionally, more upregulated genes were associated with the MAPK pathway in M3 plants during short-term low-temperature stress (CK vs. 2 h), and more upregulated genes were involved in secondary metabolic synthesis in M3 plants than in the WT during the long-time low-temperature stress treatment (CK vs. 12 h), such as fatty acid biosynthesis and elongation, glutathione metabolism, flavonoid biosynthesis, carotenoid biosynthesis, and zeatin biosynthesis. Moreover, the interaction proteins of IbMPK3 related to photosynthesis, or encoding CaM, NAC, and ribosomal proteins, were identified using yeast two-hybrid (Y2H). This study may provide a valuable resource for elucidating the sweet potato low-temperature stress resistance mechanism, as well as data to support molecular-assisted breeding with the IbMPK3 gene.

Gene-coexpression network analysis identifies specific modules and hub genes related to cold stress in rice

BackgroundWhen plants are subjected to cold stress, they undergo a series of molecular and physiological changes to protect themselves from injury. Indica cultivars can usually withstand only mild cold stress in a relatively short period. Hormone-mediated defence response plays an important role in cold stress. Weighted gene co-expression network analysis (WGCNA) is a very useful tool for studying the correlation between genes, identifying modules with high phenotype correlation, and identifying Hub genes in different modules. Many studies have elucidated the molecular mechanisms of cold tolerance in different plants, but little information about the recovery process after cold stress is available.ResultsTo understand the molecular mechanism of cold tolerance in rice, we performed comprehensive transcriptome analyses during cold treatment and recovery stage in two cultivars of near-isogenic lines (9311 and DC907). Twelve transcriptomes in two rice cultivars were determined. A total of 2509 new genes were predicted by fragment splicing and assembly, and 7506 differentially expressed genes were identified by pairwise comparison. A total of 26 modules were obtained by expression-network analysis, 12 of which were highly correlated with cold stress or recovery treatment. We further identified candidate Hub genes associated with specific modules and analysed their regulatory relationships based on coexpression data. Results showed that various plant-hormone regulatory genes acted together to protect plants from physiological damage under short-term low-temperature stress. We speculated that this may be common in rice. Under long-term cold stress, rice improved the tolerance to low-temperature stress by promoting autophagy, sugar synthesis, and metabolism.ConclusionThrough WGCNA analysis at the transcriptome level, we provided a potential regulatory mechanism for the cold stress and recovery of rice cultivars and identified candidate central genes. Our findings provided an important reference for the future cultivation of rice strains with good tolerance.

Effects of Soil Covering on Growth and Physio-Biochemical Indices of Sugar Beet Seedlings Under Short-Term Low-Temperature Stress

Effects of Soil Covering on Growth and Physio-Biochemical Indices of Sugar Beet Seedlings Under Short-Term Low-Temperature Stress

An R2R3-MYB Transcription Factor RmMYB108 Responds to Chilling Stress of Rosa multiflora and Conferred Cold Tolerance of Arabidopsis

A sudden cooling in the early spring or late autumn negatively impacts the plant growth and development. Although a number of studies have characterized the role of the transcription factors (TFs) of plant R2R3-myeloblastosis (R2R3-MYB) in response to biotic and abiotic stress, plant growth, and primary and specific metabolisms, much less is known about their role in Rosa multiflora under chilling stress. In the present study, RmMYB108, which encodes a nuclear-localized R2R3-MYB TF with a self-activation activity, was identified based on the earlier published RNA-seq data of R. multiflora plants exposed to short-term low-temperature stress and also on the results of prediction of the gene function referring Arabidopsis. The RmMYB108 gene was induced by stress due to chilling, salt, and drought and was expressed in higher levels in the roots than in the leaves. The heterologous expression of RmMYB108 in Arabidopsis thaliana significantly enhanced the tolerance of transgenic plants to freezing, water deficit, and high salinity, enabling higher survival and growth rates, earlier flowering and silique formation, and better seed quantity and quality compared with the wild-type (WT) plants. When exposed to a continuous low-temperature stress at 4°C, transgenic Arabidopsis lines–overexpressing RmMYB108 showed higher activities of superoxide dismutase and peroxidase, lower relative conductivity, and lower malondialdehyde content than the WT. Moreover, the initial fluorescence (Fo) and maximum photosynthetic efficiency of photosystem II (Fv/Fm) changed more dramatically in the WT than in transgenic plants. Furthermore, the expression levels of cold-related genes involved in the ICE1 (Inducer of CBF expression 1)-CBFs (C-repeat binding factors)-CORs (Cold regulated genes) cascade were higher in the overexpression lines than in the WT. These results suggest that RmMYB108 was positively involved in the tolerance responses when R. multiflora was exposed to challenges against cold, freeze, salt, or drought and improved the cold tolerance of transgenic Arabidopsis by reducing plant damage and promoting plant growth.

Transcriptome changes triggered by a short-term low temperature stress in winter wheat

Transcriptome changes triggered by a short-term low temperature stress in winter wheat

Short-Term Low Temperature Induces Nitro-Oxidative Stress that Deregulates the NADP-Malic Enzyme Function by Tyrosine Nitration in Arabidopsis thaliana.

Low temperature (LT) negatively affects plant growth and development via the alteration of the metabolism of reactive oxygen and nitrogen species (ROS and RNS). Among RNS, tyrosine nitration, the addition of an NO2 group to a tyrosine residue, can modulate reduced nicotinamide-dinucleotide phosphate (NADPH)-generating systems and, therefore, can alter the levels of NADPH, a key cofactor in cellular redox homeostasis. NADPH also acts as an indispensable electron donor within a wide range of enzymatic reactions, biosynthetic pathways, and detoxification processes, which could affect plant viability. To extend our knowledge about the regulation of this key cofactor by this nitric oxide (NO)-related post-translational modification, we analyzed the effect of tyrosine nitration on another NADPH-generating enzyme, the NADP-malic enzyme (NADP-ME), under LT stress. In Arabidopsis thaliana seedlings exposed to short-term LT (4 °C for 48 h), a 50% growth reduction accompanied by an increase in the content of superoxide, nitric oxide, and peroxynitrite, in addition to diminished cytosolic NADP-ME activity, were found. In vitro assays confirmed that peroxynitrite inhibits cytosolic NADP-ME2 activity due to tyrosine nitration. The mass spectrometric analysis of nitrated NADP-ME2 enabled us to determine that Tyr-73 was exclusively nitrated to 3-nitrotyrosine by peroxynitrite. The in silico analysis of the Arabidopsis NADP-ME2 protein sequence suggests that Tyr73 nitration could disrupt the interactions between the specific amino acids responsible for protein structure stability. In conclusion, the present data show that short-term LT stress affects the metabolism of ROS and RNS, which appears to negatively modulate the activity of cytosolic NADP-ME through the tyrosine nitration process.

Rice mutants deficient in ω-3 fatty acid desaturase (FAD8) fail to acclimate to cold temperatures

To investigate the role of ω-3 fatty acid (FA) desaturase (FAD8) during cold acclimation in higher plants, we characterized three independent T-DNA insertional knock-out mutants of OsFAD8 from rice (Oryza sativa L.). At room temperature (28 °C), osfad8 plants exhibited significant alterations in fatty acid (FA) unsaturation for all four investigated plastidic lipid classes. During a 5-d acclimation period at 4 °C, further changes in FA unsaturation in both wild-type (WT) and mutant plants varied according to the type of lipid. We also monitored the fluidity of the thylakoid membrane using a threshold temperature to represent the change in fluorescence. The values were altered significantly by both FAD8 mutation and cold acclimation, suggesting that factors other than FAD8 are involved in C18 FA unsaturation and fluctuations in membrane fluidity. Similarly, significant changes were noted for both the mutant and WT samples in terms of their FA compositions as well as activities related to photosystem (PS) I, PSII, and photoprotection. This included the development of non-photochemical quenching and increased zeaxanthin accumulation. Despite the relatively small changes in FA composition during cold acclimation, cold-inducible FAD8 knock-out mutants displayed strong differences in photoprotective activities and a further drop in membrane fluidity. The mutants were more sensitive than WT to short-term low-temperature stress that resulted in increased production of reactive oxygen species after 5 d of chilling. Taken together, our findings suggest that FA unsaturation by OsFAD8 is crucial for the acclimation of higher plants to low-temperature stress.

Changes in the fatty acid composition and regulation of antioxidant enzymes and physiology of juvenile genetically improved farmed tilapia Oreochromis niloticus (L.), subjected to short-term low temperature stress

We evaluated the effect of cold shock on the fatty acid composition, antioxidant enzymes, and physiological responses of genetically improved farmed tilapia (GIFT). Experimental GIFT tilapia, a warm-water teleost, were initially acclimated at 28°C and then transferred directly to 13°C. Stress responses were monitored for 120h. There was a significant change in all parameters in response to the cold stressor (P<0.05). Serum cortisol levels increased from 336.93ng/ml to a peak of 1165.31ng/ml 24h after the initial cold shock, and declined rapidly thereafter. Serum glucose and cholesterol levels were significantly lower in the low temperature group than the control group at 120h (P<0.05). Acute low temperature stress enhanced superoxide dismutase, glutathione peroxidase, catalase, and glutathione levels in the liver of GIFT tilapia. The GIFT tilapia were able to selectively metabolize fatty acids for energy needs during the early period of exposure to low-temperature stress. During this time, they primarily used saturated fatty acids for energy. However as the duration of the stressor and loss of muscle fat increased, the fish began to metabolize long-chain polyunsaturated fatty acids. Increased malondialdehyde was produced by oxidation of these fatty acids leading to oxidative damage. Our results provide insight into the changes in fatty acid metabolism physiology that allow GIFT tilapia juveniles to adapt to short-term cold stress.

Drought priming at vegetative stage improves the antioxidant capacity and photosynthesis performance of wheat exposed to a short-term low temperature stress at jointing stage

To study the effects of multi-occurring stresses and the underlying mechanisms of cross-stress tolerance. Wheat plants were exposed to moderate drought priming (the leaf water potential reached ca. -0.8 MPa) at the 5-6th leaf stage for 2 weeks, and the physiological and yield responses of the drought-primed and non-primed plants to low temperature stress (4/ 2 °C in the day/ night for 2 days) at jointing stage were investigated. Under low temperature stress, the primed plants possessed higher leaf relative water content and much activated antioxidant system and hence reduced the oxidative injury to the photosynthetic apparatus, resulting in greater photosynthetic rate and higher grain yield as compared with the non-primed plants. In addition, increase of the ABA concentrations in leaf was found closely associated with the enhanced anti-oxidant enzymes activity in the primed plants. Drought priming at vegetative stage improves cold tolerance of wheat at jointing stage via sustaining ROS homeostasis, during which ABA plays a key role in alleviating the effects of low temperature stress in the drought-primed plants.

Consequences of Short Term Low Temperature Stress on Physiological and Biochemical Aspects of Rice (Oryza sativa L.)

Consequences of Short Term Low Temperature Stress on Physiological and Biochemical Aspects of Rice (Oryza sativa L.)

Selenium Modifies the Effect of Short-Term Chilling Stress on Cucumber Plants

The objective of this study was to investigate the effect of selenium (Se) supply (0, control; 2.5, 5, 10, or 20 μM) on cucumber (Cucumis sativus L.) cv. Polan F1 plants grown under short-term low temperature stress. About 14-16 day-old seedlings, grown at an optimal temperature (25/20°C; day/night), were exposed to short-term chilling stress with a day/night temperature of 10°C/5°C for 24 h, for a further 24 h at 20°C/15°C, and then transferred to 25/20°C (re-warming) for 7 days. Se did not affect the fresh weight (FW) of plants at a concentration of 2.5-10 μM, but in the presence of 20 μM Se, the biomass of shoots significantly decreased. The contents of chlorophylls and carotenoids witnessed no significant change after Se supplementation. Compared with the control, the Se-treated plants showed an increase of proline content in leaves, once after chilling and again after 7 days of re-warming. However, proline levels were much higher immediately after chilling than after re-warming. The malondialdehyde (MDA) content in the root of plants treated with 2.5-10 μM Se decreased directly after stress. This was in comparison with the plants grown without Se, whereas it increased in roots and leaves of plants exposed to 20 μM Se. Seven days later, the MDA level in the root of plants grown in the presence of Se was still lower than those of plants not treated with Se and generally witnessed no significant change in leaves. Although Se at concentrations of 2.5-10 μM modified the physiological response of cucumber to short-term chilling stress, causing an increase in proline content in leaves and diminishing lipid peroxidation in roots, the resistance of plants to low temperature was not clearly enhanced, as concluded on the basis of FW and photosynthetic pigments accumulation.

Difference between the temperature of non-hardened and hardened winter wheat seedling shoots during cold stress

The influence of cold hardening on heat generation in winter wheat seedling shoots during cold stress is discussed. In seedling winter wheat shoots, the temperature of “living” plants was higher, than in the ones “killed” during short-term low-temperature stress (cold shock). The study of the influence of cold shock intensity has shown that an increase in cold shock intensity caused a larger temperature difference between “living” and “killed” seedling shoots and resulted from an increase in thermogenesis in winter wheat seedling shoots. Cold hardening reduced this temperature difference between “living” and “killed” winter wheat seedling shoots. Treatment of winter wheat shoots by an uncoupling agent (CCCP) caused both an increase in temperature and survival during cold shock.

The effect of hypothermia on the content of 310 kD stress protein in seedlings of winter rye and wheat

The effect of hypothermia on the content of 310 kD stress protein in seedlings of winter rye and wheat was studied by rocket-immunoelectrophoresis and radioactive label. The 1-h low-temperature stress was found to result in an increase in the content of this protein at both above- and below-zero temperatures. It was found that the increase in the relative content of the protein with mol. wt 310 kD, under the effect of short-term low-temperature stress, occurs due to induction of its synthesis. It has been found that during cold hardening of winter wheat the content of this protein decreases up to 64% compared to the control during the first day of hardening but starts to rise by the third day and reaches 179% by the seventh day, which is well correlated with the increase in cold resistance of winter wheat plants during cold hardening.

Effect of Short-term and Long-term Low Temperature Stress on Polyamine Biosynthesis in Wheat Genotypes with Varying Degrees of Frost Tolerance

Summary Two series of experiments were carried out to examine the short- and long-tetm effects of low temperature on polyamine biosynthesis in wheat. In the first series, studies were made on the polyamine accumulation in the leaves, crowns and roots of winter wheat varieties with varying degrees of frost toletance subjected to short-term low temperature stress (6h, -2 °C). A marked accumulation of Put was observed. Agm accumulation was also examined and found comparable to that of Put. This suggests that Agm, which is an intermediate product of Put synthesis only in higher plants, may play an important role during short-term cold treatment. The second series of experiments was aimed at discovering the effect of wheat chromosomes 5A and 7A, which contain major genes responsible for frost resistance, on the polyamine synthesis taking place in various parts of the seedlings during long periods of cold treatment, and especially on the alternative metabolic pathway present only in higher plants.