Very-long-chain fatty acids (VLCFA) are involved in a number of important plant physiological functions. Disorders in the expression of genes involved in the synthesis of VLCFA lead to a number of phenotypic consequences, ranging from growth retardation to the death of embryos. The elongation of VLCFA in the endoplasmic reticulum (ER) is carried out by multiple elongase complexes with different substrate specificities and adapted to the synthesis of a number of products required for a number of metabolic pathways. The information about the enzymes involved in the synthesis of VLCFA with more than 26 atoms of Carbon is rather poor. Recently, genes encoding enzymes involved in the synthesis of both regular-length fatty acids and VLCFA have been discovered and investigated. Polyunsaturated VLCFA in plants are formed mainly by 20:1 elongation into new monounsaturated acids, which are then imported into chloroplasts, where they are further desaturated. The formation of saturated VLCFA and their further transformation into a number of aliphatic compounds included in cuticular waxes and suberin require the coordinated activity of a large number of different enzymes.

Changes in the activity of superoxide dismutase (SOD), ascorbate peroxidase (APX), and catalase (CAT) for the plants Nicotiana tabacum L. and Secale cereale L. during low-temperature hardening have been investigated. It was determined that a transient increase in the content of MDA occurs in tobacco plants at the beginning of the hardening period with subsequent activation of SOD, APX, and CAT. Unlike tobacco, winter rye has an MDA content and activity of SOD, APX, and CAT throughout the hardening period that remains at the level of unhardened plants. In tobacco cells, the majority of SOD activity was detected in chloroplasts (90%), while other cellular structures accounted for only 10% of the total activity identified in PAGE. In winter rye cells, SOD activity was distributed evenly between chloroplasts and other cellular compartments (46% in chloroplasts and 54% outside of chloroplasts). The distribution of APX and CAT activity in tobacco and winter rye cells was identical: all APX was concentrated in chloroplasts, while all catalase activity was manifested outside of chloroplasts. It is concluded that tobacco and winter rye plants showed different antioxidant defense strategies under conditions of low hardening temperatures. Tobacco plants provided protection against oxidative damage during low-temperature hardening by increasing activity of SOD, APX, and CAT. Winter rye plants were able to avoid the development of oxidative stress during hardening due to uniform distribution of SOD activity in cells, maintaining the constitutive activity of SOD, APX, and CAT, and, probably, due to the accumulation of nonenzymatic antioxidants.

Changes in the fatty acid (FA) composition of leaf and root lipids of heat-loving tobacco (Nicotiana tabacum L., cv. Samsun) plants during low-temperature hardening (8°C for 6 days) were studied. Hardening could improve leaf but not root cold tolerance. As this took place, the relative content of polyunsaturated (18:2n-6 and 18:3n-3) FAs increased and the proportion of saturated and monounsaturated FAs decreased. In contrast, in the roots hardening slightly increased the concentration of saturated FAs (16:0 and 18:0) and reduced the level of unsaturated FAs (18:1n-9, 18:2n-6, and 18:3n-3). At the same time, root lipids contained much C_20–24 FAs, and their content increased during hardening. It was suggested that an increased FA saturation and elevated proportion of C_20–24 FAs in the root lipids resulting in the lower membrane fluidity could be a reason for incapability of heat-loving tobacco plant roots of hardening and plant death at the lowtemperature stress.

Specific features of low-temperature hardening (6 days at 8°C) of cold-sensitive tobacco plants (Nicotiana tabacum, cv. Samsun) related to changes in the cell-wall invertase activity were studied. During cold hardening, oppositely directed changes in this enzyme activity occurred in tobacco leaves and roots. In the leaves, cell-wall invertase was activated (approximately by 30%), the content of sugars increased (approximately by 25%), and the content of sucrose, the main transport form of sugars, in the apoplast reduced by three times; all these changes indicate that assimilate outflow from leaves to roots was inhibited. In contrast, in the root system, enzyme activity was decreased almost twice and the content of sugars in them was essentially unchanged. It is suggested that a strategy of low-temperature adaptation of cold-sensitive tobacco plants aimed at creating the high cold tolerance of aboveground parts, even at the expense of the root system, which, under conditions of native vegetation, is not practically exposed to damaging low temperatures.

One of the unresolved questions in stress-response biology is how plants coordinate expression levels between the response and adaptation. In this work, we proposed a two-level analysis that examines both transcriptional and translational profiles of Solanum lycopersicum under conditions of short-term cold stress, hardening, and their combination. By combining polysome profiling and total transcriptome analysis, we revealed that expression under cold stress is not a simple linear process but a structurally distinct system with two coordinated regulation centres. Hardening triggers a strong transcriptional program focused on biogenesis, light signalling, and structural adaptations. In contrast, acute stress prompts selective translation of metabolic and defence proteins without prior transcriptional increase. Modular analysis (WGCNA) showed little overlap between transcriptional and translational networks, indicating functional differences between regulation levels. This work demonstrates that the cold response involves a strategic reallocation of resources between transcription and translation based on the type of signal. It bridges basic biology and applied breeding, providing targets promising for improving plant stress tolerance and advancing bioengineering of adaptive agriculture.

Changes in fatty acid composition of chloroplast membrane lipids were investigated using tobacco (Nicotiana tabacum L., cv. Samsun) plants subjected to cold hardening for 6 days at 8°C. Under optimal growing temperature (22°C), the lipids of thylakoid membranes were characterized by elevated content of 16:3n-3 and 18:3n-3 fatty acids (FA). Compared to the lipids of chloroplast envelope membranes, the thylakoid lipids were less rich in the content of saturated, mono- and diunsaturated FA. The relative content of unsaturated FA in chloroplast membranes increased substantially during cold hardening, which was mainly due to the accumulation of 18:3n-3 FA. It is concluded that the observed changes in FA composition of chloroplast lipids during cold hardening adjust the fluidity of these membranes to the level sufficient for functioning of tobacco photosynthetic apparatus, which is a prerequisite for accumulation of assimilates and allows the hardened tobacco plants to survive under conditions of hypothermia.