Germination and Growth Characteristics of nud Knockout and win1 Knockout Barley Lines under Salt Stress.

Hordeum vulgare genes NUD (HvNUD) and WIN1 (HvWIN1) play a regulatory role in cuticle organization. Because the cuticle is a key evolutionary acquisition of plants for protection against environmental factors, a knockout (KO) of each gene may alter their ability to adapt to unfavorable conditions. A potential pleiotropic effect of HvNUD or HvWIN1 gene mutations can be assessed under salt stress. Initial developmental stages are the most sensitive in living organisms; therefore, we evaluated salt tolerance of nud KO and win1 KO barley lines at the seedling stage. Air-dried barley grains of the KO lines and of a wild-type (WT) line were germinated in NaCl solutions (50, 100, or 150 mM). Over 30 physiological and morphological parameters of seedlings were assessed. Potential pleiotropic effects of the HvNUD gene KO under salt stress included the stimulation of root growth (which was lower under control conditions) and root necrosis. The pleiotropic effects of the HvWIN1 gene KO under the stressful conditions manifested themselves as maintenance of longer root length as compared to the other lines; stable variation of most of morphological parameters; lack of correlation between root lengths before and after exposure to NaCl solutions, as well as between shoot lengths; and the appearance of twins. Salt tolerance of the analyzed barley lines could be ranked as follows: nud KO > win1 KO ≈ WT, where nud KO lines were the most salt-tolerant. A comparison of effects of salinity and ionizing radiation on nud KO and win1 KO barley lines indicated differences in tolerance of the lines to these stressors.

Multiannual Assessment of Quality of Plantago major L. Seed Progeny from Kyshtym Radiation Accident Area: Weather-Dependent Effects.

The effects of low-dose radiation that are observed in plant populations in radioactively contaminated areas are variable. One of the reasons is the influence of fluctuating weather conditions and the interaction of radiation with weather factors. This article summarizes results of 12-year research on the viability and radioresistance of greater plantain (Plantago major L.) seed progeny growing in the East Ural Radioactive Trace (EURT) zone and in control (nonradioactive) areas, with consideration of weather conditions' variability. The EURT was formed by the Kyshtym accident, which occurred in 1957 at the Mayak Production Association. Absorbed dose rates of P. major parental plants in the pollution gradient were 14.5-165.9 µGy h-1, which correspond to a low-dose range. Seed progeny quality was evaluated as seed weight, the survival rate, and root length of 21-day seedlings. Interannual variability in the studied parameters was high, and their ranges overlapped between EURT groups of seeds and control groups in most cases. The number of significant correlations between the parameters of seed quality and weather conditions was higher in EURT groups than in control populations. In the control groups of seeds, 88.9% of correlations were negative, whereas in the EURT groups, 78.5% were positive.

Stress memory in two generations of Plantago major from radioactive and chemical contaminated areas after the cessation of exposure.

HYPOTHESIS: The differences in viability, root length, and pro/antioxidant features of Plantago major seedlings identified in seed progeny formed in areas of radioactive and chemical contamination can persist in subsequent generations after the elimination of the stress. MATERIALS AND METHODS: The seed mixtures of F1 generation were collected from P. major natural populations (P plants) growing for a long time in the East Ural Radioactive Trace, the Karabash Copper Smelter zone, and background area. The seeds of F2 generation were obtained from F1 generation plants grown on experimental plots with 'clean' agricultural background; F3 generation was grown from F2 generation on the same plots. The viability of seed progeny was estimated by survival rate and root length. Pro/antioxidant features were determined spectrophotometrically by malondialdehyde content, superoxide dismutase and catalase activities, and total content of low molecular weight antioxidants in seedlings. RESULTS AND CONCLUSIONS: The hypothesis about the persistence of effects from chronic exposure to ionizing radiation and chemical contamination in the generations' sequence of P. major after the removal of stress was confirmed only partially. The data obtained indicated that changes in the prooxidant and antioxidant features of plants in response to low doses of ionizing radiation can persist for at least in two generations after the stress removal. In the case of long-term exposure to chemical contaminants, we observed the persistence of the effect in a succession of generations only on the morphological indicator of root length.

Seedling Biometry of nud Knockout and win1 Knockout Barley Lines under Ionizing Radiation.

The genes NUD and WIN1 play a regulatory role in cuticle organization in barley. A knockout (KO) of each gene may alter plant mechanisms of adaptation to adverse environmental conditions. A putative pleiotropic effect of NUD or WIN1 gene mutations in barley can be assessed in a series of experiments in the presence or absence of a provoking factor. Ionizing radiation is widely used in research as a provoking factor for quantifying adaptive potential of living organisms. Our aim was to evaluate initial stages of growth and development of barley lines with a KO of NUD or WIN1 under radiation stress. Air-dried barley grains with different KOs and wild-type control (WT) were exposed to γ-radiation at 50, 100, or 200 Gy at a dose rate of 0.74 R/min. Approximately 30 physiological parameters were evaluated, combined into groups: (1) viability, (2) radiosensitivity, and (3) mutability of barley seed progeny. Seed germination, seedling survival, and shoot length were similar among all barley lines. Naked nud KO lines showed lower weights of seeds, roots, and seedlings and shorter root length as compared to win1 KO lines. The shoot-to-root length ratio of nud KO lines' seedlings exceeded that of win1 KO and WT lines. In terms of the number of seedlings with leaves, all the KO lines were more sensitive to pre-sowing γ-irradiation. Meanwhile, the radioresistance of nud KO lines (50% growth reduction dose [RD50] = 318-356 Gy) and WT plants (RD50 = 414 Gy) judging by seedling weight was higher than that of win1 KO lines (RD50 = 201-300 Gy). Resistance of nud KO lines to radiation was also demonstrated by means of root length (RD50 = 202-254 Gy) and the shoot-to-root length ratio. WT seedlings had the fewest morphological anomalies. In nud KO lines, mainly alterations of root shape were found, whereas in win1 KO lines, changes in the color and shape of leaves were noted. Thus, seedlings of nud KO lines are characterized mainly by changes in the root system (root length, root number, and root anomalies). For win1 KO lines, other parameters are sensitive (shoot length and alterations of leaf shape and color). These data may indicate a pleiotropic effect of genes NUD and WIN1 in barley.

Accumulation of 90SR by Betula pendula within the East Ural Radioactive Trace zone.

This study was conducted in 2010-2020 at the head of the East Ural Radioactive Trace (EURT), which was formed in 1957 as a result of the Kyshtym accident at the Mayak Production Association. The main contaminant in this zone is the long-lived radionuclide Strontium-90 (90Sr). Secondary forests dominated by silver birch (Betula pendula) occupy 45% of the EURT area. Concentration of 90Sr in birch leaves and small branches was higher than that in the trunks. The 90Sr content in birch sapwood varied slightly in the radial direction and did not depend on tree age. This was due to the dynamic equilibrium of the migration processes responsible for the accumulation and horizontal transfer of 90Sr. The 90Sr concentration increases in false heartwood, which is formed as a result of the secondary metabolism of dying parenchyma in the inner part of sapwood and is characterised by a high content of ash elements. The concentration of radionuclides in the aboveground organs of birch increased and the aggregated transfer factors (Tag) decreased with an increase in the soil contamination density, in accordance with the power function. The reasons for these patterns are also discussed.

Genetic polymorphism of Plantago major populations from the radioactive and chemical polluted areas.

The variability of nine microsatellite loci was studied for Plantago major L. populations from radioactive (East-Ural Radioactive Trace, EURT) and chemical (Karabash Copper Smelter, KCS) contaminated areas (Urals, Russia). The absorbed dose rates in the EURT area were 178-1455 times higher than background, and the indices of the total toxic load in the KCS area were 13-42 times higher than background values. In total, 65 alleles were identified in P. major populations, while the number of alleles per locus in the EURT and KCS samples was lower than in the background samples. The expected heterozygosity in all loci significantly exceeded the observed, indicating a high level of inbreeding. The largest number of rare alleles (11-21) was found in background samples, of which 3-7 alleles were private. In the technogenically disturbed zones, 8-11 rare alleles (1-2 private) were noted. The Bayesian analysis (K = 3) showed that no unique groups were found in any of the areas; descendants of all founders (pioneers) were represented in each population, but in different proportions. However, only 4.1% of the variability was distributed between local P. major populations (FST = 0.041) and 95.9% was concentrated within the samples. A pairwise comparison revealed genetic differentiation between all EURT samples. In the KCS area, there was no significant differentiation in pairs of samples that were at a distance of 3-4 km from each other. For samples from the KCS and background sites, the Mantel test showed a statistically significant relationship between geographical and genetic distances, therefore, the intensity of migration flows between these areas is high. For samples from the EURT and background areas, no such dependence was found. In both impact zones, P. major populations showed reduced genetic diversity. This article discusses the causes of this phenomenon.