Effects of long-term chronic exposure to radionuclides in plant populations.

The results of field studies carried out on different plant species (winter rye and wheat, spring barley, oats, Scots pine, wild vetch, crested hairgrass) in various radioecological situations (nuclear weapon testing, the Chernobyl accident, uranium and radium processing) to investigate the effects of long-term chronic exposure to radionuclides are discussed. Plant populations growing in areas with relatively low levels of pollution are characterized by an increased level of both cytogenetic disturbances and genetic diversity. Although ionizing radiation causes primary damage at the molecular level, there are emergent effects at the level of populations, non-predictable from the knowledge of elementary mechanisms of cellular effects formation. Accumulation of cellular alterations may afterward influence biological parameters important for populations such as health and reproduction. Presented data provide evidence that in plant populations inhabiting heavily contaminated territories cytogenetic damage could be accompanied by a decrease in reproductive capacity. However, in less contaminated sites, because of the scarcity of data available, a steady relationship between cytogenetic effects and reproductive capacity was not revealed. Under radioactive contamination of the plant's environment, a population's resistance to exposure may increase. However, there are radioecological situations where an enhanced radioresistance has not evolved or has not persisted.

Strong light-induced reorganization of pigment-protein complexes of thylakoid membranes in rye (spectroscopic study).

The supramolecular reorganization of LHCII complexes within the thylakoid membrane in Secale cereale leaves under low and high light condition was examined. Rye seedlings were germinated hydroponically in a climate chamber with a 16 h daylight photoperiod, photosynthetic photon flux density (PPFD) of 150 µmo lm(-2)s(-1) and 24/16°C day/night temperature. The influence of pre-illumination of the plants with high light intensity on the PSII antenna complexes was studied by comparison of the structure and function of the LHCII complexes and organization of thylakoid membranes isolated from 10-day-old plants illuminated with low (150 µmo lm(-2)s(-1)) or high (1200 µmo lm(-2)s(-1)) light intensity. Aggregated and trimeric with monomeric forms of LHCII complexes were separated from the whole thylakoid membranes using non-denaturing electrophoresis. Analyses of fluorescence emission spectra of these different LHCII forms showed that the monomer was the most effective aggregating antenna form. Moreover, photoprotection connected with LHCII aggregation was more effective upon LHCII monomers in comparison to trimer aggregation. Light stress induced specific organization of neighboring LHCII complexes, causing an increase in fluorescence yield of the long-wavelength bands (centered at 701 and 734 nm). The changes in the organization of the thylakoid membrane under light stress, observed by analysis of absorbance spectra obtained by Fourier transform infrared spectroscopy, also indicated light-induced LHCII aggregation.

The xanthophyll cycle pigments in Secale cereale leaves under combined Cd and high light stress conditions.

Leaves of Secale cereale seedlings were exposed to high light illumination (1200micromolm(-2)s(-1)) and Cd ions at 5 or 50microM concentrations. Influence of these stress factors on violaxanthin cycle pigments content was analysed chromatographically. Chlorophyll a fluorescence induction was used to analyse response of PSII to stress conditions and contribution of light-harvesting complex (LHCII) in non-photochemical quenching of excitation energy. The Cd-induced all-trans violaxanthin isomerization was analysed by HPLC technique in acetonitrile:methanol:water (72:8:3, v/v) solvent mixture. Interestingly, in the control and Cd-treated leaves subjected to high light, photochemical utilization of absorbed energy increased. This indicates plant adaptation to high light stress. In control plants high light caused zeaxanthin formation, however, the presence of Cd in the nutrient solution resulted in reduction of the second step of violaxanthin de-epoxidation process and anteraxanthin accumulation. In this study we have also shown, that non-photochemical quenching can be independent of anteraxanthin and zeaxanthin content. The particular increase in the cis isomers fraction in Cd-treated leaves has been explained in terms of a direct metal-pigment interaction as confirmed by Cd-induced all-trans violaxanthin isomerization in organic solvent, leading to formation of 13-cis, 9-cis and 15-cis isomers.

Mode of translational activation of the catalase (cat1) mRNA of rye leaves (Secale cereale L.) and its control through blue light and reactive oxygen.

The enzyme catalase (EC 1.11.1.6) is inactivated by light and must be continuously replaced by new synthesis in order to maintain a constant enzyme activity in leaves. In winter rye leaves (Secale cereale L.) posttranscriptional mechanisms determine the rate of new catalase synthesis, including a light-controlled reversible modification of the catalase cat1 mRNA by methylation which greatly enhanced its translation efficiency. The specificity and regulation of this mRNA activation were further investigated. The translation efficiency of the rye cat1 mRNA was much more enhanced by N-7 methylation of the cap than that of an lhcb transcript. Investigations with truncated rye cat1 mRNAs indicated that the translational enhancement resulting from N-7 cap methylation did not require the presence of specific sequences of cat1 5'- and 3'-untranslated regions. Translational activation of the cat1 mRNA in rye leaves was independent of photosynthesis and most effectively induced by blue light. Peroxides (H(2)O(2), tertiary butyl hydroperoxide) and conditions enforcing an H(2)O(2) accumulation in the leaves (aminotriazole, paraquat) also caused an activation of the cat1 mRNA. A search for further signalling systems controlling the replenishment of inactivated catalase in light suggested that an inositol-1,4,5-triphosphate-mediated liberation of Ca(2+) from internal stores and a protein phosphatase played some role. However, these signalling systems did not affect the activation of the cat1 mRNA. After removal of Ca(2+) by EGTA the cat1 mRNA was rapidly degraded.

The biophysical basis of cell elongation and organ maturation in coleoptiles of rye seedlings: implications for shoot development.

The relationships between changes in irreversible and reversible organ length, turgor (P), osmotic pressure (pi), and metabolic activity of the cells were investigated in intact coleoptiles of rye seedlings ( Secale cereale L.) that were either grown in darkness or irradiated with continuous white light. Cessation of growth at day 4 after sowing was associated with an apparent mechanical stiffening of the cell walls. Turgor pressure was measured in epidermal and mesophyll cells with a miniaturized pressure probe. No gradient of turgor was found between the peripheral and internal cells. In juvenile (growing) coleoptiles, average turgor was 0.60 MPa and a negative water potential (P - pi) was established in these cells. Upon emergence of the primary leaf, turgor declined, but P was maintained at values of 0.43 and 0.52 MPa in 7-day-old light- and dark-grown coleoptiles, respectively. Water potential in non-growing cells approached zero. The rate of dark respiration and elongation growth were not correlated. Surgical removal of the mature coleoptile revealed that the erect position of the 7-day-old shoot was dependent on the presence of this sturdy, turgid organ sheath. It is concluded that, during the first week of seedling development, the pierced, metabolically active coleoptile fulfills an essential function as an elastic basal tube for the juvenile shoot.

Genetic consequences of radioactive contamination by the Chernobyl fallout to agricultural crops.

The genetic consequences of radioactive contamination by the fallout to agricultural crops after the accident at the Chernobyl NPP in 1986 have been studied. In the first, acute, period of this accident, when the absorbed dose was primarily due to external beta- and gamma-irradiation, the radiation injury of agricultural crops, according to the basic cytogenetic tests, resembled the effect produced by acute gamma-irradiation at comparable doses. The yield of cytogenetic damage in leaf meristem of plants grown in the 10-km zone of the ChNPP in 1987-1989 (the period of chronic, lower level radiation exposure) was shown to be enhanced and dependent on the level of radioactive contamination. The rate of decline with time in cytogenetic damage induced by chronic exposure lagged considerably behind that of the radiation exposure. Analysis of genetic variability in three sequential generations of rye and wheat revealed increased cytogenetic damage in plants exposed to chronic irradiation during the 2nd and 3rd years.

The gravitropic setpoint angle of dark-grown rye seedlings and the role of ethylene.

The orientation growth of coleoptiles of dark-grown seedlings of rye (Secale cereale L. cv. Marder II), when grown under various conditions, was analysed with respect to the gravivector ('gravitropic setpoint angle', GSA). Coleoptiles growing through moist vermiculite attain and maintain a GSA with an average of about 180 degrees, i.e. a vertical orientation. Seedlings growing uncovered either on the surface of vermiculite or positionally fixed on filter paper attain and maintain a GSA of 140-150 degrees (i.e. deviating from the vertical by an average of 30-40 degrees ). Changing the position of the embryo relative to the horizontally fixed seed kernel or of the angle of the seed with respect to gravity during germination (+/-40 degrees relative to the horizontal) had no significant effect on the subsequent GSA of both covered and uncovered seedlings. The GSA of uncovered coleoptiles could be restored close to 180 degrees by treatment of the seedlings with ethylene, either applied via ethephon or 1-aminocyclopropane-1-carboxylic acid (ACC) as well as by fruit-released ethylene. The results are discussed with respect to the mechanism of the regulation of gravitropic growth of grass seedlings.

Temperature/light dependent development of selective resistance to photoinhibition of photosystem I.

Exposure of winter rye leaves grown at 20 degrees C and an irradiance of either 50 or 250 micromol m(-2) s(-1) to high light stress (1600 micromol m(-2) s(-1), 4 h) at 5 degrees C resulted in photoinhibition of PSI measured in vivo as a 34% and 31% decrease in deltaA820/A820 (P700+). The same effect was registered in plants grown at 5 degrees C and 50 micromol m(-2) s(-1). This was accompanied by a parallel degradation of the PsaA/PsaB heterodimer, increase of the intersystem e- pool size as well as inhibition of PSII photochemistry measured as Fv/Fm. Surprisingly, plants acclimated to high light (800 micromol m(-2) s(-1)) or to 5 degrees C and moderate light (250 micromol m(-2) s(-1)) were fully resistant to photoinhibition of PSI and did not exhibit any measurable changes at the level of PSI heterodimer abundance and intersystem e- pool size, although PSII photochemistry was reduced to 66% and 64% respectively. Thus, we show for the first time that PSI, unlike PSII, becomes completely resistant to photoinhibition when plants are acclimated to either 20 degrees C/800 micromol m(-2) s(-1) or 5 degrees C/250 micromol m(-2) s(-1) as a response to growth at elevated excitation pressure. The role of temperature/light dependent acclimation in the induction of selective tolerance to PSI photoinactivation is discussed.

[An analysis of the genetic sequelae of the contamination of winter rye crops by the radioactive fallout from the Chernobyl Atomic Electric Power Station]. / Analiz geneticheskikh posledstvii zagriazneniia posevov ozimoi rzhi radioaktivnymi vypadeniiami Chernobyl'skoi AES.

Changes of important cytogenetical indices, like an yield of aberrant cells and their burden by damages, with dose value was shown at the study of genetical consequences of winter rye contamination with radioactive Chernobyl NPP fallout at the acute phase (the vegetation period of 1986). Larger genetical efficiency was demonstrated for low dose rate radiation. Stimulation of morphometric indices in rye seedlings characterizing their survival on the earlier stages of ontogenesis was observed together with significant increase of the cytogenetical damages.

[A cytological study of plants growing under exposure to different radiation levels]. / Tsitologicheskoe issledovanie rastenii, proizrastavshikh pod vozdeistviem raznykh urovnei radiatsii.

Genetically significant consequences of emergency at the Chernobyl Atomic Power Plant have been studied on wheat and rye plants. Plants grown in the 30-km zone of the plant after its emergency are determined to have high frequency of chromosome aberrations reaching 2.666% in rye and 1.075-2.572% in wheat and 2.235-3.187% as dependent on the variety, biotype (awnless, semi-awned and awned) and places of occurrence. These levels of aberrations are 5.34, 2.42-5.78 and 4.07-6.71 times higher than their frequency in control plants (0.499, 0.444 and 0.475%, respectively).

Utilisation of rye by chickens: effect of microbial status, diet gamma irradiation and sodium taurocholate supplementation.

Several experiments were conducted to determine the influence of intestinal microflora when rye diets are fed to young chicks. In a conventional environment dietary rye severely depressed growth, amino acid and fat retention, and metatarsal bone ash. These conditions could be effectively alleviated by gamma irradiation of the diet and maintenance in a germ-free environment. In gnotobiotic chicks associated with streptococci dietary rye caused effects similar to those seen in a conventional environment, however the responses were less severe. Since addition of sodium taurocholate improved fat retention from a rye diet to a greater extent than from a wheat diet, a deficiency of conjugated bile salts might contribute to the steatorrhea observed in rye-fed chicks.

Improvement of the nutritive value of rye for broiler chickens by gamma irradiation-induced viscosity reduction.

A study was undertaken with chickens to determine the effect of irradiation treatment on the viscosity-inducing properties of polysaccharide components of rye. Three-week body weight, food utilisation, fat retention, and bone ash of chicks were substantially improved by irradiating rye (0 to 10 Mrad) and were maximal at 6 Mrad. Irradiation of the entire diet resulted in no further improvement over irradiating the rye fraction alone. The improved performance coincided with radiation-induced damage to rye polysaccharides, as indicated by reduced viscosity and increased concentrations of reducing sugars.

Effect of gamma irradiation, penicillin, and/or pectic enzyme on chick growth depression and fecal stickiness caused by rye, citrus pectin, and guar gum.

The effects of gamma irradiation of rye, corn, pectin, and guar gum and supplementation of procaine penicillin and/or pectic enzyme (Irgazyme-100) were studied. Addition of 62.5% rye, 4% pectin, or 2% guar gum in place of corn significantly reduced chick growth (P < .05). Exposure to gamma irradiation, procaine penicillin, or pectic enzyme supplementation improved the growth of chicks fed rye. Growth response on the rye diet to one or more combinations of gamma irradiation, procaine penicillin, and pectic enzyme was greater than that of gamma irradiation, procaine penicillin, or pectic enzyme alone. Gamma irradiation or pectic enzyme supplementatin of pectin or guar gum improved chick growth and almost eliminated their growth depressing properties. A combination of gamma irradiation and pectic enzyme failed to give a further significant increase in growth over that of either alone. Addition of procaine penicillin to diets containing rye, irradiated pectin, or guar gum elicited a significant growth response, but the response was greatly reduced when diets contained irradiated guar gum or irradiated pectin. Feces of chicks fed diets containing rye, pectin, or guar gum adhered to screen floors in much greater amounts than for corn-fed chicks. Gamma irradiation, procaine penicillin, or pectic enzyme had no significant effect on fecal condition of birds fed diets containing rye or guar gum. In contrast, fecal condition of birds fed guar gum was significantly improved by a combination of gamma irradiation and pectic enzyme supplement. Gamma irradiation almost eliminated and pectic enzyme supplementation completely eliminated properties of pectin causing sticky feces. Results indicate that the component of rye that causes sticky feces was not changed by gamma irradiation and/or pectic enzyme, and, therefore, is different from the growth depressing factor and from citrus pectin.

Effect of gamma irradiation, fractionation, and penicillin supplementation on the rachitogenic activity of rye for chicks.

Experiments were conducted to study the effect of gamma irradiation on the rye-vitamin D antagonism in broiler chicks. In an initial study, the irradiated grain was exposed to gamma rays for 6 hr (2 Mrad) and was fed to rachitic chicks for only 12 hr before a single oral dose of vitamin D3. In another trial, birds were fed during the entire experiment (12 days) on the different experimental diets, and the irradiated grain was exposed to gamma rays for 20 hr. Results indicate that the factor in rye that interferes with vitamin D utilization was largely inactivated by exposing this grain to gamma rays for 20 hr. Additional information is reported in this paper related to the alleviation by extraction or antibiotic supplementation on the rachitogenic properties of rye. These studies were conducted by feeding to rachitic chicks the test materials for only a short period of time (12 hr) before a vitamin D3 oral dose and measuring the bone mineralization 48 hr later. Results indicate that the rachitogenic factor in rye is not present in the ash portion of the grain, that it can be largely overcome by water extraction and penicillin supplementation, and that an organic solvent extraction has no effect. Cooked beans which depress growth and increase the growth response to antibiotics are not rachitogenic.