Repair of single-strand DNA breaks and recovery of chromosomal and chromatid aberrations after treatment of plant seeds with propyl methanesulfonate in vivo.

Repair of single-strand breaks of DNA and simultaneous recovery of chromosomal aberrations were studied after treatment of barley seeds with the monofunctional alkylating chemical mutagen, propyl methanesulfonate in vivo. In soaked seeds the diminution of single-strand breaks of DNA induced by PMS was correlated with the decrease of chromosomal aberrations, whereas in dried seeds the repair of DNA breaks was depressed and, in accord with this, the frequency of chromosomal aberrations increased. The prolonged storage of seeds led to a more delayed repair of chromosomal aberrations in dry seeds and a more delayed accelerated repair in soaked seeds.     

Radioprotection by caffeine pre- and post-treatment in the bone marrow chromosomes of mice given whole-body gamma-irradiation.

The effect of caffeine given as pre- and post-treatment in mice exposed to whole-body gamma-irradiation (1.5 Gy 60Co gamma-rays) was studied. The pre-treatment was either acute or chronic. The acute dose (5 mg/kg and 15 mg/kg body weight) was in the form of an injection given intraperitoneally, 30 min before irradiation. The chronic administration was in the form of caffeine solution (4.208 x 10(-3) M and 7.72 x 10(-4) M) contained in the drinking water that mice had had ad libitum access to instead of plain drinking water for 5 weeks prior to radiation exposure. The acute pre-treatment with caffeine reduced the radiation-induced frequency of chromosomal aberrations discernibly, whereas the chronic pre-treatment afforded a much more significant degree of radioprotection. The caffeine post-treatment (5 mg/kg and 15 mg body weight) was given in the form of an intraperitoneal injection to the mice immediately following whole-body gamma-irradiation. It is noted that both post-treatment concentrations of caffeine also significantly reduced the frequency of chromosomal aberrations induced by gamma-rays. These data are briefly discussed in terms of possible mechanistic considerations.     

A Comparison of the Quantitative Effects of Seed Moisture Content and Temperature on the Accumulation of Chromosome Damage and Loss of Seed Viability in Lettuce

The rate of accumulation of cells containing chromosome aberrationsin lettuce (Lactuca sativa L.) seeds is a positive functionof temperature and moisture content. It may be described byan equation similar to that for loss of seed viability. Therelative effect of temperature on the rates of loss of viabilityand accumulation of chromosome aberrations is the same. In contrast,the relative effect of moisture on the rate of loss of viabilityis greater than that for the rate of accumulation of aberrations.Hence considerably more chromosome damage accumulates beforedeath in drier lettuce seeds. Lactuca sativa, lettuce, seed storage, seed viability, seed longevity, chromosomal aberrations temperature, moisture content     

Studies on the Mutagenic Effect of the Chromosome by 5 MeV Electron

In this experiment, electron and y-ray-radiation were used to irradiate dry seeds of Vicia faba. Experimental results show that electron just as affective as γ-ray has significant mutagenic effect on cells. In the range of the dose, the frequency of chromosomal aberrations on the root tip cells, there are singnificantly dose-effect. But the relationship between the dose and frequency of aberrations is close to straight line and redplication. An analysis from the type of chromosomal aberrations by electron also leads to different results, in which there are more bridges and rings by electron than by γ-ray. Therefore, electron possibly have stronger two hit aberrations, and the frequency of aberration increased in proporti on with the square of dose.     

Chronic irradiation with low doses can be characterized by significant biological efficacy

Chromosomal aberrations (ChA) level was analyzed in the onion root meristem after the chronic irradiation with different dose capacities. It was shown that after the chronic irradiation with doses of 0.87 cGy, 2.61 cGy and 4.35 cGy the level of chromosomal aberrations depended on the dose capacity. Its value may also correspond to those which have been induced with accute irradiation. Biological efficacy of chronic irradiation may be from 20 to 1000 time folder in order to compare it with accute irradiation and this value depends on the irradiation regime.     

Artificial ageing of Vicia faba L. seeds caused by prolongation of G-1 phase

Broad bean (Vicia faba L. “Inovec”) seeds were artificially aged by means of storage at 30 %, resp. 25 % water content at 25 °C for 7-days to study the consequences on germination, root length and frequency of chromosomal aberrations. Under these conditions, significant changes in all parameters were observed. An increase of frequency of chromosomal aberrations in ana-telophase cells was confirmed by evaluation of c-metaphase cells. Synergic effect of artificial seed ageing was studied on different harvests of old seeds. Possible principles of this effect on cell level are discussed.     

Antagonism in the action of kinetin and actinomycin D on the cytogenetic effect of radiation

A study was made of the effect of kinetin when administered separately or in combination with actinomycin D on the chromosomal aberrations in Crepis capillaris L. irradiated by X-rays. Dry seeds were treated after irradiation or seeds soaked, prior to irradiation. Significant decrease in the frequency of chromosomal aberrations was shown when actinomycin D was administered separately and prior to kinetin. The latter used separately and prior to actinomycin D slightly increased the level of the aberrations, as compared with the control. Noticeable radioprotective effect was observed after treatment of seeds with kinetin, prior to irradiation.     

Seed aging: chromosome stability and extended viability of seeds stored fully imbided

Increase in moisture content of seeds of Lactuca sativa L. and Fraxinus americana L. in air-dry storage caused a rapid decline in longevity and an increase in the rate of accumulation of chromosome aberrations. Storage of seeds fully imbibed but unable to germinate allowed a high germination capacity to be maintained for long periods, together with a very low incidence of chromosome aberrations. Seedlings grown from dry-stored seeds showed an increase in morphological abnormalities with length of storage, whereas seedlings from imbibed-stored seeds appeared normal. It is suggested that in dry tissues, enzyme-controlled turnover and repair may be temporarily suspended, and that this may be an important factor in the loss of seed viability in storage. The effect of increasing seed longevity by lowering the moisture content of dry-stored seeds is discussed in relation to this hypothesis. The relevance of the proposal is also discussed in relation to ecological studies.     

Bimodal changes in germinability of pea seeds under the influence of low doses of gamma-radiation

Pea seeds (cv. 'Nemchinovskii-85', harvest of 2002, 80%-germination percentage) were exposed to gamma-radiation with doses ranging from 19 cGy to 100 Gy. One week after the irradiation with doses of 19 cGy and 3 Gy. the germination percentage decreased to 58 and 45%, respectively; at doses of 7 and 10 Gy it was 73 and 70% respectively. At greater doses (25, 50, and 100 Gy), germination percentage decreased in proportion. Anomalous changes in seed germination percentage (as a function of irradiation dose) were caused by the redistribution of irradiated seeds between fractions I and II. The measurements of room temperature phosphorescence in air-dry seeds and the phosphorescence of endogenous porphyrines of imbibing seeds showen that the germination decrease after the irradiation with low doses (19 cGy and 3 Gy) was caused by the increase in the number of weak seeds of fraction II, which had high rates of water uptake and suffered from hypoxia under seed coat. Some of these seeds suffocated from hypoxia, and other seeds produced seedlings with morphological defects (such seeds were considered incapable of germination). During storage of seeds irradiated at doses 19 cGy-10 Gy, the recovery of germination percentage (after its initial decrease) was caused by the decrease in seed number in fraction II. The subsequent germination decrease was caused by seed death. The higher was the irradiation dose, the faster were changes in germination percentage during storage of irradiated seeds. Bimodal changes in pea seed germination with the increase of y-irradiation dose has apparently the same origin as the changes in seed germination during accelerated ageing.     

Dynamics of the induced chromosomal instability in welsh onion (Allium fistulosum L.): gamma irradiation of the seeds of different storage periods

The chromosome aberrations in root meristem cells of welsh onion (Allium fistulosum L.) seeds after gamma-irradiation (5 and 10 Gy) of different-aged seeds (7, 19, 31, 43 and 55 months of storage) were studied. The irradiation dose of 5 Gy significantly increased the frequency of aberrant anaphases (FAA) for 31- and 43-months seeds; the dose of 10 Gy significantly increased the FAA in seeds of all age groups. The irradiation of young (7 months) seeds resulted in decreasing of the fraction of bridges to the control level of the old (55-months) seeds for the dose of 5 Gy and below the control level of the old seeds--for the dose of 10 Gy. Some peculiarities of cytogenetic parameters of genome instability and the germinating capacity of the seeds made it possible to suppose that the third year of storage is a critical period for the welsh onion seeds.     

Chromosome damage induced by artificial seed aging in barley

Summary Barley (Hordeum vulgare L. Himalaya) seeds were artificially aged under two storage conditions (32 °C/12% moisture content (m.c.) and 38 °C/18% m.c.) to study the behavior of induced chromosomal aberrations during plant growth. The frequencies of aberrant anaphases at first mitosis in root tips were correlated with loss of germinability. However, after 3 and 5 weeks' growth, aberration frequency declined. In plants grown from artificially aged seeds, the frequency of aberrant anaphases appeared to be stabilized at about 1% after 5 weeks' growth, in spite of the large differences in the frequencies at first mitosis. This suggests that because of their genetic imbalance, cells with chromosomal aberrations induced by seed aging were being excluded during plant growth. Meiotic chromosome configurations at MI were normal (7 II) in all plants studied, although a few precocious separations were found. Meiotic aberrations were found at AI-TI, AII-TII and the tetrad stages in the pollen mother cells of plants grown from the control and artificially aged seeds. However, there were no clear differences among the control and the two aging treatments. It was obvious that some cells with meiotic chromosomal aberrations were lost between the AI-TI and AII-TII stages, and still more between the AII-TII and tetrad stages. The frequency of tetrads with micronuclei in plants produced from artificially aged seeds was the same as in the control. The plants grown from artificially aged seeds showed high pollen fertility (95.2 to 97.0%) and seed fertility (90.1 to 97.2%) which was comparable to the control values (97.4 and 97.9%) respectively, indicating no special effects of seed aging. Anaphase cells of the first mitosis in the next (A₂) generation were analyzed to study the transmission of chromosomal aberrations through mitotic and meiotic cell divisions in the A₁ generation. Aberrant anaphases in the progeny from the artificially aged seeds were not higher than those of the control progeny. This indicates that the chromosomal aberrations induced by seed aging are not transmitted to the next generation.Published with the approval of the Director of the Colorado state Experiment Station as Scientific Series No. 2776     

The effect of continuous gamma irradiation on formation of sister chromatid exchanges (SCEs) and chromosomal aberrations in meristematic cells ofVicia faba

Germinated seeds ofVicia faba were continuously irradiated at low dose rate of gamma rays (0.05 Gy h-¹) up to a total accumulated dose of 2 Gy. The FPG (fluorescence plus Giemsa) technique of differential chromatid staining was used to monitor the frequency of sister chromatid exchanges (SCEs) in irradiated root tip meristem cells. The results of the experiments have demonstrated that SCE frequency is raised by continuous gamma irradiation only in plant cells containing BrdU in the chromosomal DNA. No effect concerning SCE formation was recorded at continuous irradiation of meristematic cells of Vicia faba with native, i. e. BrdU-nonsubstituted, DNA. In contrast to SCEs, a significant increase was found in the yield of chromosomal aberrations in all variants of irradiation.     

Mutations and chromosomal aberrations in hMTH1-transfected and non-transfected TK6 cells after exposure to low dose rates of gamma radiation

The aim of the present study was to analyse the dose rate effect of gamma radiation at the level of mutations, chromosomal aberrations, and cell growth in TK6 cells with normal as well as reduced levels of hMTH1 protein. TK6 cells were exposed to gamma radiation at dose rates ranging from 1.4 to 30.0 mGy/h (chronic exposure) as well as 24 Gy/h (acute exposure). Cell growth, frequency of thymidine kinase mutants, and of chromosomal aberrations in painted chromosomes 2, 8, and 14 were analysed. A decline in cell growth and an increase in unstable-type chromosomal aberrations with increasing dose rate were observed in both cell lines. A dose rate effect was not seen on mutations or stable-type chromosomal aberrations in any of the two cell lines. Reduction in the hMTH1 protein does not influence the sensitivity of TK6 cells to gamma radiation. This result fits well with data of others generated with the same cell line.     

Loss of Viability in Lettuce Seeds and the Accumulation of Chromosome Damage under Different Storage Conditions

Loss of seed viability in lettuce (Lactuca sativa L.) duringstorage is associated with an increase in the frequency of cellsin the surviving seeds showing chromosome damage during firstmitoses. The relation is linear when probit of the frequencyof aberrant cells is plotted as a function of probit percentagenormal germination. The slope of the relation, however, variesaccording to moisture content so that the proportion of aberrantcells for any given loss of germination increases with decreasein moisture content over the range 13.0–5.5 per cent.At 3.3 per cent moisture content, however, the proportion ofaberrations was no greater than at 5.5 per cent moisture content;and at 18.1 per cent moisture content the proportion was noless than at 13.0 per cent moisture content. Despite these differences,the increase in chromosomal aberrations per unit time for agiven temperature was always less the lower the moisture content.Diplontic selection markedly reduced the frequency of chromosomalaberrations and eliminated the differences in these frequenciesbetween the different storage treatments. But even after fiveweeks' growth, root tips from aged seed still contained abouttwice as many aberrant cells as compared with similar root tipsderived from the original seed stock. Studies on the frequencyof recessive mutations indicated that excessive amounts of heritablemutations were not present in the progenies of aged seed, evenwhen stored at moisture contents as low as 5.5 per cent. Allthis and other evidence reinforces the view that orthodox seedsfor genetic conservation should be stored at not more than about5 per cent moisture content, and that even lower moisture contentsare worth considering. The results also emphasise the need formaintaining a high regeneration standard, i.e. the percentageto which seed viability is allowed to fall during storage beforethe seed stock is regenerated. Lactuca sativa, lettuce, seed storage, seed viability, chromosomal aberrations, phenotypic mutations     

Genome‐wide association mapping and biochemical markers reveal that seed ageing and longevity are intricately affected by genetic background and developmental and environmental conditions in barley

Globally, over 7.4 million accessions of crop seeds are stored in gene banks, and conservation of genotypic variation is pivotal for breeding. We combined genetic and biochemical approaches to obtain a broad overview of factors that influence seed storability and ageing in barley (Hordeum vulgare). Seeds from a germplasm collection of 175 genotypes from four continents grown in field plots with different nutrient supply were subjected to two artificial ageing regimes. Genome‐wide association mapping revealed 107 marker trait associations, and hence, genotypic effects on seed ageing. Abiotic and biotic stresses were found to affect seed longevity. To address aspects of abiotic, including oxidative, stress, two major antioxidant groups were analysed. No correlation was found between seed deterioration and the lipid‐soluble tocochromanols, nor with oil, starch and protein contents. Conversely, the water‐soluble glutathione and related thiols were converted to disulphides, indicating a strong shift towards more oxidizing intracellular conditions, in seeds subjected to long‐term dry storage at two temperatures or to two artificial ageing treatments. The data suggest that intracellular pH and (bio)chemical processes leading to seed deterioration were influenced by the type of ageing or storage. Moreover, seed response to ageing or storage treatment appears to be significantly influenced by both maternal environment and genetic background.     

Environmental impact on age-related dynamics of karyotypical instability in plants

The dynamics of karyotypical instability of Allium fistulosum L. (Welsh onion) during aging of genetically homogenous seeds from plants grown in three different areas was studied. We analyzed the frequency of anaphase cells with chromosomal aberrations "damage", as a number of chromosomal aberrations per cell with aberrations, and germinating capacity, as an indicator of the 'toxic' influence of age. The seeds' aging was accompanied by an increase in karyotypical instability (increasing frequency of anaphases with aberrations) and with certain changes in the spectrum of chromosome aberrations. The clearest distinctions between old and young seeds were found for the frequency of anaphase cells with chromosome aberrations. The general level of karyotypical instability positively correlates with the age of the seeds. The regression coefficient (b) corresponds to the general tendency of karyotypical instability during seeds' senescence under storage. For 'good' (A), 'normal' (B) and 'bad' (C) conditions, the coefficients (b's) are b(A)=0.22, b(B)=0.46 and b(C)=0.84 (p<0.05 for C, and p<0.001 for A and B). It was found that different ecological conditions of plant vegetation strongly influence age-related dynamics of chromosomal instability in the seeds obtained from these plants. Possible mechanisms of the transgenerational impact of this effect are discussed.     

Storage of ethyl methanesulphonate-treated barley seeds with low moisture contents

Barley seeds were treated with ethyl methanesulphonate (EMS) for 3 h at 25° C, washed with tap water for 24 h at 25° C, redried at 40° C to different moisture contents below 15% and stored at 25° C in desiccators or in sealed plastic bags. The criteria used for expressing the effect of storage were the M₁ seedling height and the frequency of chromosomal aberrations. With 14·9% seed moisture a strong increase of biological injury occurred in the course of a 2-week storage, while storage of seeds having an initial moisture content of 11·7% led to a significant increase of injury only after 6 weeks. Superdry EMS-treated seeds with 5% or less moisture can be stored at 25° C without any changes in the biological effects. A method is recommended to avoid the EMS-storage effects.     

The Influence of Pre and Post-storage Hydration Treatments on Chromosomal Aberrations, Seedling Abnormalities, and Viability of Lettuce Seeds

A 2 h soaking treatment in distilled water, or in aqueous solutionsof cysteine, potassium iodide, or sodium thiosulphate, had nosignificant effect (P > 0.25) on the subsequent longevityof lettuce seeds (Lactuca sativa L.) in two different storageenvironments. Neither did these treatments influence relationsbetween loss in germination and the frequency of chromosomalaberrations observed during first mitoses after storage. Incontrast partial hydration of lettuce seeds after storage byexposure to moist air (humidification) or to an osmoticum (priming)reversed some of the damage which resulted from ageing. Mostof the benefits occurred during the first 3 d of humidificationduring which seed moisture content rose to 34 per cent, or duringthe first 7 d of priming when seed moisture content increasedto 44 per cent. Both post-storage hydration treatments reducedthe frequency of chromosomal aberrations, increased the rateof root growth, and decreased the frequency of morphologicallyabnormal seedlings. Either treatment could be of practical use,but it is suggested that humidification is more convenient.Consideration should be given to adopting a humidification treatmentas standard practice following long-term seed storage for geneticconservation. Lactuca sativa, lettuce, seed storage, seed viability, chemical pre-treatment, seed longevity, seed humidification (conditioning), seed priming, chromosome repair, seedling abnormalities     

Comportamento Alla Germinazione e Mutabilità Cromosomica Spontanea in Triticum Durum Desf. Durante I Primi Due Anni di Vita del Seme

Abstract

Germination and spontaneous chromosome mutations in Triticum durum Desf. during the first two years of storage of the seed. — Spontaneous chromosome aberrations were recently observed in the meristem of radicles of seedlings raised from seeds (caryopses) of Triticum durum cv. Cappelli erop 1966. Germination and frequency of chromosomal aberrations, starting from the ripening of the caryopses to the end of the second year of their storage at room temperature were investigated. Cytological analysis and germination tests were accomplished at monthly intervals. It was shown that: 1) soon after ripening, the germination process was very slow, but chromosomal damage was high; 2) at the end of the dormancy period the percentage of aberrant anaphases and of chromatid breaks fell to a «minimum». This low chromosomal damage lasted for almost the whole of the two years of storage; 3) the nuclear damage was esclusively of chromatide type. This result and fact that the cytological damage is decreasing with time, are interpreted as evidence that the mutagenic agent(s) responsible is (are) more active during the germination period than during the quiescence of the embryo.     

Nature of size–number trade–off: test of the terminal-stream-limitation model for seed production of Cardiocrium cordatum

We tested the prediction of the terminal-stream-limitation model using Cardiocrium cordatum . This model predicts that the total offspring mass increases with offspring number, whereas it decreases with offspring size, because the loss of resources via maintenance respiration decreases with offspring number but increases with offspring size. We traced the growth curve of seeds and harvested seeds when they matured. The maximum gross growth rate of a seed had a strong positive effect on final seed dry mass, whereas the respiration cost had a strong negative effect on such mass. The total seed mass produced by a plant decreased with (or was independent of) an increase in the mean seed dry mass of the plant, whereas it increased with an increase in the number of seeds produced by the plant. An increase in seed number resulted in a decrease in the loss of resources due to respiration during seed growth, whereas an increase in the mean seed dry mass did not result in a decrease in the loss of resources due to respiration. Thus, we concluded that these results are consistent overall with the prediction of the model and that an increase in seed number rather than an increase in individual seed size is advantageous in terms of resource use efficiency.