Production of in vitro haploid plants from in situ induced haploid embryos in winter squash (<Emphasis Type="Italic">Cucurbita maxima</Emphasis> Duchesne ex Lam.) via irradiated pollen

The influence of pollen irradiation on the production of in vitro haploid plants from in situ induced haploid embryos was investigated in winter squash (Cucurbita maxima Duchesne ex Lam.). Pollen were irradiated at different gamma-ray doses (50, 100, 200 and 300 Gray) and durations (9, 11, 15, 21, and 28 July). Production of in vitro haploid plantlets was influenced by irradiation dose, irradiation duration, genotype, and embryo type and embryo stage. Embryos were only obtained from lower irradiation doses (50 Gray and 100 Gray) and earlier irradiation durations (9, 11, and 15 July). The greatest embryo number per fruit was procured from “G14” and “55SI06” genotypes at 50 Gray gamma-ray dose. Necrotic embryos were higher than normal embryos at delayed harvest times (5 and 6 weeks after the pollination). The convenient harvest time for embryo rescue was observed about 4 weeks (between 25 and 30 days) after pollination. All cotyledon and amorphous embryos had only diploid plants while late-torpedo, arrow-tip, and pro-cotyledon embryos produced 33.3, 50.0, and 66.7% haploid plant. The frequency of haploid plantlets was 0.11, 1.17, 10.96 and 0.28 per 100 seeds, 100 embryos, 100 plantlets and a fruit at 50 Gray gamma-ray dose, respectively.     

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.     

Changes in vascular permeability following thorax irradiation in the rat

A double isotope technique was used to measure changes in the vascular permeability surface area product (PS) for albumin after irradiation. PS was measured in several tissues of the rat during the first 38 days following 11, 13.5, 18, or 25 Gy whole thorax irradiation. After 18 and 25 Gy most irradiated and nonirradiated (shielded) tissues showed elevated permeability at 1 day after radiation, which declined to control levels by Day 4. All irradiated tissues showed a second wave of increased permeability between 14 and 38 days after radiation that varied in onset and extent depending upon tissue and dose. Lung and heart showed a direct response to dose between 11 and 18 Gy during this period. Peak lung values averaged three times control values at 19 days after 18 Gy. Peak heart values averaged twice control values at the same time and dose. The double isotope technique has proven to be a reliable means of quantitatively determining vascular permeability response to radiation over time.     

The disruption of aquaporin function in cell membranes as a cause of changes in germinability of pea seeds after exposure to low doses of gamma-radiation

Pea seeds (Pisum sativum L.) from the seed lot with 80% germinability were separated in fractions according to room temperature phosphorescence: strong seeds were assigned to fraction I, and weak seeds formed to fraction II. During imbibition, the seeds of fraction II exhibited twofold higher rates of water uptake and experienced hypoxia. Some of these seeds suffocated from hypoxia, and other seeds produced seedlings with morphological defects (such seeds were considered incapable of germination). One week after irradiation with the dose of 3 Gy, germination percentage decreased to 45%, which was caused by the increase of number of weak seeds. The germinability of seeds subjected to gamma-irradiation at doses of 7 and 10 Gy was similar to that of control seeds. In these sub-lots, there appeared so-called "improved" seeds, which were similar to non-irradiated seeds in terms of phosphorescence level, the rate of water uptake and germination percentage. It was shown with the use of PCMB that the difference in the rates of water uptake by seeds of fraction I and II depended on the permeability of cell membranes. The permeability was determined by the state of aquaporins ("open"-"closed"). The experiments with phosphatase inhibitor (NaF) shown that in seeds irradiated with dose of 3 Gy (fraction II), the mechanism of aquaporins closing was broken (phosphatase was inactivated). In "improved" seeds (after irradiation with dose of 10 Gy), aquaporins were closed irreversibly in air-dry state, when aquaporin dephosphorylation was unlikely. It was concluded that the abnormal increase (following the initially decrease) in germination of pea seeds after irradiation can be explained without invoking the hypothesis on hyper function of reparatory mechanism of at low doses of irradiation.     

Induction of phenotypically altered mammary epithelial cells by neutrons and gamma rays.

These studies have examined alterations in the in vivo growth properties of mammary epithelial cells isolated at 1, 4, and 16 weeks after in vivo irradiation with -137Cs gamma rays or fission-spectrum neutrons. Altered in vitro growth potential was characterized by the proliferation of epithelial foci (EF) from irradiated animals under conditions in which mammary cells from nonexposed animals senesced. These EF were further characterized based on their ability to be subcultured. Both gamma and neutron irradiation resulted in the appearance of cells capable of forming EF. Further, with increased time in situ between irradiation and cell isolation, the frequency of EF which were capable of being subcultured indefinitely (EFs) increased. Reducing the gamma-ray dose rate resulted in fewer EFs while reducing the neutron dose rate resulted in increased frequencies of EFs. These data confirm earlier observations following gamma irradiation and show these cellular changes are also observed following neutron irradiation. In addition, these data indicate that changes in dose rate primarily influence the emergence of immortalized cell populations.     

Retrospective dosimetry after criticality accidents using low-frequency EPR: a study of whole human teeth irradiated in a mixed neutron and gamma-radiation field

In the context of accidental or intentional radiation exposures (nuclear terrorism), it is essential to separate rapidly those individuals with substantial exposures from those with exposures that do not constitute an immediate threat to health. Low-frequency electron paramagnetic resonance (EPR) spectroscopy provides the potential advantage of making accurate and sensitive measurements of absorbed radiation dose in teeth without removing the teeth from the potential victims. Up to now, most studies focused on the dose-response curves obtained for gamma radiation. In radiation accidents, however, the contribution of neutrons to the total radiation dose should not be neglected. To determine how neutrons contribute to the apparent dose estimated by EPR dosimetry, extracted whole human teeth were irradiated at the SILENE reactor in a mixed neutron and gamma-radiation field simulating criticality accidents. The teeth were irradiated in free air as well as in a paraffin head phantom. Lead screens were also used to eliminate to a large extent the contribution of the gamma radiation to the dose received by the teeth. The EPR signals, obtained with a low-frequency (1.2 GHz) spectrometer, were compared to dosimetry measurements at the same location. The contribution of neutrons to the EPR dosimetric signal was negligible in the range of 0 to 10 Gy and was rather small (neutron/gamma-ray sensitivity in the range 0-0.2) at higher doses. This indicates that the method essentially provides information on the dose received from the gamma-ray component of the radiation.     

Mechanism of changes in irradiated thymocytes detected by anilinonaphthalenesulfonate

In experiments on rat thymocytes obtained 15 min, 2 h and 4 h after irradiation of animals with the dose of 10 Gy it was found that 15 min and 4 h following irradiation changes occurred in their membranes increasing the lifetime of the excited state of anilinonaphthalene sulfonate and hydrofobicity and viscosity of its microenvironment. The indicated parameters did not vary from the controls 2 h following irradiation. The analysis of the data obtained prompted a suggestion that the observed changes were associated with the structural rearrangements in the plasma membrane rather than with its destruction. Model experiments on irradiated thymocyte suspensions showed that there was a correlation between changes in the cell membrane and meabolic processes.     

Examples of high-frequency EPR studies in bioinorganic chemistry

Low-temperature EPR spectroscopy with frequencies between 95 and 345 GHz and magnetic fields up to 12 T has been used to study metal sites in proteins or inorganic complexes and free radicals. The high-field EPR method was used to resolve g-value anisotropy by separating it from overlapping hyperfine couplings. The presence of hydrogen bonding interactions to the tyrosyl radical oxygens in ribonucleotide reductases were detected. At 285 GHz the g-value anisotropy from the rhombic type 2 Cu(II) signal in the enzyme laccase has its g-value anisotropy clearly resolved from slightly different overlapping axial species. Simple metal site systems with S>1/2 undergo a zero-field splitting, which can be described by the spin Hamiltonian. From high-frequency EPR, the D values that are small compared to the frequency (high-field limit) can be determined directly by measuring the distance of the outermost signal to the center of the spectrum, which corresponds to (2 S-1)* mid R: Dmid R: For example, D values of 0.8 and 0.3 cm(-1) are observed for S=5/2 Fe(III)-EDTA and transferrin, respectively. When D values are larger compared to the frequency and in the case of half-integer spin systems, they can be obtained from the frequency dependence of the shifts of g(eff), as observed for myoglobin in the presence ( D=5 cm(-1)) or absence ( D=9.5 cm(-1)) of fluoride. The 285 and 345 GHz spectra of the Fe(II)-NO-EDTA complex show that it is best described as a S=3/2 system with D=11.5 cm(-1), E=0.1 cm(-1), and g(x)= g(y)= g(z)=2.0. Finally, the effects of HF-EPR on X-band EPR silent states and weak magnetic interactions are demonstrated.     

Effects of gamma-ray irradiation on metabolic changes in potato tubers in response to cutting

Potato tubers were subjected to cobalt-60 gamma-ray irradiationand stored at room temperature for from 10 days to 4 months.Effects of this irradiation on metabolic changes in potato tubersin response to cutting were investigated. The quantities ofpolyphenols, such as chlorogenic acid and isochlorogenic acid,in the tissue increased as compared with the non-irradiatedsample. Although the polyphenol content and activities of o-diphenoloxidase, peroxidase and phenylalanine ammonialyase increasedafter cutting, increases were lower in the irradiated sample.On the other hand, in the dose range between 5,000 and 12,400rad, the irradiated sample showed a larger defense action againstinfection by the black rot fungus, Ceratocystis fimbriata thandid the non-irradiated sample. (Received April 16, 1968; )     

Autoradiographically detected impairment of RNA synthesis pattern in 8-16 cell bovine embryos after irradiation

Early preimplantation bovine embryos at 8- or 16-cell stage were analysed by [5-3H]uridine autoradiography for distribution of newly synthesized RNA after 60Co irradiation with a single dose of 1 Gy, 2 Gy or 4 Gy gamma rays, respectively. Embryos irradiated with a single dose of 1 Gy showed equally decreased synthesis of RNA in nucleoplasma as well as in nucleolus. In embryos irradiated with a single dose of 2 Gy or 4 Gy, RNA synthesis was decreased and localized mostly on the periphery of the nucleus; in both cases of irradiation, the nucleus center being without labelling. In most of embryos irradiated with a dose of 4 Gy, the nucleoli were not labelled, and an increasing occurrence appeared of various nucleus chromatin segregation forms, mainly as its marginalization.     

Ultraviolet irradiation effects on serotinous shape Leucadendron laureolum seeds: altered seed physiology and ultrastructure,and seedling performance

Dry seeds of Leucadendron laureolum (Lam.) Fourc. (Proteaceae) were exposed for different intervals (range: 7 to 84 days) to visible, UV-A and UV-B radiation of different biologically effective dose (range: 0 to 11.43 kJ m^-2 d^-1). Changes in seed germination, physiology and ultrastructure, and residual UV effects on seedling performance, were examined. Germination was depressed in seeds following short (7-day) exposures to UV radiation. This depression was intensified with increased UV exposure dose, and most pronounced at shorter UV-B wavelengths. Also glutathione reductase (GR) activities increased in seeds exposed to shorter UV-B wavelengths, but these were unaffected by irradiation dose level in the UV-B range. Electrolyte leakage rates from UV-irradiated seeds were unaltered, which indicated that germination depression did not result from intrinsic membrane damage. The reversal of germination depression (UV-induced dormancy) in UV-irradiated seeds by red light pointed to the possible involvement of phytochrome in this photo-response. Germination depression disappeared in seeds after 56-days irradiation, possibly due to photoreceptor damage by excess UV light. At this stage, all UV irradiated seeds, irrespective of treatment wavelength or dose level, exhibited increased electrolyte leakage rates, which indicated membrane perturbation. Also, increased GR activities were observed in irradiated seeds, but these were proportionately smaller in seeds exposed to shorter wavelength UV-B radiation (9.1 to 35.8% increase) than longer wavelength UV-A (73.4% increase) and visible (97.7% increase) radiation. This implied a metabolic limitation for scavenging of free radicals and peroxides in aging seeds exposed to UV-B radiation, which pointed to accelerated seed deterioration. It was indirectly supported by ultrastructural evidence of sub-cellular damage (lipid coagulation and plasmalemma withdrawal from cell walls) in embryonic tissues of seeds after 84 days UV-B exposure, and reflected in decreased leaf numbers, photochemical efficiencies, and foliar chlorophyll a and carotenoid levels in seedlings cultured from these seeds.     

SOME EFFECTS OF GAMMA IRRADIATION ON SEEDS AND RHIZOMES OF MUSA

The tolerance of Musa balbisiana Colla seeds to gamma irradiation was found to be considerably greater than that of rhizomes of the parthenocarpic variety ‘Gros Michel': e.g., 11.8 krad reduced the germination of rhizomes 92% and of seeds approximately 15%. Intact seeds exposed to doses higher than 48 krad did not germinate in non-sterile soil, but, when scarified and cultured axenically after irradiation, seeds which received doses as high as 70 krad germinated. Embryos excised from seeds exposed to doses as high as 285 krad formed callus, indicating that not all metabolic processes were inhibited by these extremely high doses. There was considerable variation in radiation tolerance between seed lots which was not related to their age, moisture content, or pre-exposure viability. Germination of intact seeds appeared to be stimulated by doses of 3 or 9 krad. No lasting differences attributable to the level of irradiation were apparent in the development of seedlings derived from either intact or scarified seeds nor of plantlets derived from excised embryos. Conversely, there was a significant reduction, proportional to irradiation dose, in the growth of plants developing from rhizomes, emphasizing the greater radiation sensitivity of the vegetative propagule. The radiation tolerance of seed-borne microorganisms was considerably higher than that of the plant materials, indicating that gamma irradiation is not effective as a means of obtaining pathogen-free rhizomes or surface-sterilizing seeds of M. balbisiana.     

Effects of γ-ray treatment onCannabis saliva pollen viability

The viability and thein vitro germination capability of hemp pollen (cv. Carmagnola) were studied. Viability tests were based on the microscopic observation of the fluorescence of loaded fluorescein diacetate (FDA), while, for germinability tests, five different media were tested. The effects of irradiation with γ-rays on pollen viability and germination and on seed set were also studied, at three different irradiation doses (20, 60 and 100krad). The results show that in one of the media tested, about 85–90% of the pollen grains are viable and able to germinate in control samples, and that while viability measured by FDA test is not affected by increased γ-ray doses, the pollenin vitro germinability drops to about one-half of the controls at the maximum γ-ray dose employed, 100krad. Seed set of hemp plants pollinated with the irradiated pollen dropped to less than 1% of that of plants pollinated by untreated pollen for the higher dose used. The different media suitable forin vitro germination of hemp pollen, and the observed lack of correspondence between viability and germination capacity tests are discussed.     

Alanine radicals,part 4: relative amounts of radical species in alanine dosimeters after exposure to 6-19 MeV electrons and 10 kV-15 MV photons

The amino acid l-alpha-alanine can be used for high-precision dosimetry over a wide dose range, using EPR spectroscopy for monitoring radical concentrations. It is important, however, to understand the underlying composition of the observed EPR spectrum. In previous work, it was shown that the EPR signal from irradiated alanine consists of at least three different radical species, with the relative importance of each of these being almost independent of absorbed dose. However, it was not known whether the relative importance of each radical is independent of the radiation quality responsible for the EPR signal. In the present work, the relative contributions of the different radical species to the total EPR signal from alanine dosimeters irradiated with 6-19 MeV electrons and 10 kV-15 MV photons at a dose of 10 Gy were examined. By spectrum reconstruction using benchmark spectra generated from a simulation procedure, the relative amounts of the three different radical species were shown to be virtually independent of these radiation beam qualities.     

低能离子注入短命植物多伞阿魏(Ferula ferulaeoides)抗性生理特征

以能量30 keV、不同剂量N 离子注入多伞阿魏(Ferula ferulaeoides)种子用以研究多伞阿魏抗性生理指标的变化,以期为保护短命植物多伞阿魏提供理论依据.试验结果表明:随离子注入剂量增大,多伞阿魏种子发芽率和发芽指数逐渐下降;超氧化物歧化酶(SOD)活性增加、过氧化物酶(POD)、和过氧化氢酶(CAT)活性逐渐升高.剂量过大时,SOD、POD和CAT活性下降;游离脯氨酸含量逐渐降低,但在6×1016N /cm2剂量时含量升高;可溶性蛋白质含量先升高再降低,6×1016 N / cm2剂量时为最高值.通过对发芽率、发芽指数的结果进行方差分析,得出低剂量N 离子注入可破除多伞阿魏种子休眠,促进种子萌发;适当剂量N 离子注入可激活保护酶、脯氨酸和可溶性蛋白的表达.     

Phase transitions and permeability changes in dry membranes during rehydration

Dry phospholipid bilayers are known to undergo transient changes in permeability during rehydration. In this review, we present evidence from which we suggest that this permeability change is due to a gel to liquid-crystaline phase transition accompanying rehydration. If the transition is avoided, as in lipids that remain in gel phase whether dry or rehydrated, the problem of leakage during rehydration is obviated, at least in part. Further, the evidence that the transition temperature for dry bilayers can be depressed by certain sugars is discussed. Finally, we show that these principles can be extended to intact cells. Using pollen grains as a model, we have measured the transition temperature for membrane phospholipids and show that the transition is correlated with physiological measurements including permeability changes and subsequent germination. From theT _m values taken from pollen grains at different water contents, we have constructed a phase diagram for the intact pollen that has high predictive value for physiological properties.     

Relationship between ultra‐weak bioluminescence and vigour or irradiation dose of irradiated wheat

Ultra‐weak luminescent analysis is a new way to detect the irradiation dose and the vigour of irradiated wheat. Wheat grain and wheat flour were used in this research for ultra‐weak luminescent analysis. The experimental data showed that the bioluminescence intensity of wheat grain sample was different with increasing storage time and increasing dose, and a similar trend appeared in the germination rates of irradiated wheat grain. It was found that the differences in bioluminescence intensities and germination rates of irradiated wheat grain at different doses and storage times were due to the effect of irraditation on the wheat embryo and self‐repair during storage. As a result, ultra‐weak luminescent analysis cannot be used to detect the irradiation dose of irradiated wheat, but it can be used to determine vigour. Experiments showed that the irradiation dose had a highly significant effect on the bioluminescence intensities of wheat flour when cane sugar was added. Copyright © 2009 John Wiley & Sons, Ltd.     

γ-irradiation of PEGd,lPLA and PEG-PLGA Multiblock Copolymers: II. Effect of Oxygen and EPR Investigation

The purpose of this research was to evaluate how the presence of oxygen can affect irradiation-induced degradation reactions of PEGd,lPLA and PEG-PLGA multiblock copolymers submitted to gamma irradiation and to investigate the radiolytic behavior of the polymers. PEGd,lPLA, PEG-PLGA, PLA, and PLGA were irradiated by using a ⁶⁰Co irradiation source in air and under vacuum at 25 kGy total dose. Mw and Mn were evaluated by gel permeation chromatography. The stability study was carried out on three samples sets: (a) polymer samples irradiated and stored in air, (b) polymer samples irradiated and stored under vacuum, and (c) polymer samples irradiated under vacuum and stored in air. The thermal and radiolytic behavior was investigated by differential scanning calorimetry and electron paramagnetic resonance (EPR), respectively. Samples irradiated in air showed remarkable Mw and Mn reduction and Tg value reduction due to radiation-induced chain scission reactions. Higher stability was observed for samples irradiated and stored under vacuum. EPR spectra showed that the presence of PEG units in multiblock copolymer chains leads to: (a) decrease of the radiolytic yield of radicals and (b) decrease of the radical trapping efficiency and faster radical decay rates. It can be concluded that the presence of oxygen during the irradiation process and the storage phase significantly increases the entity of irradiation-induced damage.     

Ultrastructural alterations in ciliary cells exposed to ionizing radiation

Summary Early effects of ionizing radiation were investigated in an experimental in vitro system using the ciliary cells of the tracheal mucous membrane of the rabbit, irradiated at 30° C and at more than 90% humidity. The changes in physiological activities of the ciliary cells caused by irradiation were continuously registered during the irradiation. The specimens were examined immediately after irradiation electron microscopically. The morphological changes in irradiated material after 10–70 Gy are compared with normal material. After 40–70 Gy, scanning electron microscopy revealed the formation of vesicles on cilia, and club-like protrusions and adhesion of their tips. After 30–70 Gy, a swelling of mitochondrial membranes and cristae was apparent transmission electron microscopically. The membrane alterations caused by irradiation are assumed to disturb the permeability and flow of ATP from the mitochondria, which in turn leads to the recorded changes in the activity of the ciliated cells.This investigation was supported by grants from Konung Gustaf V:s Jubileumsfond, John and Augusta Perssons Stiftelse, B. Kamprads Fond, the Faculty of Medicine, University of Lund, Sweden and the Swedish Medical Research Council (No. B77-17X-03897-05)The authors are greatly indebted to Miss Inger Norling, Miss Marianne Palmegren and Miss Birgitta Sandström for their excellent technical assistance     

Taurine for EPR dosimetry

EPR dosimetry is characterized by its non-destructive read-out and the possibility of dose archival. Here, taurine is proposed as a radiation dosimeter using EPR spectroscopy. The EPR spectrum of taurine was studied and assigned, and changes in the taurine EPR spectrum as a result of the change in both modulation amplitude and microwave power were quantified. For gamma radiation, the energy absorption coefficient and the collision mass stopping power of taurine were compared to the corresponding values of soft tissue and alanine, in addition to calculation of effective atomic numbers. The response of taurine to gamma radiation doses in the range from 0.1 to 50 kGy was investigated, as well as that in the range from 1.0 to 20.0 Gy using numerically enhanced EPR taurine spectra. Both response curves showed a linear behavior. In addition, the time dependence of radiation-induced radicals was studied for short (during the first 6 h after irradiation) and long (during about 3 months after irradiation) time periods, and a reasonable degree of stability of the taurine radicals was observed. It is concluded that taurine is a promising dosimeter, which is characterized by its simple spectrum, radical stability, and wide range of linear response to gamma radiation.