Protein oxidative damage and redox imbalance induced by ionising radiation in CHO cells

Reactive oxygen species (ROS) are important mediators of the cytotoxicity induced by the direct reaction of ionising radiation (IR) with all critical cellular components, such as proteins, lipids, and nucleic acids. The derived oxidative damage may propagate in exposed tissues in a dose- and spatiotemporal dependent manner to other cell compartments, affecting intracellular signalling, and cell fate. To understand how cell damage is induced, we studied the oxidative events occurring immediately after cell irradiation by analysing the fate of IR-derived ROS, the intracellular oxidative damage, and the modification of redox environment accumulating in Chinese hamster ovary (CHO) within 1?h after cell irradiation (dose range 0–10?Gy). By using the immuno-spin trapping technique (IST), spectrophotometric methods, and electron paramagnetic resonance (EPR) spectroscopy, we showed that IR-derived ROS (i) induced an IST-detectable, antioxidant-inhibitable one-electron oxidation of specific intracellular proteins; (ii) altered the glutathione (GSH) content (which was found to increase below 2?Gy, and decrease at higher doses, leading to a redox imbalance); (iii) decreased glutathione peroxidase and glutaredoxin activity; (iv) modified neither glutathione reductase nor thioredoxin reductase activity; (v) were detected by spin trapping technique, but adduct intensity decreased due to cell competition for ROS; and (vi) induced no EPR-detectable radicals assignable to oxidised cellular components. In conclusion, our results showed that IR generated an early high oxidising potential (protein radical intermediates, redox imbalance, modified redox enzyme activity) in irradiated cells potentially able to propagate the damage and induce oxidative modification of secondary targets.     

兔眼越桔花粉的超微结构和营养成分与其育性关系研究

运用透射电镜观察5个兔眼越桔品种的花粉结构并分别测定分析了花粉的营养成分,以探讨兔眼越桔花粉育性与其超微花粉结构和营养成分的关系.结果表明,兔眼越桔花粉结构中,内膜系统清晰可见,发育程度因品种而异.开花前的花粉细胞核仁较大,蛋白质合成旺盛.对花粉超微结构与营养成分间关系的比较分析表明:花粉细胞的全糖和脯氨酸含量与花粉发芽率呈正相关,兔眼越桔花粉壁的厚度、花粉细胞的蛋白质含量与花粉的萌发率无相关性,而内质网发育程度与蛋白质含量呈正相关趋势.推测兔眼越桔花粉细胞的全糖和脯氨酸含量对兔眼越桔花粉的发育及其育性具有密切的关系.     

In the right place at the right time: Parnassia resolves the herkogamy dilemma by accurate repositioning of stamens and stigmas

Background and Aims

Spatial (herkogamy) and temporal (dichogamy) separation of pollen presentation and stigma receptivity have been interpreted as reducing interference between male and female functions in hermaphroditic flowers. However, spatial separation leads to a potential conflict: reduced pollination accuracy, where pollen may be placed in a location on the pollinator different from the point of stigma contact.

Methods

To understand better how herkogamous flowers resolve this conflict, a study was made of a subalpine herb, Parnassia epunctulata, the nectariferous flowers of which exhibit sequential anther dehiscence (staggered pollen presentation) and stamen movements; usually one newly dehisced anther is positioned each day over the central gynoecium, while the older stamens bend away from the central position.

Key Results

The open flowers were visited by a variety of pollinators, most of which were flies. Seed set was pollinator-dependent (bagged flowers set almost no seeds) and pollen-limited (manual pollination increased seed set over open pollination). Analyses of adaptive accuracy showed that coordinated stamen movements and style elongation (movement herkogamy) dramatically increased pollination accuracy. Specifically, dehiscing anthers and receptive stigmas were positioned accurately in the vertical and horizontal planes in relation to the opposite sexual structure and pollinator position. By contrast, the spatial correspondence between anthers and stigma was dramatically lower before the anthers dehisced and after stamens bent outwards, as well as before and after the period of stigmatic receptivity.

Conclusions

It is shown for the first time that a combination of movement herkogamy and dichogamy can maintain high pollination accuracy in flowers with generalized pollination. Staggered pollen and stigma presentation with spatial correspondence can both reduce sexual interference and improve pollination accuracy.     

The manifold characters of orbicules: structural diversity,systematic significance,and vectors for allergens

In the anthers of flowering plants, gymnosperms, and seed ferns, tiny (±1?μm) granules might occur on the radial and innermost tangential wall of secretory tapetum cells. These sporopollenin granules develop simultaneously with the pollen exine and are called orbicules or Ubisch bodies. The present paper focuses on two quite different topics associated with orbicules.

The morphological and ultrastructural diversity of orbicules in the order Gentianales is summarized, and it is demonstrated that orbicules are a plesiomorphic feature in the order. Furthermore, orbicule characters seemed to be correlated with evolutionary trends in pollen dispersal unit and tapetum type features.

In the second part, we report on our investigation of Corylus avellana L. (Hazel) pollen, using immunogold electron microscopy to gain an insight into the possible role that orbicules may play as a vector of pollen allergens. During the pollen season orbicules are dispersed into the atmosphere along with Hazel pollen grains. The localisation of homologues of the new birch pollen allergen Bet v 7 was studied at the subcellular level in Hazel anthers. The results of this study indicate that orbicules and pollen of Hazel might act as very effective vectors for homologues of Bet v 7 and that debris of Hazel anthers represent vectors of allergens after the pollen season.     

Influence of genotype-temperature interaction on pollen performance

Pollen competition and selection have significant evolutionary consequences, but very little is known about how they can be modulated. We have examined in cherry (Prunus avium L.) how pollen performance is affected by the genotype of the pollen and by the environmental conditions under which it grows, namely the pistilar tissue and temperature. The different pollen donor genotypes tested in this work differed in their behaviour both in vitro and in vivo and this behaviour was modulated depending on the female recipient they grew on. Furthermore, there was a significant temperature-genotype interaction that affected the pollen tube population census that succeeded in reaching the base of the style. The combination of these three factors, while enabling a capacity of response to variations in environmental pressures, could maintain variability in pollen performance avoiding the fixation of the genes that control pollen tube growth rate.     

Brassinosteroids promote development of rice pollen grains and seeds by triggering expression of Carbon Starved Anther,a MYB domain protein

Transport of photoassimilates from leaf tissues (source regions) to the sink organs is essential for plant development. Here, we show that a phytohormone, the brassinosteroids (BRs) promotes pollen and seed development in rice by directly promoting expression of Carbon Starved Anther (CSA) which encodes a MYB domain protein. Over‐expression of the BR‐synthesis gene D11 or a BR‐signaling factor OsBZR1 results in higher sugar accumulation in developing anthers and seeds, as well as higher grain yield compared with control non‐transgenic plants. Conversely, knockdown of D11 or OsBZR1 expression causes defective pollen maturation and reduced seed size and weight, with less accumulation of starch in comparison with the control. Mechanically, OsBZR1 directly promotes CSA expression and CSA directly triggers expression of sugar partitioning and metabolic genes during pollen and seed development. These findings provide insight into how BRs enhance plant reproduction and grain yield in an important agricultural crop.     

Recombinant expression of Ole e 6, a Cys-enriched pollen allergen, in Pichia pastoris yeast: detection of partial oxidation of methionine by NMR

Olive pollen is one of the main causes of allergy in Mediterranean countries. Ole e 6, an olive pollen allergen, is a small (5.8 kDa) and acidic protein (pI 4.2) and no homologous proteins have been isolated or characterized so far. Ole e 6 has been efficiently expressed in the methylotrophic yeast Pichia pastoris. The cDNA encoding Ole e 6 was inserted into the plasmid vector pPIC9 and overexpressed in GS115 yeast cells. The recombinant product was purified by size-exclusion chromatography followed by reverse-phase HPLC. N-terminal sequencing, amino acid composition analysis, CD, NMR, and IgG-binding experiments were employed to characterize the purified protein. NMR data revealed the oxidation of the methionine at position 28 in approximately 50% of the recombinant protein but, although this alters its electrophoretic behavior, it did not affect folding or IgG-binding properties of rOle e 6. The recombinant form of Ole e 6 expressed in P. pastoris can be employed for structural and biochemical studies.     

Pollen morphology of Solanurn L. section Solanum

J. M. EDMONDS, 1984. Pollen morphology of Solanum L. section Solnnum . The pollen morphology of both dried and fresh, fixed material was examined using SEM. The work confirmed thc spheroidal to sub-prolate shape, the tricolporate nature and the granular surface sculpturing, typical of Solanurn pollen, but failed to demonstrate the occurrence of exine patterns which could bc of practical taxonomic use is differentiating the species belonging to the section Solanurn . Quantification of the exine sculpturing, by means of granule density counts, indicated a possible relationship between this feature and the morpho-genetic diversity of certain species.     

The systematic relationships ofAegialitis (Plumbaginaceae) as revealed by pollen morphology

Light and first SEM observations on the pollen ofAegialitis indicate that the two species of the genus are clearly distinguishable from all other genera of the tribeStaticeae to whichAegialitis presently is relegated. Intraspecific exine or aperture dimorphism which appears frequently in this tribe, is not recorded in this genus.A. annulata andA. rotundifolia have distinct monomorphic pollen and show a great resemblance in exine features with the tribePlumbagineae, particularly species ofPlumbago. Considering these and other features, separate subtribal status forAegialitis within thePlumbagineae is proposed.