Amelioration of the effects of ageing in onion seeds by osmotic priming and associated changes in oxidative metabolism

Osmotic priming of aged onion seeds with 25% polyethylene glycol-8000 for 5 d resulted in a marked increase in the rate of germination and early seedling growth. Priming reduced electrolyte leakage as well as lipid peroxidation in seeds implying the activation of membrane repair processes. Priming was also associated with increased levels of antioxidants,i.e. ascorbic acid and tocopherols particularly the latter and the activities of catalase and peroxidase involved in the mitigation of oxidative damage. In comparison with the priming of unaged seeds, the aged seeds experienced a diminution of response in terms of changes in the levels of antioxidants and scavenging enzymes.     

Effects of osmotic priming and ageing on onion seed germination

Onion seeds were osmotically primed in polyethylene glycol solution (342 g/kg water) either for 14 days before accelerated ageing at 40°C. 18% m.c. for 0, 24, 48, 72 or 96 h, or for 10, 14 or 17 days after ageing. Priming improved the rate of germination compared with non-primed seed. Priming before ageing delayed the loss of viability due to ageing, but priming after ageing had no effect on viability. Primed and dried onion seed was stored for 18 months at 10°C, 9% m.c. with no effect on viability; improvements in germination rate due to priming were maintained over the storage period. Conductivity measurements of seed leachates were not a consistently reliable indicator of germination performance.     

Effect of post-irradiation ageing on onion seeds

Seeds of onion (Allium cepa) cv. Ailsa Craig were exposed to various doses of gamma radiation (0, 10, 20, 40, 80 and 100 krad) and subjected to accelerated ageing (RH 100%, 42°C) for 12 h. Radiation doses significantly affected the percentage of normal seedlings, abnormality types (%) and seedling growth. Seed viability, conductivities of seed leachates, final germination (%), germination speed and time to reach 50% germination (T ₅₀) were not affected by the seed irradiation. Accelerated ageing after irradiation had significant influence on seed viability, conductivities of seed leachates, final germination (%) and percentage of normal seedlings. Germination speed, T ₅₀ and seedling growth were not affected by the accelerated ageing. However, all the parameters studied were significantly influenced by the interaction of radiation doses and accelerated ageing. Accelerated ageing generally enhanced the damaging effects of irradiation on seeds. Therefore, it was concluded that onion seeds should not be exposed to adverse environmental conditions after irradiation.     

Improving germination and vigour of aged and stored onion seeds by matriconditioning

Germination and vigour of accelerated aged (AA) and naturally stored onion seeds were examined. Accelerated ageing was conducted at 40 °C and 100 % relative humidity (RH). Non aged seeds were stored for 34 months at 3 or 15 °C and 40, 60 or 90 % RH. To restore seed viability, stored and aged seeds were matriconditioned with Micro-Cel E. A distinct loss of germination was observed after 5 days of accelerated ageing. Naturally stored seeds maintained high viability for 34 months, when stored at 3 °C and 40, 60 and 90 % RH or at 15 °C and 40 %. An increase of RH to 60 and 90 % at 15 °C caused loss of germination and vigour. Matriconditioning improved germination and increased endogenic ethylene release and in vivo ACC oxidase activity of both aged and stored seeds.     

Changes in endogenous cytokinins of celery (Apium graveolens L.) seeds following an osmotic priming or growth regulator soak treatment

Celery seeds were less thermoinhibited when dried back after a seed soak treatment with the gibberellins A₄ and A₇ (GA_4/7) plus ethephon (G+E) or an osmotic priming treatment in the light with polyethylene glycol (PEG). At temperatures between 18 and 25° in the dark, 50 percent of the PEG-treated seeds germinated after 3 days whereas G+E-treated seeds required 7 days and untreated seeds did not germinate at all.Irrespective of treatment, dry control and dried-back, treated seeds contained very little detectable cytokinin activity. However, when such seeds were imbibed for 18 h in the dark and then analysed immediately with the soybean callus bioassay, less cytokinin activity was detected in both G+E and PEG seeds than in the untreated seeds. In particular, cytokinins with the HPLC properties of zeatin and its riboside were decreased in G+E seeds and virtually absent from PEG seeds. Conversely, extracts from PEG and to a lesser extent G+E seeds contained activity which chromatographically resembled cytokinin glucosides whereas this was absent from untreated seeds.     

Molecular basis of peptide recognition by the TCR: affinity differences calculated using large scale computing

Free energy calculations of the wild-type and the variant human T cell lymphotropic virus type 1 Tax peptide presented by the MHC to the TCR have been performed using large scale massively parallel molecular dynamics simulations. The computed free energy difference (-1.86 +/- 0.44 kcal/mol) using alchemical mutation-based thermodynamic integration agrees well with experimental data (-2.9 +/- 0.2 kcal/mol). Our simulations exploit state-of-the-art hardware and codes whose algorithms have been optimized for supercomputing platforms. This enables us to simulate larger, more realistic biological systems for longer durations without the imposition of artificial constraints.     

Fast large scale oligonucleotide selection using the longest common factor approach

We present a fast method that selects oligonucleotide probes (such as DNA 25-mers) for microarray experiments on a truly large scale. For example, reliable oligos for human genes can be found within four days, a speedup of one to two orders of magnitude compared to previous approaches. This speed is attained by using the longest common substring as a specificity measure for candidate oligos. We present a space- and time-efficient algorithm, based on a suffix array with additional information, to compute matching statistics (lengths of longest matches) between all candidate oligos and all remaining sequences. With the matching statistics available, we show how to incorporate constraints such as oligo length, melting temperature, and self-complementarity into the selection process at a postprocessing stage. As a result, we can now design custom oligos for any sequenced genome, just as the technology for on-site chip synthesis is becoming increasingly mature.     

Reliable computational design of biological-inorganic materials to the large nanometer scale using Interface-FF

Abstract

The function of nanomaterials and biomaterials greatly depends on understanding nanoscale recognition mechanisms, crystal growth and surface reactions. The Interface Force Field (IFF) and surface model database are the first collection of transferable parameters for inorganic and organic compounds that can be universally applied to all materials. IFF uses common energy expressions and achieves best accuracy among classical force fields due to rigorous validation of structural and energetic properties of all compounds in comparison to perpetually valid experimental data. This paper summarises key aspects of parameterisation, including atomic charges and transferability of parameters and current coverage. Examples of biomolecular recognition at metal and mineral interfaces, surface reactions of alloys, as well as new models for graphitic materials and pi-conjugated molecules are described. For several metal–organic interfaces, a match in accuracy of computed binding energies between of IFF and DFT results is demonstrated at ten million times lower computational cost. Predictive simulations of biomolecular recognition of peptides on phosphate and silicate surfaces are described as a function of pH. The use of IFF for reactive molecular dynamics is illustrated for the oxidation of Mo₃Si alloys at high temperature, showing the development of specific porous silica protective layers. The introduction of virtual pi electrons in graphite and pi-conjugated molecules enables improvements in property predictions by orders of magnitude. The inclusion of such molecule-internal polarity in IFF can reproduce cation–pi interactions, pi-stacking in graphite, DNA bases, organic semiconductors and the dynamics of aqueous and biological interfaces for the first time.     

Solving the coastal eutrophication problem by large scale seaweed cultivation

Eutrophication is becoming a serious problem in coastal waters in many parts of the world. It induces the phytoplankton blooms including `Red Tides', followed by heavy economic losses to extensive aquaculture area. Some cultivated seaweeds have very high productivity and could absorb large quantities of N, P, CO₂, produce large amount of O₂ and have excellent effect on decreasing eutrophication. The author believes that seaweed cultivation in large scale should be a good solution to the eutrophication problem in coastal waters. To put this idea into practice, four conditions should be fulfilled: (a) Large-scale cultivation could be conducted within the region experiencing eutrophication. (b) Fundamental scientific and technological problems for cultivation should have been solved. (c) Cultivation should not impose any harmful ecological effects. (d) Cultivation must be economically feasible and profitable. In northern China, large-scale cultivation of Laminaria japonica Aresch. has been encouraged for years to balance the negative effects from scallop cultivation. Preliminary research in recent years has shown that Gracilaria lemaneiformis (Bory) Daws. and Porphyra haitanensis Chang et Zheng are the two best candidates for this purpose along the Chinese southeast to southern coast from Fujian to Guangdong, Guangxi and Hong Kong. Gracilaria tenuistipitata var. liui Chang et Xia is promising for use in pond culture condition with shrimps and fish.     

Free radical scavenging as affected by accelerated ageing and subsequent priming in sunflower seeds

Lipid peroxidation resulting from loss of free radical scavenging is thought to be involved in deterioration of sunflower (Helianthus annuus L.) seeds during accelerated ageing. In other respects, presoaking of seeds in a solution of low water potential (osmopriming) has been demonstrated to reinvigorate aged seeds. The aim of the present work was to study the effect of osmopriming on the germination of aged sunflower seeds and to investigate whether this effect was associated with the restoration of antioxidant defence systems. Seeds were aged for 5 days at 45°C and 100% relative humidity and then primed for various durations up to 7 days at 15°C in a solution of polyethylene glycol 6000 at ?2 MPa. Lipid peroxidation was estimated by measuring malondialdehyde (MDA) and conjugated diene contents, and the activities of superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), ascorbate peroxidase (APX) and dehydroascorbate reductase (DHAR) were measured throughout the treatments. Accelerated ageing resulted in a marked decrease in the germination rate, and was associated with an increase in the levels of MDA and conjugated dienes, thus indicating lipid peroxidation. Ageing was also characterized by a decrease in the activities of CAT and GR. The activities of SOD and DHAR were much less altered. No APX activity was detected whatever the seed treatment. Priming of aged seeds progressively restored the initial germinative ability and resulted in a marked decrease in the levels of MDA and conjugated dienes, indicating a fall in lipid peroxidation processes. These effects of priming were also well correlated to the recovery of SOD, CAT and GR activities. Priming treatment for 7 days led to full restoration of the cell detoxifying mechanisms which were strongly altered during ageing. Glutathione content showed the same changes as GR activity. There existed a clear-cut relationship between seed germinative energy, expressed as the germination rate, and the efficiency of free radical scavenging systems, in particular CAT and GR activities and glutathione content. The results suggest that the antioxidant defence systems might play a key role in seed vigour.     

Vesicle formation in the membrane of onion cells (Allium cepa) during rapid osmotic dehydration

Background and Aims

Optimization of osmotic dehydration in different plant cells has been investigated through the variation of parameters such as the nature of the sugar used, the concentration of osmotic solutions and the processing time. In micro-organisms such as the yeast, Saccharomyces cerevisiae, the exposure of a cell to a slow increase in osmotic pressure preserves cell viability after rehydration, while sudden dehydration involves a lower rate of cell viability, which could be due to membrane vesiculation. The aim of this work is to study cytoplasmic vesicle formation in onion epidermal cells (Allium cepa) as a function of the kinetics of osmotic pressure variation in the external medium.

Methods

Onion epidermal cells were submitted either to an osmotic shock or to a progressive osmotic shift from an osmotic pressure of 2 to 24 MPa to induce plasmolysis. After 30 min in the treatment solution, deplasmolysis was carried out. Cells were observed by microscopy during the whole cycle of dehydration–rehydration.

Key Results

The application of an osmotic shock to onion cells, from an initial osmotic pressure of 2 MPa to a final one of 24 MPa for <1 s, led to the formation of numerous exocytotic and osmocytic vesicles visualized through light and confocal microscopy. In contrast, after application of a progressive osmotic shift, from an initial osmotic pressure of 2 MPa to a final one of 24 MPa for 30 min, no vesicles were observed. Additionally, the absence of Hechtian strand connections led to the bursting of vesicles in the case of the osmotic shock.

Conclusions

It is concluded that the kinetics of osmotic dehydration strongly influence vesicle formation in onion cells, and that Hechtian strand connections between protoplasts and exocytotic vesicles are a prerequisite for successful deplasmolysis. These results suggest that a decrease in the area-to-volume ratio of a cell could cause cell death following an osmotic shock.     

The osmotic response of large unilamellar vesicles studied by quasielastic light scattering

Large unilamellar vesicles of two phosphatidylcholines, one saturated (DMPC) and the other unsaturated (DOPC), prepared by the reverse-phase evaporation method were studied using the quasielastic light scattering technique. The accurate sizing obtained by this technique showed an osmotic response for the two kinds of vesicles when the salinity of the external medium was diluted. The elastic moduli of lipid vesicles bilayers in the liquid phase were then estimated according to the elasticity theory of spherical shells taking into account salt leakage data known from the literature.     

Structural characterization of genomes by large scale sequence-structure threading

Background  

Using sequence-structure threading we have conducted structural characterization of complete proteomes of 37 archaeal, bacterial and eukaryotic organisms (including worm, fly, mouse and human) totaling 167,888 genes.     

Mass propagation of shoots of Stevia rebaudiana using a large scale bioreactor

A procedure for the mass propagation of multiple shoots of Stevia rebaudiana is described. Isolated shoot primordia were used as the inoculum to obtain clusters of shoot primordia. Such clusters were grown in a 500 liter bioreactor to obtain shoots. A total of 64.6 Kg of shoots were propagated from 460 g of the inoculated shoot primordia. These shoots were easily acclimatized in soil.     

Fluctuation of oxidative stress indicators in Salix nigra seeds during priming

Salix nigra seeds subjected to increased humidification show a decrease in normal germination (NG) during early imbibition followed by a recovery in that parameter at increasing imbibition times. Since photo-oxidized seeds contain high levels of reactive oxygen species (ROS), it is possible to infer that the atypical decrease in NG is a consequence of a higher ROS mobilization at early imbibition and the subsequent recovery from an increase in antioxidant activity. In this study, several oxidative stress indicators were evaluated in photo-oxidized seeds subjected to priming. ROS production was studied using electronic spin resonance spectroscopy, spontaneous chemiluminescence (SCL), spectrophotometry (with XTT), and histochemical (with DAB and NBT) and cytochemical (with CeCl(3)) techniques. Four indicators of molecular damage were monitored: lipid peroxidation, pigment destruction, protein oxidation, and membrane integrity. Antioxidant activity was evaluated by changes in the enzymes SOD, CAT, APX, and POX. The results revealed that the decrease in NG at the beginning of priming occurs by an oxidative burst, as determined by increases in both SCL and superoxide anion radical (O2(·-)) Such oxidative burst generates lipid peroxidation, protein oxidation, and a decrease in both pigment content and enzyme activities. With increasing hydration, damages are progressively reversed and NG restored, which coincides with the increased activity of antioxidant defences. It is proposed that these novel observations regarding the occurrence of an oxidative burst are related to the high basal ROS levels and the high membrane content retained in the mature embryo tissues.     

Assessment of river condition at a large spatial scale using predictive models

1. RIVPACS-type predictive models were developed at a relatively large spatial scale for the Australian state called New South Wales (NSW, 801 428 km²). Aquatic macroinvertebrate samples and physical and chemical data were collected from 250 reference sites (little affected by human activities) and 23 test sites (with known human impacts) throughout NSW in autumn and spring 1995 and identified mostly to family level. Reference sites were grouped based on their macroinvertebrate data using classification (UPGMA) and ordination techniques. Relationships between macroinvertebrate and environmental data were established using principal axis correlations and stepwise multiple discriminant function analysis. models for predicting invertebrate assemblages were developed separately for edge and riffle habitats for autumn and spring data sets and for combined autumn and spring data sets. 2. Sites in the lowland sections of the western flowing rivers were characterized by low taxonomic richness and were distinct from the sites in the eastern part of the state. Sites on the western slopes of the Great Dividing Range in southern and northern NSW mostly fell into separate groups. In eastern NSW, site groups did not represent a north, central and south division. Sites on highland streams, coastal fringe streams and large rivers mostly formed distinct groups, but most of the sites on east-flowing rivers fell into large site groups that did not have clear geographic boundaries. 3. Environmental variables that were strongly correlated with ordinations of macroinvertebrate presence/absence at sites were water temperature, altitude, longitude and maximum distance from source. The predictor variables determined by DFA for the six models created included alkalinity, altitude, location (longitude and/or latitude), stream size and substratum composition. These are generally in common with the variables determined for other large geographic areas in Australia and the United Kingdom. 4. Model outputs from reference sites suggest that, among the six models, the riffle model combining autumn and spring is likely to give the most reliable predictions. The combined edge model also performed well but refinements are needed for single season models to provide reliable outputs. 5. Combined season models both for riffles and for edges detected biological impairment at all but one of the test sites. Single season riffle models also detected impairment while single season edge models characterized sites as unimpaired despite other models’ indications of impaired fauna. Riffle models may be more sensitive than edge models but the sampling of riffles is often limited by flow. Edge habitats are available at most sites but there may be few riffles in floodplain rivers. Available resources, desired model sensitivity, and river type should be considered jointly to determine the most useful habitat to sample.     

On the dynamics of flying insects populations controlled by large scale information

We present a diffusion model adapted to an operational control of flying insects. The operational objective entails working with non-homogeneous regions and the corresponding non-uniform diffusion tensors. It the becomes necessary to study the problem of scale in order to justify the diffusion equation. We show that in typical cases, the spatial resolution must be coarser than 20 m and we have specified the appropriated time scales. We propose a method for computing the tensors at the given resolution, in relation to the landscape and the insects' species and a method for the numerical solution of the equations and present two applications.