The Response Difference of Mitochondria in Recalcitrant Antiaris toxicaria Axes and Orthodox Zea mays Embryos to Dehydration Injury

Long-term preservation of recalcitrant seeds is very difficult because the physiological basis on their desiccation sensitivity is poorly understood. Survival of Antiaris toxicaria axes rapidly decreased and that of immature maize embryos very slowly decreased with dehydration. To understand their different responses to dehydration, we examined the changes in mitochondria activity during dehydration. Although activities of cytochrome (Cyt) c oxidase and malate dehydrogenase of the A. toxicaria axis and maize embryo mitochondria decreased with dehydration, the parameters of maize embryo mitochondria were much higher than those of A. toxicaria, showing that the damage was more severe for the A. toxicaria axis mitochondria than for those of maize embryo. The state I and III respiration of the A. toxicaria axis mitochondria were higher than those of maize embryo, the former rapidly decreased, and the latter slowly decreased with dehydration. The proportion of Cyt c pathway to state III respiration for the A. toxicaria axis mitochondria was low and rapidly decreased with dehydration, and the proportion of alternative oxidase pathway was high and slightly increased with dehydration. In contrast, the proportion of Cyt c pathway for maize embryo mitochondria was high, and that of alternative oxidase pathway was low. Both pathways decreased slowly with dehydration.     

The Response Difference of Mitochondria in Recalcitrant Antiaris toxicaria Axes and Orthodox Zea mays Embryos to Dehydration Injury

Long-term preservation of recalcitrant seeds is very difficult because the physiological basis on their desiccation sensitivity is poorly understood. Survival of Antiaris toxicaria axes rapidly decreased and that of immature maize embryos very slowly decreased with dehydration. To understand their different responses to dehydration, we examined the changes in mitochondria activity during dehydration. Although activities of cytochrome （Cyt） c oxidase and malate dehydrogenase of the A. toxicaria axis and maize embryo mitochondria decreased with dehydration, the parameters of maize embryo mitochondria were much higher than those of A. toxicaria, showing that the damage was more severe for the A. toxicaria axis mitochondria than for those of maize embryo. The state I and III respiration of the A. toxicaria axis mitochondria were higher than those of maize embryo, the former rapidly decreased, and the latter slowly decreased with dehydration. The proportion of Cyt c pathway to state III respiration for the A. toxicaria axis mitochondria was low and rapidly decreased with dehydration, and the proportion of alternative oxidase pathway was high and slightly increased with dehydration. In contrast, the proportion of Cyt c pathway for maize embryo mitochondria was high, and that of alternative oxidase pathway was low. Both pathways decreased slowly with dehydration.     

Possible Involvement of Reactive Oxygen Species Scavenging Enzymes in Desiccation Sensitivity of Antiaris toxicaria Seeds and Axes

The relationships among desiccation sensitivities of Antiaris toxicaria seeds and axes, changes in activities of superoxide dismutase (SOD), ascorbate peroxidase (APX), catalase (CAT), glutathione reductase (GR) and dehydroascorbate reductase, (TBA)-reactive substance were studied. Desiccation tolerance of seeds and axes decreased with dehydration. Desiccation tolerance of axes was higher than that of seeds, and that of epicotyls was higher than radicles. Activities of SOD, CAT and DHAR of seeds increased during the initial phase of dehydration, and then decreased with further dehydration, whereas activities of APX and GR decreased with dehydration. These five enzyme activities of axes, however, increased during the initial phase of dehydration, and then decreased with further dehydration. The rate of superoxide radical production, and the contents of H2O2 and TBA-reactive products of seeds and axes gradually increased with dehydration. These results show that the A. toxicaria seed is a typical recalcitrant seed. Loss of desiccation tolerance in seeds and axes was correlated with activities of seeds and axes.     

见血封喉种子中的强心苷类化合物及其细胞毒活性

从海南产见血封喉(Antiaris toxicaria (Pers.) Lesch.)种子的乙醇提取物中分离得到7个强心苷类化合物,通过波谱分析(NMR, MS, IR等),鉴定其结构分别为：杠柳苷元(1)、见血封喉阿洛糖苷(2)、见血封喉去氧阿洛糖苷(3)、毒毛旋花子苷元(4)、毒毛旋花子阿洛糖苷(5)、毒毛旋花子爪哇糖苷(6)和去葡萄糖桂竹香毒苷(7)。化合物7为首次从见血封喉种子中分离得到。细胞毒活性测试结果表明,化合物1~7均对慢性髓原白血病细胞(K562)、人胃癌细胞(SGC-7901)和人肝癌细胞(SMMC-7721)的增殖有较强的生长抑制活性。     

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     

Nitric oxide enhances desiccation tolerance of recalcitrant Antiaris toxicaria seeds via protein S-nitrosylation and carbonylation

The viability of recalcitrant seeds is lost following stress from either drying or freezing. Reactive oxygen species (ROS) resulting from uncontrolled metabolic activity are likely responsible for seed sensitivity to drying. Nitric oxide (NO) and the ascorbate-glutathione cycle can be used for the detoxification of ROS, but their roles in the seed response to desiccation remain poorly understood. Here, we report that desiccation induces rapid accumulation of H(2)O(2), which blocks recalcitrant Antiaris toxicaria seed germination; however, pretreatment with NO increases the activity of antioxidant ascorbate-glutathione pathway enzymes and metabolites, diminishes H(2)O(2) production and assuages the inhibitory effects of desiccation on seed germination. Desiccation increases the protein carbonylation levels and reduces protein S-nitrosylation of these antioxidant enzymes; these effects can be reversed with NO treatment. Antioxidant protein S-nitrosylation levels can be further increased by the application of S-nitrosoglutathione reductase inhibitors, which further enhances NO-induced seed germination rates after desiccation and reduces desiccation-induced H(2)O(2) accumulation. These findings suggest that NO reinforces recalcitrant seed desiccation tolerance by regulating antioxidant enzyme activities to stabilize H(2)O(2) accumulation at an appropriate concentration. During this process, protein carbonylation and S-nitrosylation patterns are used as a specific molecular switch to control antioxidant enzyme activities.     

从见血封喉的分布看广东省热带与亚热带的界线划分

广东省的热带与亚热带的分界线按不同的划分标准,大致已有4条分界线。见血封喉(Antiaris toxicaria)是一个典型的热带种类,它在广东的分布北限,一直被认为是广东热带的最北界限。根据最近在广东发现的见血封喉4个新分布点,通过分析见血封喉的生态学特性和地理分布特征,同时参考热带界线划分的标准与原则,讨论并尝试重新界定广东省的热带与亚热带分界线。     

Harpagophytum procumbens Prevents Oxidative Stress and Loss of Cell Viability In Vitro

Harpagophytum procumbens, popularly known as devil’s claw, is a plant commonly used in the treatment of diseases of inflammatory origin. The anti-inflammatory effects of H. procumbens have been studied; however, the mechanism of action is not elucidated. It is known that excess of reactive oxygen and nitrogen species may contribute to increasing tissue damage due to inflammation. In the present study, we examined the effects of H. procumbens infusion, crude extract and fractions on lipid peroxidation (brain homogenates) induced by different pro-oxidants (Fe²⁺ or sodium nitroprusside) and the effects of ethyl acetate fraction (rich in phenolic compounds) on antioxidant defenses (catalase activity and thiol levels) and cell damage (brain cortical slices) induced by different pro-oxidants. All tested extracts of H. procumbens inhibited lipid peroxidation in a concentration-dependent manner. Furthermore, the ethyl acetate fraction had the highest antioxidant effects either by decreasing lipid peroxidation and cellular damage or restoring thiols levels and catalase activity. Taken together, our results showed that H. procumbens acts either by preventing oxidative stress or loss of cell viability. Thus, the previously reported anti-inflammatory effect of H. procumbens could also be attributed to its antioxidant activity.     

The effect of frugivory on postdispersal seed removal and germination in the pantropical forest tree Antiaris toxicaria Leschenault

The quality of seed treatment by frugivores has an effect on seed removal after dispersal, seed germination and tree recruitment. We provide information on postdispersal seed removal, germination and subsequent recruitment in tropical forest tree species Antiaris toxicaria in Ghana. We tested whether postdispersal seed removal and germination rates were differentially affected by the following seed treatments: seeds that were spat out by monkeys with all fruit pulp removed and spitting seeds with fruit pulp partially removed as observed in some birds and bats. We used seeds of intact ripened fruits as control. Frugivore seed treatment and distance from bole affected seed removal patterns, whereas intact seeds were significantly removed from all seed stations. The germination success was greater for seeds that were spat out by monkeys and poor for seeds with fruit pulp partially removed and intact fruits. More recruits were recorded at the edge of the adult A. toxicaria canopy radius. There was weak relationship (r² = 0.042) between the number of recruits and distance away from the adult tree. Results suggest that the subsequent recruitment in tropical forest tree species may be enhanced by some frugivore fruit‐handling behaviour where fruit pulp is removed from the seeds without destroying the seeds.     

小麦种子在不同保存条件下的生活力丧失特性研究

近年来种子在低温种质库和室温保存条件下的生活力丧失特性研究日益受到关注,但是与其重要性相比,目前很少见有文献报道此方面进展。本文中我们监测了在中期库、室温和人工加速老化条件下小麦种子的生活力变化,以了解种子在这些保存条件下的生活力变化特性。结果表明,三种条件下所有种子的存活曲线均呈反S形,这种特点不受保存条件的影响。随着保存温度的降低,种子生活力丧失速度降低,反S形曲线的平台期和种子寿命延长。我们采用T检验来判断平台期和种子生活力快速下降期之间的转折点（Pt）,发现小麦种子在不同条件下保存时,其Pt均十分接近,在82.2%到83.6%之间。本文讨论了将Pt以及其他生理指标作为种子保存过程中生活力快速下降的预警指标的可能性。     

Response of Chinese Wampee Axes and Maize Embryos to Dehydration at Different Rates

Survival of wampee (Clausena lansium Sksels) axes and maize (Zea mays L.) embryos decreased with rapid and slow dehydration. Damage of wampee axes by rapid dehydration was much less than by slow dehydration, and that was contrary to maize embryos. The malondialdehyde contents of wampee axes and maize embryos rapidly increased with dehydration, those of wampee axes were lower during rapid dehydration than during slow dehydration, and those of maize embryos were higher during rapid dehydration than during slow dehydration. Activities of superoxide dismutsse (SOD), ascorbate peroxidase (APX) and catalase (CAT) of wampee axes markedly increased during the sady phase of dehydration, and then rapidly decreased, and those of rapidly dehydrated axes were higher than those of slow dehydrated axes when they were dehydrated to low water contents. Activities of SOD and APX of maize embryos notable decreased with dehydration. There were higher SOD activities and lower APX activities of slowly dehydrated maize embryos compared with rapidly dehydrated maize embryos. CAT activities of maize embryos markedly increased during the eady phase of dehydration, and then decreased, and those of slowly dehydrated embryos were higher than those of rapidly dehydrated embryos during the late phase of dehydration.     

Response of Chinese Wampee Axes and Maize Embryos to Dehydration at Different Rates

Survival of wampee （Clausena lansium Skeels） axes and maize （Zea mays L.） embryos decreased with rapid and slow dehydration. Damage of wampee axes by rapid dehydration was much less than by slow dehydration, and that was contrary to maize embryos. The malondialdehyde contents of wampee axes and maize embryos rapidly increased with dehydration, those of wampee axes were lower during rapid dehydration than during slow dehydration, and those of maize embryos were higher during rapid dehydration than during slow dehydration. Activities of superoxide dismutase （SOD）, ascorbate peroxidase （APX） and catalase （CAT） of wampee axes markedly increased during the early phase of dehydration, and then rapidly decreased, and those of rapidly dehydrated axes were higher than those of slow dehydrated axes when they were dehydrated to low water contents. Activities of SOD and APX of maize embryos notable decreased with dehydration. There were higher SOD activities and lower APX activities of slowly dehydrated maize embryos compared with rapidly dehydrated maize embryos. CAT activities of maize embryos markedly increased during the early phase of dehydration, and then decreased, and those of slowly dehydrated embryos were higher than those of rapidly dehydrated embryos during the late phase of dehydration.     

箭毒木种子蛋白质样品制备及双向电泳改良方法

建立箭毒木（Antiaris toxicaria）种子总蛋白的提取方法,以及可以对其蛋白质组进行分析的双向电泳条件。通过各种条件的优化与组合,建立了以TCA-丙酮为基础的Tris—HCl提取法提取总蛋白,第1向电泳为固相pH梯度等电聚焦,第2向电泳为垂直平板SDS-PAGE的双向电泳体系。通过对样品制备、样品溶解、等电聚焦电泳、SDS-聚丙烯酰胺凝胶电泳以及染色方法等关键步骤进行分析,获得了满意的双向电泳图谱。在探索适合箭毒木种子蛋白质组学研究双向电泳方法中,比较了三氯乙酸-丙酮沉淀法、和Tris—HCl法,以及对双向电泳过程中的关键步骤的改良,认为Tris—HCl法为最适方法,所得图谱背景清晰,蛋白质信息量最大,为箭毒木属植物的差异蛋白质组学的后续研究打下了坚实的基础。     

Effects of Hydrogen Peroxide on Wound Healing in Mice in Relation to Oxidative Damage

It has been established that low concentrations of hydrogen peroxide (H₂O₂) are produced in wounds and is required for optimal healing. Yet at the same time, there is evidence that excessive oxidative damage is correlated with poor-healing wounds. In this paper, we seek to determine whether topical application of H₂O₂ can modulate wound healing and if its effects are related to oxidative damage. Using a C57BL/6 mice excision wound model, H₂O₂ was found to enhance angiogenesis and wound closure at 10 mM but retarded wound closure at 166 mM. The delay in closure was also associated with decreased connective tissue formation, increased MMP-8 and persistent neutrophil infiltration. Wounding was found to increase oxidative lipid damage, as measured by F₂-isoprostanes, and nitrative protein damage, as measured by 3-nitrotyrosine. However H₂O₂ treatment did not significantly increase oxidative and nitrative damage even at concentrations that delay wound healing. Hence the detrimental effects of H₂O₂ may not involve oxidative damage to the target molecules studied.     

Generation of Hydroxyl Radicals and Its Relation to Cellular Oxidative Damage in Plants Subjected to Water Stress

The hydroxyl radical ('OH) is one of the roost reactive mdieales known to chemistry and is believed to be a major active free radicle responsible for modifications of macmmolecules and cellular damage. Two lines of evidence strongly indicate that 'OH radicals are generated in a Fenton-type Haber-Weiss reactions in plants subjected to water stress. Firstly, water stress causes an increase in the concentration of catalytic metals, which are critical for Fenton-like reactions to proceed in vivo. Furthermore, subrmillimolar concentrations of H2O2 and ascorbic acid(or O2－ ) in the drought-stressed plants are large enough to support the Fentontype Haber-Weiss reactions. Secondly, there is oxidation of proteins and lipids in the drought-stressed plants; a process that requires a catalytic metal and that, at least for protein oxidation, is mediated by the 'OH radicals. Protein oxidation is thought to involve binding of metal ions to the proteins and subsequent site-specific attack by the 'OH radicals arising from the roetal-catalysed decomposition of H2O2. It has been proposed that protein oxidation may be a better index than lipid peroxidation because the latter fields many different products and these only appear after a lag period. The validity of malondialdehyde (MDA), an early product of lipid peroxidation, as an index of lipid peroxidation has been argued by the non-specific method of its measurement. The 'OH radicals are not the only necessary initiator for lipid peroxidation and lipid peroxidation is not usually involved in plants exposed to water stress.     

Oxidative Damage to DNA under the Action of an Alternating Magnetic Field

The content of damaged 8-hydroxy-2-deoxyguanosine nitrogenous bases in the blood DNA of healthy donors and patients with epidermolysis bullosa after exposure to an alternating magnetic field of 550 ± 30 A/m in the frequency range from 3 to 60 Hz in vitro was determined using an enzyme immunoassay. The degree of oxidative DNA damage in epidermolysis bullosa was almost twice as high as in healthy donors. It was shown that there was a significant increase in the level of 8-oxoguanine in the DNA of both groups after the magnetic field treatment, which depended in a complex way on the frequency. The resulting effect is explained by the generation of reactive oxygen species under the influence of a magnetic field and the disruption of DNA repair processes.

     

Exposure to the Proton Scavenger Glycine under Alkaline Conditions Induces Escherichia coli Viability Loss

Our previous work described a clear loss of Escherichia coli (E. coli) membrane integrity after incubation with glycine or its N-methylated derivatives N-methylglycine (sarcosine) and N,N-dimethylglycine (DMG), but not N,N,N-trimethylglycine (betaine), under alkaline stress conditions. The current study offers a thorough viability analysis, based on a combination of real-time physiological techniques, of E. coli exposed to glycine and its N-methylated derivatives at alkaline pH. Flow cytometry was applied to assess various physiological parameters such as membrane permeability, esterase activity, respiratory activity and membrane potential. ATP and inorganic phosphate concentrations were also determined. Membrane damage was confirmed through the measurement of nucleic acid leakage. Results further showed no loss of esterase or respiratory activity, while an instant and significant decrease in the ATP concentration occurred upon exposure to either glycine, sarcosine or DMG, but not betaine. There was a clear membrane hyperpolarization as well as a significant increase in cellular inorganic phosphate concentration. Based on these results, we suggest that the inability to sustain an adequate level of ATP combined with a decrease in membrane functionality leads to the loss of bacterial viability when exposed to the proton scavengers glycine, sarcosine and DMG at alkaline pH.