长期低浓度SO2对大豆生长和产量的影响

随着化石燃料消耗量不断增加,由此产生的主要大气污染物之一SO_2的浓度和影响范围也日趋增大。SO_2对植物,特别是对农作物的影响已受到普遍重视。本研究选择我国北方种植面积大、分布广的大豆为供试作物,在野外开顶式熏气装置中进行低浓度SO_2长期暴露试验,观察SO_2对大豆生长发育及产量的影响,以期为制订农田大气环境质量标准提供有一定参考价值的生态学基准。     

Rising ozone concentrations decrease soybean evapotranspiration and water use efficiency whilst increasing canopy temperature

? Here, we investigated the effects of increasing concentrations of ozone ([O(3)]) on soybean canopy-scale fluxes of heat and water vapor, as well as water use efficiency (WUE), at the Soybean Free Air Concentration Enrichment (SoyFACE) facility. ? Micrometeorological measurements were made to determine the net radiation (R(n)), sensible heat flux (H), soil heat flux (G(0)) and latent heat flux (λET) of a commercial soybean (Glycine max) cultivar (Pioneer 93B15), exposed to a gradient of eight daytime average ozone concentrations ranging from approximately current (c. 40 ppb) to three times current (c. 120 ppb) levels. ? As [O(3)] increased, soybean canopy fluxes of λET decreased and H increased, whereas R(n) and G(0) were not altered significantly. Exposure to increased [O(3)] also resulted in warmer canopies, especially during the day. The lower λET decreased season total evapotranspiration (ET) by c. 26%. The [O(3)]-induced relative decline in ET was half that of the relative decline in seed yield, driving a 50% reduction in seasonal WUE. ? These results suggest that rising [O(3)] will alter the canopy energy fluxes that drive regional climate and hydrology, and have a negative impact on productivity and WUE, key ecosystem services.     

Increased protein carbonylation in leaves of Arabidopsis and soybean in response to elevated [CO2]

While exposure of C3 plants to elevated [CO2] would be expected to reduce production of reactive oxygen species (ROS) in leaves because of reduced photorespiratory metabolism, results obtained in the present study suggest that exposure of plants to elevated [CO2] can result in increased oxidative stress. First, in Arabidopsis and soybean, leaf protein carbonylation, a marker of oxidative stress, was often increased when plants were exposed to elevated [CO2]. In soybean, increased carbonyl content was often associated with loss of leaf chlorophyll and reduced enhancement of leaf photosynthetic rate (Pn) by elevated [CO2]. Second, two-dimensional (2-DE) difference gel electrophoresis (DIGE) analysis of proteins extracted from leaves of soybean plants grown at elevated [CO2] or [O3] revealed that both treatments altered the abundance of a similar subset of proteins, consistent with the idea that both conditions may involve an oxidative stress. The 2-DE analysis of leaf proteins was facilitated by a novel and simple procedure to remove ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) from soluble soybean leaf extracts. Collectively, these findings add a new dimension to our understanding of global change biology and raise the possibility that oxidative signals can be an unexpected component of plant response to elevated [CO2].     

Perturbations of amino acid metabolism associated with glyphosate-dependent inhibition of shikimic acid metabolism affect cellular redox homeostasis and alter the abundance of proteins involved in photosynthesis and photorespiration

The herbicide glyphosate inhibits the shikimate pathway of the synthesis of amino acids such as phenylalanine, tyrosine, and tryptophan. However, much uncertainty remains concerning precisely how glyphosate kills plants or affects cellular redox homeostasis and related processes in glyphosate-sensitive and glyphosate-resistant crop plants. To address this issue, we performed an integrated study of photosynthesis, leaf proteomes, amino acid profiles, and redox profiles in the glyphosate-sensitive soybean (Glycine max) genotype PAN809 and glyphosate-resistant Roundup Ready Soybean (RRS). RRS leaves accumulated much more glyphosate than the sensitive line but showed relatively few changes in amino acid metabolism. Photosynthesis was unaffected by glyphosate in RRS leaves, but decreased abundance of photosynthesis/photorespiratory pathway proteins was observed together with oxidation of major redox pools. While treatment of a sensitive genotype with glyphosate rapidly inhibited photosynthesis and triggered the appearance of a nitrogen-rich amino acid profile, there was no evidence of oxidation of the redox pools. There was, however, an increase in starvation-associated and defense proteins. We conclude that glyphosate-dependent inhibition of soybean leaf metabolism leads to the induction of defense proteins without sustained oxidation. Conversely, the accumulation of high levels of glyphosate in RRS enhances cellular oxidation, possibly through mechanisms involving stimulation of the photorespiratory pathway.     

20S proteasome and accumulation of oxidized and ubiquitinated proteins in maize leaves subjected to cadmium stress

In order to examine the possible involvement of the 20S proteasome in degradation of oxidized proteins, the effects of different cadmium concentrations on its activities, protein abundance and oxidation level were studied using maize (Zea mays L.) leaf segments. The accumulation of carbonylated and ubiquitinated proteins was also investigated. Treatment with 50 microM CdCl(2) increased both trypsin- and PGPH-like activities of the 20S proteasome. The incremental changes in 20S proteasome activities were probably caused by an increased level of 20S proteasome oxidation, with this being responsible for degradation of the oxidized proteins. When leaf segments were treated with 100 microM CdCl(2), the chymotrysin- and trypsin-like activities of the 20S proteasome also decreased, with a concomitant increase in accumulation of carbonylated and ubiquitinated proteins. With both Cd(2+) concentrations, the abundance of the 20S proteasome protein remained similar to the control experiments. These results provide evidence for the involvement of this proteolytic system in cadmium-stressed plants.     

Quantitative analysis of redox-sensitive proteome with DIGE and ICAT

Oxidative modifications of protein thiols are important mechanisms for regulating protein functions. The present study aimed to compare the relative effectiveness of two thiol-specific quantitative proteomic techniques, difference gel electrophoresis (DIGE) and isotope coded affinity tag (ICAT), for the discovery of redox-sensitive proteins in heart tissues. We found that these two methods were largely complementary; each could be used to reveal a set of unique redox-sensitive proteins. Some of these proteins are low-abundant signaling proteins and membrane proteins. From DIGE analysis, we found that both NF-kappaB-repressing protein and epoxide hydrolase were sensitive to H 2O 2 oxidation. In ICAT analysis, we found that specific cysteines within sacroplasmic endoplamic reticulum calcium ATPase 2 and voltage-dependent anion-selective channel protein 1 were sensitive to H 2O 2 oxidation. From these analyses, we conclude that both methods should be employed for proteome-wide studies, to maximize the possibility of identifying proteins containing redox-sensitive cysteinyl thiols in complex biological systems.     

Chronic vs. short-term acute O3 exposure effects on nocturnal transpiration in two Californian oaks

We tested the effect of daytime chronic moderate ozone (O3) exposure, short-term acute exposure, and both chronic and acute O3 exposure combined on nocturnal transpiration in California black oak and blue oak seedlings. Chronic O3 exposure (70 ppb for 8 h/day) was implemented in open-top chambers for either 1 month (California black oak) or 2 months (blue oak). Acute O3 exposure (approximately 1 h in duration during the day, 120-220 ppb) was implemented in a novel gas exchange system that supplied and maintained known O3 concentrations to a leaf cuvette. When exposed to chronic daytime O3 exposure, both oaks exhibited increased nocturnal transpiration (without concurrent O3 exposure) relative to unexposed control leaves (1.8x and 1.6x, black and blue oak, respectively). Short-term acute and chronic O3 exposure did not further increase nocturnal transpiration in either species. In blue oak previously unexposed to O3, short-term acute O3 exposure significantly enhanced nocturnal transpiration (2.0x) relative to leaves unexposed to O3. California black oak was unresponsive to (only) short-term acute O3 exposure. Daytime chronic and/or acute O3 exposures can increase foliar water loss at night in deciduous oak seedlings.     

Ozone-Induced Alterations in the Accumulation of Newly Synthesized Proteins in Leaves of Maize

We examined the response of leaves of 3-week-old maize (Zea mays L.) to short-term (5 h) fumigation with O3-enriched air (0, 0.12, 0.24, or 0.36 [mu]L/L). Older leaves and leaf tissue developed more severe visible damage at higher external O3 concentrations. To investigate the immediate effect of O3 exposure on the accumulation of newly synthesized leaf proteins, leaves were labeled with [35S]methionine after 2 h and fumigated for an additional 3 h. O3-induced alterations of leaf proteins were observed in a concentration-dependent manner. There was a significant decrease in [35S]methionine incorporation into protein at the highest O3 concentration. Developmental differences in accumulation of de novo-synthesized leaf proteins were observed when the leaf tip, middle, and basal sections were labeled under 0 [mu]L/L O3, and additional changes were apparent upon exposure to increasing O3 concentrations. Changes in leaf protein synthesis were observed in the absence of visible leaf injury. Subcellular fractionation revealed O3-induced alterations in soluble and membrane-associated proteins. A number of thylakoid membrane-associated proteins showed specific increases in response to O3 fumigation. In contrast, the synthesis of a 32-kD polypeptide associated with thylakoid membranes was reduced in response to O3 fumigation in parallel with reduced incorporation of [35S]methionine into protein. Immunoprecipitation identified this polypeptide as the D1 protein of photosystem II. A reduction in the accumulation of newly synthesized D1 could have consequences for the efficiency of photosynthesis and other cellular processes.     

Comparative proteomics analysis reveals an intimate protein network provoked by hydrogen peroxide stress in rice seedling leaves

Hydrogen peroxide (H2O2) plays a dual role in plants as the toxic by-product of normal cell metabolism and as a regulatory molecule in stress perception and signal transduction. However, a clear inventory as to how this dual function is regulated in plants is far from complete. In particular, how plants maintain survival under oxidative stress via adjustments of the intercellular metabolic network and antioxidative system is largely unknown. To investigate the responses of rice seedlings to H2O2 stress, changes in protein expression were analyzed using a comparative proteomics approach. Treatments with different concentrations of H2O2 for 6 h on 12-day-old rice seedlings resulted in several stressful phenotypes such as rolling leaves, decreased photosynthetic and photorespiratory rates, and elevated H2O2 accumulation. Analysis of approximately 2000 protein spots on each two-dimensional electrophoresis gel revealed 144 differentially expressed proteins. Of them, 65 protein spots were up-regulated, and 79 were down-regulated under at least one of the H2O2 treatment concentrations. Furthermore 129 differentially expressed protein spots were identified by mass spectrometry to match 89 diverse protein species. These identified proteins are involved in different cellular responses and metabolic processes with obvious functional tendencies toward cell defense, redox homeostasis, signal transduction, protein synthesis and degradation, photosynthesis and photorespiration, and carbohydrate/energy metabolism, indicating a good correlation between oxidative stress-responsive proteins and leaf physiological changes. The abundance changes of these proteins, together with their putative functions and participation in physiological reactions, produce an oxidative stress-responsive network at the protein level in H2O2-treated rice seedling leaves. Such a protein network allows us to further understand the possible management strategy of cellular activities occurring in the H2O2-treated rice seedling leaves and provides new insights into oxidative stress responses in plants.     

Interactive effects of ozone and climate on tree growth and water use in a southern Appalachian forest in the USA

* A lack of data on responses of mature tree growth and water use to ambient ozone (O(3)) concentrations has been a major limitation in efforts to understand and model responses of forests to current and future changes in climate. * Here, hourly to seasonal patterns of stem growth and sap flow velocity were examined in mature trees from a mixed deciduous forest in eastern Tennessee (USA) to evaluate the effects of variations in ambient O(3) exposure and climate on patterns of stem growth and water use. * Ambient O(3) caused a periodic slowdown in seasonal growth patterns that was attributable in part to amplification of diurnal patterns of water loss in tree stems. This response was mediated by statistically significant increases in O(3)-induced daily sap flow and led to seasonal losses in stem growth of 30-50% for most species in a high-O(3) year. * Decreased growth and increased water use of mature forest trees under episodically high ambient O(3) concentrations suggest that O(3) will amplify the adverse effects of increasing temperatures on forest growth and forest hydrology.     

Root and shoot gas exchange respond additively to moderate ozone and methyl jasmonate without induction of ethylene: ethylene is induced at higher O3 concentrations

The available literature is conflicting on the potential protection of plants against ozone (O(3)) injury by exogenous jasmonates, including methyl jasmonate (MeJA). Protective antagonistic interactions of O(3) and MeJA have been observed in some systems and purely additive effects in others. Here it is shown that chronic exposure to low to moderate O(3) concentrations (4-114 ppb; 12 h mean) and to MeJA induced additive reductions in carbon assimilation (A (n)) and root respiration (R (r)), and in calculated whole plant carbon balance. Neither this chronic O(3) regime nor MeJA induced emission of ethylene (ET) from the youngest fully expanded leaves. ET emission was induced by acute 3 h pulse exposure to much higher O(3) concentrations (685 ppb). ET emission was further enhanced in plants treated with MeJA. Responses of growth, allocation, photosynthesis, and respiration to moderate O(3) concentrations and to MeJA appear to be independent and additive, and not associated with emission of ET. These results suggest that responses of Pima cotton to environmentally relevant O(3) are not mediated by signalling pathways associated with ET and MeJA, though these pathways are inducible in this species and exhibit a synergistic O(3)×MeJA interaction at very high O(3) concentrations.     

Proteomic analysis of early-responsive redox-sensitive proteins in Arabidopsis

Regulation of protein function through oxidative modification has emerged as an important molecular mechanism modulating various biological processes. Here, we report a proteomic study of redox-sensitive proteins in Arabidopsis cells subjected to H(2)O(2) treatment. Four gel-based approaches were employed, leading to the identification of four partially overlapping sets of proteins whose thiols underwent oxidative modification in the H(2)O(2)-treated cells. Using a method based on differential labeling of thiols followed by immunoprecipitation and Western blotting, five of the six selected putative redox-sensitive proteins were confirmed to undergo oxidative modification following the oxidant treatment in Arabidopsis leaves. Another method, which is based on differential labeling of thiols coupled with protein electrophoretic mobility shift assay, was adopted to reveal that one of the H(2)O(2)-sensitive proteins, a homologue of cytokine-induced apoptosis inhibitor 1 (AtCIAPIN1), also underwent oxidative modification in Arabidopsis leaves after treatments with salicylic acid or the peptide elicitor flg22, two inducers of defense signaling. The redox-sensitive proteins identified from the proteomic study are involved in various biological processes such as metabolism, the antioxidant system, protein biosynthesis and processing, and cytoskeleton organization. The identification of novel redox-sensitive proteins will be helpful toward understanding of cellular components or pathways previously unknown to be redox-regulated.     

Induction of stress proteins and MMP-9 by 0.8 ppm of ozone in murine skin

Ozone (O(3)) is among the most reactive environmental oxidant pollutants to which cutaneous tissues are exposed. O(3) exposure has been shown to induce antioxidant depletion as well as the oxidation of lipids and proteins within the outermost skin layer, the stratum corneum. However, relatively little is known regarding the potential effects of O(3) on the cellular constituents of the underlying skin epidermis and dermis. In the present study, hairless mice exposed for 6 h to 0.8 ppm O(3) showed increases in lipid peroxidation, as quantitated by increases in 4-hydroxynonenal-protein adducts. O(3) exposure caused an induction of the stress proteins HSP27 and heme oxygenase-1 (HO-1), starting at 6 h and increasing up to 18 h after O(3) exposure. This was accompanied by an increase in matrix metalloproteinase-9 (MMP-9) mRNA and activity levels, indicative of possible injurious-reparative processes. Collectively, our data demonstrate that skin exposure to O(3) not only affects antioxidant levels and oxidation markers in the outermost stratum corneum layer, but also induces cellular stress responses in the deeper cellular layers of the skin.     

Bottom water oxygenation history in southeastern Arabian Sea during the past 140 ka: Results from redox-sensitive elements

The concentrations of multiple redox-sensitive elements such as Re, U, Mo, Cd, V, Sb, and Tl were determined in sediments from the southeastern Arabian Sea (9°21′N: 71°59′E) to understand the bottom water oxygenation history throughout the past 140 ka. The enrichment of redox-sensitive elements (Re, U, Cd and Sb) above average crustal abundances suggests that the Last Glacial Maxima (17.48 ka), stadials of Marine Isotope Stage (MIS)-5 (5b and 5d) and Glacial Termination (GT)-II (133 ka) were associated with suboxic bottom water conditions. Sediments deposited during these suboxic conditions show the highest Re content (up to 54 ppb normalized to a carbonate free basis) which is highly enriched over average continental crust (0.4 ppb) and these sediments appear to be the major sink for the global mass balance estimation. Marine Isotope Stages 1, 3, 4 and interstadials of MIS-5 (5a, 5c & 5e) were all associated with near-oxic conditions. Overall, the lack of enrichment of Mo and V above crustal abundance, and a high Re/Mo (ppm/ppm) ratio (avg. 18.2 × 10^− 3) suggest that sediments of the southeastern Arabian Sea never contained free H₂S during the last 140 ka. These changes in the bottom water oxygen content can be related to the oceanic circulation pattern during this time and in part are reflected in relationships between the timing of redox changes and paleoproductivity proxies.     

Supplemental ultraviolet-B and ozone: impact on antioxidants, proteome and genome of linseed (Linum usitatissimum L. cv. Padmini)

The present investigation used Linum usitatissimum L. cv. Padmini (linseed), under field conditions in open-top chambers, to evaluate the interactive effects of supplemental ultraviolet-B (sUV-B; ambient +7.2 kJ · m(-2) · d(-1)) and ozone (O(3); ambient +10 ppb). Treatment of plants with sUV-B and O(3) , individually or in combination, caused several changes in enzymatic and non-enzymatic components of the antioxidant defence system. Photo-oxidative damage caused by sUV-B and O(3) , included lipid peroxidation, changed protein profiles and caused DNA strand breakage. One-dimensional gel electrophoresis revealed that proteins of 222.24 and 50.5 kDa are specific and appear after sUV-B and O(3) exposure, and could be used as indicator proteins. Effects of sUV-B and O(3) given separately are more detrimental as compared to combined treatment. Mutational and structural alterations in linseed DNA after these stresses were also examined using RAPD with ten different primers. The study concluded that both stresses, i.e. sUV-B and O(3) , are phytotoxic, causing significant changes in metabolites, antioxidants, the leaf proteome and the genome of linseed, but their interactive effect was always less than additive.     

Regulation of growth response to water stress in the soybean primary root. I. Proteomic analysis reveals region‐specific regulation of phenylpropanoid metabolism and control of free iron in the elongation zone

In water‐stressed soybean primary roots, elongation was maintained at well‐watered rates in the apical 4 mm (region 1), but was progressively inhibited in the 4–8 mm region (region 2), which exhibits maximum elongation in well‐watered roots. These responses are similar to previous results for the maize primary root. To understand these responses in soybean, spatial profiles of soluble protein composition were analysed. Among the changes, the results indicate that region‐specific regulation of phenylpropanoid metabolism may contribute to the distinct growth responses in the different regions. Several enzymes related to isoflavonoid biosynthesis increased in abundance in region 1, correlating with a substantial increase of isoflavonoid content in this region which could contribute to growth maintenance via various potential mechanisms. In contrast, caffeoyl‐CoA O‐methyltransferase, which is involved in lignin synthesis, was highly up‐regulated in region 2. This response was associated with enhanced accumulation of lignin, which may be related to the inhibition of growth in this region. Several proteins that increased in abundance in both regions of water‐stressed roots were related to protection from oxidative damage. In particular, an increase in the abundance of ferritin proteins effectively sequestered more iron and prevented excess free iron in the elongation zone under water stress.