UV-B辐射增强对两种不同抗性水稻叶片光合生理及超显微结构的影响

在UV-B辐射增强条件下,研究了两个不同水稻品种叶片光合作用系统的变化。结果表明：（1）UV-B辐射胁迫使两个水稻品种叶片总叶绿素含量,叶绿素a与叶绿素b（Chla／Chlb）比值下降,叶绿素a荧光诱导动力学参数改变,光系统Ⅱ活性受抑制,光合作用效率降低,其中Dular受抑制的程度较Lemont大。（2）利用扫描电镜（SEM）和透射电镜（TEM）进一步研究表明,UV-B辐射胁迫使水稻叶片气孔器受破坏,叶绿体结构变形,基粒片层排列稀疏紊乱,两个供试品种结构上受破坏的程度与它们光合生理受抑制的程度一致。（3）叶片边缘受破坏的程度较主脉两侧轻,这可能与硅质乳突密度较大有关。（4）两个供试品种叶片表面主脉两侧的硅质乳突数量及其受UV-B辐射影响的特性存在明显的差异,Lemont叶表面的乳突分布密度较大,且在UV-B辐射胁迫下有增加的趋势,而Dular则相反。这说明硅质体的累积特性可能是水稻对UV-B辐射胁迫的适应机制之一。     

紫外光B辐射增强对水稻叶片内IAA和ABA含量的影响

 两个水稻品种CK46和Dular生长在人工气候箱的条件下,在0.0和13.0kJm-2 day-1(模拟臭氧浓度下降20％的UVB强度)的紫外光B(UV-B 280 nm-320 nm)下进行4周的照射处理,研究UV-B对水稻体内内源IAA和ABA含量的影响。结果表明：随着UV—B处理时间的延长,CK46和Dular叶片内的IAA含量下降。相反,UV-B辐射增强使两个品种叶片内的ABA含量上升。     

UV B-irradiation enhances the racemization and isomerizaiton of aspartyl residues and production of Nε-carboxymethyl lysine (CML) in keratin of skin

UV-B irradiation is one of the risk factors in age-related diseases. We have reported that biologically uncommon D-β-Asp residues accumulate in proteins from sun-exposed elderly human skin. A previous study also reported that carboxymethyl lysine (CML; one of the advanced glycation end products (AGEs)) which is produced by the oxidation of glucose and peroxidation of lipid, also increases upon UV B irradiation. The formation of D-β-Asp and CML were reported as the alteration of proteins in UV B irradiated skin, independently. In this study, in order to clarify the relationship between the formation of D-β-Asp and CML, immunohistochemical analysis using anti-D-β-Asp containing peptide antibodies and anti-CML antibodies was performed in UV B irradiated mice. Immunohistochemical analyses clearly indicated that an anti-D-β-Asp containing peptide antibody and anti-CML antibody reacted at a common area in UV B irradiated skin. Western blot analyses of the proteins isolated from UV B irradiated skin demonstrated that proteins of 50-70 kDa were immunoreactive towards antibodies for both D-β-Asp containing peptide and CML. These proteins were identified by proteomic analysis as members of the keratin families including keratin-1, keratin-6B, keratin-10, and keratin-14.     

Proteomics analysis of A375 human malignant melanoma cells in response to arbutin treatment

Although the toxicogenomics of A375 human malignant melanoma cells treated with arbutin have been elucidated using DNA microarray, the proteomics of the cellular response to this compound are still poorly understood. In this study, we performed proteomic analyses to investigate the anticancer effect of arbutin on the protein expression profile in A375 cells. After treatment with arbutin (8 microg/ml) for 24, 48 and 72 h, the proteomic profiles of control and arbutin-treated A375 cells were compared, and 26 differentially expressed proteins (7 upregulated and 19 downregulated proteins) were identified by MALDI-Q-TOF MS and MS/MS. Among these proteins, 13 isoforms of six identical proteins were observed. Bioinformatic tools were used to search for protein function and to predict protein interactions. The interaction network of 14 differentially expressed proteins was found to be correlated with the downstream regulation of p53 tumor suppressor and cell apoptosis. In addition, three upregulated proteins (14-3-3G, VDAC-1 and p53) and five downregulated proteins (ENPL, ENOA, IMDH2, PRDX1 and VIME) in arbutin-treated A375 cells were validated by RT-PCR analysis. These proteins were found to play important roles in the suppression of cancer development.     

Response of oxidative stress defense systems in rice (Oryza sativa) leaves with supplemental UV-B radiation

The impact of elevated ultraviolet-B radiation (UV-B, 280–320 nm) on membrane systems and lipid peroxidation, and possible involvement of active oxygen radicals was investigated in leaves of two UV-B susceptible rice cultivars (Oryza sativa L. cvs IR74 and Dular). Rice seedlings were grown in a greenhouse for 10 days and then treated with biologically effective UV-B (UV-B_BE) radiation for 28 days. Oxidative stress effects were evaluated by measuring superoxide anion (O₂) generation rate, hydrogen peroxide (H₂O₂) content, malondialdehyde (MDA) concentration and relative electrolyte conductivity (EC) for IR74 and Dular at 0 (control), 6 or 13 kJ m^?2 day^?1 UV-B_BE. Significant increases in these parameters were found in rice plants grown at 13 vs 0 kJ m^?2 day^?1 UV-B_BE after 28 days; indicating that disruption of membrane systems may be an eventual reason for UV-B-induced injury in rice plants. There was a positive correlation between O₂^? generation and increases in EC or MDA in leaves. Activities of enzymatic and nonenzymatic free radical scavengers were measured for IR74 after 7, 14, 21 and 28 days of exposure to 13 or 0 UV-B_BE to evaluate dynamics of these responses over time. Activities of catalase and superoxide dismutase (but not ascorbate peroxidase) and concentrations of ascorbic acid and glutathione were enhanced by 13 vs 0 UV-B_BE after 14 days of UV-B exposure. Further exposure to 28 days of UV-B was associated with a decline in enzyme activities and ascorbic acid, but not glutathione. It is suggested that UV-B-induced injury may be associated with disturbance of active oxygen metabolism through the destruction and alteration of both enzymatic and nonenzymatic defense systems in rice.     

Influence of supplemental ultraviolet-B on indoleacetic acid and calmodulin in the leaves of rice (Oryza sativa L.)

IR68 and Dular rice cultivars were grown under ambient, 13.0 (simulating 20% ozone depletion) and 19.1 (simulating 40% ozone depletion) kJ m^-2 day^-1 of biologically effective ultraviolet-B (UV-B_BE) for 4 weeks. Plant height and leaf area were significantly reduced by supplemental UV-B_BE radiation. Greater reduction in leaf area than of plant height was observed. A decrease in indole-3-acetic acid (IAA) content and increase in peroxidase and IAA oxidase activities of UV-B treated plants in both cultivars were observed compared with ambient control. Calmodulin content also decreased after plants were treated with high supplemental UV-B for two weeks and medium UV-B treatment for four weeks. The results indicated that peroxidase and IAA oxidase activities in rice leaves were stimulated by supplemental UV-B, resulting in the destruction of IAA which in turn may cause inhibition of rice leaf growth. Although the mechanism is unclear, calmodulin is most likely involved in leaf growth.     

Cytokinin oxidase/dehydrogenase in Pisum sativum plants during vegetative development. Influence of UV-B irradiation and high temperature on enzymatic activity

Cytokinin oxidase/dehydrogenase (EC 1.5.99.12) specific activity was determined in leaves and roots of two P. sativum cultivars (cv. Scinado and cv. Manuela) during vegetative development and the effect of UV-B irradiation or elevated temperature was assessed. The measurement of CKX activity during development showed localisation of this enzyme to roots. The reduction in CKX activity in leaves after UV-B irradiation and the increased levels of the enzyme in high temperature-treated plants suggests that the enzymes from the CKX gene family have a different expression during stress responses provoked by different factors and probably are tissue specific. Differences regarding cytokinin oxidase/dehydrogenase activity stress response were observed between the two pea cultivars.     

Inhibitory effects of ambient levels of solar UV-A and UV-B radiation on growth of cucumber

The influence of solar UV-A and UV-B radiation at Beltsville, Maryland, on growth and flavonoid content in four cultivars of Cucumis sativus L. (Ashley, Poinsett, Marketmore, and Salad Bush cucumber) was examined during the summers of 1994 and 1995. Plants were grown from seed in UV exclusion chambers consisting of UV-transmitting Plexiglas, lined with Llumar to exclude UV-A and UV-B, polyester to exclude UV-B, or cellulose acetate to transmit UV-A and UV-B. Despite previously determined differences in sensitivity to supplemental UV-B radiation, all four cultivars responded similarly to UV-B exclusion treatment. After 19–21 days, the four cultivars grown in the absence of solar UV-B (polyester) had an average of 34, 55, and 40% greater biomass of leaves, stems, and roots, respectively, 27% greater stem height, and 35% greater leaf area than those grown under ambient UV-B (cellulose acetate). Plants protected from UV-A radiation as well (Llumar) showed an additional 14 and 22% average increase, respectively, in biomass of leaves and stems, and a 22 and 19% average increase, respectively, in stem elongation and leaf area over those grown under polyester. These findings demonstrate the extreme sensitivity of cucumber not only to present levels of UV-B but also to UV-A and suggest that even small changes in ozone depletion may have important biological consequences for certain plant species.     

紫外线B对水稻叶组织中活性氧代谢及膜系统的影响

增强ＵＶ－Ｂ处理下,水稻叶片的Ｏ净产生速率、Ｈ２Ｏ２和ＭＤＡ含量以及膜透性都显著增加,敏感性弱的品种Ｏ２净产生速率和膜伤害程度较低。在ＵＶ－Ｂ处理初期活性氧清除系统的水平增高,但随着处理时间的延长,ＳＯＤ、ＣＡＴ和ＡＰ活性以及ＡＳＡ含量降低,其中ＡＰ和ＳＯＤ活性的下降最为明显,而敏感性弱的品种ＳＯＤ活性始终高于敏感性强的品种。处理１４ｄ后去掉ＵＶ－Ｂ,再经则１４ｄ上述各指标均恢复到与对照相近的水平。根据这些结果推测,水稻的ＵＶ－Ｂ伤害可能主要是由于ＳＯＤ活性的降低而导致Ｏ增生和膜脂过氧化。     

Comparative proteomic analysis of male and female plants in Jojoba (<Emphasis Type="Italic">Simmondsia chinensis</Emphasis>) leaves revealed changes in proteins involved in photosynthesis,metabolism, energy,and biotic and abiotic stresses

Jojoba is a dioecious shrub with female and male flowers in separated individuals. The plant native to North and Central American deserts, it’s cultivated in many other places worldwide for its valuable liquid wax. The male-biased ratio in cultivated jojoba affects the yield. To develop protein molecular markers for early gender differentiation, comparative proteomic study been conducted on male and female leaves. Using gel-based proteomic, 45 proteins were identified representing 19 different proteins with 18 known functions. The identified proteins were involved in photosynthesis, energy, metabolism and the respond to biotic and abiotic stress. Ribulose-1,5-bisphosphate carboxylase (Rubisco) and ATP synthase were the most abundant proteins in both male and female of jojoba leaves, both were upregulated in male compared to female. Both proteins have the potential to serve as protein biomarkers for early differentiation between male and female in jojoba plant. These results could help in better understanding the molecular mechanism of gender differentiation in jojoba.     

水稻苗期低温失绿的遗传分析及基因定位

在早季低温条件下, 籼稻品种Dular的幼苗表现出白化失绿, 而粳稻品种Lemont幼苗表现正常绿色。以Lemont和Dular作亲本构建一个F2群体,通过该群体在早季低温条件下性状的表现,发现Lemont和Dular苗期耐冷性的差异受单个主基因控制,低温下白化失绿等位基因为隐性。将该基因暂时命名为cisc(t)。利用该F2群体,采用集团分离分析(BSA)法将cisc(t)定位在9号染色体上。经过对F2群体中100个典型的白化单株的简单序列长度多态性分析,将该基因定位在5.5 cM的区间内,分别与微卫星标记RM257和RM242相距3.9 cM和1.6 cM。     

Proteomic analysis of papaya (Carica papaya L.) displaying typical sticky disease symptoms

Papaya (Carica papaya L.) hosts the only described laticifer-infecting virus (Papaya meleira virus, PMeV), which is the causal agent of papaya sticky disease. To understand the systemic effects of PMeV in papaya, we conducted a comprehensive proteomic analysis of leaf samples from healthy and diseased plants grown under field conditions. First, a reference 2-DE map was established for proteins from healthy samples. A total of 486 reproducible spots were identified, and MALDI-TOF-MS/MS data identified 275 proteins accounting for 159 distinct proteins from 231 spots that were annotated. Second, the differential expression of proteins from healthy and diseased leaves was determined through parallel experiments, using 2-DE and DIGE followed by MALDI-TOF-MS/MS and LC-IonTrap-MS/MS, respectively. Conventional 2-DE analysis revealed 75 differentially expressed proteins. Of those, 48 proteins were identified, with 26 being upregulated (U) and 22 downregulated (D). In general, metabolism-related proteins were downregulated, and stress-responsive proteins were upregulated. This expression pattern was corroborated by the results of the DIGE analysis, which identified 79 differentially expressed proteins, with 23 identified (17 U and 6 D). Calreticulin and the proteasome subunits 20S and RPT5a were shown to be upregulated during infection by both 2-DE and DIGE analyses. These data may help shed light on plant responses against stresses and viral infections.     

Changes in leaf protein and pigment contents and photosynthetic activities during senescence of detached maize leaves: influence of different ultraviolet radiations

Senescence induced loss in pigments and proteins of detached maize (Zea mays L. cv. Col) leaves was significantly enhanced on the exposure of leaves to different ranges of ultraviolet (UV) radiation. Compared to UV-A (320-400 nm) and UV-B (280-320 nm), the UV-C (200-320 nm) was the most damaging for the pigments and macromolecules. A severe decline in photosystem (PS) 2 mediated photoreduction during senescence of detached leaves exposed to UV irradiation suggested a damage of the system. The PS1 mediated photoreduction of methylviologen with 2,6-dichlorophenol indophenol as electron donor was stimulated by UV-A and UV-B radiations, suggesting a reorganisation of the PS1 complex. These results were fortified by the values of fast and slow kinetics of chlorophyll (Chl) a fluorescence transients.     

Changes in leaf protein and pigment contents and photosynthetic activities during senescence of detached maize leaves: influence of different ultraviolet radiations

Senescence induced loss in pigments and proteins of detached maize (Zea mays L. cv. Col) leaves was significantly enhanced on the exposure of leaves to different ranges of ultraviolet (UV) radiation. Compared to UV-A (320-400 nm) and UV-B (280-320 nm), the UV-C (200-320 nm) was the most damaging for the pigments and macromolecules. A severe decline in photosystem (PS) 2 mediated photoreduction during senescence of detached leaves exposed to UV irradiation suggested a damage of the system. The PS1 mediated photoreduction of methylviologen with 2,6-dichlorophenol indophenol as electron donor was stimulated by UV-A and UV-B radiations, suggesting a reorganisation of the PS1 complex. These results were fortified by the values of fast and slow kinetics of chlorophyll (Chl) a fluorescence transients. This revised version was published online in June 2006 with corrections to the Cover Date.     

Quantitative Analysis of the Global Proteome in Peripheral Blood Mononuclear Cells from Patients with New‐Onset Psoriasis

Psoriasis is a common chronic autoimmune skin disease involving the activation of T cells. To explore the proteomic signature of peripheral blood mononuclear cells, a quantitative analysis of their global proteome was conducted in samples from Chinese patients with new‐onset psoriasis (n = 31) and healthy controls (n = 32) using an integrated quantitative approach with tandem mass tag labeling and LC–MS/MS. Protein annotation, unsupervised hierarchical clustering, functional classification, functional enrichment and cluster, and protein–protein interaction analyses were performed. A total of 5178 proteins were identified, of which 4404 proteins were quantified. The fold‐change cutoff was set at 1.2 (patients vs controls); 335 proteins were upregulated, and 107 proteins were downregulated. The bioinformatics analysis indicated that the differentially expressed proteins were involved in processes related to the activation of immune cells including the nuclear factor kappa‐light‐chain‐enhancer of activated B cells (NF‐κB) pathway, cellular energy metabolism, and proliferation. Three upregulated proteins and two phosphorylated proteins in the NF‐κB pathway were verified or identified by Western blotting. These results confirm that the NF‐κB pathway is critical to psoriasis. In addition, many differentially expressed proteins identified in this study have never before been associated with psoriasis, and further studies on these proteins are necessary.