Protective Antioxidant Effects of Amentoflavone and Total Flavonoids from Hedyotis diffusa on H2O2‐Induced HL‐O2 Cells through ASK1/p38 MAPK Pathway

In this study, total flavonoids and total triterpenoid acid were extracted with ethyl acetate from Hedyotis diffusa Willd, and hepatoprotective activities of them and five compounds from total flavonoids against H₂O₂ induced hepatocyte damage on HL‐02 cells were determined. In particular, amentoflavone and total flavonoids had influence on the leakage of ALT, AST, LDH, the activities of SOD and the content of MDA. They effectively reduced the loss of MMP, the release of Cyt C, and then inhibited activation of caspase‐3/caspase‐9 cascade in hepatotoxic cells. The contents of ROS were significantly reduced to inhibit p38 in amentoflavone and flavonoids groups which decreased ASK1 and p‐p38 levels through increasing thioredoxin Trx1 and reductase TrxR1. These results suggesting that the antioxidant protection of amentoflavone and flavonoids might be reducing ROS to inhibit the H₂O₂‐induced upstream of pathway via increasing levels of Trx1 and TrxR1, which were pivotal in blocking the down streaming effectors of ASK1/p38 MAPK pathway and alleviating hepatotoxicity.     

A nuclear export signal and phosphorylation regulate Dok1 subcellular localization and functions

Dok1 is believed to be a mainly cytoplasmic adaptor protein which down-regulates mitogen-activated protein kinase activation, inhibits cell proliferation and transformation, and promotes cell spreading and cell migration. Here we show that Dok1 shuttles between the nucleus and cytoplasm. Treatment of cells with leptomycin B (LMB), a specific inhibitor of the nuclear export signal (NES)-dependent receptor CRM1, causes nuclear accumulation of Dok1. We have identified a functional NES (348LLKAKLTDPKED359) that plays a major role in the cytoplasmic localization of Dok1. Src-induced tyrosine phosphorylation prevented the LMB-mediated nuclear accumulation of Dok1. Dok1 cytoplasmic localization is also dependent on IKKbeta. Serum starvation or maintaining cells in suspension favor Dok1 nuclear localization, while serum stimulation, exposure to growth factor, or cell adhesion to a substrate induce cytoplasmic localization. Functionally, nuclear NES-mutant Dok1 had impaired ability to inhibit cell proliferation and to promote cell spreading and cell motility. Taken together, our results provide the first evidence that Dok1 transits through the nucleus and is actively exported into the cytoplasm by the CRM1 nuclear export system. Nuclear export modulated by external stimuli and phosphorylation may be a mechanism by which Dok1 is maintained in the cytoplasm and membrane, thus regulating its signaling functions.     

Seed-to-seed potential allelopathic effects between <Emphasis Type="Italic">Ligularia virgaurea</Emphasis> and native grass species of Tibetan alpine grasslands

Allelopathy is an important process in plant communities, but the role of seed allelopathy in natural ecosystems remains poorly understood. In the present study, we examined the potential allelopathic effects of Ligularia virgaurea (a dominant species in degraded Tibetan grasslands) seeds on the germination of four native grass species (Festuca sinensis, Agrostis gigantean, Bromus inermis, and Elymus nutans). The results showed that L. virgaurea seeds can have potential allelopathic effects on seed germination, mean time to germination and root growth rates of native grass species. We further demonstrate that these effects are driven by a water-soluble seed leachate. Species with smaller seeds were generally more sensitive than larger seeded species. The results suggest that seed-to-seed allelopathic potential may be an important mechanism driving the dominance of L. virgaurea in degraded alpine grasslands on the Tibetan Plateau. Further studies are required to demonstrate effects of seed-to-seed potential allelopathy in a field setting.     

Computational investigation of the enzymatic mechanisms of phosphothreonine lyase

SpvC, a virulence effector injected through type III secretion system by some Salmonella serovars, belongs to the newly discovered enzyme family, phosphothreonine lyase. Previous experimental studies have demonstrated that SpvC irreversibly inactivates mitogen-activated protein kinases by removing the phosphate group from phosphothreonine-containing substrate through a β-elimination mechanism, and results in a β-methyldehydroalanine product. Interestingly, further biochemical investigations also indicated a secondary reaction occurring other than elimination, where a covalently bound complex is formed. Here, we employed molecular dynamics simulations and quantum mechanics calculations to gain insights on the microscopic details of such novel reaction mechanisms. Our theoretical results are consistent with the experimental observations, in which the critical stages of SpvC catalyzed reaction are revealed and the roles of several important binding site residues are reconciled. The deprotonation and precise position of the catalytic base K136 are facilitated by the formation of the fully desolvated active site upon substrate binding. The abstraction of the alpha hydrogen by K136 and the elimination of the phosphate group occur nearly simultaneously, promoted by the proton donation from the catalytic acid H106, and thus strongly supports an E2-like mechanism. K104, which is not directly involved in the enzymatic reaction, stabilizes the transition state and facilitates the reaction to occur. Remarkably, the subsequent deprotonation of K136 happens to be a natural sequel of the primary elimination reaction, restores its nucleophile capacity to attack the double bond containing elimination product, and leads to a covalently bound complex via a Michael-addition mechanism. The reaction mechanism used by phosphothreonine lyases might serve as a method of programmed regulation to fine tune their enzymatic activity.     

七种云南地衣植物的化学成分

本文报道分属于三个科的七种云南产地衣植物的化学成分。这些植物是亚洲树发(Alectoria asiatica DR.)、沟树发(Alectoria sulcata Nyl.)、长茎松萝(Usnea longissimaAch.)、胡子松萝(Usnea comosa(Ach.)Rohl.)、林石蕊(Cladonia arbuscula(Rabh.)Rabh.)、砖孢发(Oropogon loxensis(Fee.)Th.Fr.)和卷梢雪花衣(Anaptychiaboryi(Fee.)Mass.)。根据光谱数据测定,其化学成分为:松萝酸(Usnic acid)(1),维任西酸(Virensic acid)(2),赤星衣酸乙酯(Ethyl haematommate)(3),瑞藏酸(Rhizonic acid)(4),赤星衣酸(Haematommic acid)(5),扁枝衣酸乙酯(Ethyleverninate)(6),黑茶渍素(Atranorin)(7)和泽屋萜(Zeorin)(8)。上述资料,对开发利用这些植物资源提供了科学依据。     

大豆初生根凯氏带对铜离子的通透性

采用在根内生成有色铜沉淀的方法研究大豆(Glycine max)初生根凯氏带对铜离子的通透性。用真空泵抽取浓度为200 μmol·L^–1 的CuSO₄溶液进入根中, 然后在重力作用下从根基部灌注400 μmol·L^–1的K₄[Fe(CN)₆]溶液, 两种物质在根内相遇即可产生棕色的Cu₂[Fe(CN)₆]沉淀, 根据沉淀的位置来确定铜离子所经过的途径。结果表明: Cu²⁺可以穿过内皮层凯氏带, 在木质部导管壁以及凯氏带至木质部之间的细胞壁处产生棕色沉淀, 侧根发生的部位也产生了大量的沉淀; 当抽取K₄[Fe(CN)₆]溶液后再灌注CuSO₄溶液, 发现Cu²⁺仍然可以穿过凯氏带, 并在凯氏带外侧以及外皮层细胞的细胞壁处产生棕色沉淀。研究结果证明凯氏带并不是一个可以完全阻止离子进出的完美屏障。     

外来植物入侵的全境性研究进展与展望

由于外来种入侵地一般都远离其自然分布区(原产地), 如果只在其入侵地或者原产地进行研究, 很难真正发现其入侵性形成和成功入侵的根本原因。目前, 许多学者开始关注和倡导对入侵种在原产地和入侵地的表现同时进行研究, 即入侵种的全境性研究(whole-range studies), 为入侵生物现有地理分布格局的形成原因和入侵机制等提供解释。本文结合国内外关于入侵植物全境性研究的进展和成果, 分别针对研究的主要目的、内容、意义等进行了全面的阐述, 探讨了存在的问题与不足, 并对未来相关研究进行了展望。目前已有的全境性研究主要是通过野外直接观测和同质种植园实验来比较入侵种在入侵地和原产地的生长、繁殖和生理生态等表型性状的差异, 以及应用分子标记方法比较入侵地种群和原产地种群遗传多样性的差异, 进行入侵植物的分子系统地理学研究, 从而有效检验生物入侵机制的理论和假说, 深入阐明植物入侵的机制, 为制定入侵植物的防控策略提供指导。值得注意的是, 由于外来植物入侵的全境性研究起步较晚, 现有研究的方法和内容还不够完善, 今后需要在加强国际合作的基础上进一步改善。     

Precise mapping of a quantitative trait locus interval for spike length and grain weight in bread wheat (Triticum aestivum L.)

The spike characteristics length, spikelet density and fertile floret number are related yield components and are important in cereal improvement. QSpl.nau-2D is a major quantitative trait locus controlling spike length (SPL) detected in the recombinant inbred line population developed by crossing wheat (Triticum aestivum) cultivars Nanda2419 with Wangshuibai. In this study, to validate its genetic effect and determine its precise location, QSpl.nau-2D’s near-isogenic line (NIL) was developed using Mianyang99-323 as the recurrent parent through marker-assisted selection. Field trials showed that the NIL not only had significantly longer spikes on average than the recurrent parent but also had significantly higher grain weight, but did not differ in spikelet number and kernel number per spike. In the F2 population derived from a cross of the NIL with Mianyang99-323, QSpl.nau-2D functioned like a single gene and conditioned the SPL in a partially dominant manner, and was thus designated as HL1 (for head length). To precisely map HL1, 89 recombinants, consisting of 11 genotypes, were identified in the NIL-derived F2 population of 674 plants by using markers in the Xwmc25–Xgpw4080 interval. Phenotyping these lines showed that the introduction of a 0.9-cM interval flanked by Xcfd53 and DG371 in Nanda2419 resulted in longer spikes and a higher grain weight in the NIL. The availability of markers closely linked to HL1 could facilitate its use in breeding programs.