Hydrogen sulfide is involved in the regulation of ascorbate and glutathione metabolism by jasmonic acid in <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis>

This study investigated the role of hydrogen sulfide (H₂S) in the regulation of the ascorbate (AsA) and glutathione (GSH) metabolism by jasmonic acid (JA) in the leaves of Arabidopsis thaliana by using H₂S scavenger hypotaurine (HT) and H₂S synthetic mutant (SALK_041918, designated Atl-cdes). The results showed that JA significantly increased the H₂S content, the activities of L-cysteine desulfhydrase (L-CDes), D-cysteine desulfhydrase (D-CDes), ascorbate peroxidase (APX), glutathione reductase (GR), monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR), L-galactono-1,4-lactone dehydrogenase (GalLDH) and γ-glutamylcysteine synthetase (γ-ECS), the ratio of AsA to dehydroascorbate (DHA), and decreased the content of malondialdehyde (MDA) and H₂O₂ in the wild type of A. thaliana, compared to control. The above effects of JA except the increased activities of L-CDes and D-CDes were suppressed by addition of HT. However, JA and HT+JA had no significant effects on the ratio of reduced GSH to oxidized GSH (GSSG) in the wild type of A. thaliana. Application of HT to the control decreased H₂S content, AsA/DHA ratio, and activities of APX, GR, DHAR, MDHAR, γ-ECS, and GalLDH, but had no effects on MDA content, activities of L-CDes and D-CDes, and GSH/GSSG ratio. In the H₂S synthetic mutant, JA had no obvious effects on above mentioned parameters except the D-CDes activity compared with the control. Our results suggest that JA-induced H₂S, which is a signal that leads to the up-regulation of the AsA and GSH metabolism.     

Inhibition of in vivo Slicer activity of Argonaute protein 1 by the viral 2b protein independent of its dsRNA‐binding function

The 2b protein of Cucumber mosaic virus (CMV) has several unique properties, such as targeting to the nucleolus and interaction with both Argonautes (AGOs) and short and long double‐stranded RNA (dsRNA). We have recently uncoupled the domain requirements for dsRNA binding and nucleolar targeting from the physical interactions with AGO proteins, and have found that the direct 2b–AGO interaction is sufficient to inhibit the in vitro AGO1 Slicer function independent of the other biochemical properties of 2b. Because the AGO binding activity of 2b is not required for its suppressor function in vivo, this raises the question of whether in vivo 2b–AGO interaction is possible to inhibit the in vivo AGO Slicer function. In this study, by taking advantage of a technology for the production of artificial trans‐acting small interfering RNA (tasiRNA), a process uniquely associated with AGO1‐mediated in vivo Slicer activity, we demonstrated that the expression of the 2b protein in planta interfered with the production of tasiRNA. Through further detailed analysis with deletion mutants of 2b proteins, we found that the inhibition of in vivo AGO1 Slicer function required the nucleolar localization signal (NoLS), in addition to the AGO‐binding domain, of the 2b protein. Our finding demonstrates that in vivo 2b–AGO1 interaction is sufficient to inhibit AGO1 Slicer function independent of the dsRNA‐binding activity of the 2b protein.     

黄渤海甲壳类的分类多样性

为了解黄渤海甲壳类的分类多样性特征, 我们统计了2010-2015年中国水产科学研究院黄海水产研究所调查捕获的黄渤海甲壳类(软甲纲: 十足目与口足目)物种名录。结合历史文献, 进一步系统整理得到黄渤海甲壳类物种总名录。基于这2个名录, 应用分类阶元包含指数(the inclusion index at taxonomic level, TINCL_i)、平均分类差异指数(average taxonomic distinctness index, Δ⁺)和分类差异变异指数(variation in taxonomic distinctness index, Λ⁺)研究了其分类多样性特征。结果显示: 2010-2015年调查名录中, 甲壳类共93种, 隶属于2目39科66属, 其中10种为新分布种; 对虾科、藻虾科、长臂虾科、梭子蟹科和弓蟹科的物种数最多, 合计占总物种数的38.71%; TINCL_i分别为1.41种/属和2.38种/科; Δ⁺和Λ⁺分别为50.25和35.20。总名录中, 甲壳类共228种, 隶属于2目53科123属, 其中藻虾科、豆蟹科、对虾科、弓蟹科和鼓虾科的物种数最多, 合计占总物种数的30.70%; TINCL_i分别为1.85种/属和4.30种/科, Δ⁺和Λ⁺分别为50.18和30.87。对虾科的相对丰富度指数(the relative richness index, R_r)最高(100), 其次是梭子蟹科(71.43)和长臂虾科(62.50), 豆蟹科最低(6.25)。黄渤海甲壳类的平均分类差异指数(Δ⁺)明显小于鱼类(P < 0.05)。2010-2015年调查的Δ⁺计算值高于理论值, 且在理论值的95%置信区间内, 说明黄渤海甲壳类群落正处在中等程度的干扰中。     

硫酸乙酰肝素3-O硫酸基转移酶5的表达、纯化及酶学性质研究

经过PCR克隆得到硫酸乙酰肝素3-O硫酸基转移酶5(3-OST-5)的基因,将其与大肠杆菌表达载体pET-15b连接后,在大肠杆菌BL21(DE3)中诱导表达,使用镍亲和层析柱纯化得到具有活性的3-OST-5。经测定纯化后的3-OST-5比活达到0.58 U/mg,是纯化前的5.27倍,回收率达80.4%。在此基础上,研究了该酶的酶学性质,酶反应的最适温度为35℃,稳定范围为20-40℃;最适pH为7.0,在pH7.0-9.0范围内稳定。在反应液中加入终浓度为1 mmol/L的K+、Ca2+、Ba2+对酶促反应有一定的促进作用。     

宁夏灵武市沟眶象发生特点

系统调查沟眶象Eucryptorrhynchus chinensis(Olivier)在宁夏灵武市的发生情况和特点发现:沟眶象以成虫和幼虫在土壤中越冬,成虫有2个发生高峰期,越冬结束后以成虫越冬的沟眶象出土出现第1个高峰期,以幼虫越冬的沟眶象经化蛹羽化出土出现第2个高峰期;幼虫危害臭椿根部形成的瘤状物可以作为鉴别沟眶象危害的特征;沟眶象幼虫在土壤中垂直分布范围集中在0～45 cm深土层,约占80%;成虫出土孔集中在距臭椿树干基部75 cm范围内,占88%。据此建议土壤施药化学防治沟眶象时,施药深度不宜低于45 cm,施药点距树干基部水平距离不得少于75 cm,且树周各个方向均需施药。每年10月末沟眶象开始"入土"、翌年4月末开始"出土",在沟眶象"入土"结束后但未冻土之前的11中旬和土壤解冻之后但尚未"出土"前的4月中旬,这两个时期土壤施药防治效果较好;防治沟眶象的同时,还应适时采取打孔注药、熏蒸等措施防治臭椿(Ailanthus altissima(Mill.)Swingle)树干内的臭椿沟眶象Eucryptorrhynchus brandti(Harold)。     

Phosphatidylinositol 3-kinase/Akt activation by integrin-tumor matrix interaction suppresses Fas-mediated apoptosis in T cells

It has recently become apparent that the microenvironment made up of the extracellular matrix may affect cell signaling. In this study, we evaluated Fas-triggered apoptosis in T cells in contact with tumor cells, which resembles the cell-to-cell interactions found in tumor regions. Jurkat cells were less susceptible to the Fas-mediated apoptosis when cocultured with U118, HeLa, A549, and Huh-7 tumor cells. This was indicated by less plasma membrane alteration, an amelioration of the loss of mitochondria membrane potential, a decrease in caspase-8 and caspase-3 activation, a decrease in DNA fragmentation factor-45/35 cleavage, and a reduction in the breakage of DNA when compared with Jurkat cells cultured alone. In contrast, the tumor cell lines MCF-7 and HepG2 produced no such protective effect. This protective event was independent of the expression of Fas ligand on the tumor cells. Interrupting the beta integrins-matrix interaction diminished the coculture effect. In Jurkat cells, cell matrix contact reduced the assembly of the Fas death-inducing signaling complex and Bcl-x(L) cleavage, but enhanced the phosphorylation of ERK1/2, p38 MAPK, and Akt. Only PI3K inhibitor, but not kinase inhibitors for MEK, ERK1/2, p38 MAPK, JNK, protein kinase C, and protein kinase A, completely abolished this tumor cell contact-associated protection and in parallel restored Fas-induced Bcl-x(L) cleavage as well as decreasing the phosphorylation of Bad at serine 136. Together, our results indicate that stimulation of the beta integrin signal of T cells by contact with tumor cells may trigger a novel protective signaling through the PI3K/Akt pathway of T cells against Fas-mediated apoptosis.     

Structural and functional effects of tryptophans inserted into the membrane-binding and substrate-binding sites of human group IIA phospholipase A2

Phospholipase A(2) (PLA(2)) enzymes become activated by binding to biological membranes and hydrolyze phospholipids to free fatty acids and lyso-phospholipids, the precursors of inflammatory mediators. To understand the functional significance of amino acid residues at key positions, we have studied the effects of the substitution of Val(3) (membrane binding surface) and Phe(5) (substrate binding pocket) of human group IIA PLA(2) by tryptophan on the structure and function of the enzyme. Despite the close proximity of the sites of mutations, the V3W mutation results in substantial enhancement of the enzyme activity, whereas the F5W mutant demonstrates significantly suppressed activity. A structural analysis of all three proteins free in buffer and bound to membranes indicates that large differences in activities result from distinct conformational changes in PLA(2)s upon membrane binding. Although PLA(2) and the V3W mutant demonstrate a decrease in helical content and an increase in helix flexibility, the F5W mutant experiences partial distortion of the alpha-helical structure presumably resulting from the tendency of Trp(5) to insert into the membrane. Furthermore, whereas the PLA(2) and the V3W mutant bind to the membrane at similar and apparently productive-mode orientation, the F5W mutant binds to membranes with a distinctly different orientation. It is suggested that both the stimulatory effect of the V3W mutation and the inhibitory effect of the F5W mutation result from the high affinity of Trp for the membrane-water interface. Although Trp(3) at the membrane binding face of PLA(2) facilitates the proper membrane binding of the enzyme, Trp(5) in the internal substrate binding site causes partial unwinding of the N-terminal helix in order to interact with the membrane.     

Design,synthesis and algicides activities of thiourea derivatives as the novel scaffold aldolase inhibitors

By using a new Fragment-Based Virtual Screen strategy, two series of novel FBA-II inhibitors (thiourea derivatives) were de novo discovered based on the active site of fructose-1, 6-bisphosphate aldolase from Cyanobacterial (CyFBA). In comparison, most of the N-(2-benzoylhydrazine-1-carbonothioyl) benzamide derivatives (L14～L22) exhibit higher CyFBA-II inhibitory activities compared to N-(phenylcarbamothioyl) benzamide derivatives (L1～L13). Especially, compound L14 not only shows higher CyFBA-II activity (K_i?=?0.65?μM), but also exhibits most potent in vivo activity against Synechocystis sp. PCC 6803 (EC₅₀?=?0.09?ppm), higher (7-fold) than that of our previous inhibitor (EC₅₀?=?0.6?ppm). The binding modes of compound L14 and CyFBA-II were further elucidated by jointly using DOX computational protocol, MM-PBSA and site-directed mutagenesis assays. The positive results suggest that strategy adopted in this study was promising to rapidly discovery the potent inhibitors with novel scaffolds. The satisfactory algicide activities suggest that the thiourea derivatives is very likely to be a promising lead for the development of novel specific algicides to solve Cyanobacterial harmful algal blooms (CHABs).     

耐盐性不同的小麦根和根茎结合部的拒Na~ 作用

NaCl处理引起小麦幼苗干重下降 ,盐敏感品种比耐盐品种下降幅度要大。盐敏感品种的拒Na 能力为 35 %左右 ,耐盐品种的拒Na 能力为 5 0 %～ 6 0 %左右 ,这表明耐盐品种的拒Na 能力远远大于盐敏感品种。不同耐盐性小麦的拒Na 部位不同 ,盐敏感品种的拒Na 部位主要在根茎结合部 ,而耐盐品种的拒Na 部位主要在根部     

Photosynthetic activity and proteomic analysis highlights the utilization of atmospheric CO2 by Ulva prolifera (Chlorophyta) for rapid growth

Free‐floating Ulva prolifera is one of the causative species of green tides. When green tides occur, massive mats of floating U. prolifera thalli accumulate rapidly in surface waters with daily growth rates as high as 56%. The upper thalli of the mats experience environmental changes such as the change in carbon source, high salinity, and desiccation. In this study, the photosynthetic performances of PSI and PSII in U. prolifera thalli exposed to different atmospheric carbon dioxide (CO₂) levels were measured. Changes in photosynthesis within salinity treatments and dehydration under different CO₂ concentrations were also analyzed. The results showed that PSII activity was enhanced as CO₂ increased, suggesting that CO₂ assimilation was enhanced and U. prolifera thalli can utilize CO₂ in the atmosphere directly, even when under moderate stress. In addition, changes in the proteome of U. prolifera in response to salt stress were investigated. Stress‐tolerance proteins appeared to have an important role in the response to salinity stress, whereas the abundance of proteins related to metabolism showed no significant change under low salinity treatments. These findings may be one of the main reasons for the extremely high growth rate of free‐floating U. prolifera when green tides occur.