Cloning of aroG, the gene coding for phospho-2-keto-3-deoxy-heptonate aldolase(phe), in Escherichia coli K-12, and subcloning of the aroG promoter and operator in a promoter-detecting plasmid

Defective transducing phages carrying aroG, the structural gene for phenylalanine (phe)-inhibitable phospho-2-keto-heptonate aldolase (EC 4.1.2.15; previously known as 3-deoxy-D-arabinoheptulosonate-7-phosphate synthetase[phe]), have been isolated, and DNA from two of these phages has been used to construct a restriction map of the region from att lambda to aroG. A 7.6-kb PstI-HindIII fragment from one of these phages was cloned into pBR322 and shown to contain aroG. The location of aroG within the 7.6 kb was established by subcloning and Tn3 transpositional mutagenesis. A fragment carrying the aroG promoter and operator has been cloned into a high copy number promoter-cloning vector (pMC489), and the resulting aroGpo-LacZ' (alpha) fusion subcloned in a low copy number vector. Strains with this fusion on the low copy number vector exhibit negative regulation of beta-galactosidase expression by both phenylalanine and tryptophan and positive regulation by tyrosine in a tyrR+ background.     

转OvDREB2B基因陆地棉鉴定及其对水分渗透胁迫的分析

通过花粉管通道技术,以该实验室自育陆地棉品系TH₁和TH₂为材料,将诸葛菜(Orychophragmus vidaceus)抗逆转录因子OvDREB2B基因构建到植物表达载体后,导入棉花基因组,经卡那霉素筛选和分子鉴定表明目的基因已整合到棉花基因组中并表达。将T₁代转基因植株和受体对照在温室中栽培,待植株生长至四叶一心时,用不同渗透势的PEG-6000水溶液进行渗透胁迫处理,分析探讨转基因植株的抗旱效果及其抗旱机理。结果显示:当渗透势为0和0.5 MPa处理时,转基因植株和对照无明显差异;当渗透势为0.8 MPa和1.1 MPa处理时转基因植株较对照抗旱性明显提高。当渗透势为1.1 MPa处理96 h时,对照植株F_v/F_m降至0.2左右,而转基因植株仍正常生长,F_v/F_m值约为0.51,而且初始荧光（F₀）值、净光合速率（P_n）、胞间CO₂浓度（C_i）、蒸腾速率（T_r）等一系列参数转基因植株都明显优于对照,表明DREB2B基因能够提高棉花对水分胁迫的耐受性。     

Study of the serum albumin-polyethyleneglycol interaction to predict the protein partitioning in aqueous two-phase systems

The theoretical framework based only on the excluded volume forces is not enough to explain the bovine serum albumin partitioning behaviour in aqueous biphasic systems. The goal of this work is to look at the phase separation via the polymer effect on the water structure. Our findings suggest that polyethyleneglycol 600-protein interaction is conducted by van der Waals forces between the hydrophobic surfaces from PEG and protein molecules, which implies the rupture of hydrogen bonds from the structured water in their neighbours. Therefore, the protein will concentrate in the most water-structured phase (polyethyleneglycol) in order to reach the minimal free energy condition. When polyethyleneglycol molecular weight increases, its exclusion from protein surface prevails, thus pushing the bovine serum albumin to the bottom phase.     

The different effects of PEG 6000 and NaCI on leaf development are associated with differential inhibition of root water transport

The inhibitory effects of PEG on whole-plant growth can exceed the effects of other osmolytes such as NaCI, and this has been ascribed to toxic contaminants, or to reduced oxygen availability in PEG solutions. We investigated another possibility, namely that PEG has an additional inhibitory effect on root water transport which in turn affects leaf development. The effects on first-leaf growth of applications of PEG 6000 or isoosmotic NaCI to the roots were determined using hydroponically grown maize (Zea mays L.) seedlings. Leaf growth rates were inhibited within minutes of PEG application to the roots and remained inhibited for days. The inhibitory effects on growth of NaCI, and also of KCl and mannitol, were much smaller. The comparative effects of NaCI and PEG on root water transport were determined by assaying pressurized flow through excised roots. PEG induced a 7-fold greater inhibition of flow through live roots than NaCI. Killing of the roots by heat treatment, to reduce cell membrane resistances to solute penetration, nearly doubled the flow rate for roots in NaCI, but not for roots in PEG. We suggest that the greater viscosity of PEG solutions, as compared with NaCI, may be a primary factor contributing to the additional inhibition of water flow through live and killed roots. PEG did not have additional effects on leaf turgor but had a 3 times greater inhibitory effect than NaCI on the irreversible extensibility of the leaves and induced 16 times more leaf accumulation of the growth inhibitory stress hormone abscisic acid (ABA). We conclude that greater inhibition of root water transport by PEG 6000, as compared with NaCI, leads to additional reductions in extensibility, additional ABA accumulation, and a greater inhibition of leaf growth.     

New coupled model used inversely for reconstructing past terrestrial carbon storage from pollen data: validation of model using modern data

The knowledge of potential impacts of climate change on terrestrial vegetation is crucial to understand long-term global carbon cycle development. Discrepancy in data has long existed between past carbon storage reconstructions since the Last Glacial Maximum by way of pollen, carbon isotopes, and general circulation model (GCM) analysis. This may be due to the fact that these methods do not synthetically take into account significant differences in climate distribution between modern and past conditions, as well as the effects of atmospheric CO₂ concentrations on vegetation. In this study, a new method to estimate past biospheric carbon stocks is reported, utilizing a new integrated ecosystem model (PCM) built on a physiological process vegetation model (BIOME4) coupled with a process-based biospheric carbon model (DEMETER). The PCM was constrained to fit pollen data to obtain realistic estimates. It was estimated that the probability distribution of climatic parameters, as simulated by BIOME4 in an inverse process, was compatible with pollen data while DEMETER successfully simulated carbon storage values with corresponding outputs of BIOME4. The carbon model was validated with present-day observations of vegetation biomes and soil carbon, and the inversion scheme was tested against 1491 surface pollen spectra sample sites procured in Africa and Eurasia. Results show that this method can successfully simulate biomes and related climates at most selected pollen sites, providing a coefficient of determination ( R ) of 0.83–0.97 between the observed and reconstructed climates, while also showing a consensus with an R -value of 0.90–0.96 between the simulated biome average terrestrial carbon variables and the available observations. The results demonstrate the reliability and feasibility of the climate reconstruction method and its potential efficiency in reconstructing past terrestrial carbon storage.     

Effect of water stress on germination of some Hungarian wheat landraces varieties

In order to examine germination characters, seedling parameters, water relative content, tolerance index and enzyme activities of seven Hungarian wheat landraces varieties (Tiszadadai, Riscsei, Komloi, Leweucei, Mateteleki, Mikebudai and Nyiradi) under five concentrations of water stress (0, 6, 12, 18, and 24%) of polyethylene glycol (PEG-6000). A laboratory experiment has been conducted through Factorial Experiment in Randomized Complete Block Design (RCBD) with four repetitions at Research Institute of Nyiregyhaza, Hungary. From the obtained results, Leweucei variety was surpassed other studied verities under study and recorded the highest values of all studied characters followed by Mateteleki, Komloi, Nyiradi, Riscsei, Tiszadadai and Mikebudai. Increasing water stress (PEG-6000) from 0 to 6, 12, 18 and 24% significantly reduced germination characters, seedlings parameters, water relative content (WRC), tolerance index and α and β-amylases activities. Generally, under water stress condition, Leweucei and Mateteleki varieties were recorded the highest values of water relative content (WRC), tolerance index (TI) and α and β-amylases activities as well as able to prompt better drought tolerance and could be suggested as a good resource for breeding programs and cultivation under drought stress conditions compared with other wheat landraces varieties.     

Tocopherol and plastoquinone synthesis in spinach chloroplasts subfractions

Subfractions isolated from intact purified spinach chloroplasts are able to prenylate the aromatic moiety of α-tocopherol and plastoquinone-9 precursors. The biosynthesis of α-tocopherol and plastoquinone-9 is a compartmentalized process. The chloroplast envelope membranes are the only site of the enzymatic prenylation in α-tocopherol synthesis whereas the thylakoid membrane is also involved in the prenylation and methylation sequence of plastoquinone-9 biosynthesis. A very active kinase which forms phytyl-PP is localized in the stroma. Phytol but not geranylgeraniol is the polyprenol precursor of the side chain of α-tocopherol in spinach chloroplasts.     

青海高原春小麦PEG胁迫与复水后叶片差异蛋白质组学研究

春小麦是青海省的主要粮食作物,青海高原干旱频繁且严重,尤以春旱为首,对小麦生长发育造成严重影响．为了从蛋白质组学水平分析小麦对干旱胁迫的应答特征,探讨小麦可能的抗旱机制,本研究对青海主栽春小麦品种青春38幼苗进行聚乙二醇 (polyethylene glycol,PEG) 6000胁迫和复水处理,采用IEF/SDS PAGE双向凝胶电泳技术,对PEG胁迫和复水处理的小麦叶片总蛋白质分别与正常浇水的对照进行差异蛋白质组学研究,经考马斯亮蓝G 250染色获得清晰度和重复性较好的双向电泳图谱．PD Quest软件处理分析各对照和处理图谱,在等电点4.0～7.0线性范围内,均可识别650个以上清晰蛋白质点,获得比较明显的差异表达蛋白点43个,其中8个蛋白点重复.从35个差异蛋白点中选取24个差异点进行MALDI-TOF-TOF-MS肽质量指纹图谱分析,应用Mascot软件在NCBInr数据库中搜索鉴定蛋白质,得到22个阳性结果．对鉴定得到的差异表达蛋白进行功能分析,它们分别参与了光合作用、蛋白质合成、能量代谢途径、细胞防御、氧化还原、运输、信号转导等过程,而且根据差异表达蛋白功能分类所占比例,发现干旱胁迫与光合作用关系最为紧密.     

Allosteric modulation of adenosine receptors

Allosteric modulators for adenosine receptors may have potential therapeutic advantage over orthosteric ligands. Allosteric enhancers at the adenosine A₁ receptor have been linked to antiarrhythmic and antilipolytic activity. They may also have therapeutic potential as analgesics and neuroprotective agents. A₃ allosteric enhancers are postulated to be useful against ischemic conditions or as antitumor agents. In this review, we address recent developments regarding the medicinal chemistry of such compounds. Most efforts have been and are directed toward adenosine A₁ and A₃ receptors, whereas limited or no information is available for A_2A and A_2B receptors. We also discuss some findings, mostly receptor mutation studies, regarding localization of the allosteric binding sites on the receptors.