弓形虫蛋白质翻译后修饰研究进展

弓形虫是一种复杂的单细胞原生动物寄生虫,通过入侵宿主细胞、分裂和诱导宿主细胞破裂等一系列的过程,在温血动物体内进行增殖。弓形虫的生物学特征受基因组、表观遗传及转录等多种因素的调控。蛋白质翻译后修饰（protein post-translational modifications,PTMs）,如磷酸化、泛素化、巴豆酰化、棕榈酰化、琥珀酰化、乙酰化、甲基化、糖基化和二羟基异丁酰化等,是弓形虫调节对细胞外刺激的反应和生命周期转变的主要机制之一。弓形虫蛋白质翻译后修饰通过改变靶蛋白的定位、结构、活性、蛋白质-蛋白质相互作用等增加蛋白质的复杂性和多样性,从而在虫体生命周期的任何时间点都可以发挥至关重要的作用。在这篇综述中,作者重点对弓形虫蛋白质翻译后修饰进行简要总结,以期为深入研究弓形虫的生物学特征奠定基础。     

Fishway entrance modifications enhance fish attraction

Two Denil fishways on the Grand River, Ontario, were monitored annually since 1994 for activity by several dozen fish species. Fishway entrances were enlarged and repositioned approximately 2 m closer to the weir face, in areas where fish were attracted by weir discharge. These simple modifications resulted in increased attraction efficiency for pumpkinseed, Lepomis gibbosus (L.). After modifications, annual relative rate of recapture was 39% (95% confidence interval [CI]=32–46%), representing a 2.6–3-fold increase in fishway use relative to pre-modification conditions. Median daily recapture rates also increased significantly from 0% at both fishways to approximately 2%. These results suggest that fishway entrances should be located as close to a dam or weir face as possible, but velocity barriers from spillway or tailrace discharge must not compromise access.     

乙酰化蛋黄果多糖及其抗氧化活性研究

蛋黄果（Lucuma nervosa A. DC）乙酰化修饰多糖的抗氧活性优于未修饰多糖,通过分析乙酰化蛋黄果多糖的乙酰化度、红外光谱和形貌特征,阐明引起抗氧活性提升的原因。试验采用超声波辅助法提取蛋黄果果肉粗多糖,用三氯乙酸法除去蛋白质得到多糖。选取3份0.50 g多糖加入20% NaOH溶液溶解,分别加入1、3、5 mL乙酸酐,得到3种取代度（0.237±0.05、0.275±0.07、0.228±0.04）的乙酰化蛋黄果果肉多糖,评价其清除DPPH自由基、OH自由基和ABTS自由基的能力。结果表明,乙酸酐添加量由少到多时,蛋黄果多糖乙酰基取代度先显著升高,后趋于平缓下降。乙酰化蛋黄果多糖的红外光谱显示了3417 cm^-1（-OH）、2950 cm^-1（-CH）处的伸缩振动和1636 cm^-1处的（-OH）转动振动,1738 cm^-1处出现酯基C=O的伸缩振动吸收峰,表明乙酰化已经成功接入。扫描电镜结果显示,未经乙酰化的蛋黄果多糖表面相对光滑平整,呈片状结构;乙酰化修饰多糖分子间交联增强,构象发生了变化,变为表面形状不规则,出现很多空隙且表面粗糙。3种乙酰化蛋黄果多糖中DHG-Ac2（3 mL醋酐）清除效果最好,在浓度1.0 mg/mL时,DPPH清除能力、ABTS自由基清除率、OH自由基清除率分别为91.37%、58.73%、56.36%,分别高于蛋黄果多糖10.0%、43.7%、25.3%。这是引入的乙酰基能活化多糖链中的异头碳,使多糖的供氢能力提升,继而提升乙酰化蛋黄果多糖的抗氧化作用。本研究为海南产蛋黄果多糖的深入开发与抗氧化应用提供了基础研究数据。     

Multiple Seasonal Modifications in Enzymatic Flavour Formation in Tea

Carotenoid cleavage enzymes isolated from Japanese Camellia sinensis leaves （cultivar Yabukita） were used for investigating the structural patterns of carotenoid cleavage enzymes.Fresh tea leaves were used for the isolation of active enzymes and purified to single band stage in SDS PAGE gels after isoelectric focusing.The specific activity of the carotenoid cleavage enzymes was tested and the active fractions selected for further analysis.The sugar content and the amount of phosphate present in the purified enzymes were elucidated by the following methods：Phosphates were detected by phosphatase assays,fluorescence marker kits and ammoniumheptamolybdate complex measurements after incineration of the samples.Sugars were detected in gels using PAS reagent （periodide acid/ Schiff reagent） staining and by GC-MS after hydrolysation of the proteins with trifluoric acid.Phosphorylations as well as glycosylations of the samples could be detected in all cases,thus giving evidence for an increasing phosphorylation level of proteins in Camellia sinensis from spring （1.84 g/mg） to autumn （2.39 g/rg） as well as the presence of at least four different sugars （arabinose,xylose,galactose and ribose）.These secondary modifications of the carotenoid cleavage enzymes and their dependency on the harvesting season may well correspond to the changes on the functional level which were detected between spring （Michaelis Constant （Km） =9.45 mol/1） and autumn （K =17.16 mol/l） harvests.     

植物多倍化过程中小分子RNA调控基因表达机制研究进展

多倍体在植物界中广泛存在,因其往往具有显著超出双亲的生长优势和适应性而成为植物学研究的热点之一。越来越多的研究表明小分子RNA介导的基因激活或沉默及染色质修饰在植物多倍化过程中的基因表达调控方面发挥重要作用。本文结合本实验室在小麦多倍化研究中的观察,着重综述近几年来小分子RNA层面的植物多倍化过程中基因表达调控的机制,以期对多倍体作物的研究和利用提供有益的借鉴。     

Monitoring the development of thermoregulation in poultry embryos and its influence by incubation temperature

The general purpose of this review is to show the stage of the development of peripheral and central nervous thermoregulatory mechanisms in poultry embryos at the end of incubation, and the impact of long-term changes in incubation temperature. Methods are described which (a) allow continuous measurement of peripheral thermoregulatory mechanisms simultaneously with the body temperature of the embryo, and (b) can be used for identification of changes in the sensitivity of the central controller of body temperature during the development as well as after prenatal temperature experiences. Further, a method for characterisation of ‘critical periods’ in the development of the respective body function is introduced.The results of our investigations were discussed in relation to the following general rules:(a) The development of peripheral and central nervous thermoregulatory mechanisms begins in the course of the prenatal ontogeny. At the end of incubation poultry embryos have all the prerequisites to react to changes in incubation temperature. Regarding the peripheral thermoregulatory mechanisms the most sensitive parameter for characterization of the developmental level of embryonic thermoregulation is the deep body temperature.(b) Functional systems of the organism develop from open loop system without feedback control into closed system controlled by feedback mechanism. Acute changes in the environmental conditions (e.g. incubation temperature) induce as a rule, initially uncoordinated and immediately non-adaptive reactions. Later the uncoordinated (immediately non-adaptive) reactions change into coordinated (adaptive) reactions, probably with closing of the regulatory system (‘critical period’). Environmental manipulation of immature physiological mechanisms could be used for characterization of ‘critical periods’ of the respective system. Monitoring of changes in the reactions of thermoregulatory mechanisms on the applied changes in incubation temperature during different perinatal time windows could help to limit ‘critical periods’ in the development of the thermoregulatory system.(c) During this ‘critical periods’, the actual environment modulates the development of the respective physiological control systems for the entire life period. Perinatal epigenetic temperature adaptation could be a tool to adapt poultry embryos or hatchlings to later climatic conditions. For detection of immediate and long-term effects of perinatal epigenetic temperature adaptation (‘imprinting’ of the thermoregulatory system) recordings of changes in neuronal hypothalamic thermosensitivity as well as in neuronal response on temperature stress are useful and have to be verified by identification of the respective effector genes and epigenetic changes in its expression.     

油菜直播机组无人播种作业远程监测系统设计

为解决油菜直播机组无人播种作业过程中田间播种质量信息难以实时直观展示的问题,本研究以雷沃804拖拉机及其搭载的2BYQ-8型气送式油菜直播机为试验平台,设计一套油菜直播机组无人播种作业远程监测系统。该系统由油菜直播机组无人播种作业平台、无人播种作业数据采集系统和播种质量监测云平台三部分构成,通过对雷沃804拖拉机档位、离合、动力输出装置（power take off,PTO）、悬挂机构进行电控液压改装,设计相应控制策略实现直播机组的无人播种作业;利用车载路由器组建播种监测终端和车载计算机之间的局域网,实现对播种数据与导航数据的融合同步,并通过网络连接传输给云平台进行数据存储与实时展示;云平台计算播种质量数据及其对应的田间位置数据,基于网页端高精度地图生成田间作业区域的播种状态图。结果显示,直播机组无人播种作业段横向平均偏差0.037 m,最大偏差0.125 m,电控液压改装系统运行稳定、可靠,满足直播机组无人作业要求,4G网络条件下,云平台通信最大数据传输时延不超过100 ms,云存储数据完整无遗漏,各播种通道田间播量检测准确率不低于96.16%,满足远程监测系统实时性和准确性要求。研...     

利用MSAP分析18个芥蓝齐口期的表观遗传多样性

本研究利用MSAP检测18个芥蓝齐口期DNA甲基化水平,分析了表观遗传多样性,探讨DNA甲基化模式对齐口期的影响。结果表明,18个芥蓝齐口期平均为50d,叶片数平均为10片,齐口期和叶片数不相关（相关系数为0.296）;变异系数分别为21%和18%;遗传距离分布在0~40,平均值为12.2276,在10.62处分为3类。MSAP分析表明,5对引物组合扩增得到432条多态性条带,201条片段表现出多态性,多态性比率为47%;Nei遗传距离分布在0.004~0.467,平均值为0.0958,表明遗传多样性水平较低;在0.04处分为3类。Mantel测验表明两种分析的遗传距离相关系数为-0.1366,显示齐口期、叶片数与DNA甲基化多态性没有相关性。DNA甲基化模式分析表明,非甲基化片段为110条,甲基化多态性片段为322条,分为3种带型,类型一为非甲基化带型（110条）,类型二为甲基化带型（110条）,类型三为半甲基化带型（152条）,非甲基化片段和半甲基化片段在不同品种之间呈现多态性,甲基化片段在不同品种之间呈现多态性与单态性相差不大,显示MSAP多态性主要来源于非甲基化和半甲基化片段,芥蓝甲基化模式以半甲基化为主。本文推测DNA甲基化水平降低参与芥蓝齐口期调控,MSAP分析既可用于基因组结构研究,又可用于基因组水平上性状的功能研究。     

Characterization of a Novel Conus bandanus Conopeptide Belonging to the M-Superfamily Containing Bromotryptophan

A novel conotoxin (conopeptide) was biochemically characterized from the crude venom of the molluscivorous marine snail, Conus bandanus (Hwass in Bruguière, 1792), collected in the south-central coast of Vietnam. The peptide was identified by screening bromotryptophan from chromatographic fractions of the crude venom. Tandem mass spectrometry techniques were used to detect and localize different post-translational modifications (PTMs) present in the BnIIID conopeptide. The sequence was confirmed by Edman’s degradation and mass spectrometry revealing that the purified BnIIID conopeptide had 15 amino acid residues, with six cysteines at positions 1, 2, 7, 11, 13, and 14, and three PTMs: bromotryptophan, γ-carboxy glutamate, and amidated aspartic acid, at positions “4”, “5”, and “15”, respectively. The BnIIID peptide was synthesized for comparison with the native peptide. Homology comparison with conopeptides having the III-cysteine framework (–CCx₁x₂x₃x₄Cx₁x₂x₃Cx₁CC–) revealed that BnIIID belongs to the M-1 family of conotoxins. This is the first report of a member of the M-superfamily containing bromotryptophan as PTM.