脂质组学研究方法及其应用

脂质不仅是生物膜的骨架成分和能量贮存物质,越来越多的证据表明,脂质也参与细胞的许多重要功能。脂质组学是代谢组学的一个重要分支,主要研究生物体内所有的脂质分子的特性以及它们在蛋白质表达和基因调控过程中的作用。脂质组学是依赖技术驱动的科学。近年来,随着人们对脂质研究的重视,脂质组学研究方法和策略有了突破性进展,在动物上开发出的脂质组学分析方法已经扩展应用到植物上。该文重点介绍脂质组学的研究方法及其应用,以期推动脂质组学,特别是植物脂质组学的进一步发展。     

植物脂质应答逆境胁迫生理功能的研究进展

脂质是生命有机体中一类重要的化合物,可以参与并调节多种生命活动,并且在植物应答非生物胁迫(盐胁迫、干旱胁迫和温度胁迫等)过程中发挥着重要生理功能。但长期以来,对于脂质的研究多集中于动物细胞和医学领域,却疏于关注植物研究领域。借助于"组"学思想和生物技术的快速发展,脂质组学由于可以深层次、全面地揭示脂质的组分与功能,近年来备受关注。基于此,文中通过对脂质的功能与分类、脂质组学技术进展、植物脂质响应干旱胁迫、盐胁迫和温度胁迫生理功能进展等的国内外现有研究进行了归纳与总结,并提出了不足与展望,为探索脂质在植物抗逆过程的生理功能和脂质组学等领域深入研究提供一定的基础。     

脂质组学研究方法及其应用

脂质不仅是生物膜的骨架成分和能量贮存物质, 越来越多的证据表明, 脂质也参与细胞的许多重要功能。脂质组学是代谢组学的一个重要分支, 主要研究生物体内所有的脂质分子的特性以及它们在蛋白质表达和基因调控过程中的作用。脂质组学是依赖技术驱动的科学。近年来, 随着人们对脂质研究的重视, 脂质组学研究方法和策略有了突破性进展, 在动物上开发出的脂质组学分析方法已经扩展应用到植物上。该文重点介绍脂质组学的研究方法及其应用, 以期推动脂质组学,特别是植物脂质组学的进一步发展。     

拟南芥SRO蛋白家族的结构及功能分析

许多生物及非生物胁迫都会引起植物的氧化胁迫, 参与植物氧化胁迫反应组分的鉴定备受人们的关注。拟南芥SRO家族成员包括AtRCD1、AtSRO1、AtSRO5等, 调节植物对氧化胁迫的反应。AtSROs参与植物正常的生长发育, 同时在植物应对干旱、盐、重金属等胁迫反应中扮演重要角色。AtSROs存在保守的PARP、RST等特殊功能区, 推测其可能具备蛋白的转录、调节、修饰等功能。文章就拟南芥SRO家族成员的基本状况, 在植物生长发育及应对非生物胁迫反应中的作用进行概述, 为进一步研究AtSROs的生物学功能提供理论基础。     

一氧化氮调控植物缺铁响应的研究进展

一氧化氮(nitric oxide,NO)是植物体内一种重要的信号分子,不仅对植物的生长发育具有重要的调控作用,而且在植物应答缺铁胁迫中同样扮演着关键角色。近年来,有关NO介导的植物缺铁响应调控机制研究取得了一系列重要进展。本文重点针对植物体内NO的合成及其信号转导途径在缺铁胁迫应答中的作用和NO与其他信号分子互作介导植物缺铁响应调控研究进行系统综述与展望,以加深NO在植物缺铁响应调控功能的认识。     

基质辅助激光解吸电离飞行时间质谱在微生物鉴定中的研究进展

与传统的微生物鉴定技术相比，基质辅助激光解吸电离飞行时间质谱(matrix-assisted laser desorption ionization time-of-flight mass spectrometry, MALDI-TOF MS)是一种准确、可靠和快速的鉴定和分型的技术。本文通过检索近年来国内外相关研究论文，总结最新的研究进展，发现MALDI-TOF MS在临床病原微生物、食源性微生物以及环境微生物等鉴定中有较大的优势，加快了微生物鉴定的进程，同时探索该技术在新领域的最新进展和面临的挑战，以期为我国基质辅助激光解吸电离飞行时间质谱技术的发展提供参考。     

植物应答逆境胁迫的蛋白质组学研究进展

逆境胁迫是制约植物生长发育、影响作物产量和质量的关键因子,揭示植物应答胁迫的分子机理一直是人们长期探索的重大课题.随着拟南芥、水稻等模式植物基因组测序的完成,植物基因组学的研究重点已经转变为功能基因组学研究,蛋白质组学是后基因组时代的新兴研究领域,它有助于人们从分子水平上了解植物耐受胁迫的机制.介绍了植物应答非生物胁迫,如盐胁迫、温度胁迫、干旱胁迫、营养胁迫和机械伤害等,以及生物胁迫,如病菌侵害的蛋白质组学最新研究进展,并探讨了利用蛋白质组学技术研究植物抗逆性方面的优势和前景.     

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.     

茉莉素与植物生物胁迫反应

植物固定生长不能移动,时刻面临着害虫咬噬、病原微生物侵染等多种外界环境胁迫。为了应对这些胁迫,植物进化出了复杂且被精细调控的防御系统,包括利用植物激素调控抗性基因的表达以及抗性相关次生代谢产物的积累等。有趣的是,许多昆虫和病原微生物被发现能够采用各种策略来逃避、克服甚至操控植物防御系统,以促进其对宿主植物的利用。茉莉素是一种重要的脂质植物激素,调控生长发育的诸多方面,同时也在植物应对多种生物胁迫和非生物胁迫的防御反应中发挥重要作用。近年来,茉莉素的生物合成、信号转导和生理功能等均得到了广泛研究,并取得了一系列重要进展。概述了茉莉素的生物合成调控与信号转导途径,同时也探讨了茉莉素对植物生物胁迫反应的调控机制,并介绍了昆虫和病原微生物对植物茉莉素途径的操控策略,以期为深入理解茉莉素介导的植物与病原生物之间的相互作用提供参考。     

植物甾醇在植物逆境胁迫中的研究进展

植物甾醇是一类重要的生理活性物质,对植物的生长发育具有重要作用,对响应植物逆境胁迫也具有重要功能.植物甾醇是细胞膜和脂质筏的重要组分,与膜的稳定性密切相关,主要通过甾醇含量的相对变化维持膜的稳定性及影响脂质筏的生物功能响应逆境胁迫.植物甾醇作为信号分子参与逆境胁迫中的信号传导,油菜素内酯类(BRs)是植物甾醇合成途径的重要产物,作为一种重要的信号分子调控植物甾醇合成酶基因的表达以响应逆境胁迫.     

Spatial mapping of lipids at cellular resolution in embryos of cotton

Advances in mass spectrometry (MS) have made comprehensive lipidomics analysis of complex tissues relatively commonplace. These compositional analyses, although able to resolve hundreds of molecular species of lipids in single extracts, lose the original cellular context from which these lipids are derived. Recently, high-resolution MS of individual lipid droplets from seed tissues indicated organelle-to-organelle variation in lipid composition, suggesting that heterogeneity of lipid distributions at the cellular level may be prevalent. Here, we employed matrix-assisted laser desorption/ionization-MS imaging (MALDI-MSI) approaches to visualize lipid species directly in seed tissues of upland cotton (Gossypium hirsutum). MS imaging of cryosections of mature cotton embryos revealed a distinct, heterogeneous distribution of molecular species of triacylglycerols and phosphatidylcholines, the major storage and membrane lipid classes in cotton embryos. Other lipids were imaged, including phosphatidylethanolamines, phosphatidic acids, sterols, and gossypol, indicating the broad range of metabolites and applications for this chemical visualization approach. We conclude that comprehensive lipidomics images generated by MALDI-MSI report accurate, relative amounts of lipid species in plant tissues and reveal previously unseen differences in spatial distributions providing for a new level of understanding in cellular biochemistry.     

Lipidomics in tissues, cells and subcellular compartments

Mass spectrometry (MS) advances in recent years have revolutionized the biochemical analysis of lipids in plants, and made possible new theories about the structural diversity and functional complexity of lipids in plant cells. Approaches have been developed to profile the lipidome of plants with increasing chemical and spatial resolution. Here we highlight a variety of methods for lipidomics analysis at the tissue, cellular and subcellular levels. These procedures allow the simultaneous identification and quantification of hundreds of lipids species in tissue extracts by direct-infusion MS, localization of lipids in tissues and cells by laser desorption/ionization MS, and even profiling of lipids in individual subcellular compartments by direct-organelle MS. Applications of these approaches to achieve improved understanding of plant lipid metabolism, compartmentation and function are discussed.     

An Arabidopsis lipid map reveals differences between tissues and dynamic changes throughout development

Mass spectrometry is the predominant analytical tool used in the field of plant lipidomics. However, there are many challenges associated with the mass spectrometric detection and identification of lipids because of the highly complex nature of plant lipids. Studies into lipid biosynthetic pathways, gene functions in lipid metabolism, lipid changes during plant growth and development, and the holistic examination of the role of plant lipids in environmental stress responses are often hindered. Here, we leveraged a robust pipeline that we previously established to extract and analyze lipid profiles of different tissues and developmental stages from the model plant Arabidopsis thaliana. We analyzed seven tissues at several different developmental stages and identified more than 200 lipids from each tissue analyzed. The data were used to create a web-accessible in silico lipid map that has been integrated into an electronic Fluorescent Pictograph (eFP) browser. This in silico library of Arabidopsis lipids allows the visualization and exploration of the distribution and changes of lipid levels across selected developmental stages. Furthermore, it provides information on the characteristic fragments of lipids and adducts observed in the mass spectrometer and their retention times, which can be used for lipid identification. The Arabidopsis tissue lipid map can be accessed at http://bar.utoronto.ca/efp_arabidopsis_lipid/cgi-bin/efpWeb.cgi .     

磷脂酶Dδ缺失加剧UV-B诱导的膜伤害

检测了拟南芥野生型（WS）及磷脂酶D8缺失突变体在uV-B辐射下的膜脂分子变化,并比较了二者在紫外辐射下的膜脂含量、双键指数及碳链长度的差异。结果发现,紫外辐射导致植株膜脂发生了降解,其中叶绿体膜脂MGDG和DGDG是膜伤害的主要作用靶点,而且突变体中的膜脂降解比野生型剧烈。上述结果说明磷脂酶D8的缺失会加剧紫外辐射诱导的膜伤害,导致植株对紫外辐射更加敏感。     

Lipid profiling of lipoproteins by electrospray ionization tandem mass spectrometry

Lipoproteins are of fundamental importance for the lipid transport and cardiovascular disease. The function and metabolism of lipoproteins is intimately linked to the biophysical properties of their surface lipids. Although a number of disease associations were found for lipid species in plasma, only a few studies reported lipid profiles of lipoproteins. Here, we provide an overview of techniques for lipoprotein separation, methods for lipid species analysis based on electrospray ionization tandem mass spectrometry (ESI-MS/MS) as well as data from recent lipidomic studies on lipoprotein fractions. We also discuss the different analytical strategies and how lipid profiling can expand our understanding of the biology and structures of lipoproteins.     

磷脂酶Dδ缺失加剧UV-B诱导的膜伤害

检测了拟南芥野生型（WS）及磷脂酶Dδ缺失突变体在UV-B辐射下的膜脂分子变化,并比较了二者在紫外辐射下的膜脂含量、双键指数及碳链长度的差异。结果发现,紫外辐射导致植株膜脂发生了降解,其中叶绿体膜脂MGDG和DGDG是膜伤害的主要作用靶点,而且突变体中的膜脂降解比野生型剧烈。上述结果说明磷脂酶Dδ的缺失会加剧紫外辐射诱导的膜伤害,导致植株对紫外辐射更加敏感。     

Deep-lipidotyping by mass spectrometry: recent technical advances and applications

In-depth structural characterization of lipids is an essential component of lipidomics. There has been a rapid expansion of mass spectrometry methods that are capable of resolving lipid isomers at various structural levels over the past decade. These developments finally make deep-lipidotyping possible, which provides new means to study lipid metabolism and discover new lipid biomarkers. In this review, we discuss recent advancements in tandem mass spectrometry (MS/MS) methods for identification of complex lipids beyond the species (known headgroup information) and molecular species (known chain composition) levels. These include identification at the levels of carbon-carbon double bond (C=C) location and sn-position, as well as characterization of acyl chain modifications. We also discuss the integration of isomer-resolving MS/MS methods with different lipid analysis workflows and their applications in lipidomics. The results showcase the distinct capabilities of deep-lipidotyping in untangling the metabolism of individual isomers and sensitive phenotyping by using relative fractional quantitation of the isomers.     

Application of the accurate mass and time tag approach in studies of the human blood lipidome

We report a preliminary demonstration of the accurate mass and time (AMT) tag approach for lipidomics. Initial data-dependent LC-MS/MS analyses of human plasma, erythrocyte, and lymphocyte lipids were performed in order to identify lipid molecular species in conjunction with complementary accurate mass and isotopic distribution information. Identified lipids were used to populate initial lipid AMT tag databases containing 250 and 45 entries for those species detected in positive and negative electrospray ionization (ESI) modes, respectively. The positive ESI database was then utilized to identify human plasma, erythrocyte, and lymphocyte lipids in high-throughput LC-MS analyses based on the AMT tag approach. We were able to define the lipid profiles of human plasma, erythrocytes, and lymphocytes based on qualitative and quantitative differences in lipid abundance.     

Lipid species profiling: a high-throughput approach to identify lipid compositional changes and determine the function of genes involved in lipid metabolism and signaling

The development of electrospray ionization mass spectrometry has provided the foundation for the development of strategies to identify and quantify complex lipids from unfractionated extracts of small biological samples. In the 1990s, the feasibility of detailed lipid profiling was demonstrated; in the past two years, analytical strategies have been extended to include classes of lipids that are unique to plants. High-throughput lipid profiling by electrospray ionization tandem mass spectrometry, in combination with forward- or reverse-genetics approaches, has recently been utilized to identify lipid metabolic pathways that are involved in plant development and stress responses, to specify the roles of particular genes and enzymes in plant responses to environmental cues, to determine the lipid species that serve as the substrates and products of specific enzymes, and to identify lipid-metabolizing enzymes that are involved in varied plant processes.