Metabolomics: creating new potentials for unraveling the mechanisms in response to salt and drought stress and for the biotechnological improvement of xero-halophytes

Breeders have long been interested in understanding the biological function and mechanism of xero-halophytes and their ability for growth in drought-stricken and salinized environments. However, the mechanisms in response to stress have been difficult to unravel because their defenses require regulatory changes to the activation of multiple genes and pathways. Metabolomics is becoming a key tool in comprehensively understanding the cellular response to abiotic stress and represents an important addition to the tools currently employed in genomics-assisted selection for plant improvement. In this review, we highlight the applications of plant metabolomics in characterizing metabolic responses to salt and drought stress, and identifying metabolic quantitative trait loci (QTLs). We also discuss the potential of metabolomics as a tool to unravel stress response mechanisms, and as a viable option for the biotechnological improvement of xero-halophytes when no other genetic information such as linkage maps and QTLs are available, by combining with germplasm-regression-combined marker-trait association identification.     

基于代谢组的厌氧氨氧化菌群对温度的响应机制

【背景】在环境工程领域中,大多数耐冷菌从活性污泥中分离得到。了解活性污泥菌群对低温的响应有助于耐冷菌的驯化培养。【目的】以厌氧氨氧化污泥菌群作为研究对象,研究温度对厌氧氨氧化菌群代谢通路与代谢产物的影响,以期初步阐释厌氧氨氧化菌群低温响应机理。【方法】在25°C与35°C条件下驯化培养厌氧氨氧化污泥,研究温度对反应器脱氮效能、菌群活性与生长以及群落结构的影响,通过代谢组学比较两个温度下厌氧氨氧化菌群代谢物丰度以及代谢通路活性。【结果】虽然低温导致厌氧氨氧化菌群CO_2固定、TCA循环与丙酮酸代谢的下调,进而导致氮去除活性以及增长速率显著下降,但菌群RNA合成水平、腐胺与信号分子合成上调,从而通过转录调控、调控膜脂组成与改变膜结构的方式,调控菌群代谢以适应低温环境。【结论】从分子机理的角度探究了厌氧氨氧化污泥菌群适应低温环境的生理机制,首次阐释了腐胺与信号分子在污泥菌群适应低温过程中的重要作用。     

两种野生灌木幼苗对干旱和盐交叉胁迫的生理响应

长枝木蓼(Ateaphaxis viegata)和刺叶锦鸡儿(Caragana acanthophulla)是乌鲁木齐周边植被组成的重要种,在植被恢复中具有潜在价值。该文通过盆栽控水控盐法研究两种野生灌木幼苗在不同程度的干旱和盐交叉胁迫下的生理反应。结果表明,干旱和盐交叉胁迫下,长枝木蓼可溶性糖增幅较刺叶锦鸡儿大;轻度交叉胁迫时,两种灌木的可溶性糖含量高于仅干旱胁迫或盐胁迫下的。中度和重度交叉胁迫下,长枝木蓼叶片丙二醛含量增幅高于刺叶锦鸡儿。交叉胁迫下,两种灌木叶片叶绿素含量下降,且长枝木蓼叶片的叶绿素含量降幅大于刺叶锦鸡儿。因膜系统的过氧化作用,使MDA含量升高。轻度干旱胁迫提高了两种野生灌木对盐胁迫的耐受能力。两种灌木能很好地适应中度交叉胁迫,重度交叉胁迫对两种灌木的伤害很大,但也能生存。综合比较各参数发现,刺叶锦鸡儿更耐早,而长枝木蓼更耐盐;综合比较各生理参数发现,刺叶锦鸡儿对干旱和盐胁迫的耐受能力较长枝木蓼强。     

Protein kinases in plant responses to drought,salt, and cold stress

Protein kinases are major players in various signal transduction pathways. Understanding the molecular mechanisms behind plant responses to biotic and abiotic stresses has become critical for developing and breeding climate-resilient crops. In this review,we summarize recent progress on understanding plant drought, salt, and cold stress responses, with a focus on signal perception and transduction by different protein kinases, especially sucrose nonfermenting1(SNF1)-related protein kinases(Sn RKs),mitogen-activated protein kinase(MAPK) cascades,calcium-dependent protein kinases(CDPKs/CPKs),and receptor-like kinases(RLKs). We also discuss future challenges in these research fields.     

A parallel proteomic and metabolomic analysis of the hydrogen peroxide- and Sty1p-dependent stress response in Schizosaccharomyces pombe

Using an integrated approach incorporating proteomics, metabolomics and published mRNA data, we have investigated the effects of hydrogen peroxide on wild type and a Sty1p-deletion mutant of the fission yeast Schizosaccharomyces pombe. Differential protein expression analysis based on the modification of proteins with matched fluorescent labelling reagents (2-D-DIGE) is the foundation of the quantitative proteomics approach. This study identifies 260 differentially expressed protein isoforms from 2-D-DIGE gels using MALDI MS and reveals the complexity of the cellular response to oxidative stress and the dependency on the Sty1p stress-activated protein kinase. We show the relationship between these protein changes and mRNA expression levels identified in a parallel whole genome study, and discuss the regulatory mechanisms involved in protecting cells against hydrogen peroxide and the involvement of Sty1p-dependent stress-activated protein kinase signalling. Metabolomic profiling of 29 intermediates using 1H NMR was also conducted alongside the protein analysis using the same sample sets, allowing examination of how the protein changes might affect the metabolic pathways and biological processes involved in the oxidative stress response. This combined analysis identifies a number of interlinked metabolic pathways that exhibit stress- and Sty1-dependent patterns of regulation.     

Genetic mapping of the early responses to salt stress in Arabidopsis thaliana

Salt stress decreases plant growth prior to significant ion accumulation in the shoot. However, the processes underlying this rapid reduction in growth are still unknown. To understand the changes in salt stress responses through time and at multiple physiological levels, examining different plant processes within a single set-up is required. Recent advances in phenotyping has allowed the image-based estimation of plant growth, morphology, colour and photosynthetic activity. In this study, we examined the salt stress-induced responses of 191 Arabidopsis accessions from 1 h to 7 days after treatment using high-throughput phenotyping. Multivariate analyses and machine learning algorithms identified that quantum yield measured in the light-adapted state (F_v′/F_m′) greatly affected growth maintenance in the early phase of salt stress, whereas the maximum quantum yield (QY_max) was crucial at a later stage. In addition, our genome-wide association study (GWAS) identified 770 loci that were specific to salt stress, in which two loci associated with QY_max and F_v′/F_m′ were selected for validation using T-DNA insertion lines. We characterized an unknown protein kinase found in the QY_max locus that reduced photosynthetic efficiency and growth maintenance under salt stress. Understanding the molecular context of the candidate genes identified will provide valuable insights into the early plant responses to salt stress. Furthermore, our work incorporates high-throughput phenotyping, multivariate analyses and GWAS, uncovering details of temporal stress responses and identifying associations across different traits and time points, which are likely to constitute the genetic components of salinity tolerance.     

Metabolomics integrated with transcriptomics and proteomics: Evaluation of systems reaction to nitrogen deficiency stress in microalgae

Microalgae have higher productivity of biomass than the conventional crops of fuel and are therefore, considered a potential biofuel source. Lipid, an important precursor of biodiesel, can be overproduced in microalgae by nitrogen deprivation. During nitrogen deficiency, radicals are overproduced, and the antioxidant levels are insufficient to counteract the radicals. Thus, the increase in cellular oxidative stress level, consequently acts as a stimulus for lipid accumulation. Lipid accumulation requires an excess of acetyl CoA and NADPH that is made possible by the following mechanism. Glycolysis upregulation overproduces pyruvate, which could be further transformed into acetyl CoA by the pyruvate dehydrogenase complex; while the upregulation of the oxidative pentose phosphate cycle generates a high amount of NADPH. In addition to lipid overproduction, the lack of nitrogen often causes the accumulation of carbohydrates in selected species of microalgae, which could be used to generate biogas and bioethanol from the defatted biomass. By providing details on the differential regulation of the biochemical pathways leading to lipid and carbohydrate accumulation in nitrogen starved microalgae, the review opens up new possibilities in the microalgal biofuel production.     

Ectopic overexpression of AtHDG11 in tall fescue resulted in enhanced tolerance to drought and salt stress

Tall fescue (Festuca arundinacea Schreb.) is a cool-season perennial grass, which has been conventionally grown in the temperate area. However, as a major type of cool-season turf grass, its growth has been extended to the sub-tropical climate or even to the transitional climate between the sub-tropical and the tropical, and, in some cases, to heavily salinized lands. The extended growth imposes a serious challenge to its tolerance to the abiotic stress, particularly to drought, salt and high temperature. Here, we report a successful introduction of Arabidopsis AtHDG11 into the tall fescue via Agrobacterium-mediated transformation. The ectopic overexpression of AtHDG11 under the control of CaMV 35S promoter with four enhancers resulted in significantly enhanced tolerance to drought and salt stress. No obvious adverse effects on growth and development were observed in the transgenic plants. The enhanced stress tolerance was associated with a more extensive root system, a lower level of malondialdehyde, a nearly normal Na⁺/K⁺ ratio, a higher level of proline and a kinetically accelerated induction of SOD and CAT activities observed in the transgenic plants during drought and/or salt stress, indicating that an enhanced ROS scavenging capability might play a significant role in the acquired tolerance to the abiotic stress. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. Ya-Jun Cao and Qiang Wei contributed equally to this work.     

植物对盐碱胁迫的响应机制研究进展

盐碱胁迫是制约植物生长发育的主要非生物胁迫之一,也是制约农作物生产和生态环境建设的严峻问题。研究作物的耐盐碱机理,对开发和有效利用盐碱地有重要的现实意义。许多研究将盐碱胁迫笼统称为盐胁迫,实际上这是两种不同的非生物胁迫,且碱胁迫对植物的伤害要大于盐胁迫。总结性阐述了盐碱胁迫对植物的危害。从生物量、光合作用、离子平衡和膜透性等方面分析了植物对盐碱胁迫的响应机制,并结合最新研究从多角度综述了植物的抗盐碱机理,包括合成渗透调节物质、提高抗氧化酶活性、对离子的选择性吸收及p H平衡和诱导抗盐碱相关基因表达。提出了抗盐碱性的途径,即外源物质的加入、与真菌的协同效应、利用生物技术手段、培育耐盐碱品种和抗性锻炼。最后针对植物适应盐碱逆境方面的研究进行了展望,提出了当前研究需要解决的问题和突破口,旨在为提高植物耐盐碱能力、增加作物产量提供一定的理论依据。     

Heterologous expression of the AtDREB1A gene in chrysanthemum increases drought and salt stress tolerance

DNA cassette containing an AtDREB1A cDNA and a nos terminator, driven by a cauliflower mosaic 35S promoter, or a stress-inducible rd29A promoter, was transformed into the ground cover chrysanthemum (Dendranthema grandiflorum) ‘Fall Color’ genome. Compared with wild type plants, severe growth retardation was observed in 35S:DREB1A plants, but not in rd29A:DREB1A plants. RT-PCR analysis revealed that, under stress conditions, the DREB1A gene was over-expressed constitutively in 35S:DREB1A plants, but was over-expressed inductively in rd29A:DREB1A plants. The transgenic plants exhibited tolerance to drought and salt stress, and the tolerance was significantly stronger in rd29A:DREB1A plants than in 35S:DREB1A plants. Proline content and SOD activity were increased inductively in rd29A:DREB1A plants than in 35S:DREB1A plants under stress conditions. These results indicate that heterologous AtDREB1A can confer drought and salt tolerance in transgenic chrysanthemum, and improvement of the stress tolerance may be related to enhancement of proline content and SOD activity.     

Heterologous expression of the AtDREB1A gene in chrysanthemum increases drought and salt stress tolerance

DNA cassette containing an AtDREB1A cDNA and a nos terminator,driven by a cauli- flower mosaic 35S promoter,or a stress-inducible rd29A promoter,was transformed into the ground cover chrysanthemum(Dendranthema grandiflorum)'Fall Color'genome.Compared with wild type plants,severe growth retardation was observed in 35S:DREB1A plants,but not in rd29A:DREB1A plants.RT-PCR analysis revealed that,under stress conditions,the DREB1A gene was over-expressed constitutively in 35S:DREB1A plants,but was over-expressed inductively in rd29A:DREB1A plants.The transgenic plants exhibited tolerance to drought and salt stress,and the tolerance was significantly stronger in rd29A:DREB1A plants than in 35S:DREB1A plants.Proline content and SOD activity were increased inductively in rd29A:DREB1A plants than in 35S:DREB1A plants under stress conditions.These results indicate that heterologous AtDREB1A can confer drought and salt tolerance in transgenic chrysanthemum,and improvement of the stress tolerance may be related to enhancement of proline content and SOD activity.     

Co-culture with Tetragenococcus halophilus changed the response of Zygosaccharomyces rouxii to salt stress

Zygosaccharomyces rouxii and Tetragenococcus halophilus exhibit remarkable salt tolerance and play roles in high-salt fermented food production. This study investigated the effect of co-culture with T. halophilus on Z. rouxii based on analysis of the viability of Z. rouxii in high-salt environments, the plasma membrane integrity, Na⁺, K⁺-ATPase activity, amino acid content of Z. rouxii cell after salt stress and organic acids assay. The results showed both T. halophilus broth supernatant and intracellular component of T. halophilus increased the viability of Z. rouxii in the 12 % environment. Co-cultured Z. rouxii cells maintained better plasma membrane integrity and lowered Na⁺, K⁺-ATPase activity than single-cultured after salt stress. Co-cultured Z. rouxii cells exhibited higher contents of aspartic acid, threonine, serine, asparagine, glutamic acid, alanine, α-amino-n-butyric acid, methionine, homo-cystine, arginine and proline compared with single-cultured after salt stress. More contents of propionic acid, lactic acid and L-pyroglutamic acid and lower contents of L-malic acid and citric acid were detected in co-culture broth. This study shows preculture of T. halophilus and then co-culture with Z. rouxii enhanced the viability of Z. rouxii in high-salt environment. The results may contribute to further understand the interactions between Z. rouxii and T. halophilus in high-salt environments.     

Field response of wheat to arbuscular mycorrhizal fungi and drought stress

Mycorrhizal plants often have greater tolerance to drought than nonmycorrhizal plants. This study was conducted to determine the effects of arbuscular mycorrhizal (AM) fungi inoculation on growth, grain yield and mineral acquisition of two winter wheat (Triticum aestivum L.) cultivars grown in the field under well-watered and water-stressed conditions. Wheat seeds were planted in furrows after treatment with or without the AM fungi Glomus mosseae or G. etunicatum. Roots were sampled at four growth stages (leaf, tillering, heading and grain-filling) to quantify AM fungi. There was negligible AM fungi colonization during winter months following seeding (leaf sampling in February), when soil temperature was low. During the spring, AM fungi colonization increased gradually. Mycorrhizal colonization was higher in well-watered plants colonized with AM fungi isolates than water-stressed plants. Plants inoculated with G. etunicatum generally had higher colonization than plants colonized with G. mosseae under both soil moisture conditions. Biomass and grain yields were higher in mycorrhizal than nonmycorrhizal plots irrespective of soil moisture, and G. etunicatum inoculated plants generally had higher biomass and grain yields than those colonized by G. mosseae under either soil moisture condition. The mycorrhizal plants had higher shoot P and Fe concentrations than nonmycorrhizal plants at all samplings regardless of soil moisture conditions. The improved growth, yield and nutrient uptake in wheat plants reported here demonstrate the potential of mycorrhizal inoculation to reduce the effects of drought stress on wheat grown under field conditions in semiarid areas of the world.     

植物水通道蛋白的干旱应答机制研究进展

干旱胁迫是严重影响全球作物生产的非生物胁迫之一,研究植物耐旱机制已成为一个重要领域。水通道蛋白是一类特异、高效转运水及其它小分子底物的膜通道蛋白,在植物中具有丰富的亚型,参与调节植物的水分吸收和运输。近10年来,水通道蛋白在植物不同生理过程中的作用,一直受到研究人员的关注,特别是在非生物胁迫方面,而研究表明水通道蛋白在干旱胁迫下对植物的耐旱性起着至关重要的作用,能维持细胞水分稳态和调控环境胁迫快速响应。水通道蛋白在植物耐旱过程中的调控机制及功能较复杂,而关于其应答机制和不同亚型功能性研究的报道甚少。该文综述了植物水通道蛋白的分类、结构、表达调控和活性调节,分别从植物水通道蛋白响应干旱表达调控机制、水通道蛋白基因表达的时空特异性、水通道蛋白基因的表达与蛋白丰度,水通道蛋白基因的耐旱转化四个方面阐明干旱胁迫下植物水通道蛋白的表达,重点阐述其参与植物干旱胁迫应答的作用机制,并提出水通道蛋白研究的主要方向。     

Accumulation of trehalose in response to desiccation and salt stress in the terrestrial cyanobacterium Nostoc commune

Changes in photosynthetic activity and trehalose levels in field‐isolated, natural colonies of the terrestrial cyanobacterium Nostoc commune responding to desiccation and salt stress were investigated. As the water content decreased in N. commune colonies during desiccation, photosynthetic O₂‐evolving activity decreased and no activity was detected in desiccated colonies. A high level of O₂ evolution was restored in the colonies as they absorbed atmospheric moisture, indicating that only a small amount of water is required for reactivation of photosynthesis. No detectable trehalose was found in fully hydrated N. commune colonies; however, trehalose accumulation occurred in response to water loss during desiccation and high levels of trehalose were detected in the air‐dried colonies. Moreover, a 0.2 M NaCl treatment also induced trehalose accumulation to a level equivalent to that by desiccation. Photosynthetic O₂ evolution was inhibited by 0.2 M NaCl, indicating that N. commune can tolerate only low levels of salt. These results suggest that cessation of photosynthesis and trehalose accumulation occur in response to both matric water stress (desiccation) and osmotic water stress (high salt concentration), and that while trehalose may be a less effective osmoprotective compound than others, it is important for the extreme tolerance to desiccation observed in terrestrial cyanobacterium.     

干旱胁迫下菊芋各器官生物量及物质分配规律

以红皮和白皮菊芋为研究材料,通过设置不同的灌水梯度,研究了干旱胁迫对菊芋各器官生物量及物质分配规律的影响。试验结果表明,干旱胁迫降低了红皮和白皮菊芋各器官的生物量;其中,50%灌水条件下两种菊芋的总生物量、地上生物量和地下生物量显著高于10%、20%、30%和40%;红皮菊芋根冠比随着灌水量的减少呈现逐渐增加的变化趋势,白皮菊芋呈现"升-降-升"的变化趋势,且在相同水分处理条件下红皮菊芋茎叶比和根冠比均高于白皮;红皮菊芋随着灌水量的降低根系贡献率呈现逐渐增加的变化趋势,叶片贡献率、块茎贡献率、花贡献率、叶柄贡献率、茎秆贡献率均呈现逐渐降低的变化趋势,白皮菊芋块茎贡献率、花贡献率随灌水量的降低呈现逐渐降低的变化趋势,其他各器官的贡献率随灌水量降低没有规律可循。