枇杷果皮响应高温强光胁迫的蛋白质组分析

为探讨枇杷[Eriobotrya japonica(Thunb.)Lindl.]果皮在高温强光胁迫下的蛋白质组分变化,采用蛋白质组学方法分析了果实日灼抗性差的枇杷种质‘WDYDB’果皮蛋白质对高温强光胁迫的应答反应。结果表明,在自然高温强光胁迫与遮光处理(对照)下,枇杷果皮蛋白质双向电泳图谱中表达量差异在2倍以上的蛋白点共有31个;通过MALDI-TOF-TOF/MS质谱分析成功鉴定出26个差异蛋白点,包括11个下调蛋白和15个上调蛋白。根据这些蛋白功能,可将其分为防御应答、碳水化合物和能量代谢、光合作用、其它等4类蛋白。同时,对这些蛋白质在高温强光胁迫下的功能和作用进行了讨论。这些差异蛋白质参与了枇杷对高温强光胁迫的响应。     

水胁迫下沙漠小球藻蛋白质组学分析

通过研究小球藻(Chlorella sp.)在干旱胁迫下的蛋白质组变化,从蛋白质表达水平解释小球藻对干旱胁迫的响应机理。以20%的PEG 6000胁迫处理0、6、12、18、24和30 d,提取小球藻总蛋白,利用双向电泳和质谱鉴定技术分析差异蛋白。共鉴定29个差异蛋白,按功能可分为7类:光合作用、能量合成和转化、物质代谢、抗氧化、转运和细胞结构、抗逆和功能未知蛋白质。这些蛋白的表达变化影响着沙漠小球藻的油脂积累,对干旱胁迫应答起到了至关重要的作用,并直接或间接地参与了沙漠小球藻细胞内的光合作用和油脂合成。     

干旱胁迫下蒙古沙冬青叶片蛋白质组学研究

通过对蒙古沙冬青[Ammopiptanthus mongolicus(Maxim)Cheng.f]叶组织干旱胁迫下蛋白质组变化的研究,从蛋白表达水平阐释其应答干旱胁迫的分子机制。以20%PEG 6000胁迫处理1 h和72 h,以0 h为对照,提取叶片总蛋白,利用双向电泳和质谱鉴定技术分析差异表达蛋白。共鉴定了40个差异表达蛋白,按功能可分为9类:光合作用,ROS清除,蛋白的合成、加工与降解,物质运输,防御相关,RNA加工,氨基酸代谢,其他相关蛋白和功能未知蛋白质。蒙古沙冬青叶片应答干旱胁迫的核心是叶绿体结构和光合作用的维持。     

水稻叶片对镉胁迫响应的蛋白质差异表达

为揭示水稻镉抗性的分子机理,以抗镉水稻品种P1312777和镉敏感水稻品种IR24为材料,在镉离子浓度为0(对照)、50和100 μmol·L-1条件下水培处理7 d,应用蛋白质组学方法分析了2种水稻叶片对镉胁迫响应的蛋白质差异表达.结果表明:镉胁迫下水稻PI312777叶片中共检测到差异表达蛋白质点31个,通过MALDI-TOF/MS分析,鉴定了其中的24个蛋白质(包括20个不同蛋白质,4个重复检出蛋白质);IR24叶片中共检测到差异表达蛋白质点19个,其中15个蛋白质得到鉴定.PI312777叶片鉴定出的20个蛋白质覆盖了IR24叶片鉴定的15个蛋白质,前者有4个与光合作用相关,11个与细胞防御代谢相关,3个与其他代谢相关,2个为功能未知蛋白.与对照相比,不同浓度镉胁迫下,抗镉水稻PI312777叶片中热激蛋白、谷胱甘肽还原酶、蛋白酶体α亚基6型、果糖1,6-二磷酸醛缩酶、硫氧还蛋白和DNA重组修复蛋白均上调表达;镉敏感水稻IR24叶片中热激蛋白、谷胱甘肽还原酶、蛋白酶体α亚基6型的表达无显著差异,果糖1,6-二磷酸醛缩酶和硫氧还蛋白则下调表达.此外,DNA重组修复蛋白仅在镉胁迫的PI312777叶片中表达.水稻PI312777比IR24具有更强的镉抗性与这些差异表达的蛋白质密切相关.     

干旱胁迫对华北绣线菊和金山绣线菊光合能力的影响

分别采取轻度、中度、重度干旱胁迫和复水处理,研究华北绣线菊和金山绣线菊的光合能力动态变化;利用二维双向电泳与质谱鉴定等技术,分析鉴定干旱胁迫前后2种绣线菊蛋白质的差异表达,以及引起其光合能力改变的生理机制.结果表明:干旱胁迫处理显著影响了2种绣线菊的光合能力,最大光合速率、光补偿点和光饱和点逐渐下降,其干旱胁迫反应为渐进效应.轻度和中度干旱胁迫后,2种绣线菊的恢复能力较强;而重度干旱胁迫后的恢复能力较弱.经干旱胁迫诱导后,抗旱能力弱的金山绣线菊有6处蛋白点消失、11处新增蛋白点、13处蛋白点上调表达、4处蛋白点下调表达,均为低分子量酸性蛋白;其中由干旱诱导表达的3种差异蛋白分别为放氧增强蛋白因子1、2和1,5-二磷酸核酮糖羧化酶/加氧酶大亚基的降解片断.绣线菊抗旱能力的差异与干旱胁迫期间光合能力的变化有关.     

杜仲叶片干旱胁迫响应相关差异蛋白的筛选与鉴定

为研究干旱胁迫下杜仲幼苗生理生化及分子响应机制,利用盆栽试验,通过持续（3、6、9、12、15 d）干旱胁迫处理和复水处理,研究杜仲幼苗的生理响应特性。同时,通过研究对照与处理15 d后的杜仲幼苗差异蛋白质组,分析杜仲幼苗对干旱胁迫的分子响应机制。结果表明,随着干旱处理时间的延长,杜仲叶片的水分饱和亏逐渐增加;光合速率、蒸腾速率、胞间二氧化碳浓度、气孔导度均逐渐减小;SOD、POD、CAT活性呈先上升后降低的趋势;丙二醛含量则呈现先上升,然后下降,最后又上升的变化特点;脯氨酸和可溶性糖含量的变化趋势与SOD等活性变化一致,前期上升,后期下降。在复水后,杜仲叶片的所有指标均有所恢复,但未达到干旱处理之前的水平。表明干旱胁迫影响了杜仲叶片的正常生长代谢。通过对干旱处理15 d后杜仲叶片总蛋白进行双向电泳分离和MALDI-TOF-TOF生物质谱鉴定,成功鉴定出36个差异表达蛋白,其中22个上调表达,14个下调表达。对36个差异蛋白进行功能分析发现,这些差异蛋白主要涉及信号传导、光合作用、碳代谢、能量代谢、次级代谢物合成、抗氧化保护酶、氨基酸代谢和蛋白质代谢。推测杜仲为适应干旱胁迫,首先是感应干旱胁迫信号,并传导至细胞内,影响杜仲叶片中光合作用、次级代谢物合成和蛋白质的生物合成;同时,通过过氧化物保护酶的作用,将过多活性氧加以清除;另一方面,则是通过增强糖酵解,磷酸戊糖途径,产生能量供杜仲正常生长所需。从生理机制来看,杜仲叶片同过增加胞内脯氨酸、可溶性糖含量,降低胞内渗透势,减少叶片中水分损失,与氨基酸合成和糖代谢相关蛋白的表达量上升的结果一致。     

盐芥叶片响应干旱胁迫的蛋白质组学初步分析

盐芥是新兴起的植物非生物逆境研究模式植物,研究盐芥叶片蛋白质组对于干旱胁迫的响应,以推进对植物干旱耐受机制的认识。该研究应用双向电泳技术分析了干旱胁迫对于盐芥叶片蛋白质组的影响,结果共鉴定了63个干旱胁迫差异表达蛋白,包括丰度上调的31个,新出现的蛋白点14个,丰度下调的15个,消失的蛋白点3个。应用生物质谱分析技术确定了包括硫氧还蛋白,铁蛋白-1和凝集素在内的9个干旱胁迫响应蛋白的身份,对这些干旱胁迫响应蛋白的功能分类分析表明,盐芥的耐旱机制可能涉及自由基清除能力的增强、能量代谢的调整以及光合作用的维持。     

干旱胁迫下不同抗旱水平陆地棉的叶片蛋白质组学比较研究

为了探讨陆地棉品种抗旱机理,以陆地棉抗旱品种‘中H177’和不抗旱品种‘中S9612’为材料,运用双向电泳结合质谱技术,分析干旱胁迫下不同陆地棉三叶期叶片蛋白质组分差异变化。结果表明:干旱胁迫下,不同陆地棉叶片蛋白表达差异较大;‘中H177’出现30个差异表达蛋白质点,‘中S9612’出现47个差异表达蛋白质点,只在‘中H177’表达差异的蛋白点11个,只在‘中S9612’表达差异的蛋白点28个,差异表达一致蛋白点8个,表达不一致蛋白点11个。质谱共鉴定出43个差异表达蛋白;功能分类分析表明,干旱胁迫蛋白参与光合作用、物质与能量代谢、抗逆相关蛋白、物质运输和活性氧清除;Rubisco活化酶和能量代谢相关蛋白ATP合成酶类表达差异最大。研究结果可以初步为陆地棉抗旱机理的探讨提供一定的理论基础。     

闽楠叶片响应干旱胁迫的蛋白质组分析

闽楠对土壤水分要求较高,为了提高闽楠造林的成活率,该研究以正常供水为对照,测定了不同干旱胁迫时间(7d、14d)及复水后7d的叶片蛋白质组及生理生化指标变化,以探讨闽楠响应干旱胁迫的分子生理机制。结果表明:(1)不同干旱胁迫处理下闽楠叶片蛋白质双向电泳(2-DE)分析结果共发现51个差异表达蛋白;采用MALDI-TOF/TOF成功鉴定到45个蛋白点;这些鉴定出的差异蛋白与光合作用、碳水化合物和能量代谢、胁迫响应与防御、翻译后修饰、蛋白质转换与分子伴侣功能等生理代谢过程密切相关。(2)检测不同干旱处理时间闽楠叶片膜脂过氧化相关MDA含量,防御相关酶SOD、CAT、POD活性和糖代谢相关酶PFK、AGPase、PK及PDH活性,发现各指标在14d持续干旱胁迫时主要呈下降趋势,且变化均达到极显著水平(P0.01)。研究认为,持续干旱胁迫下,光合作用和植物防御系统以及能量和糖代谢的降低是闽楠不耐干旱的重要生理生化原因,研究结果为今后耐旱闽楠的分子育种提供了理论依据。     

花叶木薯变种和木薯栽培种ZM-Seaside叶片光合参数及蛋白组学分析

为研究木薯叶片光合效率对块根产量的影响,本研究利用蛋白质组学方法分析花叶木薯变种(低产种质)和栽培种ZM-Seaside(高产种质)光合作用能力的差异,揭示其鲜薯产量差异原因,为选育高产木薯品种提供基础数据。采用便携式LI-6400光合作用测定仪测定叶片净光合速率(Pn)、气孔导度(Cs)、胞间CO2浓度(Ci)和蒸腾速率(Tr),表明栽培种ZM-Seaside和花叶木薯变种叶片Cs、Ci和Tr没有显著差异,但ZM-Seaside的Pn显著高于花叶木薯变种;利用Western Blot技术分析叶片蛋白质表达水平,结果显示ZM-Seaside叶片与光合作用相关蛋白质Rubisco、OEC和PRXQ的表达水平显著高于花叶木薯变种;采用苯酚法提取叶片全蛋白质,并进行双向电泳分离,及Delta2D软件确定差异蛋白质点,以花叶木薯为对照,在ZM-Seaside叶片蛋白质双向电泳图谱上得到20个差异蛋白质点,其中上调表达15个,下调表达5个;通过MALDI-TOF-MS鉴定差异蛋白质,结合KEGG数据库将其按照功能进行分类,成功鉴定到其中16个涉及光合作用、碳和能量代谢、分子伴侣、结构蛋白、保护蛋白、解毒和抗氧化及未知功能蛋白质;利用String在线软件构建蛋白质互作网络,推测Ribulose-5-phosphate-3-epimerase和chloroplast latex aldolase-like protein是影响木薯叶片光合效率的关键蛋白质,由于它们的上调表达,对木薯块根产量提高有一定促进作用。     

Coronatine Alleviates Water Deficiency Stress on Winter Wheat Seedlings

With the aim to determine whether coronatine(COR) alleviates drought stress on wheat,two winter wheat(Triticum aestivum L.) cultivars,ChangWu134(drought-tolerant) and Shan253(drought-sensitive) were studied under hydroponic conditions.Seedlings at the three-leaf stage were cultured in a Hoagland solution containing COR at 0.1 μM for 24 h,and then exposed to 20% polyethylene glycol 6000(PEG6000).Under simulated drought(SD),COR increased the dry weight of shoots and roots of the two cultivars significantly;the root/shoot ratio also increased by 30% for Shan253 and 40% for ChangWu134.Both cultivars treated with COR under SD(0.1COR+PEG) maintained significantly higher relative water content,photosynthesis,transpiration,intercellular concentration of CO2 and stomatal conductance in leaves than those not treated with PEG.Under drought,COR significantly decreased the relative conductivity and malondialdehyde production,and the loss of 1,1-diphenyl-2-picrylhydrazyl scavenging activity in leaves was significantly alleviated in COR-treated plants.The activity of peroxidase,catalase,glutathione reductase and ascorbate peroxidase were adversely affected by drought.Leaves of plants treated with COR under drought produced less abscisic acid(ABA) than those not treated.Thus,COR might alleviate drought effects on wheat by reducing active oxygen species production,activating antioxidant enzymes and changing the ABA level.     

An integrative proteome analysis of different seedling organs in tolerant and sensitive wheat cultivars under drought stress and recovery

Roots, leaves, and intermediate sections between roots and leaves (ISRL) of wheat seedlings show different physiological functions at the protein level. We performed the first integrative proteomic analysis of different tissues of the drought‐tolerant wheat cultivar Hanxuan 10 (HX‐10) and drought‐sensitive cultivar Chinese Spring (CS) during a simulated drought and recovery. Differentially expressed proteins (DEPs) in the roots (122), ISRLs (146), and leaves (163) showed significant changes in expression in response to drought stress and recovery. Numerous DEPs associated with cell defense and detoxifications were significantly regulated in roots and ISRLs, while in leaves, DEPs related to photosynthesis showed significant changes in expression. A significantly larger number of DEPs related to stress defense were upregulated in HX‐10 than in CS. Expression of six HSPs potentially related to drought tolerance was significantly upregulated under drought conditions, and these proteins were involved in a complex protein–protein interaction network. Further phosphorylation analysis showed that the phosphorylation levels of HSP60, HSP90, and HOP were upregulated in HX‐10 under drought stress. We present an overview of metabolic pathways in wheat seedlings based on abscisic acid signaling and important protein expression patterns.     

Response of wheat canopy CO2 and water gas-exchange to soil water content under ambient and elevated CO2

The nature of the interaction between drought and elevated CO₂ partial pressure (pC_a) is critically important for the effects of global change on crops. Some crop models assume that the relative responses of transpiration and photosynthesis to soil water deficit are unaltered by elevated pC_a, while others predict decreased sensitivity to drought at elevated pC_a. These assumptions were tested by measuring canopy photosynthesis and transpiration in spring wheat (cv. Minaret) stands grown in boxes with 100 L rooting volume. Plants were grown under controlled environments with constant light (300 µmol m^?2 s^?1) at ambient (36 Pa) or elevated (68 Pa) pC_a and were well watered throughout growth or had a controlled decline in soil water starting at ear emergence. Drought decreased final aboveground biomass (?15%) and grain yield (?19%) while elevated pC_a increased biomass (+24%) and grain yield (+29%) and there was no significant interaction. Elevated pC_a increased canopy photosynthesis by 15% on average for both water regimes and increased dark respiration per unit ground area in well‐watered plants, but not drought‐grown ones. Canopy transpiration and photosynthesis were decreased in drought‐grown plants relative to well‐watered plants after about 20–25 days from the start of the drought. Elevated pC_a decreased transpiration only slightly during drought, but canopy photosynthesis continued to be stimulated so that net growth per unit water transpired increased by 21%. The effect of drought on canopy photosynthesis was not the consequence of a loss of photosynthetic capacity initially, as photosynthesis continued to be stimulated proportionately by a fixed increase in irradiance. Drought began to decrease canopy transpiration below a relative plant‐available soil water content of 0.6 and canopy photosynthesis and growth below 0.4. The shape of these responses were unaffected by pC_a, supporting the simple assumption used in some models that they are independent of pC_a.     

外源甜菜碱对干旱胁迫下小麦幼苗膜脂过氧化作用的影响

用PEG4000溶液对小麦（Triticum aestivum L.heimang)幼苗进行胁迫处理和加入外源甜菜碱（Betaine)作同样处理,6、12、18和24h测定生理变化。结果表明,与对照相比,外源甜菜碱导致小麦幼苗叶片含水量下降,相对膜透性增大,MDA含量升高,SOD和POD活性先降后增,同时脯氨酸大量积累。说明外源甜菜碱对小麦幼苗受伤害程度有明显加深作用。     

Identification of changes in Triticum aestivum L. leaf proteome in response to drought stress by 2D-PAGE and MALDI-TOF/TOF mass spectrometry

Drought is an abiotic stress that strongly influences plant growth, development and productivity. To gain a better understanding of the drought-stress responses at physiological and molecular level in wheat plants (Triticum aestivum cv. KTC86211), we performed a comparative physiological and proteomics analysis. Eight-day-old wheat seedlings were treated with polyethylene glycol-simulated drought stress for 0, 24, 48 and 72 h. Drought treatment resulted in alterations of morphology, increased relative electrolyte leakage and reduced length and weight on leaf and root. Stress-induced proteome changes were analyzed by two-dimensional gel electrophoresis in conjunction with MALDI-TOF/TOF. Twenty-three spots differed significantly between control and treated plants following 48 h of drought stress, with 19 upregulated, and 4 downregulated, in leaf tissues. All of the differentially expressed protein spots were identified, revealing that the majority of proteins altered by drought treatment were involved in reactive oxygen species scavenging enzymes and photosynthesis. Other proteins identified were involved in protein metabolism, cytoskeleton structure, defense response, acid metabolism and signal transduction. All proteins might contribute cooperatively to reestablish cellular homeostasis under drought stress. The present study not only provides new insights into the mechanisms of acclimation and tolerance to drought stress in wheat plants, but also provides clues for improving wheat’s drought tolerance through breeding or genetic engineering.     

Identification of Leaf Proteins Differentially Accumulated between Wheat Cultivars Distinct in Their Levels of Drought Tolerance

The drought-tolerant ‘Ningchun 47’ (NC47) and drought-sensitive ‘Chinese Spring’ (CS) wheat (Triticum aestivum L.) cultivars were treated with different PEG6000 concentrations at the three-leaf stage. An analysis on the physiological and proteomic changes of wheat seedling in response to drought stress was performed. In total, 146 differentially accumulated protein (DAP) spots were separated and recognised using two-dimensional gel electrophoresis. In total, 101 DAP spots representing 77 unique proteins were identified by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry. These proteins were allocated to 10 groups according to putative functions, which were mainly involved in carbon metabolism (23.4%), photosynthesis/respiration (22.1%) and stress/defence/detoxification (18.2%). Some drought stress-related proteins in NC47, such as enolase, 6-phosphogluconate dehydrogenase, Oxygen-evolving enhancer protein 2, fibrillin-like protein, 2-Cys peroxiredoxin BAS1 and 70-kDa heat shock protein, were more upregulated than those in CS. Multivariate principal components analysis revealed obvious differences between the control and treatments in both NC47 and CS, while cluster analysis showed that the DAPs displayed five and six accumulation patterns in NC47 and CS, respectively. Protein–protein interaction network analysis showed that some key DAPs, such as 2-Cys peroxiredoxin BAS1, RuBisCO large subunit-binding protein, 50S ribosomal protein L1, 6-phosphogluconate dehydrogenase, glyceraldehyde 3-phosphate dehydrogenase isoenzyme and 70-kDa heat shock protein, with upregulated accumulation in NC47, had complex interactions with other proteins related to amino acid metabolism, carbon metabolism, energy pathway, signal transduction, stress/defence/detoxification, protein folding and nucleotide metabolism. These proteins could play important roles in drought-stress tolerance and contribute to the relatively stronger drought tolerance of NC47.     

Shotgun proteomic analysis of long-distance drought signaling in rice roots

Rice (Oryza sativa L. cv. IR64) was grown in split-root systems to analyze long-distance drought signaling within root systems. This in turn underpins how root systems in heterogeneous soils adapt to drought. The approach was to compare four root tissues: (1) fully watered; (2) fully droughted and split-root systems where (3) one-half was watered and (4) the other half was droughted. This was specifically aimed at identifying how droughted root tissues altered the proteome of adjacent wet roots by hormone signals and how wet roots reciprocally affected dry roots hydraulically. Quantitative label-free shotgun proteomic analysis of four different root tissues resulted in identification of 1487 nonredundant proteins, with nearly 900 proteins present in triplicate in each treatment. Drought caused surprising changes in expression, most notably in partially droughted roots where 38% of proteins were altered in level compared to adjacent watered roots. Specific functional groups changed consistently in drought. Pathogenesis-related proteins were generally up-regulated in response to drought and heat-shock proteins were totally absent in roots of fully watered plants. Proteins involved in transport and oxidation-reduction reactions were also highly dependent upon drought signals, with the former largely absent in roots receiving a drought signal while oxidation-reduction proteins were strongly present during drought. Finally, two functionally contrasting protein families were compared to validate our approach, showing that nine tubulins were strongly reduced in droughted roots while six chitinases were up-regulated, even when the signal arrived remotely from adjacent droughted roots.