两种绣线菊低温锻炼与脱锻炼处理对蛋白质表达的影响

通过2维双向电泳法,研究抗寒能力与光合能力不同的金山绣线菊与华北绣线菊,低温锻炼处理对蛋白质表达所产生的影响。结果表明,低温锻炼处理对2种绣线菊蛋白质表达的影响显著不同,在低温锻炼与脱锻炼处理中,金山绣线菊产生低温诱导特异蛋白,华北绣线菊没有低温诱导特异蛋白表达;蛋白质双向电泳图谱分析表明,华北绣线菊与金山绣线菊经过低温锻炼后发生蛋白质差异表达,经质谱鉴定其中的3种显著差异表达的蛋白质分别为2种未知新蛋白,2种未知蛋白的部分氨基酸序列分别为VSHLAGFSSNNPK、LKADKPTLLSEAK;DPNDHPNPFTVK、DGNGFFLYLLDPDSSK、NGDGMFLYLLDGLESK,另一种差异蛋白为抗坏血酸过氧化物酶;低温锻炼处理使绣线菊中与光合作用密切相关的蛋白质抗坏血酸过氧化物酶发生了显著的变化。     

两种绣线菊耐弱光能力的光合适应性

研究彩色叶花灌木金山绣线菊(Spiraea bunmalba‘Goldmound’)、金焰绣线菊(Spiraea bunmalba‘Goldflame’)对弱光胁迫的光合适应能力,分析其弱光处理及恢复过程中光合特性的变化,探讨其对弱光环境的光合生理适应性变化,为绣线菊在城市园林不同光照环境中得到科学应用提供参考.2种绣线菊具明显光合日变化规律,光强为自然光照60％-65％、40％-45％时光合速率日变化为双峰型曲线,第1峰值高于第2峰值,有明显光合“午休”现象,光强为自然光照20％-25％时光合速率日变化为单峰型曲线,无明显光合“午休”现象;2种绣线菊最大净光合速率、光补偿点、光饱和点、最大表观量子效率和暗呼吸速率随处理光强减弱而减小;叶绿素(a+b)、类胡萝卜素含量随处理光强减弱而增加,叶绿素a/b值随处理光强减弱而降低;随处理光强减弱2种绣线菊叶片变薄,上、下表皮细胞变小,栅栏组织、海绵组织厚度变薄,海绵组织细胞间隙变大,2者比值减小;2种绣线菊弱光胁迫均产生低分子量(45.0-66.2 kDa)特异表达蛋白,推测为弱光胁迫诱导蛋白.2种绣线菊经60％-65％、40％-45％弱光处理后具一定恢复能力,光强仅为自然光照20％-25％处理时其恢复能力较弱.栅栏组织与海绵组织厚度比值、叶绿素a/b值和最大净光合速率是评价2种绣线菊耐弱光能力的重要指标,叶绿素a/b值与耐弱光能力显著负相关,栅栏组织与海绵组织厚度比值、最大净光合速率与耐弱光能力显著正相关,2种绣线菊耐弱光能力按隶属函数值排序为金焰绣线菊＞金山绣线菊.     

玉米抗旱突变体edt1抗旱机制的初步解析

为了选育玉米抗旱新品种,并进一步探究玉米应对干旱胁迫的分子调控机制,本研究采用田间抗旱筛选得到的抗旱突变体edt1,分别进行中度和重度2种干旱胁迫处理,并测定相关参数。结果表明:在中度干旱胁迫下,edt1的净光合速率(P_n)显著高于野生型‘郑58’,PSII基因psb A、lhcb3和lhcb4的表达与‘郑58’相比显著增强;在重度干旱胁迫下,edt1的质膜受损程度较轻,PSⅡ的最大光化学效率(F_v/F_m)、叶片相对含水量及活性氧清除均显著高于‘郑58’,抗氧化酶活基因Mn-sod3、apx、cat2及cat3的表达较‘郑58’中均显著增加。以上结果说明,干旱胁迫下,edt1具有较强的光合能力与活性氧清除能力,这与PSII反应中心、捕光复合体及抗氧化酶基因的表达调控密切相关。     

干旱-复水对桂西北喀斯特地区青冈栎叶片光合能力、叶绿素荧光和显微结构的影响

为了探讨喀斯特地区适生种青冈栎幼苗对“干旱－复水”环境的适应机制,以当年生青冈栎实生苗为材料,通过盆栽控水试验,研究了4 种土壤干旱胁迫强度[对照（–0.1 MPa）、轻度干旱（–0.5 MPa））、中度干旱（–0.9 MPa）和重度干旱（–1.5 MPa）]及复水处理对叶片的水分状况、光合、叶绿素荧光和解剖结构参数的影响。结果表明：（1）随干旱胁迫加剧,叶片相对含水率、水势、净光合速率（Pn）、蒸腾速率（Tr）、气孔导度（Gs）和胞间CO2浓度（Ci）均显著降低,而气孔限制值（Ls）显著增加;轻度胁迫下各光合参数以及轻中度胁迫下瞬时水分利用效率（WUE）均不受显著影响。复水后,各干旱处理叶片水分参数、Pn、Tr、Gs、Ci、WUE均比复水前提高,Ls比复水前降低;轻度胁迫复水后叶片水分和光合参数均优于对照,中度胁迫仅Ls未恢复到对照,重度胁迫复水后叶片水分和光合参数均未恢复。（2）随干旱胁迫加剧,叶片初始荧光（Fo）显著增加,而最大荧光（Fm）、最大光化学量子产量（Fv/Fm）和潜在光化学效率（Fv/Fo）均显著下降,且在轻度胁迫下均与对照显著差异。复水后,各干旱胁迫Fm、Fv/Fm和Fv/Fo比复水前提高,而Fo均略低于复水前,轻度胁迫复水后各叶绿素荧光参数均恢复到或优于对照,中度和重度胁迫复水后Fo未恢复到对照,且重度胁迫复水后Fv /Fm仅为0.75。（3）随干旱胁迫加剧,叶片厚度、上下表皮厚度、气孔密度、主脉导管直径均显著增加,叶片气孔器长度、宽度、开口面积、海绵组织厚度均显著降低,而栅栏组织厚度、栅海比和主脉厚度均表现为中度＞轻度＞对照＞重度。复水后,仅各干旱胁迫处理的气孔开口面积和主脉厚度比复水前显著提高;轻度胁迫复水后叶片结构参数也均恢复到或优于对照,中度胁迫复水后气孔开口面积仍显著低于对照,重度胁迫复水后气孔开口未能恢复打开,主脉厚度也低于对照。因此,青冈栎幼苗有耐旱性和旱后恢复能力,适合作为喀斯特地区的生态恢复树种,但在青冈栎幼苗抚育阶段应免受中度以上干旱胁迫（–0.9 MPa）,以利于其旱后恢复生长。     

持续干旱胁迫下外源精胺对红椿幼苗的形态与生理调节效应的影响

为了掌握持续干旱胁迫下外源精胺(Spm)对红椿幼苗的形态和生理调节效应,本研究以2年生红椿盆栽幼苗为试验对象,在人工遮雨棚中开展干旱胁迫及外源精胺调节试验,试验设置4个持续干旱胁迫处理:(1)对照(土壤相对含水量45%~50%,每天浇水至饱和状态,CK);(2)轻度干旱胁迫(土壤相对含水量30%~38%,持续干旱7 d);(3)中度干旱胁迫(土壤相对含水量25%~30%,持续干旱14 d);(4)重度干旱胁迫(土壤相对含水量20%~25%,持续干旱21 d)。然后,连续3天对轻度、中度和重度干旱胁迫处理喷施外源Spm(浓度为1 mmol·L-1)进行修复调节,并正常管护5天。结果表明:喷施Spm对重度干旱胁迫下植株形态损伤的修复速度远低于轻度和中度干旱胁迫植株;在对照、轻度干旱胁迫(或中度、重度)以及喷施外源Spm调节三者之间,红椿幼苗叶片相对含水量和叶绿素含量均差异极显著(P0.01);在对照、中度干旱胁迫(或重度)以及喷施外源Spm调节三者之间,红椿幼苗叶片相对电导率均差异极显著(P0.01);在对照、轻度干旱胁迫以及喷施外源Spm调节三者之间,红椿幼苗叶片MDA含量和POD含量差异显著(P0.05);在对照、重度干旱胁迫以及喷施外源Spm调节三者之间,红椿幼苗叶片MDA含量和POD含量差异极显著(P0.01);在对照、重度干旱胁迫以及喷施外源Spm调节三者之间,红椿幼苗叶片游离脯氨酸含量差异极显著(P0.01);喷施1 mmol·L-1Spm可修复红椿在遭受干旱胁迫时造成的形态和生理损伤并提高植株的持续抗旱能力,其修复作用对轻度和中度干旱胁迫更为显著。     

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。     

辣椒开花结果期对干旱胁迫的形态与生理响应

在遮雨网室选用抗旱性较强的农城椒二号和抗旱性较弱的陕蔬2001,研究辣椒在轻度、中度和重度干旱胁迫下不同时间的生长、产量、渗透调节物质、保护酶活性的变化规律及其生理调节机制。结果表明:随干旱胁迫时间的延长,辣椒的株高、分枝数、叶面积、单位面积产量、叶绿素含量和叶片相对含水量的抗旱系数呈下降趋势,下降速率与干旱胁迫程度呈正相关,与品种的抗旱性呈负相关;脯氨酸、丙二醛含量和细胞膜透性相对值随干旱胁迫时间的延长呈上升趋势;POD、SOD、CAT活性和可溶性蛋白相对值随着干旱胁迫时间的延长先升高后下降,抗旱性强的材料增加幅度低于抗旱性弱的材料;可溶性糖含量的相对值在轻度和中度干旱胁迫下呈上升趋势,在重度干旱胁迫下呈上升—下降趋势,且抗旱性强的材料上升速度大于抗旱性弱的材料。相关分析表明,干旱胁迫下,产量与株高、分枝数、叶片叶绿素含量、叶面积、叶片相对含水量抗旱系数呈显著正相关;与细胞膜透性、CAT活性和可溶性蛋白含量抗旱系数呈显著负相关。主成分分析表明,用作辣椒抗旱性鉴定的主要指标是单株产量、株高、叶面积、分枝数、可溶性蛋白、可溶性糖、MDA、叶绿素含量和细胞膜透性及叶片相对含水量,叶片POD、SOD、CAT活性、脯氨酸含量可做为辣椒抗旱性鉴定的次要鉴选指标。     

大青杨对不同干旱胁迫强度的形态和生理响应

为了研究大青杨对干旱环境的耐受性,通过聚乙二醇(PEG-6000)模拟干旱胁迫,对不同胁迫强度下大青杨的生长状态、生物量分配、生理生化应答特征进行测定分析。结果显示,随着干旱胁迫时间的延长和强度的增加,大青杨幼苗的苗高、地径生长量和生物量积累明显减少,轻度、中度、重度胁迫下苗高生长量分别下降了17.71%、20.29%、45.96%,地径生长量下降了4.78%、13.62%、17.29%,根冠比上升了11.93%、32.61%、64.10%。叶片相对含水量呈下降趋势,相对电导率呈上升趋势,轻度、中度、重度胁迫下相对含水量分别比对照低23.60%、38.56%和66.78%,相对电导率比对照高106.97%、161.84%、241.75%,说明干旱胁迫对叶片造成了明显的损伤。脯氨酸、可溶性蛋白、可溶性糖含量均呈现先上升再下降的趋势,且轻度胁迫比中度、重度胁迫变化幅度小。在胁迫前期和中期均高于对照,说明大青杨幼苗有一定的抗旱能力。在胁迫末期有低于对照的趋势,此时幼苗细胞、组织受到了不可逆转的伤害。大青杨叶片的净光合速率明显呈现双峰模式,受胁迫幼苗显著低于对照且到达峰值的时间不同。胁迫后幼苗的第一个峰值出现的时间比对照提前2h。大青杨能够忍耐短时轻度的干旱胁迫环境,中度和重度干旱胁迫对大青杨幼苗期的生长和生理过程有显著影响。     

复水对旱后不同玉米品种植株生长恢复能力及其生理响应特性的影响

该研究选用2个抗旱能力相似但旱后恢复能力存在显著差异的玉米品种‘P3’和‘郑单958’为材料,采用盆栽称重控水法在苗期进行干旱及复水处理,通过测定其生长、水分状况、光合参数、叶绿素荧光参数以及叶绿素含量在干旱及复水过程中的变化规律,探讨干旱及复水过程中生理生化响应与旱后恢复能力的关系。结果发现:(1)抗旱性相同的2个玉米品种在干旱复水后的生长恢复能力表现为‘P3’显著强于‘郑单958’。(2)干旱胁迫后,‘郑单958’和‘P3’的叶片相对含水量差异不显著,但‘P3’能维持较高的叶水势、PSⅡ最大光化学效率和叶绿素含量。(3)经干旱胁迫复水后,‘P3’的净光合速率,PSⅡ最大光化学效率和气孔导度恢复速度快于‘郑单958’,说明‘P3’光合损失恢复能力高于‘郑单958’。研究表明,玉米品种‘P3’的旱后复水生长恢复能力较强,因‘P3’在干旱胁迫下能维持较高的Fv/Fm值和叶绿素含量,光系统的损伤较轻,而且复水后也能较快的恢复;在干旱过程中减轻干旱胁迫对植物光合系统的伤害是旱后复水快速恢复生长的基础,而在复水后快速修复光系统损失能够加快植物复水的恢复速度。     

盐旱交叉胁迫对柽柳幼苗渗透调节物质含量的影响

以2年生柽柳幼苗为试验材料,采用盆栽模拟试验研究不同盐分和干旱胁迫对其叶片中渗透调节物质的影响,以探讨柽柳幼苗对盐旱交叉胁迫的适应性.结果表明:(1)随盐旱胁迫的不断加剧,幼苗叶片中可溶性糖含量呈先升高后降低的趋势,且中度和重度盐旱胁迫下均显著高于对照(CK).(2)幼苗叶片中脯氨酸含量在不同盐旱胁迫下均呈逐渐上升趋势,但在重度盐分和中度、重度干旱交叉胁迫下显著高于CK.(3)幼苗叶片中Na+、Cl含量在不同干旱胁迫下,随盐胁迫的加剧呈不同的变化规律,盐旱胁迫的各个处理水平下均显著高于CK,而K+、Ca2+、SO42-含量在轻度和重度干旱胁迫下随盐胁迫增强不断降低.(4)在中度盐旱胁迫下,K+、Ca2+含量与CK无明显差异.研究表明,柽柳幼苗中渗透调节物质在其抗旱耐盐性方面具有积极的调节作用;柽柳幼苗在盐旱胁迫下表现出一定的交叉适应性,适度的干旱胁迫能增强柽柳幼苗对盐分胁迫的耐受能力.     

Protein expression patterns in two Spiraea species in response to cold treatment

We analyzed the different cold-resistance species Spiraea trichocarpa Nakai and Spiraea bumalda ‘Goldmound’ for low-temperature protein expression, protein types identification, and investigated the cold resistance mechanisms under different levels of low temperature by two-dimensional gel electrophoresis (2-DE) and mass spectrometry. An average of 668 and 559 protein spots were detected by 2-DE of S. bumalda ‘Goldmound’ and S. trichocarpa Nakai, respectively, under different low-temperature treatments. Matrix-assisted laser desorption/ionization time-of-flight mass spectroscopy identified 48 proteins, with varying expression, related to metabolism, amino acid synthesis, transportation, stress responses and oxidation–reduction reactions. The results showed that the photosynthesis of S. bumalda ‘Goldmound’ had been affected, enzymes (RuBisCO large and small subunits) involved in the Calvin cycle were up- and down-regulated, and ATP synthase in photophosphorylation was down-regulated. Cytosolic malate dehydrogenase expression weakened in the TCA cycle, while amino acid synthesis strengthened. The activity of four antioxidant enzymes (superoxide dismutase [Cu–Zn], L-ascorbate peroxidase, glutathione peroxidase and peroxidase) was reduced under varying low temperatures. Enzymes (ribulose-bisphosphate carboxylase and RuBisCO small chain precursor) involved in the photosynthesis of S. trichocarpa Nakai showed obvious up- and down-regulation under low temperatures. Cold treatment influenced the photosynthesis of S. trichocarpa Nakai and S. bumalda ‘Goldmound’, but the results showed significant differences between the two species, which were supposed to the fact that low temperature modified the metabolic mechanisms and led to the weaker cold resistance in S. bumalda ‘Goldmound’ than in S. trichocarpa Nakai.     

Study of the physiological mechanisms of two species of <Emphasis Type="Italic">Spiraea</Emphasis> during adaptation to drought treatment

The photosynthetic rate, light saturation point, light compensation point, changes in the MDA and SOD activities, and protein expression of two different drought-resistance species, Spiraea fritschiana and Spiraea trichocarpa, were assessed in this study. Furthermore, the drought-resistant physiological mechanisms of both species were analyzed at the protein level. The photosynthetic capacities of two Spiraea species decreased under drought stress, and the light saturation point and light compensation point decreased. However, their capacities to use weak light increased. Spiraea fritschiana, which demonstrated a stronger drought resistance, showed a better ability to adapt to weak light than S. trichocarpa. The content of MDA in S. fritschiana was notably lower than that in S. trichocarpa, indicating that the concentration of the membrane peroxidation products of S. fritschiana was less than those of S. trichocarpa. Compared with S. trichocarpa, S. fritschiana’s SOD activity was higher, and its ability to remove ROS was also better. Sixty-six proteins were identified with significantly different expression behavior and included regulatory, redox homeostasis, metabolism and energy, and cytoskeleton proteins. The results showed that the photosynthesis of S. trichocarpa was significantly affected by the drought stress. Enzymes in photosynthesis changed significantly; the expression of the RuBisCo large subunit decreased; and RuBisCo carboxylase, the chlorophyll a–b binding protein, ATP synthase, OEC 33 kD photosystem II protein and 23 kD OEC protein greatly increased. In addition, four antioxidant enzymes greatly increased, GroES chaperonin decreased, and eIF5A significantly increased under light stress. When S. fritschiana Schneid encountered serious drought stress, in addition to those enzymes that changed significantly under light drought stress in S. trichocarpa Nakai, NAD(P)H-quinone oxidoreductase and eIF5A were up-regulated. Specifically, three heat-shock proteins were induced. The expression of the enzymes of the two Spiraea that were related to photosynthesis, oxidation–reduction and regulation were all affected, but their species and expression patterns were different. In S. trichocarpa Nakai and S. fritschiana Schneid, there were significant changes in the proteins related to energy metabolism and the proteins related to energy transport, respectively. Thus, we considered that, in the case of protein involvement, the differences in the metabolic pathways and adjustment levels might contribute to S. trichocarpa having a weaker drought tolerance than S. fritschiana.     

Proteomic study of Carissa spinarum in response to combined heat and drought stress

Carissa spinarum is one of the secondary advantage plants grown in dry‐hot valleys in China, which can survive under stress conditions of high temperature and extreme low humidity. Here, we studied the physiological and proteomic changes of C. spinarum in response to 42°C heat stress treatment in combination with drought stress. Dynamic changes in the leaf proteome were analyzed at four time points during the stress treatment and recovery stages. Approximately, 650 protein spots were reproducibly detected in each gel. Forty‐nine spots changed their expression levels upon heat and drought treatment, and 30 proteins were identified by MS and 2‐D Western blot. These proteins were classified into several categories including HSP, photosynthesis‐related protein, RNA‐processing protein and proteins involved in metabolism and energy production. The potential roles of these stress‐responsive proteins are discussed.     

Proteome response of Elymus elongatum to severe water stress and recovery

Tall wheatgrass (Elymus elongatum Host) is a drought-tolerant, cool-season forage grass native to Iran. A proteomic approach has been applied to identify mechanisms of drought responsiveness and tolerance in plants undergoing vegetative stage drought stress and then recovery after rewatering. Uniformed clones were reproduced from a parent plant collected from Brojen (central region of Iran). Clones were grown in pots and drought was initiated by withholding water for 16 d. The leaf samples were taken in triplicate from both stressed/rewatered plants and continuously watered controls at five times: (i) 75% FC, (ii) 50% FC, (iii) 25% FC, (iv) 3 d after rewatering, and (v) 14 d after rewatering. Changes in the proteome pattern of shoots were studied using two-dimensional gel electrophoresis. Following the 16 d water stress, both shoot dry weight and leaf width decreased up to 67% compared with the well-watered plants, whereas proline content increased up to 20-fold. Leaf relative water contents (RWC) also declined from 85% to 24%. Out of about 600 protein spots detected on any given two-dimensional gel, 58 protein spots were reproducibly and significantly changed during drought stress and recovery. Only one protein (abscisic acid- and stress-inducible protein) showed significant changes in expression and position in response to severe drought. The fifty-eight responsive proteins were categorized in six clusters including two groups of proteins specifically up- and down-regulated in response to severe drought stress. Eighteen proteins belonging to these two groups were analysed by liquid chromatography tandem mass spectrometry leading to the identification of 11 of them, including the oxygen-evolving enhancer protein 2, abscisic acid- and stress-inducible protein, several oxidative stress tolerance enzymes, two small heat shock proteins, and Rubisco breakdown. The results suggest that E. elongatum may tolerate severe drought stress by accumulating proline and several proteins related to drought-stress tolerance. Recovery after rewatering might be another mechanism by which plants tolerate erratic rainfall in semi-arid regions.     

Responses of the photosynthetic electron transport system to excess light energy caused by water deficit in wild watermelon

In plants, drought stress coupled with high levels of illumination causes not only dehydration of tissues, but also oxidative damage resulting from excess absorbed light energy. In this study, we analyzed the regulation of electron transport under drought/high-light stress conditions in wild watermelon, a xerophyte that shows strong resistance to this type of stress. Under drought/high-light conditions that completely suppressed CO(2) fixation, the linear electron flow was diminished between photosystem (PS) II and PS I, there was no photoinhibitory damage to PS II and PS I and no decrease in the abundance of the two PSs. Proteome analyses revealed changes in the abundance of protein spots representing the Rieske-type iron-sulfur protein (ISP) and I and K subunits of NAD(P)H dehydrogenase in response to drought stress. Two-dimensional electrophoresis and immunoblot analyses revealed new ISP protein spots with more acidic isoelectric points in plants under drought stress. Our findings suggest that the modified ISPs depress the linear electron transport activity under stress conditions to protect PS I from photoinhibition. The qualitative changes in photosynthetic proteins may switch the photosynthetic electron transport from normal photosynthesis mode to stress-tolerance mode.     

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.     

Proteome analysis of soybean roots subjected to short-term drought stress

Drought is one of the most important constraints on the growth and productivity of many crops, including soybeans. However, as a primary sensing organ, the plant root response to drought has not been well documented at the proteomic level. In the present study, we carried out a proteome analysis in combination with physiological analyses of soybean roots subjected to severe but recoverable drought stress at the seedling stage. Drought stress resulted in the increased accumulation of reactive oxygen species and subsequent lipid peroxidation. The proline content increased in drought-stressed plants and then decreased during the period of recovery. The high-resolution proteome map demonstrated significant variations in about 45 protein spots detected on Comassie briliant blue-stained 2-DE gels. Of these, 28 proteins were identified by mass spectrometry; the levels of 5 protein spots were increased, 21 were decreased and 2 spots were newly detected under drought condition. When the stress was terminated by watering the plants for 4 days, in most cases, the protein levels tended towards the control level. The proteins identified in this study are involved in a variety of cellular functions, including carbohydrate and nitrogen metabolism, cell wall modification, signal transduction, cell defense and programmed cell death, and they contribute to the molecular mechanism of drought tolerance in soybean plants. Analysis of protein expression patterns revealed that proteins associated with osmotic adjustment, defense signaling and programmed cell death play important roles for soybean plant drought adaptation. The identification of these proteins provides new insight that may lead to a better understanding of the molecular basis of the drought stress responses.     

A root proteomics-based insight reveals dynamic regulation of root proteins under progressive drought stress and recovery in <Emphasis Type="Italic">Vigna radiata</Emphasis> (L.) Wilczek

To understand the complex drought response mechanism in crop plants, a systematic root proteomics approach was adopted to identify and analyze the expression patterns of differentially expressed major root proteins of Vigna radiata during short-term (3 days) and consecutive long-term water-deficit (6 days) as well as during recovery (6 days after re-watering). Photosynthetic gas exchange parameters of the plant were measured simultaneously during the stress treatment and recovery period. A total of 26 major protein spots were successfully identified by mass spectrometry, which were grouped according to their expression pattern during short-term stress as significantly up-regulated (9), down-regulated (10), highly down-regulated, beyond detection level of the software (2) and unchanged (5). The subsequent changes in the expression patterns of these proteins during long-term stress treatment and recovery period was analyzed to focus on the dynamic regulation of these functionally important proteins during progressive drought and recovery period. Cytoskeleton-related proteins were down-regulated initially (3d) but regained their expression levels during subsequent water-deficit (6d) while glycoprotein like lectins, which were primarily known to be involved in legume–rhizobia symbiosis, maintained their enhanced expression levels during both short and long-term drought treatment indicating their possible role in drought stress response of legumes. Oxidative stress-related proteins including Cu/Zn superoxide dismutase, oxidoreductase and aldehyde reductase were also up-regulated. The analyses of the dynamic regulation of these root proteins during short- and long-term water-deficit as well as recovery period may prove crucial for further understanding of drought response mechanisms in food legumes.     

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.