外源绿原酸缓解黑大豆SB铝毒害的机理研究

该文研究了外源绿原酸(CGA)对Al胁迫下铝敏感型黑大豆SB根生理生化指标以及根中胁迫相关基因表达的变化,探讨外源CGA缓解SB根铝毒害的效果及分子机理。以不同浓度Al和CGA处理SB,筛选出CGA缓解Al毒害的最佳浓度,测定Al含量、抗氧化系统酶活性、14-3-3蛋白与H~+-ATP酶的表达、H~+泵活性。结果表明:低浓度CGA能缓解Al胁迫下黑大豆SB根伸长抑制,并促进侧根数目增加,而高浓度CGA的缓解效果下降;0.01 g·L~(-1) CGA使Al胁迫下SB根尖Al含量与MDA含量下降,促进根系柠檬酸的分泌。RTPCR和Western Bloting分析表明0.01 g·L~(-1) CGA促进Al胁迫下SB根中14-3-3b、14-3-3m、14-3-3k和GHA2基因(质膜H~+-ATP酶)的表达,抑制MATE基因的表达。同时,0.01 g·L~(-1) CGA能促进Al胁迫下质膜H~+-ATP酶蛋白磷酸化水平以及其与14-3-3蛋白结合,且能提高质膜H~+-ATP酶和H~+泵活性。因此推测外源CGA可能通过增加侧根数,增强14-3-3蛋白和质膜H~+-ATP酶基因蛋白表达水平和互作,弥补Al胁迫下MATE表达的抑制,增加柠檬酸的分泌,增强SB对铝毒害的耐受性。     

水杨酸调控内源H_2S缓解黑大豆铝胁迫的作用机理研究

该研究以铝(Al)敏感型黑大豆(SB)根为实验材料,通过一系列生理生化和组织化学实验手段,探讨了水杨酸(SA)通过调控内源H_2S信号缓解铝胁迫的作用方式。结果表明:(1)AlCl_3处理黑大豆SB根系Al积累增加,AlCl_3与SA共处理能明显抑制Al在SB根系的累积,加入H_2S清除剂(HT)或H_2S合成抑制剂(PAG)后SB根系Al累积量增加。(2)SA使Al胁迫下黑大豆(SB)根内源H_2S水平增加1.5倍,并显著缓解Al胁迫导致的根生长抑制、活性氧(ROS)累积、氧化损伤和细胞死亡,共处理HT或PAG均能够显著降低内源H_2S水平,并可逆转上述所有SA对Al胁迫的缓解效应。(3)SA降低了Al胁迫下黑大豆(SB)根尖抗氧化酶CAT、SOD和APX活性,抑制SB根系细胞ROS的产生,用HT或PAG抑制H_2S信号可增强抗氧化酶活性。(4)在Al胁迫条件下,SA可进一步上调一系列耐Al基因的表达,包括外部解毒机制中的耐铝转录因子GmART1、柠檬酸合成酶基因GmCS、柠檬酸转运蛋白基因GmMATE,内部解毒机制中的苹果酸转运蛋白基因GmAlCT以及Al3+相关转运蛋白基因GmAlS1和GmNIP1;2,通过HT或PAG降低内源H_2S水平可逆转SA对上述基因表达的调控。(5)SA可提高Al胁迫下黑大豆(SB)根柠檬酸的分泌量,此效应亦可被HT或PAG抑制。研究发现,H_2S可作为SA的下游信号参与调控黑大豆(SB)响应Al胁迫的过程,为揭示植物Al耐受信号调控网络途径提供部分新的理论基础。     

叶片喷施甲醇对铝胁迫下黑大豆抗氧化酶活性的影响

在营养液水培条件下,分析叶片预喷施5％ (V/V)甲醇对50μmol·L-1的AlCl3胁迫下黑大豆叶和根中H2O2和MDA含量以及抗氧化酶SOD、POD和CAT活性的影响,为进一步研究甲醇及抗氧化酶在响应铝胁迫应答中的调控机制奠定基础.结果显示:(1)在铝胁迫下,随着铝处理浓度的升高根尖吸收的铝越多,且根的生长被抑制越严重.(2)铝胁迫能诱导黑大豆叶和根中可溶性总蛋白、H2O2和MDA含量的增加,POD和CAT活性增强,但SOD活性变化不明显.(3)当叶片喷施5％甲醇预处理后再进行50 μmol·L1 AlCl3胁迫,随着处理时间增加黑大豆叶和根中可溶性总蛋白含量及POD和CAT活性显著增加,而H2O2和MDA含量显著下降,SOD活性变化亦不显著.研究结果表明,在铝胁迫下叶片喷施5％甲醇能够增强黑大豆叶和根中POD和CAT活性,降低H2O2和MDA的积累,这可能是甲醇通过抗氧化酶参与铝胁迫应答的一种重要机制.     

过氧化氢预处理增强敏感型黑豆抗铝能力的生理机制

以铝敏感型黑豆(简称SB)幼苗为实验材料,在水培条件下进行不同浓度过氧化氢(H2O2)和AlCl3处理,考察其生长和相关生理指标变化,探讨H2 O2预处理缓解黑豆铝毒害的生理以及分子机理.结果显示:(1)黑豆幼苗根的相对生长量在0.1和1.0 μmol·L-1 H2O2处理下始终得到显著促进,并以后者效果更好,而在10.0和100.0 μmol·L1 H2O2处理下先表现促进后受到显著抑制.(2)经过1.0 μmol·L-1 H2O2预处理的SB幼苗在不同程度Al3+胁迫(50～400 μmol·L 1)3周后,其叶片和根中总蛋白含量分别显著增加了16.7％～41.2％和10.0％～25.0％,MDA含量减少了近50％;而其同期的SOD和POD活性显著增加.(3) H2 O2预处理可诱导SB根和叶中的SOD基因(Mg/Fe-SOD和Mn-SOD)的表达水平明显提高.研究表明,低浓度H2O2能显著促进铝胁迫下铝敏感型黑豆幼苗生长,且主要是通过增加植株抗氧化酶活性和相关基因表达水平来提高其对铝胁迫的抗性.     

土壤水分胁迫对小麦根系与旗叶衰老的影响

随土壤水分胁迫加剧,旗叶,根中的超氧化物歧化酶,过氧化氢酶活性降低,腊脂过氧化产物丙二醛含量增加,花后１４ｄ为膜脂过氧化作用加重的转折点,此时受水分胁迫愈重,旗叶与根中ＳＯＤ,ＣＡＴ活性降低愈迅速,ＭＤＡ含量迅速升高,根系活力,旗叶与根中可溶性蛋白含量骤降,加重了膜脂过氧化程度,降低了清除了自由基能力,加速植株衰老。     

干旱胁迫下牛心朴子幼苗相对含水量、质膜透性及保护酶活性变化

牛心朴子(Cynanchum komarovii)幼苗用不同浓度PEG-6000(聚乙二醇)模拟干旱处理,测定处理12d和复水24h中根、叶的RWC、质膜相对透性、MDA含量及几种保护酶(SOD、CAT、POD)活性变化情况,结果表明：高浓度胁迫后期,RWC明显下降,MDA含量增大,致使膜脂过氧化引起膜损伤;低、中浓度胁迫下,RWC下降程度低,细胞膜脂过氧化及膜透性影响小,且可能对膜脂过氧化起到一定的防御作用。复水后,低、中度胁迫下,各项指标能迅速恢复到CK水平,而高浓度胁迫下,除叶质膜透性、MDA含量、SOD活性不能恢复外,根、叶的其余指标均能达到CK水平。保护酶系统的作用,在一段时间内,可能是通过它们之间的相互协调而保持一个稳定的平衡态进行的。     

外源一氧化氮供体硝普钠对干旱胁迫下小麦幼苗叶中ATP酶活性和膜脂过氧化的影响

研究外源一氧化氮（NO）供体硝普钠（sNP）对干旱胁迫下小麦幼苗叶片ATP酶活性和膜脂过氧化影响的结果表明,15％聚乙二醇．6000（PEG-6000）模拟的干旱胁迫下小麦幼苗叶中H^＋-ATP酶和Ca^2＋．ATP酶活性显著升高后迅速下降,硫代巴比妥酸反应产物（TBARs）和质量膜透性增加;0．1mm01．L^-1 SNP可提高干旱胁迫下小麦幼苗叶中超氧化物歧化酶（SOD）、过氧化物酶（POD）和过氧化氢酶（cAT）活性,降低超氧阴离子（O2^-）和过氧化氢（H2O2）水平,缓解膜脂过氧化,稳定生物膜的结构和功能,H^＋-ATP酶和Ca^2＋-ATP酶也可以保持更高的活性。     

不同抗旱性玉米幼苗根系抗氧化系统对水分胁迫的反应

以抗旱性不同的2个玉米品种为材料,研究不同程度水分胁迫下玉米根系活性氧清除系统的变化及膜脂过氧化水平。明确了轻度水分胁迫下玉米根系POD、CAT、APX等保护酶活性明显提高;中、重度胁迫下其活性急剧下降,但几种酶对水分胁迫的敏感程度不同。SOD对水分胁迫表现最不敏感,在中度水分胁迫下仍保持上升趋势;抗氧化剂GSH含量变化趋势与保护酶相似;而AsA含量在不同程度水分胁迫下持续下降;MDA含量随水分胁迫程度加剧而增加。其中抗旱性强的鲁玉14与抗旱性弱的掖单13相比具有较高的保护酶活性和抗氧化剂含量,膜脂过氧化程度较轻,除POD外,品种间抗氧化酶活性（抗氧化剂含量）呈极显著差异,说明抗氧化能力强是抗旱性品种具有较强抗旱性的重要原因之一。     

铝对秋茄幼苗生理特性的影响

为探讨铝对秋茄幼苗生理特性的影响,实验研究了不同浓度铝盐(0-100 mmol/L AlCl3)处理后秋茄的各种生理反应,对幼苗的生长、净光合效率、膜脂过氧化作用、游离脯氨酸含量等生理指标与胁迫程度及时间的关系作了对比研究,特别分析了高浓度(25-100 mmol/L Al3+)胁迫下,秋茄叶片和根部活性氧清除系统保护酶活性的变化趋势。研究发现,在10 mmol/L浓度以下,秋茄在生理特性上表现出对铝胁迫的最大适应性,能维持正常生命生长过程。当浓度增加至25-100 mmol/L,秋茄的生理反应较为敏感,膜脂过氧化加重,MDA含量及保护酶活性随铝浓度增加的变化趋势与其在海莲中的表现基本相似。高浓度铝胁迫的极端环境使植物体内产生过量的活性氧自由基,诱导了细胞膜系统的氧化损伤,最终导致秋茄植株衰老甚至死亡。抗氧化保护酶系统SOD、CAT、POD的协同作用,在一定时间内可以维持细胞内活性氧代谢平衡;特别是POD被激活的程度最大,且持续时间最长,可以考虑作为秋茄幼苗抗铝胁迫的生理标志。秋茄叶片和根部的游离脯氨酸含量在25和100 mmol/L Al3+胁迫下均显著增加。       

轻度水分胁迫的小麦幼苗中与呼吸有关的几种酶活性变化

轻度水分胁迫使小麦幼苗叶片呼吸升高时,叶中琥珀酸去氢酶和细胞色素氧化酶活性均明显升高;而同样胁迫使根呼吸下降时,根中这两种酶活性均明显下降。叶和根中ATP酶分解活性在胁迫下都明显升高。轻度水分胁迫使叶片过氧化氢酶活性升高。叶中有明显的乙醇酸氧化酶活性,抗旱品种的酶活性较高,胁迫使此酶活性降低。     

Effects of aluminum on superoxide dismutase and peroxidase activities,and lipid peroxidation in the roots and calluses of soybeans differing in aluminum tolerance

The seedlings of two soybean genotypes, Al-tolerant PI 416937 (PI) and Al-sensitive Young, were cultured in the solution containing 0, 25 or 50 μM Al (AlCl₃·6H₂O) for 24, 36 or 48 h in the hydroponics, and the calluses induced from two genotypes were cultured in medium containing 0, 10, 50 or 100 μM Al for 5, 10 or 15 days, respectively. The effects of Al on growth of seedling roots and calluses, antioxidant enzyme activities of superoxide dismutase (SOD) and peroxidase (POD) and lipid peroxidation were investigated. Under Al stress, PI was more tolerant to Al toxicity than Young at both intact plant and tissue levels and lower concentrations of Al significantly stimulated the root and callus growth of PI. Al application enhanced the activities of SOD and POD and lipid peroxidation in both roots and calluses of two genotypes. Although the differences of SOD activities between two genotypes in response to Al toxicity depended on Al concentration and durations of treatment, SOD activities in the roots of PI were higher than those in the roots of corresponding Young in the presence of Al for 36 or 48 h. Meanwhile, the POD activities in PI roots increased as the Al levels and durations of treatment increased, significantly higher than those in the corresponding Young roots. Moreover, Al-treated PI had significantly lower lipid peroxidation than Young at both root and callus levels. These results suggest that the enhanced antioxidant-related enzyme activities and reduced lipid peroxidation in PI might be one of Al-tolerant mechanisms.     

Effect of soil salinity, soil drought, and their combined action on the biochemical characteristics of cotton roots

Stress caused by soil salinity and soil drought limits cotton productivity in China. To determine the tolerance levels of cotton, we assessed the effects of soil salinity and soil drought on the biochemical characteristics of the roots of two cotton cultivars (CCRI-44, salt-tolerant; Sumian 12, salt-sensitive). Specifically, we analyzed root biomass, fatty acid composition, antioxidative enzyme activity, lipid peroxidation, H⁺-ATPase and Ca²⁺-ATPase activities. The cotton root biomass of the two cultivars declined significantly under conditions of soil salinity, soil drought, and the two stressors combined. The antioxidant enzyme activity of the roots also decreased markedly, which caused lipid peroxidation to increase, and changed the composition of the fatty acid membrane. H⁺-ATPase, Ca²⁺-ATPase and antioxidant enzyme activity decreased more under the two stressors combined. However, H₂O₂ content and O₂ ^? generation increased under the two stressors combined, compared to each stressor separately. Overall, the combination of soil salinity and drought has a greater inhibitory effect and more harmful impact on root growth than each stressor separately. The higher tolerance of CCRI-44 to soil salinity and drought stress than Sumian 12 might be explained by differences in cotton root antioxidative enzyme activity. The lipid peroxidation levels of cotton roots might represent an important biochemical trait for stress tolerance.     

Influence of arbuscular mycorrhiza on lipid peroxidation and antioxidant enzyme activity of maize plants under temperature stress

The influence of the arbuscular mycorrhizal (AM) fungus, Glomus etunicatum, on characteristics of growth, membrane lipid peroxidation, osmotic adjustment, and activity of antioxidant enzymes in leaves and roots of maize (Zea mays L.) plants was studied in pot culture under temperature stress. The maize plants were placed in a sand and soil mixture under normal temperature for 6 weeks and then exposed to five different temperature treatments (5oC, 15oC, 25oC, 35oC, and 40oC) for 1 week. AM symbiosis decreased membrane relative permeability and malondialdehyde content in leaves and roots. The contents of soluble sugar content and proline in roots were higher, but leaf proline content was lower in mycorrhizal than nonmycorrhizal plants. AM colonization increased the activities of superoxide dismutase, catalase, and peroxidase in leaves and roots. The results indicate that the AM fungus is capable of alleviating the damage caused by temperature stress on maize plants by reducing membrane lipid peroxidation and membrane permeability and increasing the accumulation of osmotic adjustment compounds and antioxidant enzyme activity. Consequently, arbuscular mycorrhiza formation highly enhanced the extreme temperature tolerance of maize plant, which increased host biomass and promoted plant growth.     

Antioxidant responses of shoots and roots of lentil to NaCl-salinity stress

The effect of salt stress (100 mM and 200 mM NaCl) on antioxidant responses in shoots and roots of 14-day-old lentil (Lens culinaris M.) seedlings was investigated. Salt stress caused a significant decrease in length, wet-dry weight and an increase in proline content of both shoot and root tissues. In leaf tissues, high salinity treatment resulted in a 4.4 fold increase in H₂O₂ content which was accompanied by a significant level of lipid peroxidation and an increase in electrolyte leakage. Root tissues were less affected with respect to these parameters. Leaf tissue extracts exhibited four activity bands, of which two were identified as Cu/Zn-SOD and others as Fe-SOD and Mn-SOD. Fe-SOD activity was missing in root extracts. In both tissues Cu/Zn-SOD activity comprised 70–75% of total SOD activity. Salt stress did not cause a significant increase in total SOD activity of leaf tissues but a significant enhancement (88%) was observed in roots mainly due to an enhancement in Cu/ZnSOD isoforms. Compared to leaf tissues a significantly higher constitutive ascorbate peroxidase (APX) and glutathion reductase (GR) activity was observed in root tissues. Upon salt stress no significant change in the activity of APX, catalase (CAT) and GR was observed in root tissues but a higher APX activity was present when compared to leaf tissues. On the other hand, in leaf tissues, with the exception of CAT, salt stress caused significant enhancement in the activity of other antioxidant enzymes. These results suggested that, root tissues of lentil are protected better from NaCl stress induced oxidative damage due to enhanced total SOD activity together with a higher level of APX activity under salinity stress. To our knowledge this is the first report describing antioxidant enzyme activities in lentil.     

Protein accumulation in leaves and roots associated with improved drought tolerance in creeping bentgrass expressing an ipt gene for cytokinin synthesis

Cytokinins (CKs) may be involved in the regulation of plant adaptation to drought stress. The objectives of the study were to identify proteomic changes in leaves and roots in relation to improved drought tolerance in transgenic creeping bentgrass (Agrostis stolonifera) containing a senescence-activated promoter (SAG12) and the isopentyl transferase (ipt) transgene that increases endogenous CK content. Leaves of SAG12-ipt bentgrass exhibited less severe senescence under water stress, as demonstrated by maintaining lower electrolyte leakage and lipid peroxidation, and higher photochemical efficiency (F(v)/F(m)), compared with the null transformant (NT) plants. SAG12-ipt plants had higher root/shoot ratios and lower lipid peroxidation in leaves under water stress than the NT plants. The suppression of drought-induced leaf senescence and root dieback in the transgenic plants was associated with the maintenance of greater antioxidant enzyme activities (superoxide dismutase, peroxidase, and catalase). The SAG12-ipt and NT plants exhibited differential protein expression patterns under well-watered and drought conditions in both leaves and roots. Under equivalent leaf water deficit (47% relative water content), SAG12-ipt plants maintained higher abundance of proteins involved in (i) energy production within both photosynthesis and respiration [ribulose 1,5-bisphosphate carboxylase (RuBisCO) and glyceraldehyde phosphate dehydrogenase (GAPDH)]; (ii) amino acid synthesis (methionine and glutamine); (iii) protein synthesis and destination [chloroplastic elongation factor (EF-Tu) and protein disulphide isomerases (PDIs)]; and (iv) antioxidant defence system (catalase and peroxidase) than the NT plants. These results suggest that increased endogenous CKs under drought stress may directly or indirectly regulate protein abundance and enzymatic activities involved in the above-mentioned metabolic processes, thereby enhancing plant drought tolerance.     

Caspase-like enzymatic activity and the ascorbate-glutathione cycle participate in salt stress tolerance of maize conferred by exogenously applied nitric oxide

Salinity stress causes ionic stress (mainly from high Na⁺ and Cl^- levels) and osmotic stress (as a result of inhibition of water uptake by roots and amplified water loss from plant tissue), resulting in cell death and inhibition of growth and ultimately adversely reducing crop productivity. In this report, changes in root nitric oxide content, shoot and root biomass, root H₂O₂ content, root lipid peroxidation, root cell death, root caspase-like enzymatic activity, root antioxidant enzymatic activity and root ascorbate and glutathione contents/redox states were investigated in maize (Zea mays L. cv Silverking) after long-term (21 d) salt stress (150 mM NaCl) with or without exogenously applied nitric oxide generated from the nitric oxide donor 2,2′-(Hydroxynitrosohydrazano)bis-ethane. In addition to reduced shoot and root biomass, salt stress increased the nitric oxide and H₂O₂ contents in the maize roots and resulted in elevated lipid peroxidation, caspase-like activity and cell death in the roots. Altered antioxidant enzymatic activities, along with changes in ascorbate and glutathione contents/redox status were observed in the roots in response to salt stress. The detrimental effects of salt stress in the roots were reversed by exogenously applied nitric oxide. These results demonstrate that exogenously applied nitric oxide confers salt stress tolerance in maize by reducing salt stress-induced oxidative stress and caspase-like activity through a process that limits accumulation of reactive oxygen species via enhanced antioxidant enzymatic activity.     

Aluminum-induced oxidative stress and changes in antioxidant defenses in the roots of rice varieties differing in Al tolerance

The effects of aluminum (Al) on root elongation, lipid peroxidation, hydrogen peroxide (H₂O₂) accumulation, antioxidant levels, antioxidant enzymatic activity, and lignin content in the roots of the Al-tolerant rice variety azucena and the Al-sensitive variety IR64 were investigated. Treatment with Al induced a greater decrease in root elongation and a greater increase in H₂O₂ and lipid peroxidation as determined by the total thiobarbituric acid-reactive substance (TBARS) level in IR64 than in azucena. Azucena had significantly higher levels of superoxide dismutase, ascorbate peroxidase, glutathione reductase, and glutathione peroxidase GSH POD activity compared with IR64. The concentrations of reduced glutathione (GSH) and ascorbic acid, and the GSH/GSSG ratio (reduced vs. oxidized glutathione) were also higher in azucena than in IR64 in the presence of Al. The addition of 1 mg/L GSH improved root elongation in both varieties and decreased H₂O₂ production under Al stress. By contrast, treatment with buthionine sulfoximine, a specific inhibitor of GSH synthesis, decreased root elongation in azucena and stimulated H₂O₂ production in both varieties. Moreover, Al treatment significantly increased the cytoplasmic activity of peroxidase (POD) as well as the levels of POD bound ionically and covalently to cell walls in the Al-sensitive variety. The lignin content was also increased. Treatment with exogenous H₂O₂ also increased the lignin content and decreased root elongation in IR64. These results suggest that Al induces lignification in the roots of Al-sensitive rice varieties, probably through an increase in H₂O₂ accumulation.     

Biochemical responses of Hyacinth bean (<Emphasis Type="Italic">Lablab purpureus)</Emphasis> to salinity stress

Effect of salinity on Hyacinth bean, Lablab purpureus (HA-4 cultivar) was evaluated in 10-day old seedlings with 100–500 mM NaCl over 72 h of exposure. The stress reduced dry and fresh weight, leaf surface area, root and shoot length, total chlorophyll, and RWC. Oxidative stress markers, H₂O₂, glutathione, TBARS, proline, ascorbic acid, total phenols, and total soluble sugar contents were significantly elevated. Salinity enhanced antioxidant enzymes, POX, and GR activities and reduced that of CAT in concentration and time dependent manner in leaves. Antioxidant enzymes in roots showed inverse relationship with concentration and time of exposure. Metabolic enzyme β-amylase activity increased in both leaves and roots. Acid phosphatase decreased in leaves and elevated in roots. Intensity of constitutive isozymes correlated with in vitro levels under stress, but the protein band patterns differed from controls. Lablab showed reasonable tolerance up to 300 mM NaCl, but leaves and roots differed in their response.