盐胁迫下匍匐翦股颖抗氧化酶活性及基因表达机制研究

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肖国增, 滕珂, 李林洁, 晁跃辉, 韩烈保. 盐胁迫下匍匐翦股颖抗氧化酶活性及基因表达机制研究. 草业学报, 2016,25(9): 74-82
XIAO Guo-Zeng, TENG Ke, LI Lin-Jie, CHAO Yue-Hui, HAN Lie-Bao. Antioxidant enzyme activity and gene expression in creeping bentgrass under salt stress. Acta Prataculturae Sinica,2016,25(9): 74-82 复制到剪切板

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盐胁迫下匍匐翦股颖抗氧化酶活性及基因表达机制研究

肖国增^1,², 滕珂¹, 李林洁¹, 晁跃辉¹^*, 韩烈保^1,^2,^*

1.北京林业大学草坪研究所,北京 100083

2.长江大学园艺园林学院,湖北荆州 434025

*通信作者Corresponding author. E-mail: chaoyuehui@163.com, hanliebao@163.com

作者简介:肖国增(1979-),男,湖南永州人,讲师,在读博士。E-mail:x_gz99@163.com

收稿日期: 2016-03-09

基金项目:国家高技术研究发展计划(863计划)(2013AA102706),深圳市科技计划基20160093植物抗逆基因挖掘与应用研究(项目编号:JCYJ20160331151245672),湖北省教育厅科学技术项目(Q20121202)和青年人才托举工程(中国林学会)资助

摘要

为研究匍匐翦股颖在盐胁迫下抗氧化酶活性与基因表达的影响机制,模拟盐胁迫条件,用0 mol/L(CK),0.2 mol/L的NaCl溶液对两种不同耐盐性的匍匐翦股颖Penncross和SeasideⅡ进行处理,测定了超氧化物歧化酶(SOD),过氧化氢酶(CAT),抗坏血酸过氧化物酶(APX)和过氧化物酶(POD)活性;用同源克隆的方法克隆了匍匐翦股颖 CytSOD, FeSOD, CAT, APX, POD的部分片段,并设计了5个基因的qRT-PC引物,对5个抗氧化酶基因表达量进行了荧光实时定量测定,分析了两种不同耐盐性的匍匐翦股颖在盐胁迫条件下,抗氧化酶活性与基因表达的变化规律。结果表明,盐胁迫对两种不同耐盐性的匍匐翦股颖抗氧化酶活性和基因表达的影响不同,在盐胁迫初期,盐敏感的Penncross SOD活性比耐盐的SeasideⅡ高,盐处理中、后期Penncross SOD活性快速下降,SeasideⅡSOD活性下降慢而且能保持相对稳定,在这一阶段SeasideⅡSOD活性显著高于Penncross;CAT,APX和POD活性随着盐处理的延长,酶活性增加,Penncross CAT和APX活性低于SeasideⅡ,POD活性两者差异不大。盐胁迫下,Penncross CytCu/ ZnSOD, CAT、 APX 和 POD表达量上调,而 FeSOD下调;SeasideⅡ5种抗氧化酶基因表达量上调, CytCu/ ZnSOD, FeSOD, CAT和 APX的表达量显著高于Penncross,而 POD表达量差异不大。两种匍匐翦股颖 CytCu/ ZnSOD、 CAT、 APX和 POD表达量变化与酶的活性变化较一致,而盐敏感的 FeSOD的表达量变化与SOD活性变化不一致。

关键词: 盐胁迫; 匍匐翦股颖; 抗氧化酶; 基因表达

Antioxidant enzyme activity and gene expression in creeping bentgrass under salt stress

XIAO Guo-Zeng^1,², TENG Ke¹, LI Lin-Jie¹, CHAO Yue-Hui^1,^*, HAN Lie-Bao^1,^2,^*

1.Institute of Turfgrass Science, Beijing Forestry University, Beijing 100083, China

2.The College of Horticulture and Garden, Yangtze University, Jingzhou 434025, China

Abstract

The aim of this research was to investigate the antioxidant enzyme activity and gene expression mechanisms in creeping bentgrass ( Agrostis stolonifera) under salt stress. Two different salt-resistant creeping bentgrass cultivars, Penncross and SeasideⅡ, were treated with 0 (CK) and 200 mmol/L NaCl solutions. The activities of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) and peroxidase (POD) were measured. The qRT-PCR primers for the CytSOD, FeSOD, CAT, APX and POD genes of creeping bentgrass were designed using partial gene segments, which were identified with homology cloning techniques. The gene expression levels of CytSOD, FeSOD, CAT, APX, and POD were measured and the activities of antioxidant enzymes and gene expression of the two different salt-resistant creeping bentgrass cultivars were analyzed. Salt stress had different effects on antioxidase activities and gene expressions on the two creeping bentgrass cultivars. At the beginning of salt stress, SOD activity of salt-sensitive Penncross was higher than that of salt-resistant SeasideⅡ. However, in the later stages of the treatment, the activity of Penncross SOD declined dramatically while the activity of SeasideⅡ SOD declined slowly and tended to be relatively stable, during which the activity of SeasideⅡ SOD was significantly higher than Penncross. The activities of CAT, APX and POD increased with salt treatment. The CAT and APX activities of Penncross were lower than those of SeasideⅡ, while POD activity was not different. Under the salt treatment, CytSOD, CAT, APX and POD expression of Penncross increased while FeSOD expression decreased. For SeasideⅡ, gene expression of the five antioxidases increased; CytSOD, FeSOD, CAT and APX gene expression were significantly higher in SeasideⅡ than Penncross, but POD was not different. The relative expression levels of CytCu/ZnSOD, CAT, APX and POD genes changed consistently with enzyme activities, but the relative expression level of the FeSOD gene in Penncross was not consistent with enzyme activity.

Keyword: salt stress; creeping bentgrass; antioxidant enzymes; gene expression

Show Figures

随着城市规模的不断发展, 淡水资源在城市中越来越缺乏, 因此, 大多数城市当中的园林绿地、软质运动场不得不选用城市再生水来浇灌草坪^{[1, 2]}。然而, 长期用含盐量较高的再生水浇灌草坪, 不但会增加土壤的盐碱化, 也会对草坪草造成盐胁迫使草坪草受到伤害从而影响草坪质量^[3]。

盐胁迫是过多的钠离子对植物造成脱水和渗透胁迫伤害, 影响了植物的正常生长和发育^{[4, 5]}, 为应对盐胁迫对植物造成的离子毒害和渗透胁迫, 植物演化出多种机制来使这种伤害降到最低。其中, 氧化应激是植物早期快速响应盐胁迫的机制之一, 此过程形成的活性氧(ROS)包括:超氧阴离子、过氧化氢和羟基自由基等^[6]。活性氧(ROS)大量积累, 会对植物造成膜系统氧化损伤^[7], 体内激素平衡被破坏^[8], 蛋白质和核酸变性; ROS能激活质膜渗透性Ca²⁺和K⁺的离子通道蛋白, 催化Ca²⁺信号途径, 导致K⁺流失甚至引起植物细胞程序死亡^{[9, 10]}。植物为了应对ROS的伤害, 形成了一套由抗氧化酶和非抗氧化酶组成的抗氧化体系^[11]。抗氧化酶主要包括超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD)、抗坏血酸过氧化物酶(APX), 植物通过这些酶的作用减少或是清除ROS的积累, 减轻和抵御盐胁迫对植物细胞的伤害^[12]。在清除ROS的机制中SOD是第一个激活酶, 它清除超氧阴离子产生过氧化氢和氧气, 随后CAT、APX和POD酶清除过氧化氢^{[13, 14]}。

同时, 非生物胁迫也会对植物的抗氧化酶基因在转录水平上造成影响。在高温胁迫下, 水稻(Oryza sativa)开花期剑叶的SOD、CAT和POD基因表达量显著增加, 从而维持了较高的酶活性来应答高温胁迫^[15]; 过量的镉积累增强了龙葵(Solanum nigrum)SOD、POD和CAT的转录水平^[16]; 而随着镉对玉米(Zea mays)幼苗处理时间的增加, SOD的表达量受到抑制^[17], 高钙处理苹果(Malus pumila)果实在12 h后SOD和CAT表达量增加^[18]; 盐胁迫加强了CytCu/ZnSOD、CAT和POD等基因的表达^[19], 因此, 研究抗氧化酶活性与转录水平调控对理解草坪草在非生物胁迫, 尤其是盐胁迫下的抗氧化机制具有重要作用。

匍匐翦股颖(Agrostis stolonifera)是优良的冷季型草坪草, 具有极耐低修剪、成坪快、草坪质量高的优点, 被广泛应用于城市园林绿化和运动场草坪, 但是较低的耐盐性^[20]限制了其应用。目前, 在盐胁迫下对匍匐翦股颖抗氧化酶活性和基因表达方面的研究较少, 特别是抗氧化酶基因表达水平的研究还未见报道。因此, 本研究通过不同盐浓度处理两种不同耐盐性的匍匐翦股颖, 研究盐胁迫条件下匍匐翦股颖抗氧化酶活性及基因表达的变化规律, 揭示其在抗氧化酶系统及基因表达对盐胁迫的响应机制, 为草坪草在逆境生理与分子研究领域提供理论依据, 也为匍匐翦股颖在盐害方面的草坪管理提供指导。

1 材料与方法

1.1 材料培养

供试两种匍匐翦股颖Penncross和SeasideⅡ 种子来源于实验室保存, 其中Penncross对盐敏感, 而SeasideⅡ 有很好的耐盐性^[21]。试验于2014年8月-12月, 在苗圃温室和草坪实验室进行。苗圃温室气温为(22± 1) ℃/(16± 1) ℃(白天/黑夜), 相对湿度为65%~75%, 14 h平均光有效辐射(PAR)值为400 μ mol/(m²· s)。匍匐翦股颖Penncross和SeasideⅡ 按5 g/m²播种量, 均匀撒播于直径15 cm, 高13 cm的方形塑料花盆中, 基质为过筛的纯沙。种入时间为2014年8月1日, 出苗前用塑料薄膜覆盖。苗期进行间苗, 留下长势一致和分布均匀的苗株, 采用正常的养护管理, 每星期浇一次0.5强度(在原配方的基础上减半)的Hoagland营养液^[22]直至从底部流出。

1.2 材料处理与取样

1.2.1 生理实验处理与取样至2014年11月1日, 移入水培箱内培养。塑料水培箱:50 cm× 38 cm× 26 cm; 从花盆中取栽培3个月的健康草皮, 剪成直径为4 cm, 厚3 cm的草皮放于挂篮上, 再把挂篮放于黑色塑料板的孔中, 然后把黑色塑料板放于水培箱上, 使草皮与水充分接触, 挂篮直径5 cm, 一块板上平均分布20个。水培箱用增氧泵通气, 每星期换一次水, 换水时加入0.5强度的Hoagland营养液, 水培两周后进行处理。

用NaCl溶液进行盐处理, 共设两个盐梯度分别为0 mol/L(CK)和0.2 mol/L, 各处理均设4个重复, 两个匍匐翦股颖草种, 共16盆。第1次加盐时, 把相应盐浓度平均分4份, 每天加一次, 于11月16日加盐, 11月19日达到相应盐浓度。加盐前进行第1次取样, 达到相应盐浓度后每隔5 d取一次样, 取样时间分别为11月15日(0处理), 11月24日, 11月29日, 12月4日, 12月9日, 取样的叶片立即用锡箔纸包好, 编号后放入液氮中, -80 ℃保存。

1.2.2 分子实验处理与取样分子实验材料取栽培3个月的Penncross和SeasideⅡ 移入人工气候箱, 生长条件与苗圃温室一致, 预培养两周后, 进行NaCl处理, 浓度梯度与生理实验一致, 4个重复。处理时分别用0 mol/L(蒸馏水, CK)和0.2 mol/L浓度的NaCl浇灌花盆, 浇灌时尽量避免盐水接触叶片, 直至溶液从花盆底部流出。取样时间分别为0 h(盐处理前), 1 h, 3 h, 6 h, 12 h, 24 h。取样的叶片立即用锡箔纸包好, 编号后放入液氮中, -80 ℃保存。

1.3 抗氧化酶活性测定

1.3.1 酶液的提取称取0.15 g 叶片, 用液氮磨成粉末, 加2 mL 提取液研磨成匀浆, 于12000 r/min、4 ℃条件下离心20 min, 上清液即为粗酶液。提取液为含有1 mmol/L 的乙二胺四乙酸(EDTA)、1% 的聚乙烯吡咯烷酮(PVP-40)(w/v)、1 mmol/L 的二硫苏糖醇(DTT) 和1 mmol/L 的苯甲基磺酰氟(PMSF)的磷酸缓冲液(PBS 7.0)(50 mmol/L)^[23]。

1.3.2 抗氧化酶活性测定 SOD活性用Giannopolitis等^[24]的方法测定, CAT、POD和APX的活性采用Zhang等^[23]的方法进行测定, 可溶性蛋白用Bradford^[25]的方法测定。

1.4 抗氧化酶基因表达

1.4.1 匍匐翦股颖cDNA制备采用TRIzol 试剂盒提取RNA, 并使用DNase(Turbo DNA-Free Kit; Ambion, Austin, TX)处理去除DNA 污染, 用反转录酶[PrimeScript^TM RT Reagent Kit (Perfect Real Time) Takara, Japan]反转录成cDNA, 用于后续实验。

1.4.2 抗氧化酶qRT-PCR引物设计对匍匐翦股颖的SOD(CytCu/ZnSOD、FeSOD)、APX、CAT、POD 5个抗氧化酶基因采用同源克隆的方法进行基因克隆。通过NCBI中与匍匐翦股颖同源物种小麦(Triticum aestivum)、大麦(Hordeum vulgare)、二穗短柄草(Brachypodium distachyon)、玉米和水稻等相对应的抗氧化酶基因序列比对后的保守区域设计相应引物, 扩增抗氧化酶CytCu/ZnSOD(GenBank:KU966135)、FeSOD(GenBank: KU966136)、APX(GenBank:KU981301)、CAT(GenBank: KU966134)和POD(GenBank: KX034392)5个基因的部分序列。用所得基因序列和匍匐翦股颖Actin(GenBank:JX644005.1)序列设计相应基因的荧光实时定量反应(quantitativereal-time polymerase chain reaction, qRT-PCR)引物(表1), 经PCR和qRT-PCR验证后, 用于抗氧化酶基因荧光实时定量分析。

表1 qRT-PCR所用的引物 Table 1 Primers for qRT-PCR

1.4.3 抗氧化酶基因表达量测定匍匐翦股颖叶片抗氧化酶基因的表达分析采用qRT-PCR方法, 荧光定量试剂UltraSYBR Mixture (with ROX I)(CWBIO. Chain), 以两个盐处理梯度的相对应的cDNA为模版, 用Bio-RAD CFX connecttm Real-time system采用两步法, 20 μ L体系, 4个生物学重复, 以匍匐翦股颖Actin作为内参基因, 进行5个抗氧化酶基因CytCu/ZnSOD、FeSOD、APX、CAT和POD的荧光定量分析, 相对表达量用2^{-Δ Ct}法计算, 其中Δ Ct=Ct_{(目的基因)}-Ct_{(内参基因)}。

1.5 数据处理

采用SPSS 18.0进行数据单因素方差分析(ANOVA), 使用最小显著差异法(least significant difference, LSD)在0.05显著性水平进行差异显著性分析, 图表用Excel 10.0绘制。

2 结果与分析

2.1 盐胁迫对抗氧化酶活性的影响

SOD是抗氧化保护系统清除ROS机制中首先被激活的酶, 把超氧阴离子分解成过氧化氢和氧气^[13]。盐胁迫对两种不同耐盐性的匍匐翦股颖SOD活性的影响不同(图1)。盐敏感的Penncross在0.2 mol/L盐浓度处理下, SOD活性呈现先升高再降低的趋势。在第10天达到最高值, 是CK的142.3%, 差异显著。耐盐的SeasideⅡ 在0.2 mol/L盐浓度处理下的SOD活性呈现先降低, 后升高的趋势, 第10天达到最低值, 比CK值低43.8%, 其值是Penncross第10天的44.4%, 差异显著; 第20天SeasideⅡ 0.2 mol/L盐浓度处理下的SOD活性高于CK值和0.2 mol/L盐浓度处理下Penncross的SOD活性值。SeasideⅡ SOD响应盐胁迫比Penncross慢, 但随着盐胁迫时间的增加, SeasideⅡ SOD活性增强, 而Penncross SOD活性快速下降, 表明SeasideⅡ SOD比Penncross清除ROS的能力更持久, 能更好地保护细胞减轻盐害。

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	图1 盐胁迫对两种匍匐翦股颖SOD活性的影响竖线表示标准误, 不同的小写字母表示差异显著(P≤ 0.05), 下同。Fig.1 Effects of different salinities treatments on SOD ctivities in two different creeping bentgrass cultivars Vertical bars indicate mean± SE. Different lowercase letters represent significant differences (P≤ 0.05). The same below.

CAT在抗氧化保护机制中起到了重要作用, 它把SOD清除ROS产生的过氧化氢催化分解成水和氧气^[26]。盐处理下两种匍匐翦股颖CAT活性变化都呈现上升的趋势, 盐处理期间两种草0.2 mol/L盐处理的CAT活性均比对照高(图2)。第5天SeasideⅡ 0.2 mol/L盐处理的CAT活性显著低于Penncross的CAT活性; 然而, 从第10天后SeasideⅡ 0.2 mol/L盐处理的CAT活性高于Penncross, 在第15天SeasideⅡ CAT活性是Penncross CAT活性的131.6%, 两者差异显著。这表明盐敏感的Penncross CAT响应盐胁迫比耐盐的SeasideⅡ 快, 但在盐处理中、后期SeasideⅡ CAT的响应逐渐加强, 活性显著高于Penncross, 表明SeasideⅡ CAT清除过氧化氢的能力更强。

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	图2 盐胁迫对两种匍匐翦股颖CAT活性的影响Fig.2 Effects of different salinities treatments on CAT activities in two different creeping bentgrass cultivars

在植物叶绿体内谷胱甘肽循环的第一步, APX使用电子供体的抗坏血酸清除过氧化氢^[26]。在盐胁迫下, 两种匍匐翦股颖APX活性整体呈上升趋势(图3)。但是, 盐敏感的Penncross APX活性在第10天达到最高值, 是CK的149.5%, 差异显著, 随着盐处理时间的延长, APX活性缓慢下降。耐盐的SeasideⅡ APX活性盐处理后比CK和Penncross APX的活性都高, 而且呈一直上升的趋势, 在第10天达到最高值, 而后其活性趋于稳定, 在第20天其值是CK的118.2%, 是盐处理下Penncross APX的116.8%, 差异不显著; 这表明盐处理下SeasideⅡ 和Penncross APX清除过氧化氢的能力相近。

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	图3 盐胁迫对两种匍匐翦股颖APX活性的影响Fig.3 Effects of different salinities treatments on APX activities in two different creeping bentgrass cultivars

植物体内的POD通过共基质的酚类抗氧化剂或化合物氧化来分解过氧化氢^[26]。盐胁迫下, 两种匍匐翦股颖的POD活性差异不显著, 都呈先升高后下降的趋势, 在第15天POD的活性都达到最高值(图4)。两种匍匐翦股颖在盐处理下的POD活性都高于CK, 除了第0天和第20天不显著以外, 其他的盐处理时间0.2 mol/L盐处理和CK差异显著。

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	图4 盐胁迫对两种匍匐翦股颖POD活性的影响Fig.4 Effects of different salinities treatments on POD activities in two different creeping bentgrass cultivars

2.2 盐胁迫对抗氧化酶基因表达的影响

2.2.1 抗氧化酶基因qRT-PCR引物设计与验证根据确定的匍匐翦股颖CytCu/ZnSOD、FeSOD、APX、CAT和POD五个基因的序列设计相应的qRT-PCR引物, 用PCR和qRT-PCR验证扩增产物。5个基因的PCR产物都在300 bp以下, 而且目的条带单一, 明亮; qRT-PCR溶解曲线图的溶解峰单一, 同一个抗氧化酶盐处理的多个样品出现溶解峰的温度较一致。因此, 设计的5个匍匐翦股颖抗氧化酶基因的qRT-PCR引物, 可以用于荧光实时定量分析。

2.2.2 盐胁迫对抗氧化酶基因表达的影响以匍匐翦股颖内源AsActin作为内参基因, 将0 h处理设为参照, 在盐胁迫下, 两种匍匐翦股颖CytCu/ZnSOD相对表达量的变化趋势有所不同(图5)。盐敏感的Penncross随着盐处理时间的延长, CytCu/ZnSOD相对表达量先升高后降低; 在第3 h达到最高值, 是CK的169.1%, 差异显著, 而后表达量下降。耐盐的SeasideⅡ 随着盐处理时间的延长, CytCu/ZnSOD相对表达量呈上升趋势, 其表达量在第3 h盐处理与CK差异显著, 但相对表达量低于盐处理Penncross的相对表达量, 两者差异显著; 但盐处理SeasideⅡ 在第24 h达到最大值, 是Penncross的167.1%, 差异显著。

	Figure Option View Download New Window
	图5 盐胁迫下对两种匍匐翦股颖CytCu/ZnSOD相对表达量的影响Fig.5 Gene expression of CytCu/ZnSOD in two different creeping bentgrass cultivars under salt treatment

盐胁迫下, 两种匍匐翦股颖FeSOD相对表达量变化趋势不同(图6)。盐敏感的Penncross随着盐处理时间的延长, FeSOD相对表达量下调。耐盐的SeasideⅡ 随着盐处理时间的延长, FeSOD相对表达量呈先下调后上调的趋势, 其值高于CK和盐处理的Penncross; 在第12 h达到最高值, 是盐处理Penncross的324.6%, 差异显著。

	Figure Option View Download New Window
	图6 盐胁迫下对两种匍匐翦股颖FeSOD相对表达量的影响Fig.6 Gene expression of FeSOD in two different creeping bentgrass cultivars under salt treatment

盐胁迫下, 两种匍匐翦股颖CAT相对表达量变化趋势相同(图7), 都上调, 在第24 h达到最大值, SeasideⅡ CAT的表达量是Penncross 的265.3%, 是CK的214.9%, 差异显著。在盐处理的时间里, SeasideⅡ CAT表达量均高于CK和Penncross, 差异显著。

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	图7 盐胁迫下对两种匍匐翦股颖CAT相对表达量的影响Fig.7 Gene expression of CAT in two different creeping bentgrass cultivars under salt treatment

盐胁迫下, 两种匍匐翦股颖APX相对表达量变化趋势见图8, 盐敏感的Penncross随着盐处理时间的延长, APX相对表达量先升高后下降; 在第3 h达到最大值; 耐盐的SeasideⅡ 随着盐处理时间的延长, APX相对表达量呈先上升后下降的趋势, 在第12 h达到最大值, 是CK的208.9%, 是盐处理Penncross的251.7%, 差异显著。

	Figure Option View Download New Window
	图8 盐胁迫下对两种匍匐翦股颖APX相对表达量的影响Fig.8 Gene expression of APX in two different creeping bentgrass cultivars under salt treatment

盐胁迫下, 两种匍匐翦股颖POD相对表达量变化趋势大致相同(图9), 都先升高后降低。盐处理的Penncross和SeasideⅡ POD相对表达量在第3 h达到最高值, 两者差异不显著; 盐处理的两种匍匐翦股颖POD相对表达量均高于CK, 差异显著。耐盐的SeasideⅡ POD相对表达量在第12 h的表达是盐处理Penncross的225.0%, 显著高于CK和Penncross。

	Figure Option View Download New Window
	图9 盐胁迫下对两种匍匐翦股颖POD相对表达量的影响Fig.9 Gene expression of POD in two different creeping bentgrass cultivars under salt treatment

3 讨论与结论

植物体内存在ROS生成与清除机制, 在正常情况下两类机制处于平衡状态, 但植物在遭受外界环境胁迫时, 这种平衡受到破坏, 积累过多的ROS会引起植物的氧化伤害^[27], 在这种机制中抗氧化酶SOD、CAT、APX、POD在保持细胞内ROS动态平衡发挥了重要作用。本研究在盐胁迫下, 对两种耐盐性不同的匍匐翦股颖抗氧化酶SOD、CAT、APX、POD的活性和基因表达量的变化进行了较系统分析。

在0.2 mol/L盐胁迫下, 两种匍匐翦股颖SOD活性和转录水平响应不同, 盐敏感的Penncross SOD活性先升高再降低, 与刘凤歧等^[28]对盐胁迫下燕麦(Avena sativa)的SOD变化一致; 耐盐的SeasideⅡ SOD活性先降低后升高到较稳定状态。盐处理前期Penncross SOD活性比SeasideⅡ 高, 而盐处理后期Penncross SOD活性比SeasideⅡ 低。这表明盐敏感的Penncross对盐胁迫响应更快, 随着盐处理时间的增加, SOD的活性降低, ROS的动态平衡被打破, 过多的ROS导致了Penncross对盐胁迫的抵抗力下降; 而在盐处理后期, 耐盐的SeasideⅡ SOD显著高于Penncross, SOD活性高有利于清除ROS^[29], 从而增加植物对盐害的抵抗力。在0.2 mol/L盐胁迫下, 两种匍匐翦股颖抗氧化酶CAT、APX和POD的活性都增加^{[30, 31]}, 而且耐盐的SeasideⅡ 比Penncross CAT、APX和POD的活性更高, 这与Puyang等^[32]对草地早熟禾在盐胁迫下抗氧化酶的影响研究结果一致。

在0.2 mol/L盐胁迫下, 盐敏感的Penncross CytCu/ZnSOD、APX和POD先上调后下调, CAT上调; 耐盐的SeasideⅡ CytCu/ZnSOD、FeSOD、CAT、APX和POD都上调, SeasideⅡ 比Penncross抗氧化酶基因表达量高, 这与两种匍匐翦股颖在盐胁迫抗氧化酶活性的变化趋势一致。这表明在盐胁迫下转录水平上, 上调抗氧化酶基因表达量可以激活抗氧化酶的活性, 能更有效地减少ROS的积累, 维持细胞膜稳定性和生理平衡^[18]。对盐胁迫下大麦的研究表明FeSOD活性没变化, 甚至有的大麦基因型在盐处理下FeSOD活性降低^[33]。本研究的Penncross FeSOD在盐胁迫下基因表达量下调, 而SOD活性升高, 酶活性与FeSOD基因表达量不一致, 表明在盐胁迫下SOD的活性可能是受除FeSOD以外的其他的基因调控。因此, 草坪草抗氧化酶活性与基因编码调控之间的相互作用机制还有待更深入的研究。

综上所述, 本研究从生理与分子转录水平上, 分析了两种不同耐盐性的匍匐翦股颖抗氧化酶在盐胁迫下的响应机制。在盐胁迫下, 两种匍匐翦股颖除SOD外其他3个抗氧化酶活性变化较一致。0.2 mol/L盐处理匍匐翦股颖的Penncross和SeasideⅡ CAT和APX活性呈上升趋势, POD活性呈先升高后下降的趋势; Penncross SOD活性先升高后下降, 而SeasideⅡ SOD先下降后升高。在盐处理后期SeasideⅡ SOD、CAT和APX活性都比Penncross高, POD活性两者差异不大。分子水平上, 两种匍匐翦股颖CytCu/ZnSOD、CAT、APX和POD变化与酶的活性变化较一致, 而盐敏感的匍匐翦股颖FeSOD的基因表达与酶活性变化不一致。

The authors have declared that no competing interests exist.

参考文献

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[1]	Dean D E, Devitt D A, Verhick L S, et al. Turfgrass quality, growth, and water use influenced by salinity and water stress. Agronomy Journal, 1996, 88(5): 844-849. [本文引用:1]
[2]	Morris R L. Water quality changes in golf course irrigation ponds transitioning to reuse water. Hortscience A Publication of the American Society for Horticultural Science, 2005, 40(7): 2151-2156. [本文引用:1]
[3]	Qian Y L, Mecham B. Long-term effects of recycled wastewater irrigation on soil chemical properties on golf course fairways. Agronomy Journal, 2005, 97(3): 717-721. [本文引用:1]
[4]	Zhu J K. Plant salt tolerance. Trends in Plant Science, 2001, 6(2): 66-71. [本文引用:1]
[5]	Zhu J K. Regulation of ion homeostasis under salt stress. Current Opinion in Plant Biology, 2003, 6(5): 441-445. [本文引用:1]
[6]	Barry H. Reactive species and antioxidants redox biology is a fundamental theme of aerobic life. Plant Physiology, 2006, 141(2): 312-322. [本文引用:1]
[7]	Jiang M, Zhang J. Effect of abscisic acid on active oxygen species, antioxidative defence system and oxidative damage in leaves of maize [Zea mays] seedlings. Plant & Cell Physiology, 2001, 42(11): 1265-1273. [本文引用:1]
[8]	Liu Y J, Zhang L, Tian X Y, et al. The effects of salt stress on endogenous hormones, NADKase and Ca²+-ATPase in leaves of Puccinellia chinampoensis seedlings. Pratacultural Science, 2008, 25(4): 51-54. [本文引用:1]
[9]	Bethke P C, Jones R L. Cell death of barley aleurone protoplasts is mediated by reactive oxygen species. Plant Journal, 2001, 25(1): 19-29. [本文引用:1]
[10]	Demidchik V. Mechanisms of oxidative stress in plants: From classical chemistry to cell biology. Environmental and Experimental Botany, 2015, 109: 212-228. [本文引用:1]
[11]	Scand alios J G. Oxygen stress and superoxide dismutases. Plant Physiology, 1993, 101(1): 7-12. [本文引用:1]
[12]	Fan R P, Zhou Q, Zhou B, et al. Effects of salinization stress on growth and the antioxidant system of tall fescue. Acta Prataculturae Sinica, 2012, 21(1): 112-117. [本文引用:1]
[13]	Elstner E F. Oxygen activation and oxygen toxicity. Annual Review of Plant Physiology, 2003, 33(1): 73-96. [本文引用:2]
[14]	Bowler C, And M V M, Inze D. Superoxide dismutase and stress tolerance. Annual Review of Plant Physiology & Plant Molecular Biology, 1992, 43(4): 83-116. [本文引用:1]
[15]	Wang Y, Gao P, Huang M, et al. Effects of high temperature on the activity and expression of antioxidative enzymes in rice flag leaves during the flowering stage. Plant Science Journal, 2015, 33(3): 355-361. [本文引用:1]
[16]	Zhang Y X, Jin L, Feng S S, et al. Effects of Cd on activity and gene expression of antioxidant enzymes in hyperaccumulator Solanum nigrum L. Journal of Graduate University of Chinese Academy of Sciences, 2013, 30(1): 11-17. [本文引用:1]
[17]	Zhao S C, Sun J W, Ma Y Z, et al. Effects of cadmium on reactive oxygen species metabolism, activities and gene expressions of superoxide dismutase and catalase in maize (Zea mays) seedling. Scientia Agricultura Sinica, 2008, 41(10): 3025-3032. [本文引用:1]
[18]	Liang G Q, Sun J W, Zhou W, et al. Effects of calcium on activities and gene expressions of superoxide dismutase and catalas in apple (Malus pumila Mill. ) fruits. Plant Nutrition and Fertilizer Science, 2011, 17(2): 438-444. [本文引用:2]
[19]	Tao H, Li H Y, Zhang X Z, et al. Toxic effect of NaCl on ion metabolism, antioxidative enzymes and gene expression of perennial ryegrass. Ecotoxicology & Environmental Safety, 2011, 74(7): 2050-2056. [本文引用:1]
[20]	Wang N, Li H M. Research on salt tolerance of cool-season turf grass. Hubei Agricultural Sciences, 2011, 50(10): 2047-2051. [本文引用:1]
[21]	Marcum K B. Salinity tolerance of 35 bentgrass cultivars. Hort Science, 2001, 36(2): 374. [本文引用:1]
[22]	Hoagland D R, Arnon D I. The water-culture method for growing plants without soil. California Agricultural Experiment Station, 1950, 347(2): 32. [本文引用:1]
[23]	Zhang J, Kirkham M B. Antioxidant responses to drought in sunflower and sorghum seedlings. New Phytologist, 1996, 132(3): 361-373. [本文引用:2]
[24]	Giannopolitis C N, Ries S K. Superoxide dismutase: I. Occurrence in higher plants. Plant Physiology, 1977, 59(2): 309-314. [本文引用:1]
[25]	Bradford M M. A rapid method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 1976, 72(s 1/2): 248-254. [本文引用:1]
[26]	Amako K, Chen G X, Asada K. Separate assays specific for ascorbate peroxidase and guaiacol peroxidase and for the chloroplastic and cytosolic isozymes of ascorbate peroxidase in plants. Plant & Cell Physiology, 1994, 35(3): 497-504. [本文引用:3]
[27]	Wang L P, Sun J, Guo S R, et al. Salt stress tolerance of cucumber-grafted rootstocks. Chinese Journal of Applied Ecology, 2012, 23(5): 18-21. [本文引用:1]
[28]	Lu F Q, Liu J L, Zhu R F, et al. Physiological responses and tolerance of four oat varieties to salt stress. Acta Prataculturae Sinica, 2015, 24(1): 183-189. [本文引用:1]
[29]	Meng Y X, Wang S H, Wang J C, et al. Influences of CoCl₂ on the growth and seedling physiological indexes of Hordeum vulgare under NaCl stress. Acta Prataculturae Sinica, 2014, 23(3): 160-166. [本文引用:1]
[30]	Asish Kumar P, Anath Band hu D, Prasanna M. Defense potentials to NaCl in a mangrove, Bruguiera parviflora: differential changes of isoforms of some antioxidative enzymes. Journal of Plant Physiology, 2004, 161(5): 531-542. [本文引用:1]
[31]	Duan J, Li J S, Kang Y. Exogenous spermidine affects polyamine metabolism in salinity-stressed cucumis sativus roots and enhances short-term salinity tolerance. Journal of Plant Physiology, 2008, 165(15): 1620-1635. [本文引用:1]
[32]	Puyang X, An M, Han L, et al. Protective effect of spermidine on salt stress induced oxidative damage in two Kentucky bluegrass (Poa pratensis L. ) cultivars. Ecotoxicology & Environmental Safety, 2015, 117: 96-106. [本文引用:1]
[33]	Nounjan N, Nghia P T, Theerakulpisut P. Exogenous proline and trehalose promote recovery of rice seedlings from salt-stress and differentially modulate antioxidant enzymes and expression of related genes. Journal of Plant Physiology, 2012, 169(6): 596-604. [本文引用:1]
[8]	刘延吉, 张蕾, 田晓艳, 等. 盐胁迫对碱茅幼苗叶片内源激素、NAD激酶及Ca²+-ATPase的效应. 草业科学, 2008, 25(4): 51-54. [本文引用:1]
[12]	樊瑞苹, 周琴, 周波, 等. 盐胁迫对高羊茅生长及抗氧化系统的影响. 草业学报, 2012, 21(1): 112-117. [本文引用:1]
[15]	王艳, 高鹏, 黄敏, 等. 高温对水稻开花期剑叶抗氧化酶活性及基因表达的影响. 植物科学学报, 2015, 33(3): 355-361. [本文引用:1]
[16]	张玉秀, 金玲, 冯珊珊, 等. 镉对镉超累积植物龙葵抗氧化酶活性及基因表达的影响. 中国科学院研究生院学报, 2013, 30(1): 11-17. [本文引用:1]
[17]	赵士诚, 孙静文, 马有志, 等. 镉对玉米幼苗活性氧代谢、超氧化物歧化酶和过氧化氢酶活性及其基因表达的影响. 中国农业科学, 2008, 41(10): 3025-3032. [本文引用:1]
[18]	梁国庆, 孙静文, 周卫, 等. 钙对苹果果实超氧化物歧化酶、过氧化氢酶活性及其基因表达的影响. 植物营养与肥料学报, 2011, 17(2): 438-444. [本文引用:2]
[20]	王娜, 李海梅. 冷季型草坪草耐盐性研究. 湖北农业科学, 2011, 50(10): 2047-2051. [本文引用:1]
[27]	王丽萍, 孙锦, 郭世荣, 等. 黄瓜砧用白籽南瓜对不同盐胁迫的耐性评价. 应用生态学报, 2012, 23(5): 18-21. [本文引用:1]
[28]	刘凤歧, 刘杰淋, 朱瑞芬, 等. 4种燕麦对NaCl胁迫的生理响应及耐盐性评价. 草业学报, 2015, 24(1): 183-189. [本文引用:1]
[29]	孟亚雄, 王世红, 汪军成, 等. CoCl₂对NaCl胁迫下大麦生长及幼苗生理指标的影响. 草业学报, 2014, 23(3): 160-166. [本文引用:1]

1996

0.0

... 随着城市规模的不断发展,淡水资源在城市中越来越缺乏,因此,大多数城市当中的园林绿地、软质运动场不得不选用城市再生水来浇灌草坪^[1,2] ...

2005

0.0

... 随着城市规模的不断发展,淡水资源在城市中越来越缺乏,因此,大多数城市当中的园林绿地、软质运动场不得不选用城市再生水来浇灌草坪^[1,2] ...

2005

0.0

... 然而,长期用含盐量较高的再生水浇灌草坪,不但会增加土壤的盐碱化,也会对草坪草造成盐胁迫使草坪草受到伤害从而影响草坪质量^[3] ...

2001

0.0

. 2001, 6(2):66-71

Plant salt tolerance

Zhu J K

Dept Plant Sciences, University of Arizona, Tucson, AZ 85721, USA. jkzhu@ag.arizona.edu

Soil salinity is a major abiotic stress in plant agriculture worldwide. This has led to research into salt tolerance with the aim of improving crop plants. However, salt tolerance might have much wider implications because transgenic salt-tolerant plants often also tolerate other stresses including chilling, freezing, heat and drought. Unfortunately, suitable genetic model systems have been hard to find. A recently discovered halophytic plant species, Thellungiella halophila, now promises to help in the detection of new tolerance determinants and operating pathways in a model system that is not limited to Arabidopsis traits or ecotype variations.

PMID: 11173290

... 盐胁迫是过多的钠离子对植物造成脱水和渗透胁迫伤害,影响了植物的正常生长和发育^[4,5],为应对盐胁迫对植物造成的离子毒害和渗透胁迫,植物演化出多种机制来使这种伤害降到最低 ...

2003

0.0

2006

0.0

. 2006, 141(2):312-322

Reactive species and antioxidants redox biology is a fundamental theme of aerobic life

Barry H

Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117597. bchbh@nus.edu.sg

PMID: 16760481

... 其中,氧化应激是植物早期快速响应盐胁迫的机制之一,此过程形成的活性氧(ROS)包括:超氧阴离子、过氧化氢和羟基自由基等^[6] ...

2001

0.0

. 2001, 42(11):1265-1273

Effect of abscisic acid on active oxygen species, antioxidative defence system and oxidative damage in leaves of maize [Zea mays] seedlings

Jiang M , Zhang J

Department of Biology, Hong Kong Baptist University, Kowloon Tong, Hong Kong.

Leaves of maize (Zea mays L.) seedlings were supplied with different concentrations of abscisic acid (ABA). Its effects on the levels of superoxide radical (O(2)(-)), hydrogen peroxide (H(2)O(2)) and the content of catalytic Fe, the activities of several antioxidative enzymes such as superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) and glutathione reductase (GR), the contents of several non-enzymatic antioxidants such as ascorbate (ASC), reduced glutathione (GSH), alpha-tocopherol (alpha-TOC) and carotenoid (CAR), and the degrees of the oxidative damage to the membrane lipids and proteins were examined. Treatment with 10 and 100 microM ABA significantly increased the levels of O(2)(-) and H(2)O(2), followed by an increase in activities of SOD, CAT, APX and GR, and the contents of ASC, GSH, alpha-TOC and CAR in a dose- and time-dependent pattern in leaves of maize seedlings. An oxidative damage expressed as lipid peroxidation, protein oxidation, and plasma membrane leakage did not occur except for a slight increase with 100 microM ABA treatment for 24 h. Treatment with 1,000 microM ABA led to a more abundant generation of O(2)(-) and H(2)O(2) and a significant increase in the content of catalytic Fe, which is critical for H(2)O(2)-dependent hydroxyl radical production. The activities of these antioxidative enzymes and the contents of alpha-TOC and CAR were still maintained at a higher level, but no longer further enhanced when compared with the treatment of 100 microM ABA. The contents of ASC and GSH had no changes in leaves treated with 1,000 microM ABA. These results indicate that treatment with low concentrations of ABA (10 to 100 microM) induced an antioxidative defence response against oxidative damage, but a high concentration of ABA (1,000 microM) induced an excessive generation of AOS and led to an oxidative damage in plant cells.

PMID: 11726712

... 活性氧(ROS)大量积累,会对植物造成膜系统氧化损伤^[7],体内激素平衡被破坏^[8],蛋白质和核酸变性 ...

2008

0.0

... 活性氧(ROS)大量积累,会对植物造成膜系统氧化损伤^[7],体内激素平衡被破坏^[8],蛋白质和核酸变性 ...

2001

0.0

. 2001, 25(1):19-29

Cell death of barley aleurone protoplasts is mediated by reactive oxygen species

Bethke P C , Jones R L

Department of Plant and Microbial Biology, University of California, Berkeley, CA 94720-3102, USA. pcbethke@nature.berkeley.edu

The barley aleurone layer is a terminally differentiated secretory tissue whose activity is hormonally controlled. The plant hormone gibberellic acid (GA) stimulates the secretion of hydrolytic enzymes and triggers the onset of programmed cell death (PCD). Abscisic acid (ABA) antagonizes the effects of GA and inhibits enzyme secretion and PCD. Reactive oxygen species (ROS) are key players in many types of PCD, and data presented here implicate ROS in hormonally regulated death of barley aleurone cells. Incubation of aleurone layers or protoplasts in H(2)O(2)-containing media results in death of GA-treated but not ABA-treated aleurone cells. Cells that are programmed to die are therefore less able to withstand ROS than cells that are programmed to remain alive. Illumination of barley aleurone protoplasts with blue or UV-A light results in a rapid increase in intracellular H(2)O(2) production. GA-treated protoplasts die rapidly in response to this increase in intracellular H(2)O(2) production, but ABA-treated protoplasts do not die. The rate of light-induced death could be slowed by antioxidants, and incubating protoplasts in the dark with the antioxidant butylated hydroxy toluene reduces the rate of hormonally induced death. Taken together, these data demonstrate that GA-treated aleurone protoplasts are less able than ABA-treated protoplasts to tolerate internally generated or exogenously applied H(2)O(2), and strongly suggest that ROS are components of the hormonally regulated cell death pathway in barley aleurone cells.

PMID: 11169179

... ROS能激活质膜渗透性Ca²⁺和K⁺的离子通道蛋白,催化Ca²⁺信号途径,导致K⁺流失甚至引起植物细胞程序死亡^[9,10] ...

2015

0.0

... ROS能激活质膜渗透性Ca²⁺和K⁺的离子通道蛋白,催化Ca²⁺信号途径,导致K⁺流失甚至引起植物细胞程序死亡^[9,10] ...

1993

0.0

. 1993, 101(1):7-12

Oxygen stress and superoxide dismutases

Scandalios J G

Department of Genetics, Box 7614, North Carolina State University, Raleigh, North Carolina 27695-7614.

PMID: 12231660

... 植物为了应对ROS的伤害,形成了一套由抗氧化酶和非抗氧化酶组成的抗氧化体系^[11] ...

2012

0.0

. 2012, 21(1):112-117

盐胁迫对高羊茅生长及抗氧化系统的影响

樊瑞苹, 周琴, 周波

... 抗氧化酶主要包括超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD)、抗坏血酸过氧化物酶(APX),植物通过这些酶的作用减少或是清除ROS的积累,减轻和抵御盐胁迫对植物细胞的伤害^[12] ...

2003

0.0

... 在清除ROS的机制中SOD是第一个激活酶,它清除超氧阴离子产生过氧化氢和氧气,随后CAT、APX和POD酶清除过氧化氢^[13,14] ...

... 1 盐胁迫对抗氧化酶活性的影响SOD是抗氧化保护系统清除ROS机制中首先被激活的酶,把超氧阴离子分解成过氧化氢和氧气^[13] ...

1992

0.0

... 在清除ROS的机制中SOD是第一个激活酶,它清除超氧阴离子产生过氧化氢和氧气,随后CAT、APX和POD酶清除过氧化氢^[13,14] ...

2015

0.0

. 2015, 33(3):355-361

高温对水稻开花期剑叶抗氧化酶活性及基因表达的影响

王艳, 高鹏, 黄敏

... 在高温胁迫下,水稻(Oryza sativa)开花期剑叶的SOD、CAT和POD基因表达量显著增加,从而维持了较高的酶活性来应答高温胁迫^[15] ...

2013

0.0

. 2013, 30(1):11-17

镉对镉超累积植物龙葵抗氧化酶活性及基因表达的影响

张玉秀, 金玲, 冯珊珊

... 过量的镉积累增强了龙葵(Solanum nigrum)SOD、POD和CAT的转录水平^[16] ...

2008

0.0

. 2008, 41(10):3025-3032

镉对玉米幼苗活性氧代谢、超氧化物歧化酶和过氧化氢酶活性及其基因表达的影响

赵士诚, 孙静文, 马有志

... 而随着镉对玉米(Zea mays)幼苗处理时间的增加,SOD的表达量受到抑制^[17],高钙处理苹果(Malus pumila)果实在12 h后SOD和CAT表达量增加^[18] ...

2011

0.0

. 2011, 17(2):438-444

钙对苹果果实超氧化物歧化酶、过氧化氢酶活性及其基因表达的影响

梁国庆, 孙静文, 周卫

... 而随着镉对玉米(Zea mays)幼苗处理时间的增加,SOD的表达量受到抑制^[17],高钙处理苹果(Malus pumila)果实在12 h后SOD和CAT表达量增加^[18] ...

... 这表明在盐胁迫下转录水平上,上调抗氧化酶基因表达量可以激活抗氧化酶的活性,能更有效地减少ROS的积累,维持细胞膜稳定性和生理平衡^[18] ...

2011

0.0

. 2011, 74(7):2050-2056 DOI:10.1016/j.ecoenv.2011.07.013

Toxic effect of NaCl on ion metabolism, antioxidative enzymes and gene expression of perennial ryegrass

Tao H , Li H Y , Zhang X Z

Key Laboratory of Plant Germplasm Enhancement and Specialty Agriculture, Wuhan Botanical Garden, The Chinese Academy of Science, Wuhan City, Hubei 430074, PR China.

Two-month old seedlings of perennial ryegrass (Lolium perenne L.) were subjected to four different levels of salinity for 7 days. The NaCl treatments reduced turf quality and normalized transpiration rates. Both chlorophyll (Chl) a and Chl b contents decreased in the grass exposed to 255 mM relative to the control. An increase in the lipid peroxidationin was observed. The activity of leaf superoxide dismutase increased while, peroxidase and catalase activities decreased in response to NaCl treatments. The expression of Chl Cu/ZnSOD, Cyt Cu/ZnSOD, FeSOD, CAT, POD, GPX and GR was up-regulated for NaCl-treated grass. Salt stress increased accumulation of Na(+) and decreased K(+)/Na(+) ratio, Mg(2+) and P content in both shoots and roots of perennial ryegrass. The findings of this study suggest that salt stress may cause toxicity to perennial ryegrass through oxidative injury and damage to Chl and cell membrane integrity.

PMID: 21813179

... 盐胁迫加强了CytCu/ZnSOD、CAT和POD等基因的表达^[19],因此,研究抗氧化酶活性与转录水平调控对理解草坪草在非生物胁迫,尤其是盐胁迫下的抗氧化机制具有重要作用 ...

2011

0.0

... 匍匐翦股颖(Agrostis stolonifera)是优良的冷季型草坪草,具有极耐低修剪、成坪快、草坪质量高的优点,被广泛应用于城市园林绿化和运动场草坪,但是较低的耐盐性^[20]限制了其应用 ...

2001

0.0

... 有很好的耐盐性^[21] ...

1950

0.0

... 5强度(在原配方的基础上减半)的Hoagland营养液^[22]直至从底部流出 ...

1996

0.0

. 1996, 132(3):361-373 DOI:10.1111/j.1469-8137.1996.tb01856.x

Antioxidant responses to drought in sunflower and sorghum seedlings

Zhang J , Kirkham M B

Department of Agronomy, Throckmorton Plant Sciences Center, Kansas State University, Manhattan, KS 66506-5501, USA.

To determine if antioxidant responses to drought differ between C3 and C4 plants, we grew Sorghum bicolor (C4 ) and Helianthus annuus (C3 ) under either watered or dry conditions in a growth chamber. Levels of leaf enzymatic antioxidants (ascorbate peroxidase, catalase, guaiacol peroxidase, dehydroascorbate reductase, monodehydroascorbate reductase, glutathione reductase and superoxide dismutase), nonenzymatic antioxidants (ascorbate, glutathione and carotenoids) and stress parameters (Chl and malondialdehyde [MDA]) were determined. Under watered and drought conditions, Chl contents and leaf relative water contents (RWC) were higher in sorghum than in sunflower; however, MDA levels were higher in sunflower than in sorghum. Under watered conditions, inherent levels of antioxidants were not consistently higher or lower in sorghum than in sunflower. In response to drought, levels of antioxidants, Chl and MDA showed increase, decrease or remained unchanged depending on crop, duration of drought and kind of antioxidants. Duration of drought was divided arbitrarily into three stages. At an early stage of drought (watering had stopped for 3-4 d) when soil water content (WC) and leaf RWC had decreased only in sunflower, drought generally did not affect levels of antioxidants and stress parameters. At a middle stage of drought (watering had stopped for 5-6 d) when soil WC had decreased for both sorghum and sunflower but leaf WC and RWC had decreased only in sunflower, drought changed levels of some antioxidants in sunflower and sorghum. At a late stage of drought (watering had stopped for 7-8 d) when soil WC, leaf WC and RWC had decreased in sorghum and sunflower, most parameters studied were affected by drought. Because of the differential effect of drought, levels of antioxidants were not consistently higher or lower in sorghum than in sunflower under drought. These results show that, under both drought and watered conditions, sorghum does not have consistently higher or lower antioxidant levels than sunflower, and that antioxidant responses to drought differ in C3 and C4 plants.

PMID: 26763632

... 0)(50 mmol/L)^[23] ...

... 2 抗氧化酶活性测定 SOD活性用Giannopolitis等^[24]的方法测定,CAT、POD和APX的活性采用Zhang等^[23]的方法进行测定,可溶性蛋白用Bradford^[25]的方法测定 ...

1977

0.0

. 1977, 59(2):309-314

Superoxide dismutase: I. Occurrence in higher plants

Giannopolitis C N , Ries S K

Department of Horticulture, Michigan State University, East Lansing, Michigan 48824.

Shoots, roots, and seeds of corn (Zea mays L., cv. Michigan 500), oats (Avena sativa L., cv. Au Sable), and peas (Pisum sativum L., cv. Wando) were analyzed for their superoxide dismutase content using a photochemical assay system consisting of methionine, riboflavin, and p-nitro blue tetrazolium. The enzyme is present in the shoots, roots, and seeds of the three species. On a dry weight basis, shoots contain more enzyme than roots. In seeds, the enzyme is present in both the embryo and the storage tissue. Electrophoresis indicated a total of 10 distinct forms of the enzyme. Corn contained seven of these forms and oats three. Peas contained one of the corn and two of the oat enzymes. Nine of the enzyme activities were eliminated with cyanide treatment suggesting that they may be cupro-zinc enzymes, whereas one was cyanide-resistant and may be a manganese enzyme. Some of the leaf superoxide dismutases were found primarily in mitochondria or chloroplasts. Peroxidases at high concentrations interfere with the assay. In test tube assays of crude extracts from seedlings, the interference was negligible. On gels, however, peroxidases may account for two of the 10 superoxide dismutase forms.

PMID: 16659839

... 2 抗氧化酶活性测定 SOD活性用Giannopolitis等^[24]的方法测定,CAT、POD和APX的活性采用Zhang等^[23]的方法进行测定,可溶性蛋白用Bradford^[25]的方法测定 ...

1976

0.0

... 2 抗氧化酶活性测定 SOD活性用Giannopolitis等^[24]的方法测定,CAT、POD和APX的活性采用Zhang等^[23]的方法进行测定,可溶性蛋白用Bradford^[25]的方法测定 ...

1994

0.0

... CAT在抗氧化保护机制中起到了重要作用,它把SOD清除ROS产生的过氧化氢催化分解成水和氧气^[26] ...

... 在植物叶绿体内谷胱甘肽循环的第一步,APX使用电子供体的抗坏血酸清除过氧化氢^[26] ...

... 植物体内的POD通过共基质的酚类抗氧化剂或化合物氧化来分解过氧化氢^[26] ...

2012

0.0

. 2012, 23(5):18-21

黄瓜砧用白籽南瓜对不同盐胁迫的耐性评价

王丽萍, 孙锦, 郭世荣

... 3 讨论与结论植物体内存在ROS生成与清除机制,在正常情况下两类机制处于平衡状态,但植物在遭受外界环境胁迫时,这种平衡受到破坏,积累过多的ROS会引起植物的氧化伤害^[27],在这种机制中抗氧化酶SOD、CAT、APX、POD在保持细胞内ROS动态平衡发挥了重要作用 ...

2015

0.0

. 2015, 24(1):183-189

4种燕麦对NaCl胁迫的生理响应及耐盐性评价

刘凤歧, 刘杰淋, 朱瑞芬

... 2 mol/L盐胁迫下,两种匍匐翦股颖SOD活性和转录水平响应不同,盐敏感的Penncross SOD活性先升高再降低,与刘凤歧等^[28]对盐胁迫下燕麦(Avena sativa)的SOD变化一致 ...

2014

0.0

. 2014, 23(3):160-166 DOI:10.11686/cyxb20140318

CoCl 2 对NaCl胁迫下大麦生长及幼苗生理指标的影响

孟亚雄, 王世红, 汪军成

1.Gansu Key Lab of Crop Improvement & Germplasm Enhancement Gansu provincial Key Lab of Aridland Crop Science, Lanzhou 730070, China; 2. College of Agronomy, Gansu Agriculture University, Lanzhou 730070, China; 3.College of Life Sciences and Technology, Gansu Agricultural University, Lanzhou 730070, China; 4. Institute of Wheat, Gansu Academy of Agricultural Sciences, Lanzhou 730070, china

To determine the beneficial effect of cobalt on the growth of Hordeum vulgare under NaCl stress, the effects of addition of CoCl 2 on seed germination, seedling growth, endogenous hormones and some physiological indexes of barley seedlings under NaCl stress were studied. With the aggravation of NaCl stress, the germination vigor, germination capacity, shoot length and root length decreased considerably, while the contents of abscisic acid (ABA) and malondialdehyde (MDA) increased significantly, and the activities of antioxidant enzymes were enhanced. Under NaCl stress, germination vigor, germination capacity, shoot length and root length and the activities of antioxidant enzymes were elevated by the treatment of 0.1 mmol/L CoCl 2 , compared with NaCl stress alone, and it dramatically inhibited the accumulation of MDA and ABA. This effect was more significant under the more serious NaCl stress. The results showed that the addition of CoCl 2 could improve (IAA+GA 3 )/ABA, enhance the ability to clear up reactive oxygen species, reduce lipid peroxidation under NaCl stress, and promote seed germination, seedling growth and adaptive capacity of barley.

为了明确钴对NaCl胁迫的缓解作用,本研究就CoCl 2 对不同浓度NaCl胁迫下大麦种子的萌发、幼苗生长及内源激素与相关生理指标的影响进行了探讨。结果表明,随着NaCl胁迫程度的加重,大麦种子发芽势、发芽率显著降低,芽与根的生长受到显著抑制。幼苗脱落酸(ABA)与丙二醛(MDA)大量积累,抗氧化酶活性提高。加入0.1 mmol/L CoCl 2 显著提高了NaCl胁迫下大麦种子的发芽势与发芽率,促进了芽与根的生长,抑制了幼苗ABA的积累,同时提高了抗氧化酶活性,降低了MDA的含量,且在较严重NaCl胁迫下作用更明显。CoCl 2 提高了NaCl胁迫下幼苗(IAA+GA 3 )/ABA的值,减缓了活性氧(ROS)的积累与膜脂过氧化进程,对种子萌发及幼苗生长起到了促进作用,在一定程度上提高了大麦幼苗对NaCl胁迫的适应能力。

... SOD显著高于Penncross,SOD活性高有利于清除ROS^[29],从而增加植物对盐害的抵抗力 ...

2004

0.0

. 2004, 161(5):531-542

Defense potentials to NaCl in a mangrove, Bruguiera parviflora: differential changes of isoforms of some antioxidative enzymes

Asish Kumar P , Anath Bandhu D , Prasanna M

National Institute for Plant Biodiversity Conservation and Research, Nayapalli, Bhubaneswar 751015, Orissa, India.

In order to assess the role of the antioxidative defense system against salt treatment, the activities of some antioxidative enzymes and levels of antioxidants were monitored in a true mangrove, Bruguiera parviflora, subjected to varying levels of NaCl under hydroponic culture. In the leaves of B. parviflora, salt treatment preferentially enhanced the content of H2O2 as well as the activity of ascorbate peroxidase (APX), guaiacol peroxidase (GPX), glutathione reductase (GR), and superoxide dismutase (SOD), whereas it induced the decrease of total ascorbate and glutathione (GSH+GSSG) content as well as catalase (CAT) activity. Analysis of isoforms of antioxidative enzymes by native PAGE and activity staining revealed that leaves of B. parviflora had one isoform each of Mn-SOD and Cu/Zn-SOD and three isoforms of Fe-SOD. Expression of Mn-SOD and Fe-SOD-2 was preferentially elevated by NaCl. Similarly, out of the six isoforms of GPX, the GPX-1, 2, 3 and 6 were enhanced by salt treatment but the levels of GPX-4 and -5 changed minimally as compared to those of a control. Activity staining gel revealed only one prominent isoform of APX and two isoforms of GR (GR-1 and GR-2), all of these isoforms increased upon salt exposure. Four CAT-isoforms were identified, among which the prominent CAT-2 isoform level was maximally reduced, suggesting differential down regulation of CAT isoforms by NaCl. The concentrations of malondialdehyde (MDA), a product of lipid peroxidation, remained unchanged in leaves of the plant treated with different concentrations of NaCl. This suggests that plants are protected against activated oxygen species by the elevated levels of certain antioxidative enzymes, thus avoiding lipid peroxidation during salt exposure. The differential changes in the levels of the isoforms due to NaCl treatment may be useful as markers for recognizing salt tolerance in mangroves.

PMID: 15202709

... 2 mol/L盐胁迫下,两种匍匐翦股颖抗氧化酶CAT、APX和POD的活性都增加^[30,31],而且耐盐的Seaside#cod#x02161 ...

2008

0.0

. 2008, 165(15):1620-1635 DOI:10.1016/j.jplph.2007.11.006

Exogenous spermidine affects polyamine metabolism in salinity-stressed cucumis sativus roots and enhances short-term salinity tolerance

Duan J , Li J S , Kang Y

College of Horticulture, Nanjing Agricultural University, Nanjing, China.

We investigated the effects of short-term salinity stress and spermidine application to salinized nutrient solution on polyamine metabolism and various stress defense reactions in the roots of two cucumber (Cucumis sativus L.) cultivars, Changchun mici and Jinchun No. 2. Seedlings grown in nutrient solution salinized with 50mM NaCl for 8d displayed reduced relative water content, net photosynthetic rates and plant growth, together with increased lipid peroxidation and electrolyte leakage in the roots. These changes were more marked in cv. Jinchun No. 2 than in cv. Changchun mici, confirming that the latter cultivar is more salinity-tolerant than the former. Salinity stress caused an increase in superoxide and hydrogen peroxide production, particularly in cv. Jinchun No. 2 roots, while the salinity-induced increase in antioxidant enzyme activities and proline contents in the roots was much larger in cv. Changchun mici than in cv. Jinchun No. 2. In comparison to cv. Jinchun No. 2, cv. Changchun mici showed a marked increase in arginine decarboxylase, ornithine decarboxylase, S-adenosylmethionine decarboxylase and diamine oxidase activities, as well as free spermidine and spermine, soluble conjugated and insoluble bound putrescine, spermidine and spermine contents in the roots during exposure to salinity. On the other hand, spermidine application to salinized nutrient solution resulted in alleviation of the salinity-induced membrane damage in the roots and plant growth and photosynthesis inhibition, together with an increase in polyamine and proline contents and antioxidant enzyme activities in the roots of cv. Jinchun No. 2 but not of cv. Changchun mici. These results suggest that spermidine confers short-term salinity tolerance on cucumber probably through inducing antioxidant enzymes and osmoticants.

PMID: 18242770

... 2 mol/L盐胁迫下,两种匍匐翦股颖抗氧化酶CAT、APX和POD的活性都增加^[30,31],而且耐盐的Seaside#cod#x02161 ...

2015

0.0

. 2015, 117:96-106 DOI:10.1016/j.ecoenv.2015.03.023

Protective effect of spermidine on salt stress induced oxidative damage in two Kentucky bluegrass (Poa pratensis L.) cultivars

Puyang X , An M , Han L

Turfgrass Research Institute, Beijing Forestry University, Beijing 100083, PR China.

To improve the salinity tolerance of turfgrass and investigate the effect of spermidine (Spd) on antioxidant metabolism and gene expression under salinity stress condition, exogenous Spd was applied before two kentucky bluegrass (Poa pratensis L.) cultivars ('Kenblue' and 'Midnight') were exposed to 200 mM sodium chloride (NaCl) stress for 28 d. Salinity stress decreased the turfgrass quality, increased the content of malonyldialdehyde (MDA), superoxide anion (O₂(·-)) and hydrogen peroxide (H₂O₂), and enhanced activities of superoxide dismutase (SOD), catalase (CAT), guaiacol peroxidase (POD) and ascorbate peroxidase (APX) and isozymes intensity in both cultivars. In addition, the expression level of Cu/ZnSOD was down-regulated in 'Kenblue' but up-regulated in 'Midnight' after salt treatment. Salinity stress also enhanced the expression of APX but inhibited the expression of CAT and POD in both cultivars. Exogenous Spd treatment alleviated the salinity-induced oxidative stress through decreasing MDA, H₂O₂ and O₂(·-) contents in both cultivars. Besides, exogenous Spd further enhanced the activities of SOD, CAT, POD and APX accompanied with the increased intensity of specific isozymes of SOD, CAT and APX in both cultivars and POD in 'Kenblue'. Moreover, Spd further up-regulated expression levels of Cu/ZnSOD and APX, but down-regulated those of CAT and POD in both cultivars. These results indicated that exogenous Spd might improve turfgrass quality and promote the salinity tolerance in the two cultivars of kentucky bluegrass through reducing oxidative damages and increasing enzyme activity both at protein and transcriptional levels.

PMID: 25841065

... 比Penncross CAT、APX和POD的活性更高,这与Puyang等^[32]对草地早熟禾在盐胁迫下抗氧化酶的影响研究结果一致 ...

2012

0.0

... 对盐胁迫下大麦的研究表明FeSOD活性没变化,甚至有的大麦基因型在盐处理下FeSOD活性降低^[33] ...

2008

0.0

... 活性氧(ROS)大量积累,会对植物造成膜系统氧化损伤^[7],体内激素平衡被破坏^[8],蛋白质和核酸变性 ...

2012

0.0

. 2012, 21(1):112-117

盐胁迫对高羊茅生长及抗氧化系统的影响

樊瑞苹, 周琴, 周波

2015

0.0

. 2015, 33(3):355-361

高温对水稻开花期剑叶抗氧化酶活性及基因表达的影响

王艳, 高鹏, 黄敏

... 在高温胁迫下,水稻(Oryza sativa)开花期剑叶的SOD、CAT和POD基因表达量显著增加,从而维持了较高的酶活性来应答高温胁迫^[15] ...

2013

0.0

. 2013, 30(1):11-17

镉对镉超累积植物龙葵抗氧化酶活性及基因表达的影响

张玉秀, 金玲, 冯珊珊

... 过量的镉积累增强了龙葵(Solanum nigrum)SOD、POD和CAT的转录水平^[16] ...

2008

0.0

. 2008, 41(10):3025-3032

镉对玉米幼苗活性氧代谢、超氧化物歧化酶和过氧化氢酶活性及其基因表达的影响

赵士诚, 孙静文, 马有志

... 而随着镉对玉米(Zea mays)幼苗处理时间的增加,SOD的表达量受到抑制^[17],高钙处理苹果(Malus pumila)果实在12 h后SOD和CAT表达量增加^[18] ...

2011

0.0

. 2011, 17(2):438-444

钙对苹果果实超氧化物歧化酶、过氧化氢酶活性及其基因表达的影响

梁国庆, 孙静文, 周卫

... 而随着镉对玉米(Zea mays)幼苗处理时间的增加,SOD的表达量受到抑制^[17],高钙处理苹果(Malus pumila)果实在12 h后SOD和CAT表达量增加^[18] ...

... 这表明在盐胁迫下转录水平上,上调抗氧化酶基因表达量可以激活抗氧化酶的活性,能更有效地减少ROS的积累,维持细胞膜稳定性和生理平衡^[18] ...

2011

0.0

2012

0.0

. 2012, 23(5):18-21

黄瓜砧用白籽南瓜对不同盐胁迫的耐性评价

王丽萍, 孙锦, 郭世荣

2015

0.0

. 2015, 24(1):183-189

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刘凤歧, 刘杰淋, 朱瑞芬

2014

0.0

. 2014, 23(3):160-166 DOI:10.11686/cyxb20140318

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孟亚雄, 王世红, 汪军成

... SOD显著高于Penncross,SOD活性高有利于清除ROS^[29],从而增加植物对盐害的抵抗力 ...