Involvement of antioxidant defense system in chill hardening-induced chilling tolerance in Jatropha curcas seedlings

Jatropha curcas L. is a sustainable energy plant with great potential for biodiesel production, and low temperature is an important limiting factor for its distribution and production. In this present work, chill hardening-induced chilling tolerance and involvement of antioxidant defense system were investigated in J. curcas seedlings. The results showed that chill hardening at 10 or 12 °C for 1 and 2 days greatly lowered death rate and alleviated electrolyte leakage as well as accumulation of the lipid peroxidation product malondialdehyde (MDA) of J. curcas seedlings under severe chilling stress at 1 °C for 1–7 days, indicating that the chill hardening significantly improved chilling tolerance of J. curcas seedlings. Measurement of activities of the antioxidant enzymes superoxide dismutase (SOD), ascorbate peroxidase (APX), catalase (CAT), peroxidase (POD), and glutathione reductase (GR), and the levels of the antioxidants ascorbic acid (AsA) and glutathione (GSH) showed the chill hardening at 12 °C for 2 days could obviously increase the activities of these antioxidant enzymes and AsA and GSH contents in the hardened seedlings. When the hardened and non-hardening (control) seedlings were subjected to severe chilling stress at 1 °C for 1–7 days, the chill-hardened seedlings generally maintained significantly higher activities of the antioxidant enzymes SOD, APX, CAT, POD, and GR, and content of the antioxidants AsA and GSH as well as ratio of the reduced antioxidants to total antioxidants [AsA/(AsA + DHA) and GSH/(GSH + GSSG)], when compared with the control without chill hardening. All above-mentioned results indicated that the chill hardening could enhance the chilling tolerance, and the antioxidant defense system plays an important role in the chill hardening-induced chilling tolerance in J. curcas seedlings.     

外源α-萘乙酸对花期长期干旱大豆叶片抗氧化系统的影响

以不同耐旱型品种‘南农99-6’和‘科丰1号’大豆为材料,2012年在南京农业大学牌楼试验站进行为期110 d的盆栽试验,研究大豆花期叶面喷施α-萘乙酸(NAA)对长期干旱条件下大豆植株抗氧化系统的影响.结果表明: 干旱胁迫显著降低了大豆地上部干物质量,叶片中丙二醛(MDA)含量及活性氧(ROS)水平显著升高,同时,超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)、抗坏血酸过氧化物酶(APX)、单脱氢抗坏血酸还原酶（MDHAR）、谷胱甘肽还原酶(GR)和谷胱甘肽过氧化物酶(GPX)活性,还原型抗坏血酸(AsA)、还原型谷胱甘肽(GSH)含量及AsA/DHA(双脱氢抗坏血酸)和GSH/GSSG(氧化型谷胱甘肽)比值显著升高,其中‘科丰1号’大豆的抗氧化能力更高,从而维持较低的ROS水平和MDA含量.NAA可显著提高叶片中的APX、POD、CAT、MDHAR活性及AsA/DHA、GSH/GSSG比值,其中‘科丰1号’大豆叶片的脱氢抗坏血栓还原酶（DHAR）活性和AsA含量极显著增加.     

渗透胁迫对花生幼叶细胞性氧伤害的定量分析

在渗透胁迫条件下不同花生（Arachis hypogaea Linn．）品种的水势、质膜相对透性（RPMP）、超氧离子、丙二醛（MDA）含量和超氧化物歧化酶（SOD）活性两两之间都呈显著相关。而花生各品种的过氧化氢酶活性、过氧化物酶活性、谷胱甘肽含量和抗坏血酸含量与水势、RPMP、超氧离子、MDA含量和SOD活性等指标之间相关程度波动较大。水势对RPMP、MDA含量、超氧离子和SOD活性的弹性系数处于负效应阶段。SOD活性对RPMP、MDA含量和超氧离了的弹性系数的效应处于递减阶段。超氧离了对RPMP和MDA含量的生系数的效应处于递减阶段。各指标的弹性系数和相应的边际量因花生品种的抗旱能力不同而异。     

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。     

Retardation of Leaf Senescence by Triadimefon Treatment in Mung Bean Seedling

Treatment of triadimefon on detached leaves of mung bean (Phaseolus radiatus L. ) seedlings increased the levels of chlorophyll and soluble proteins. Declined activities of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT) and ascorbate-peroxidase (AsA-POD) and contents of ascorbate (ASA) and glutathione (GSH) were observed during the senescence of detached young leaves. Triadimefon at concentration of 20 mg/L promoted the activities of POD, AsA-POD and levels of AsA and GSH, but had no effect on the activities of SOD and CAT. On the other hand, the levels of malondialdehyde (MDA) were increased and the increase of which was markedly negative correlated with the activities of POD, AsA-POD and with the contents of AsA and GSH during the senescence of leaves. MDA contents were decreased by triadimefon treatment. These resuits suggested that triadimefon retarded the senescence of leaves in mung bean seedlings in terms of enhancing the protective ability of plant tissues against membrane lipid peroxidation.     

Combined action of antioxidant defense system and osmolytes in chilling shock-induced chilling tolerance in Jatropha curcas seedlings

Low non-freezing temperature is one of the major environmental factors affecting growth, development and geographical distribution of chilling-sensitive plants, Jatropha curcas is considered as a sustainable energy plants with great potential for biodiesel production. In this study, chilling shock at 5 °C followed by recovery at 26 °C for 4 h significantly improved survival percentage of J. curcas seedlings under chilling stress at 1 °C. In addition, chilling shock could obviously enhance the activities of antioxidant enzymes superoxide dismutase (SOD), ascorbate peroxidase (APX), catalase (CAT) and glutathione reductase (GR), and the levels of antioxidants ascorbic acid (AsA) and glutathione (GSH), as well as the contents of osmolytes proline and betaine in leaves of seedlings of J. curcas compared with the control without chilling shock. During the process of recovery, GR activity, AsA, GSH, proline and betaine contents sequentially increased, whereas SOD, APX and CAT activities gradually decreased, but they markedly maintained higher activities than those of control. Under chilling stress, activities of SOD, APX, CAT, GR and GPX, and contents of AsA, GSH, proline and betaine, as well as the ratio of the reduced antioxidants to total antioxidants [AsA/(AsA + DHA) and GSH/(GSH + GSSG)] in the shocked and non-shock seedlings all dropped, but shocked seedlings sustained significantly higher antioxidant enzyme activity, antioxidant and osmolyte contents, as well as ratio of reduced antioxidants to total antioxidants from beginning to end compared with control. These results indicated that the chilling shock followed by recovery could improve chilling tolerance of seedlings in J. curcas, and antioxidant enzymes and osmolytes play important role in the acquisition of chilling tolerance.     

Exogenous trehalose differentially modulate antioxidant defense system in wheat callus during water deficit and subsequent recovery

To elucidate the resistance mechanism of exogenous trehalose on water deficit further, we investigated the effect of exogenous trehalose (50 mM) in wheat callus during water deficit and subsequent recovery. Enhanced levels of endogenous trehalose were detected in calli exposed to water deficit (W) and trehalose (T) medium, moreover, W plus T treatment showed an additive effect. Water deficit elevated the accumulation of ROS (hydrogen peroxide and formation rate of O ₂ ^.? ) and the endogenous MDA (Malonaldehyde), and resulted in the decrease of cell viability and biomass. Exogenous trehalose (TW) could alleviate the damage induced by water deficit, which was involved in the decrease of MDA and the generation of ROS, and resulted in elevating cell viability and biomass. Additionally, water deficit induced activity of antioxidative enzymes (Peroxidase, POD; Catalase, CAT; Glutathione reductase, GR). Content of AsA (Reduced ascorbate) was also increased by water deficit, while the content of GSH (Glutathione) showed the opposite effect. The combined effect of T and W treatment led to a higher activity of enzymatic antioxidants including SOD (Superoxide dismutase) and GR, and elevated the content of nonenzymatic antioxidants including AsA and GSH, but had a negative effect on enzymatic antioxidants including POD and CAT in comparison to the water deficit treatment alone. During recovery, calli treated by TW showed a greater reduction in ROS resulted in enhancing a higher cell viability and biomass. The scavenging mechanism of ROS by exogenous trehalose is mainly dependent on nonenzymatic antioxidants, especially AsA-GSH cycle, rather than enzymatic mechanisms and trehalose itself.     

CPPU提高柚树苗抗旱性的研究

水分胁迫下,柚树苗叶片RWC、ψW、Pn及可溶性蛋白质、叶绿素含量下降,游离脯氨酸、MDA含量及SOD活性升高,CAT活性先上升后下降,APX活性、AsA和GSH含量明显下降,抗旱性强的品种具有较高的活性氧清除能力。CPPU处理提高柚树苗叶片RWC、Pn、叶绿素、游离脯氨酸、AsA、GSH含量,并增强SOD、APX活性,降低MDA水平,从而提高柚树苗抗旱性。实验结果表明,CPPU处理提高叶片对膜脂过氧化作用的保护能力可能是其提高柚树苗抗旱性的原因。     

CPPU提高柚树苗抗旱性的研究

水分胁迫下,柚树苗叶片RWC、ψw、Pn及可溶性蛋白质,叶绿素含量下降,游离脯氨酸、MDA含量及SOD活性升高,CAT活性先上升后下降,APX活性、AsA和GSH含量明显下降,抗旱性强的品种具有较高的活性氧清除能力。CPPU处理提高柚树苗叶片RWC、Pn、叶绿素、游离脯氨酸、AsA、GSH含量,并增加SOD、APX活性,降低MDA水平,从而提高柚树苗抗旱性。实验结果表明,CPPU处理提高叶片对膜脂过氧化作用的保护能力可能是其提高柚树苗抗旱性的原因。     

Synergistic effects of drought and ascorbic acid on growth,mineral nutrients and oxidative defense system in canola (Brassica napus L.) plants

A study was conducted to find out the role of ascorbic acid (AsA) in modulating growth and different physio-biochemical attributes of canola plants under well-watered as well as water-deficit conditions. Drought stress imposed on 60 % field capacity significantly decreased the shoot and root fresh and dry weights, leaf chlorophyll contents, shoot and root P, root K⁺, and activity of CAT enzyme, while increased chlorophyll a/b contents, MDA, NPQ, leaf total phenolics, free proline and GB contents in both canola cultivars. Foliar-applied varying levels (50, 100 and 150 mg L^?1) of AsA enhanced shoot and root fresh and root dry weights, qN, NPQ, shoot and root P, AsA as well as the activity of POD enzyme particularly under drought stress conditions. Of both canola cultivars, cv. Dunkeld was higher in shoot fresh weights, ETR and F _v /F _m, MDA, proline and GB contents, and POD activity, however, cv. Cyclone in total phenolics and qN under well-watered and water-deficit conditions. Overall, the foliar-applied AsA had a positive effect, though not marked, on salt sensitive cv. Cyclone in terms of improved growth and other attributes, whereas exogenously applied AsA had a non-significant effect on relatively salt tolerant cv. Dunkeld.     

H_2S对干旱胁迫下白三叶幼苗抗氧化防御能力的影响

以‘拉丁诺’白三叶(Trifolium repens cv.‘Ladino’)为试验材料,研究外源H2S处理对PEG6 000(聚乙二醇)模拟干旱胁迫下白三叶叶片相对含水量(RWC)、膜脂过氧化、活性氧成分、抗氧化酶、抗坏血酸-谷胱甘肽循环代谢和非酶抗氧化物质的影响,以揭示H_2S调控白三叶抗旱性的生理机制。结果显示:(1)0.2 mmol/L的外源NaHS(H_2S供体)能显著提高干旱胁迫下白三叶的叶片相对含水量,维持显著较低的电解质渗透率(EL)和丙二醛(MDA)含量。(2)与直接干旱胁迫相比,干旱胁迫下外源添加NaHS处理的白三叶叶片内超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)活性显著增强,抗坏血酸-谷胱甘肽循环代谢中关键酶抗坏血酸过氧化物酶(APX)、脱氢抗坏血酸还原酶(DHAR)、单脱水抗坏血酸还原酶(MDHAR)和谷胱甘肽还原酶(GR)活性及其抗氧化中间产物抗坏血酸(AsA)、谷胱甘肽(GSH)含量也显著提高。(3)叶片类黄酮、总酚和原花青素的含量在一定的胁迫时间范围内亦显著增加,并伴随着活性氧成分O_2~(-·)产生速率和H_2O_2水平降低。研究认为,外源H2S能通过促进干旱胁迫下白三叶体内的多重抗氧化防御能力来提高其幼苗的抗旱性。     

干旱胁迫下沙生灌木花棒的抗氧化保护响应研究

采用盆栽试验,测定了在不同土壤水分胁迫程度和胁迫时间下花棒苗抗氧化保护酶(SOD、CAT、POD、APX)活性和抗氧化物(AsA、GSH、Car)含量,并用隶属函数值法综合评价了试验条件下花棒苗的抗氧化能力。结果显示:(1)与同期适宜水分处理相比,花棒苗的抗氧化保护酶活性在中度和重度干旱胁迫30d时均显著增加;胁迫60d时,POD活性均显著降低,而其余酶活性仍增加(重度胁迫)或显著增加(中度胁迫);胁迫90d时,中度胁迫处理除APX活性持平外,其余酶活性仍显著升高,而重度胁迫处理的SOD活性持平、POD活性显著增加、CAT和APX活性均显著降低。(2)相对于同期适宜水分处理,干旱胁迫30d时,中度胁迫处理的花棒苗AsA、Car、GSH含量分别显著增加、稍有增加和显著降低,而重度胁迫的AsA显著增加,Car和GSH含量均显著降低;胁迫60d时,中度胁迫处理的AsA、Car、GSH含量分别显著增加、稍有降低和稍有提高,而重度胁迫的AsA、Car、GSH含量分别显著增加、稍有增加和显著降低;胁迫90d时,中度胁迫处理的AsA含量显著增加,Car和GSH含量均稍有提高,而重度胁迫的AsA含量稍有下降,Car和GSH含量则持平。(3)花棒苗的MDA含量在干旱胁迫过程中均比适宜水分处理增加,且中后期达到显著水平。(4)花棒苗抗氧化能力在干旱胁迫30d表现为重度干旱>中度干旱>适宜水分,胁迫60d为中度干旱>重度干旱>适宜水分,胁迫90d时为中度干旱>适宜水分>重度干旱。研究表明,花棒苗在中度干旱胁迫下能调动自身抗氧化保护酶类与抗氧化物质并提高其抗氧化能力,消除或减轻干旱诱导的活性氧毒害;重度干旱胁迫下的花棒苗在胁迫前中期亦能增强自身的抗氧化能力,但在胁迫后期因部分活性氧清除机制受到破坏而使抗氧化能力降低。     

不同海拔青海云杉与祁连圆柏叶片抗氧化系统

以祁连山寺大隆林区连续海拔梯度(2 665~3 365 m)上青海云杉(Picea crassifolia)和祁连圆柏(Sabina przewalskii)为材料, 测定叶片中抗氧化保护系统的变化, 探讨常绿木本植物抗氧化系统对高山极端环境的适应机制。结果显示, 祁连圆柏和青海云杉叶片中丙二醛(MDA)含量均与海拔高度呈正相关, 相同海拔上青海云杉MDA含量极显著高于祁连圆柏(p＜0.01)。随海拔升高, 两树种抗氧化保护酶超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)、抗坏血酸过氧化物酶(APX)、谷胱甘肽还原酶(GR)活性和非酶促抗氧化剂脯氨酸(Pro)、抗坏血酸(AsA)、还原性谷胱甘肽(GSH)含量均呈明显增加趋势。青海云杉叶片的AsA水平高于祁连圆柏, 但对海拔变化的敏感性较低; 祁连圆柏的GSH、Pro水平及其对海拔变化的敏感性均高于青海云杉。结果表明, 研究区青海云杉所受过氧化伤害较祁连圆柏更严重, 但两树种清除O₂^-·的能力相当而主要负责分解H₂O₂的酶种有所不同: 祁连圆柏中为POD, 青海云杉中则为CAT、APX和GR, AsA-GSH循环系统在青海云杉活性氧清除中的作用强于在祁连圆柏中, 祁连圆柏的活性氧清除物质可能以Pro为主。     

抗氧化系统参与循环干旱锻炼提高烟草植株抗旱性的形成

对漂浮育苗的烟草幼苗进行控水-半萎焉-复水-恢复的循环干旱锻炼。结果表明,这种干旱锻炼能显著提高烟草叶片抗氧化剂谷胱甘肽（GSH）、抗坏血酸（AsA）的含量、还原型抗氧化剂在总抗氧化剂中的比例和抗氧化酶包括超氧化物歧化酶（SOD）、抗坏血酸过氧化物酶（APX）、过氧化氢酶（CAT）、愈创木酚过氧化物酶（GPX）、谷胱甘肽还原酶（GR）的活性,降低丙二醛（MDA）的含量。当烟草植株遭受后续的干旱胁迫时,与未锻炼的对照相比,干旱锻炼过的烟草植株能保持较高的GSH和AsA含量、还原型抗氧化剂在总抗氧化剂中的比例和抗氧化酶（SOD、APx、CAT、GPX和GR）活性,以及较低的MDA含量,表明这种循环干旱锻炼提高了细胞抗氧化能力,有助于缓解烟草植株由干旱引起的氧化胁迫及其所导致的伤害,最终提高其抗旱性。这些结果表明。抗氧化系统参与了循环干旱锻炼提高烟草植株抗旱性的形成过程。     

水分胁迫对龙眼幼苗叶片膜脂过氧化及内源保护体系的影响

对龙眼（ＤｉｍｏｃａｒｐｕｓｌｏｎｇａｎａＬｏｕｒ．）实生幼苗进行人工水分胁迫,检测其抗氧化酶和抗氧化剂的变化,研究了干旱对叶片膜脂过氧化及内源保护体系的影响,结果表明,在水分胁迫下,龙眼叶片ＲＷＣ下降,电解质渗漏上升,ＲＷＣ的下降和电解质渗漏的上升呈极显著负相关;叶片中ＳＯＤ和ＰＯＤ活性及ＭＤＡ含量明显上升,且与电解质渗漏的增加呈极显著正相关;叶片中可溶性蛋白质,ＧＳＨ含量和ＡｓＡ－ＰＯＤ活性显     

Does exogenous application of ascorbic acid modulate growth,photosynthetic pigments and oxidative defense in okra (<Emphasis Type="Italic">Abelmoschus esculentus</Emphasis> (L.) Moench) under lead stress?

Seed priming increases tolerance of plants against various environmental stresses. Although ample literature is available that depicts the beneficial effects of priming under different environmental stresses, the information on induction of tolerance to Pb stress through seed priming with ascorbic acid (AsA) is limited. Therefore, this study was performed to examine the effect of seed priming with AsA (50 and 100 mg L^?1), hydropriming and without priming (control) on physiochemical processes of okra cultivars (Subz-Pari and Arka Anamika) under Pb stress (0, 100 mg L^?1). Pb stress caused a considerable decline in plant growth and photosynthetic pigments. Contrarily, Pb stress exhibited rise in the contents of total amino acids, free proline, total soluble proteins and AsA. The POD, CAT, and SOD activities were recorded highest at 100 mg L^?1 of Pb. Moreover, Pb stress markedly increased H₂O₂ and MDA levels that triggered oxidative stress. However, plants raised from seed primed with AsA and water exhibited better growth and had higher chlorophylls, free proline, total proteins, total amino acids, AsA and activities of enzymatic antioxidants. Priming with AsA (50 mg L^?1) induced better tolerance to Pb stress in okra plants. Plants of cv. Arka Anamika exhibited greater tolerance to Pb than that of cv. Subz-Pari as was evident from higher plant fresh and dry masses.     

Spermidine pretreatment enhances heat tolerance in rice seedlings through modulating antioxidative and glyoxalase systems

This study was undertaken to investigate the possible involvement of the antioxidant defense and glyoxalase systems in protecting rice seedlings from heat-induced damage in the presence of spermidine (Spd). Hydroponically grown 14-day-old seedlings were subjected to foliar spray with Spd (1 mM, 24 h) prior to heat stress (42 °C, 48 h) followed by subsequent recovery (27 °C, 48 h). Lipoxygenase activity, malondialdehyde (MDA), hydrogen peroxide (H₂O₂) and proline (Pro) content increased significantly whereas fresh weight (FW) and chlorophyll (Chl) content decreased during heat stress and after recovery, indicating unrecoverable damage to rice seedlings. Heat-induced damage was also evident in decreased levels of ascorbate (AsA), glutathione (GSH), and AsA and GSH redox ratios. Superoxide dismutase (SOD) and catalase (CAT) activities increased during heat stress but declined after recovery. Activities of glutathione peroxidase (GPX), ascorbate peroxidase (APX), monodehydroascorbate reductase, dehydroascorbate reductase (DHAR) and glutathione reductase (GR) decreased during heat stress but an opposite trend for most of these enzymes was observed after recovery. Heat stress also resulted in significant increases in the activities of glyoxalase enzymes (Gly I and Gly II). In contrast, exogenous Spd protected rice seedlings from heat-induced damage as marked by lower levels of MDA, H₂O₂, and Pro content coupled with increased levels of AsA, GSH, FW, Chl, and AsA and GSH redox status. After recovery, Spd-pretreated heat-exposed seedlings displayed higher activities of SOD, CAT, GPX, GST APX, DHAR and GR as well as of Gly I and Gly II. In addition, polyamine analysis revealed that exogenously applied Spd significantly elevated the levels of free and soluble conjugated Spd. Therefore, we conclude from our results that heat exposure provoked an oxidative burden while enhancement of the antioxidative and glyoxalase systems by Spd rendered rice seedlings more tolerant to heat stress. Further, co-induction of the antioxidative and glyoxalase systems was closely associated with Spd mediated enhanced level of GSH.     

贵州特有植物美丽红山茶水分胁迫的生理响应

采用水分抗逆性指标主成分分析法,选择4年实生苗,测定和评价了连续30 d水涝和干旱胁迫的生理响应.结果表明：(1)水涝处理的叶片生长旺盛,光泽度和观赏性无明显变化,叶片相对含水量、MDA含量、SOD活性、POD活性、CAT活性的变化差异在5％以内,土壤含水量增加3．47％,细胞膜透性减少1．59％,短期水涝对美丽红山茶伤害不明显;(2)随着干旱时间延长,新叶卷缩、老叶脱落,土壤含水量、叶片相对含水量随时间呈负相关缓慢减少,细胞膜透性呈正相关缓慢增长,但复水10 d 后可恢复正常生长,无明显生理变化响应;(3)SOD活性和MDA含量分别增加了0．66％和5．31％,POD活性随干旱时间上下波动而增加6．47％, CAT活性随干旱时间延长而增加,20 d增加33．33％达最大值20．8 U??g-1??min-1.在抗性生理中,维持膜稳定性的叶片含水量、POD和CAT的变化对抗旱能力大小起主要作用.     

Glutathione improves survival of cryopreserved embryogenic calli of <Emphasis Type="Italic">Agapanthus praecox</Emphasis> subsp. <Emphasis Type="Italic">orientalis</Emphasis>

The effect of supplementation of reduced glutathione (GSH) to cryoprotectant solution on the generation of reactive oxygen species (ROS) (e.g., H₂O₂, OH^·, and O ₂ ^·? ) and antioxidants (e.g., SOD, POD, CAT, AsA, and GSH), as well as membrane lipid peroxidation (i.e., MDA content) mitigation in cryopreserving of embryogenic calli (EC) of Agapanthus praecox subsp. orientalis was investigated. The vitrification-based cryopreservation method was used in this study. The addition of GSH at a final concentration of 0.08 mM to the cryoprotectant solution has significantly improved cryotolerance of A. praecox EC. The EC post-thaw survival rate increased by 68.34 % using the cryoprotectant solution containing 0.08 mM GSH as compared to the control (GSH-free). EC treated with GSH displayed the reduction in  OH^· generation activity and the contents of H₂O₂ and MDA, as well as enhancement in the inhibition of O ₂ ^·? generation and the antioxidant activity. Treatment with exogenous GSH also increased endogenous AsA and GSH contents after dehydration step. Expression of stress-responsive genes, e.g., peroxidase (POD), peroxiredoxin, ascorbate peroxidase (APX), monodehydroascorbate reductase (MDHAR), and glutathione peroxidase (GPX), was also increased during cryopreservation processes. The expression of DAD1 (Defender against apoptotic cell death) was elevated, while cell death-related protease SBT was suppressed. These results demonstrated that the addition of GSH to cryoprotectant solution affects the ROS level and could effectively improve survival of A. praecox EC through enhancing antioxidant enzyme activities and decreasing cell death.