Effects of mechanical damage and herbivore wounding on H2O2 metabolism and antioxidant enzyme activities in hybrid poplar leaves

The changes of hydrogen peroxide (H₂O₂) metabolism and antioxidant enzyme activities in a hybrid poplar (Populus simonii × P. pyramidalis ‘Opera 8277’) in response to mechanical damage (MD) and herbivore wounding (HW) were investigated to determine whether H₂O₂ could function as the secondary messenger in the signaling of systemic resistance. Results show that H₂O₂ was generated in wounded leaves through MD and HW treatments and systemically in unwounded leaves around the wounded leaves. The activities of antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX) were also enhanced. However, the H₂O₂ accumulation and antioxidant enzyme activities were inhibited in MD leaves through the pretreatment with DPI (which is a specific inhibitor of NADPH oxidase). The results of this study suggest that H₂O₂ could be systemically induced by MD and HW treatments, and H₂O₂ metabolism was closely related to the change in SOD, APX and CAT activities. A high level of antioxidant enzymes could decrease membrane lipid peroxidation levels and effectively induce plant defense responses. Foundation project: This research is supported by the Key Science Program of the Sate Forestry Administration of China (2006–59), and the National Key Project of Scientific and Technical Supporting Programs Funded by Ministry of Science & Technology of China (2006BAD01A15; 2006BAD24B04). Biography: AN Yu (1982–), female, Postgraduate in College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, P. R. China.     

Ionic homeostasis and reactive oxygen species control in leaves and xylem sap of two poplars subjected to NaCl stress

We investigated the effects of increasing soil NaCl concentration on intracellular compartmentalization of salt and on the activities of antioxidant enzymes (superoxide dismutase (SOD), ascorbic peroxidase (APX), catalase (CAT) and glutathione reductase (GR)) and their role in the regulation of reactive oxygen species (ROS; O(2)(-*) and H(2)O(2)) in leaves and xylem sap of salt-tolerant Populus euphratica Oliv. and salt-sensitive P. popularis cv. 35-44. Mesophyll cells of P. euphratica exhibited a high capacity for NaCl exclusion and compartmentalization of salt in vacuoles compared with P. popularis. In P. popularis, the salt treatment resulted in large accumulations of Na(+) and Cl(-) in leaves that induced significant increases in O(2)(-*) and H(2)O(2) production despite marked increases in the activities of antioxidant enzymes in leaves and xylem sap. Separation of the isoforms of leaf SOD, APX and CAT by polyacrylamide gel electrophoresis followed by in-gel activity staining revealed that the salt-induced activities of APX and CAT were the result of increases in activities of all the isoenzymes. Leaf injury and shedding of aged leaves occurred following the oxidative burst in P. popularis, indicating that the increased activities of antioxidant enzymes in P. popularis were insufficient to counter the harmful effects of ROS at high soil NaCl concentrations. Unlike P. popularis plants, P. euphratica plants did not exhibit an oxidative burst in response to the NaCl treatments, because of (1) a high salt exclusion capacity and effective compartmentalization of salt in vacuoles, and (2) up-regulation of antioxidant enzymatic activities after the onset of salt stress. We conclude that P. euphratica plants subjected to saline conditions control ROS homeostasis through two pathways: (1) by maintaining cellular ionic homeostasis and thereby limiting the NaCl-induced enhancement of ROS production under long-term saline conditions; and (2) by rapidly up-regulating antioxidant defenses to prevent oxidative damage.     

遮荫下2种热带树苗叶片光合特性和抗氧化酶系统对自然降温的响应

选取 2种遮荫条件下 (8%、2 5 %全光照 )西双版纳地区热带雨林上层树种滇南红厚壳和中层树种玉蕊的幼苗 ,测定了雨季末自然降温过程中 2种植物叶片光合特性、抗氧化酶 (SOD、CAT、APX)活性、丙二醛 (MDA)含量的变化。研究结果表明 :低温使 2种植物单位面积叶绿素含量和净光合速率明显下降 ,生长在中等光强下的幼苗比低光强下的幼苗下降的比率大 ,滇南红厚壳比玉蕊下降的比率大。在降温初期 ,即从 10月到 12月初 ,2种光强下 2种植物SOD和APX活性上升 ,CAT活性出现波动 ,黎明光化学效率和MDA含量基本没有变化 ,光合机构没有受到氧化破坏。 12月末 ,中等光强下滇南红厚壳黎明光化学效率降低 ,SOD和APX活性有所下降 ,MDA含量显著上升 ;而低光强下的滇南红厚壳和 2种光强下玉蕊SOD和APX活性仍在上升 ,MDA含量变化较小。这表明 ,2种光强下 2种热带树苗是以不同的方式响应自然降温 ,低温对中等光强下的植物比低光强下的影响大 ,上层树种滇南红厚壳受到的影响比中层树种玉蕊大。同时 ,根据降温过程中抗氧化酶活性和MDA含量的变化 ,大致可将西双版纳热带雨林幼苗对低温的响应分为 2个阶段 ,即第 1阶段为低温适应阶段 ,抗氧化酶活性上升 ,光合机构没有受到明显的氧化破坏 ;第 2阶段 ,中等光强下滇南红厚壳抗氧化酶     

CO_2浓度升高对落叶松叶片抗氧化系统的影响

在CO_2浓度升高和对照处理的两个开顶式气室(OTC-Ⅰ型)内,研究分析了高浓度CO_2对落叶松针叶抗氧化系统酶活力的影响。结果表明:短期内(2个月左右)CO_2浓度升高条件下,超氧化物歧化酶(SOD)、抗坏血酸(ASA)、抗坏血酸过氧化物酶(APX)、谷胱甘肽还原酶(GR)含量上升,且SOD与O_2·~-产生速率之间呈极显著负相关;但长期高浓度CO_2处理下,ASA含量与SOD、APX、GR活性略有下降,可见,较长期高浓度CO_2处理可能使此效果发生逆转。在CO_2浓度升高条件下,植物可能通过抗氧化能力的增强来减轻胁迫造成的细胞损伤,从而增强植物的抗性。     

钙对酸化处理平邑甜茶根系抗氧化酶活性及线粒体功能的影响

为探讨酸化处理及钙对酸化处理下平邑甜茶根系抗氧化酶活性及线粒体功能的影响,将平邑甜茶幼苗分2组分别置于pH3,4,5,6(对照)的1/2Hoagland营养液中及3个钙离子浓度梯度(10,20,30mmol·L-1)的pH值为3的1/2Hoagland营养液中,于第0,2,3,4天分别测定抗氧化酶活性及线粒体指标。结果表明:与对照相比,各处理的SOD,CAT酶活性在处理的第2天达到最大值,POD酶活性变化不一,pH3与pH4处理在第2天达到最大值,pH5处理在第3天达到最大值;根系线粒体膜通透性、膜电位及细胞色素c/a随着酸度增加和时间的延长逐渐降低;H2O2含量先降低后升高。低浓度外源钙(10,20mmol·L-1)处理能提高SOD,CAT,POD酶活性并维持一定时间,缓解线粒体各项功能的伤害,抑制H2O2的积累,主要表现在与单独酸处理相比H2O2含量降低了6.78%,13.43%,且20mmol·L-1钙离子缓解效果最好;30mmol·L-1钙离子处理却加剧了酸化伤害,H2O2含量上升了8.96%。酸化处理会对平邑甜茶根系造成伤害而营养液适量加钙可以缓解其伤害。     

蓝莓离体繁育新梢形态与抗氧化酶对营养元素的应答

为了研究营养元素硝酸铵、硝酸钾、葡萄糖酸钙对蓝莓离体繁育的影响,以蓝莓‘夏普蓝’继代增殖苗为试材,就添加了不同浓度的硝酸铵、硝酸钾、葡萄糖酸钙的改良培养基对蓝莓离体增殖的新梢形态及体内超氧化物歧化酶(Superoxide Dismutase,SOD)、过氧化物酶(Peroxidase,POD)、过氧化氢酶(catalase,CAT)活性的影响情况进行了试验研究。试验结果表明:当培养基中硝酸铵的浓度为600 mg·L~(-1)、硝酸钾的浓度为400 mg·L~(-1)、葡萄糖酸钙的浓度为600 mg·L~(-1)时,新梢植株发育良好,植株长势健壮,颜色正常;发育正常的新梢其SOD活性间不存在明显差异,其POD活性间不存在明显差异,其CAT活性间也不存在明显差异;而且其SOD、POD、CAT活性均明显高于发育不正常的新梢。分析结果表明:矿质营养元素能调节蓝莓不定芽抗氧化酶活性;而稳定、平衡的抗氧化酶活性又能促进不定芽的发育。     

土壤干旱胁迫对巨龙竹实生苗光合特性和抗氧化酶活性的影响

试验选取盆栽巨龙竹(Dendrocalamus sinicus)实生苗，通过控制土壤水分含量设置轻度干旱、中度干旱和重度干旱3个胁迫水平研究不同干旱胁迫处理对巨龙竹实生苗的光合参数和抗氧化酶活性的影响。结果表明：随干旱程度的增加，土壤含水量显著下降，巨龙竹实生苗叶片的叶绿素含量(SPAD值)、净光合速率(Pn)、蒸腾速率(Tr)、气孔导度(Gs)均呈下降趋势，而胞间CO₂浓度(Ci)在中度和重度干旱时呈上升趋势；在3种干旱强度下，SOD活性呈现先上升后下降再上升的变化趋势，APX活性和丙二醛含量均高于对照，CAT均低于对照。由研究结果可见，干旱会引起巨龙竹实生苗光合参数下降，光合速率下降的原因表现为随着干旱强度的增加由气孔限制原因转为非气孔限制原因，并且SOD和APX是巨龙竹在干旱胁迫下起主要作用的2种酶。巨龙竹实生苗在轻度干旱环境中表现出良好的适应性，但在中度和重度干旱环境中其适应性较差。     

菲胁迫下蒿柳抗氧化系统的响应

[目的]研究菲胁迫下活性氧和抗氧化物质的变化规律,探究蒿柳抗氧化系统的防御机制,为提高其对多环芳烃(PAHs)的抗性及加强植物修复的研究提供理论依据。[方法]以蒿柳扦插苗为试验材料,采用水培方式,研究其在0、1.0 mg·L^-1菲处理下活性氧、抗氧化酶、抗氧化剂以及丙二醛(MDA)的动态变化,处理时间为16 d。[结果]研究表明:(1)菲处理后第4天,H2O2含量和氧自由基(O2^·-)生成速率迅速增加,MDA含量升高,过氧化氢酶(CAT)活性显著上升;第8天超氧化物歧化酶(SOD)和过氧化物酶(POD)活性显著升高;第16天时,H2O2含量下降到与对照无显著差异,O2^·-和MDA的增加量下降。(2)还原型谷胱甘肽(GSH)和谷胱甘肽还原酶(GR)在处理后第4天即迅速上升,谷胱甘肽-S-转移酶(GST)呈缓慢上升趋势。(3)还原型抗坏血酸(AsA)含量在处理后第4天低于对照,但随着处理时间的延长呈上升趋势,在第16天时高于对照。[结论]菲胁迫下,O2^·-是造成细胞膜脂过氧化的主要活性氧,SOD活性一直高于对照,但不足以清除增加的O2^·-,CAT和POD的升高可以清除过量的H2O2;GSH是抵御菲胁迫的有效抗氧化剂,并通过GST的催化参与菲的解毒。     

Na2SO4盐胁迫对紫穗槐酶活性的影响

对紫穗槐幼苗进行不同浓度的Na2SO4处理,测定并分析了其幼苗叶中超氧化物歧化酶（SOD）、过氧化氢酶（CAT）及过氧化物酶（POD）活性的变化。结果表明：1）300 mmol/L的盐浓度是适宜紫穗槐生长的临界土壤盐浓度。2）紫穗槐幼苗体内活性氧的保护酶体系主要途径是由SOD催化超氧自由基,再由POD和CAT共同协作来清除SOD催化超氧自由基的产物。     

巨龙竹实生苗对干旱胁迫的形态及生理响应

以在云南孟连县(源生地)和昆明市(引种地)培育的2年生巨龙竹(Dendrocalamus sinicus)实生苗为对象,研究了干旱胁迫对其生理特性的影响,并采用隶属函数法对两地培育的竹苗抗旱性进行综合评价。结果表明：随着干旱胁迫的增强,孟连苗和昆明苗的叶绿素含量逐渐下降,叶片相对含水量均呈先升高后下降的变化趋势;2种竹苗的细胞膜透性、可溶性蛋白和MDA含量均随着干旱胁迫的增强而升高,说明干旱胁迫对膜系统造成了损害;在干旱胁迫下,2种竹苗的SOD、CAT、APX酶活性均有显著变化,且在不同的胁迫水平下有显著差异(P>0.05)。采用隶属函数法对2种竹苗的抗旱能力进行综合评定显示,昆明苗的抗旱性优于孟连苗。     

盐胁迫对‘南林895’杨组培再生芽生根及抗氧化能力的影响

以‘南林895’杨经组培获得的2 cm长再生芽为试验材料,研究不同浓度NaCl处理下再生芽不定根发生能力及其耐盐能力。结果表明:‘南林895’杨再生芽在75 mmol/L以下能够正常产生不定根,100 mmol/L完全抑制不定根的形成。25～50 mmol/L盐处理能够促进‘南林895’杨再生芽不定根的伸长,增加根数,提高叶绿素a及总叶绿素的含量。25 mmol/L处理的SOD、CAT、POD、GR 4种保护酶与对照之间没有显著性差异,MDA含量和电导率也低于对照;而在高浓度盐处理下(75～100 mmol/L),SOD、CAT、GR保护酶活性下降,导致膜透性加大,MDA含量增加,POD和AsA-POD酶活性的提高有助于缓解盐胁迫对‘南林895’杨造成的氧化损伤。     

The role of antioxidant system in freezing acclimation-induced freezing resistance of Populus suaveolens cuttings

We investigated the changes in the contents of H2O2, malonaldehyde (MDA) and endogenous antioxidants, the activities of protective enzymes and some critical enzymes involved in the ascorbate-glutathione (ASA-GSH) cycle as well as freezing resistance (expressed as LT50) and correlations mentioned above, in detail using Populus suaveolens cuttings. The purpose was to explore the physiological mechanism of the enhancement of freezing resistance induced by freezing acclimation at –20°C, and to elucidate the physiological mechanisms by which trees adapt to freezing. The results showed that freezing acclimation enhanced the activities of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), monodehydroascorbate reductase (MDAR), ascorbate peroxidase (APX), dehydroascorbate reductase (DHAR) and glutathione reductase (GR). And it increased the contents of reduced ascorbate (ASA), reduced glutathione (GSH), dehydroascorbate (DHA) and oxidized glutathione (GSSG). However, H2O2 and MDA contents and LT50 of cuttings were decreased. LT50 in cuttings was found to be closely correlated to the levels of SOD, POD, CAT, APX, DHAR, MDAR, GR, H2O2, MDA, ASA, GSH, DHA and GSSG during freezing acclimation. This suggested that the enhancement of freezing resistance of cuttings induced by freezing acclimation may relate to the distinct increase for the levels of SOD, POD, CAT, APX, DHAR, MDAR, GR, ASA, GSH, DHA, and GSSG. In addition, the observed levels of APX, DHAR, MDAR, GR, ASA, DHA, GSH and GSSG were higher than those of SOD, POD and CAT during freezing acclimation. It indicated that a higher capacity of the ASA-GSH cycle is required for H2O2 detoxification, and growth and development of cuttings. Based on the obtained results, it can be concluded that the ASA-GSH cycle plays an important role in enhancement of freezing resistance of P. suaveolens cuttings during freezing acclimation.     

低温锻炼对冷胁迫下油茶幼苗光合速率与抗氧化酶活性的影响

以油茶品种"杂优闽1"为试验材料,以未经低温锻炼的油茶幼苗为对照,研究了油茶幼苗在(10±0.5)℃低温锻炼、(2±0.5)℃冷胁迫和常温恢复期间的光合速率和抗氧化酶活性的变化。结果表明:低温锻炼3 d可明显提高油茶幼苗的抗寒性;冷胁迫3 d,经低温锻炼后的油茶幼苗叶片净光合速率和相对含水量明显高于对照;且其在常温下的恢复也较对照迅速。在低温锻炼和冷胁迫期间,油茶幼苗叶片脯氨酸和可溶性糖含量明显增加,且丙二醛(MDA)含量明显低于对照;经低温锻炼的油茶幼苗叶片超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和抗坏血酸过氧化物酶(APX)活性则均明显高于未经低温锻炼的油茶幼苗,说明渗透调节物质含量增加和保护酶活性增强在低温锻炼诱导油茶幼苗的抗寒性上发挥着重要的作用。     

外源苯甲酸对平邑甜茶幼苗根系膜脂过氧化的影响

大多数果树都存在连作障碍(重茬、再植病、忌地现象)现象,其中苹果(Malus)、李(Prunus salicina)、桃(Prunus)等连作障碍更为严重.杨兴洪等(1991)调查苹果重茬地成活率只有40%～60%,美国华盛顿州10年间苹果产区因重茬每公顷损失4万美元,山东省烟台大量苹果园已经表现并将面临更大面积的苹果连作问题.     

重金属胁迫对2种地被竹抗氧化酶与脂质过氧化的影响

采用盆栽法对4种重金属(Cd2+、Pb2+、Cu2+、Zn2+)胁迫下的菲黄竹与菲白竹叶片中的超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)活性及脂质过氧化产物(MDA)进行了研究。结果表明,菲黄竹的SOD、POD、及CAT活性随着重金属浓度的增加总体趋势是先增大,之后降低,最后又增大;而菲白竹则是先升高,后降低;过氧化产物(MDA)则于酶活性的变化趋势相反。表明重金属胁迫下,2种地被竹体内的抗氧化酶活性被诱导,且菲白竹比菲黄竹有更强的耐重金属胁迫性。     

毛白杨叶片保护酶活性等对干旱胁迫的响应

通过盆栽控水试验研究了不同水分胁迫条件下毛白杨(Populus tomentosa)8个1(2)-O型无性系苗木叶片中膜脂过氧化产物丙二醛(MDA)含量、超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)活性的动态变化过程.结果表明,随干旱胁迫强度的增大,毛白杨无性系叶片的MDA含量、CAT活性呈递增...     

寄主植物对芒果壮铗普瘿蚊为害的生理效应

研究芒果壮铗普瘿蚊不同程度为害下寄主植物叶片中硝态氮、可溶性糖、可溶性蛋白质、叶绿素含量以及植物保护酶系——超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)活性的变化.将叶片受害程度划分为4个等级:1～50,51～100,101～200,＞200(每叶片虫瘿数),并以无虫瘿为害的叶片为对照组(CK).结果表明:可溶性糖与硝态氮含量在各个等级之间均没有显著的差异;可溶性蛋白和叶绿素a、叶绿素b含量各为害等级叶片均显著低于CK;而类胡萝卜素含量,高密度为害等级(101～200,＞200)显著高于其他各组及CK.植物保护酶系测定结果显示:CK组活力最低,＞200等级SOD,POD,CAT活力均高于其他各组;其中,51～100与＞200等级SOD活力显著高于其他等级;为害等级量重的(＞200)POD和CAT活力显著高于其他等级;而1～50等级显著低于其他等级.因此,人侵害虫芒果壮铗普瘿蚊的不同为害程度直接影响植物的光合作用及有关营养物质;随着为害等级的加重可导致植物保护酶系的活性增大.     

干旱胁迫对黄山紫荆生长形态和生理特性的影响

采用盆栽控水法研究黄山紫荆Cercis chingii 1年生幼苗在干旱胁迫下的形态变化和生理响应机制.结果表明,随着干旱胁迫时间的增加,黄山紫荆的叶片、嫩枝逐渐出现萎蔫、枯落;在干旱胁迫15 d时,叶片中的可溶性糖、可溶性蛋白、丙二醛(MAD)含量和抗氧化酶活性开始升高;持续干旱胁迫20 d时,抗氧化酶的活性如CAT...     

俄罗斯桃叶卫矛组织培养苗对盐胁迫的生理响应

采用盆栽方法,以俄罗斯桃叶卫矛组织培养苗为材料,用不同浓度(0,40,80,120mmol/L)的NaCl溶液进行处理,对盐胁迫下俄罗斯桃叶卫矛的形态表现和叶绿素含量、膜透性、丙二醛(MDA)、超氧化物歧化酶(SOD)活性、过氧化物酶(POD)活性等生理指标进行研究。结果表明:在盐胁迫下植物的株高生长量逐渐受到抑制;叶绿素含量随盐浓度的增加呈现下降趋势;膜透性、MDA、SOD、POD活性均随盐浓度增加而增加;40—80mmol/L的NaCl低浓度溶液胁迫后促进了俄罗斯桃叶卫矛根冠比的增加,而高浓度(120mmol/L)NaCl溶液抑制了根冠比的增加。进一步酶活指标分析表明,在盐胁迫下,植株的叶片质膜结构受到一定的伤害,但MDA含量差异不明显,而SOD和POD活性随着盐浓度升高显著上升。研究认为,俄罗斯桃叶卫矛通过提高植株的抗氧化能力来抵抗盐胁迫造成的伤害,对盐胁迫具有较强的适应能力。     

昆虫取食和挥发物诱导的杨树叶片中LOX和PAL活性变化

为了探索昆虫取食诱导的木本植物体内所产生的防御反应,以合作杨（Populus simonii×P.pyramidalis,‘Opera8277’）扦插苗为实验材料,经杨扇舟蛾（Clostera anachoreta）幼虫取食后,检测叶片中茉莉酸（jasmonate,JA）途径中的关键酶——脂氧合酶（lipoxygenase,LOX）及苯基丙酸类合成途径中的限速酶——苯丙氨酸解氨酶（phenylalanine ammonia lyase,PAL）的活性变化。结果显示,LOX和PAL的活性不仅在虫咬叶片中出现增加,在虫咬叶片上部的系统叶片中也有显著升高,表明茉莉酸途径和苯基丙酸类合成途径被激活,而且防御反应被系统性诱导。并且,与虫咬植株邻近的健康杨树叶片中LOX和PAL的增加表明,杨树间存在由昆虫取食诱导挥发物介导的信息传递。熏蒸实验也证明,茉莉酸甲酯（methyl jasmonate,MeJA）能够作为气体信号诱导合作杨植株产生防御反应。