Anti-oxidant response of Cucumis sativus L. to fungicide carbendazim

Our purpose in this research was to determine the response of anti-oxidative enzymes of cucumber (Cucumis sativus L.) when carbendazim applied as soil drench at 0, 5, 50, and 100 mg kg⁻¹. The changes in the activity of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX) enzymes were measured in roots, stems, and leaves of cucumber. The plants were sampling at cotyledon phase (14 days) and florescence phase (56 days). A strong correlation between anti-oxidant content and carbendazim concentration was observed. As concentration increased, the activities of SOD and CAT increased in roots and leaves. While in stems, SOD and CAT activities were increased in florescence phase and declined in cotyledon phase. The content of GPX increased in stems, and declined in leaves. Higher levels of SOD, CAT, and GPX were observed in cucumber cotyledons than the older leaves. The present study suggested that carbendazim treatments had different effects on cucumber anti-oxidant system in different tissues. It was concluded that cotyledons might play an important role for adaptation as the carbendazim concentration increased, and the ability of mature cucumber to maintain a balance between the formation and detoxification of activated oxygen species appeared likely to enhance. On the basis of our observations, we conclude that increased SOD, CAT and GPX activity provides plant with increased carbendazim stress tolerance.     

小麦-条锈菌互作过程中活性氧及保护酶系的变化研究

 对条锈菌与小麦不同互作体系中组织学特征、活性氧产生及其相关酶系的变化进行了研究。组织学观察表明:非亲和组合相对于亲和组合存在明显差异,表现为菌丝生长受抑,吸器母细胞和吸器形成减少,寄主细胞在接种后18h左右出现过敏性坏死。生化测定结果表明:在非亲和组合中,O₂^-的产生速率和H₂O₂的含量均高于亲和组合,且O₂^-产生速率在接种后12h达到一个峰值,H₂O₂的含量在接种后20和72h出现2个高峰。而在亲和组合中,O₂^-产生速率低于对照或与对照相似,H₂O₂的含量虽然高于对照,但却普遍低于非亲和组合;SOD在亲和组合中的活性总体上要高于非亲和组合;接种24h后,CAT在2种组合中的活性均高于对照,在接种后36和48h时,亲和组合中的CAT活性高于非亲和组合,而在接种60h后又开始低于非亲和组合;POD活性在接种24h后均明显升高,但亲和组合中POD活性增幅大;MDA在非亲和组合中于接种后72h含量明显上升。结果表明,亲和与非亲和组合中条锈菌扩展、活性氧的产生及相关酶活变化都存在明显差异,这些差异与小麦抗锈性的表达可能有密切联系。     

Histological and ultrastructural comparisons of compatible, incompatible and -β-amino- n -butyric acid-induced resistance responses of pepper stems to Phytophthora capsici

The compatible interaction of pepper stems with Phytophthora capsici showed more rapid and severe disease development than did the incompatible interaction, although pathogen penetration styles of host cells in compatible and incompatible interactions were similar to each other. Treatment with -β-amino- n -butyric acid (BABA) protected the pepper plants against P. capsici infection. Reduced hyphal growth and sporangial formation were found after P. capsici infection in BABA-induced resistant and incompatible reactions. One of the most noticeable ultrastructural features of the BABA-induced resistant reaction was the formation of electron-dense wall appositions. The thick and dense wall appositions that encased the haustoria restricted haustorial development, thus leading to limitation of further pathogen penetration into inner plant tissues. A main host response in the incompatible interaction was the occlusion of cortical cells with an amorphous material. Plugging of the intercellular spaces in the cortical cells with electron opaque material was frequently observed in the incompatible interaction, but not in the compatible interaction. Another common feature of the BABA-induced resistant and incompatible reactions was degeneration of mitochondrial structure within penetrating hyphal cytoplasm. The mitochondrial structure in the BABA-induced resistant or incompatible reactions had no distinct double membrane layer and well-shaped cristae.     

Activities of antioxidant enzymes and photosynthetic responses in tomato pre-treated by plant activators and inoculated by Xanthomonas vesicatoria

The activities of antioxidant enzymes and photosynthetic responses were investigated in tomato (Lycopersicon esculentum L. var.) pre-treated by plant activators and inoculated by Xanthomonas vesicatoria. Plants were sprayed with acibenzolar-S-methyl, ASM [Bion^® 50 WG (0.2 g l⁻¹)] and aqueous extract from dry necrotic tissue flour (VLA) of ‘Lobeira’ (Solanum lycocarpum) bush. Four days later, the plants were challenged with a virulent strain of X. vesicatoria, under greenhouse conditions. Tomato leaves were then assessed to determine the activities of the main antioxidant enzymes: superoxide dismutase (SOD), catalase (CAT), peroxidase (POX), and ascorbate peroxidase (APX). A reduction of bacterial leaf spot severity was observed in plants treated by ASM (49.3%) and VLA (31.4%), without any in vitro inhibitory activity over the pathogen. Controls showed decreases in CO₂ assimilation, transpiration, photosynthetic rates, and stomatal conductance. Water use efficiency and carboxylation efficiency were strongly affected in ASM- and VLA-treated, in comparison to controls and healthy plants. The tested substances induced increases in SOD and CAT activities in a delayed enzymatic response typical in compatible plant–pathogen interactions. Measured at daily intervals, activities of APX and POX were significantly (ρ0.05) higher in treated plants than in controls, except for APX in ASM-treated plants where no difference was found when compared to controls. Only POX was clearly induced at the earlier stages after spraying the tomato plants with ASM or VLA. Our results suggest that late increases in antioxidant enzyme activities may play a role in mitigating oxidative damage in restoring the photosynthetic imbalance imposed by the expansion of bacterial lesions.     

稻瘟菌GP66激发子诱导的水稻膜脂过氧化及其保护酶活性变化

 以抗稻瘟病水稻近等基因系C101LAC及背景品系CO39为材料,研究稻瘟菌(Magnaporthe grisea)来源的GP66激发子诱导的水稻膜脂过氧化及保护酶活性变化.结果表明,激发子诱导非亲和性互作水稻超氧阴离子(O₂^·)积累和脂氧合酶(LOX)活性在早期明显高于亲和性互作水稻;O₂^·积累和LOX活性的升高进而导致了非亲和性互作水稻的膜脂过氧化,其相对电导率及丙二醛(MDA)含量出现的高峰期和强度也明显要早和高于亲和性互作水稻.超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性均趋于下降,不同亲和性互作水稻间的变化则不明显;非亲和性互作水稻过氧化物酶(POD)活性则在激发子诱导早期明显高于亲和性互作水稻,可能与其参与其它抗性有关.这些结果表明膜脂过氧化的发生是激发子诱导水稻抗性的主要生理机制之一.     

Salicylic acid alleviates growth inhibition and oxidative stress caused by zucchini yellow mosaic virus infection in Cucurbita pepo leaves

The effects of zucchini yellow mosaic virus (ZYMV) infection and pretreatments with salicylic acid (SA) on biomass accumulation of pumpkin (Cucurbita pepo cv. Eskandarani) were investigated. The response of photosynthesis, transpiration and the activities of antioxidant enzymes in leaves was also considered. Significant reductions in growth parameters (i.e. leaf area, biomass and shoot height), photosynthesis and chlorophyll a and b content were detected in ZYMV-infected leaves in comparison to healthy controls. Antioxidant enzyme activities were increased up to 3-fold for peroxidase (POD), 2-fold for ascorbate peroxidase (APX) and catalase (CAT) activities and 1.3-fold for SOD activity by virus infection. ZYMV infection also caused increases in H₂O₂ and malondialdehyde (MDA) contents. These results suggest that ZYMV infection causes oxidative stress in pumpkin leaves leading to the development of epidemiological symptoms. Interestingly, spraying pumpkin leaves with SA led to recovery from the undesirable effects of ZYMV infection. Leaves treated with 100 μM SA three days before inoculation had the appearance of healthy leaves. No distinct disease symptoms were observed on the leaves treated with 100 μM SA followed by inoculation with ZYMV. In non-infected plants, SA application increased activities of POD and superoxide dismutase (SOD) and inhibited APX and CAT activities.In contrast, SA treatment followed by ZYMV inoculation stimulated SOD activity and inhibited activities of POD, APX and CAT. In addition, MDA displayed an inverse relation, indicating inhibition of lipid peroxidation in cells under SA treatment. It is suggested that the role of SA in inducing plant defense mechanisms against ZYMV infection might have occurred through the SA-antioxidant system. Such interference might occur through inhibition or activation of some antioxidant enzymes, reduction of lipid peroxidation and induction of H₂O₂ accumulation following SA application.     

Tolerance to a non-host isolate of Verticillium dahliae in tomato

Tolerance to Verticillium spp. is a condition in which a host plant develops few symptoms despite substantial colonization by the pathogen. In the present paper we have shown that Craigella tomatoes are tolerant to a non-host isolate of V. dahliae, Dvd E6. Symptom expression was used to quantify disease and quantitative PCR to assess the amount of fungus in the stems. The classical incompatible and compatible interactions between Craigella resistant or Craigella susceptible near isolines and V. dahliae, race 1 were used for comparative purposes. Additional experiments using cytological assessment and quantitative PCR showed that in the tolerant interactions one plant defence response, vascular coating, was deployed as effectively as in resistant plants, limiting pathogen distribution. However, a second defence response, which causes the cyclical elimination of fungus from the stem in the classical interactions either does not occur or is substantially delayed in tolerant plants. Thus, the Verticillium population remains stable and substantial throughout the studied time course.     

Photochemical damage and comparative performance of superoxide dismutase and ascorbate peroxidase in sugarcane leaves exposed to paraquat-induced oxidative stress

The physiological responses of sugarcane (Saccharum officinarum L.) to oxidative stress induced by methyl viologen (paraquat) were examined with respect to photochemical activity, chlorophyll content, lipid peroxidation and superoxide dismutase (SOD) and ascorbate peroxidase (APX) activities. Thirty-day-old sugarcane plants were sprayed with 0, 2, 4, 6 and 8 mM methyl viologen (MV). Chlorophyll fluorescence was measured after 18 h and biochemical analyses were performed after 24 and 48 h. Concentrations of MV above 2 mM caused significant damage to photosystem II (PSII) activity. Potential and effective quantum efficiency of PSII and apparent electron transport rate were greatly reduced or practically abolished. Both chlorophyll and soluble protein contents steadily decreased with MV concentrations above 2 mM after 24 h of exposure, which became more pronounced after 48 h, achieving a 3-fold decrease. Insoluble protein contents were little affected by MV. Oxidative stress induced by MV was evidenced by increases in lipid peroxidation. Specific activity of SOD increased, even after 48 h of exposure to the highest concentrations of MV, but total activity on a fresh weight basis did not change significantly. Nondenaturing PAGE assayed with H₂O₂ and KCN showed that treatment with MV did not change Cu/Zn-SOD and Mn-SOD isoform activities. In contrast, APX specific activity increased at 2 mM MV but then dropped at higher doses. Oxidative damage induced by MV was inversely related to APX activity. It is suggested that the major MV-induced oxidative damages in sugarcane leaves were related to excess H₂O₂, probably in chloroplasts, caused by an imbalance between SOD and APX activities, in which APX was a limiting step. Reduced photochemical activity allowed the early detection of the ensuing oxidative stress.     

二氧化碳浓度和温度升高对烟粉虱主要保护酶和解毒酶活性的影响

为明确烟粉虱Bemisia tabaci(Gennadius)在大气CO2浓度和温度双因子胁迫下的生理响应,以CO2浓度和温度为作用因子,研究了4种不同组合处理下烟粉虱成虫体内超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)、谷胱甘肽转移酶(GST)、乙酰胆碱酯酶(Ach E)活性的变化。结果表明:常温处理下,CO2浓度升高烟粉虱体内POD和GST活性分别增加87.6%和295%,SOD和CAT活性分别降低22.4%和28.2%;高温处理下,CO2浓度升高烟粉虱体内Ach E和GST活性分别增加103.6%和167.5%,CAT活性降低31.6%;常CO2浓度处理下,温度升高烟粉虱体内POD和SOD活性分别增加46.2%和18.2%,CAT活性降低35.8%;高CO2浓度处理下,温度升高烟粉虱体内Ach E和SOD活性分别增加75.3%和40.3%,CAT活性降低38.9%。表明CO2浓度和温度升高是导致烟粉虱体内SOD、POD、GST和Ach E活性升高的主要原因,并且SOD和POD活性变化受到CO2和温度的交互影响。烟粉虱可能通过改变体内保护酶或解毒酶的活性来适应CO2浓度和温度升高的环境。     

Plant defence reactions against fusarium wilt in chickpea induced by incompatible race 0 of Fusarium oxysporum f.sp. ciceris and nonhost isolates of F. oxysporum

Germinated seeds of 'kabuli' chickpea cv. ICCV 4 were inoculated with a conidial suspension of the incompatible race 0 of Fusarium oxysporum f.sp. ciceris (Foc) or of nonhost F. oxysporum resistance 'inducers', and 3 days later were challenged by root dip with a conidial suspension of highly virulent Foc race 5. Prior inoculation with inducers delayed the onset of symptoms and/or significantly reduced the final amount of fusarium wilt caused by race 5. However, the extent of disease suppression varied with the nature of the inducing agent; the nonhost isolates of F. oxysporum were more effective at disease suppression than the incompatible Foc race 0. Inoculation with the inducers gave rise to synthesis of maackiain and medicarpin phytoalexins in inoculated seedlings; these did not accumulate in plant tissues but were released into the inoculum suspension. Inoculation with inducers also resulted in accumulation of chitinase, β-1,3-glucanase and peroxidase activities in plant roots. These defence-related responses were induced more consistently and intensely by nonhost isolates of F. oxysporum than by incompatible Foc race 0. The phytoalexins and, to a lesser extent, the antifungal hydrolases, were also induced after challenge inoculation with Foc race 5. However, in this case the defence responses were induced in both preinduced and noninduced plants infected by the pathogen. It is concluded that the suppression of fusarium wilt in this study possibly involved an inhibitory effect on the pathogen of preinduced plant defences, rather than an increase in the expression of defence mechanisms of preinduced plants following a subsequent challenge inoculation.     

Metalaxyl-induced changes in the antioxidant metabolism of Solanum nigrum L. suspension cells

The antioxidant responses of Solanum nigrum L. cell suspension cultures to metalaxyl exposure were investigated. An increase in lipid peroxidation and hydrogen peroxide content, for both concentrations tested (20 mg L⁻¹; 40 mg L⁻¹) revealed the response of oxidative metabolism of cell suspensions to metalaxyl. Superoxide dismutase (SOD; EC 1.15.1.1), catalase (CAT; EC 1.11.1.6) and ascorbate peroxidase (APX; EC 1.11.1.11) activities increased, particularly in the highest concentration of metalaxyl used. An analysis by non-denaturing polyacrylamide gel (PAGE) followed by staining for enzyme activity, revealed seven SOD isoenzymes, two CAT isoenzymes, and nine APX isoenzymes. Metalaxyl levels were quantified in the culture medium and results suggest that suspension cells were able to accumulate and/or degrade the fungicide five hours after exposure. SOD, CAT and APX isoenzymes were differently affected by the metalaxyl treatment. Results suggest that the higher concentration of metalaxyl induced oxidative stress to cell suspension cultures of S. nigrum.     

小麦抗条锈反应中活性氧及细胞质钙离子的作用

 为探讨活性氧和细胞质钙离子在小麦抗条锈病反应中的作用,以小麦品种洛夫林13与具有不同致病力的单孢锈菌CY29、CY25的互作体系为平台,对锈菌侵染后小麦叶片中活性氧(ROS)积累、保护酶系(SOD、CAT和APX)活性变化动态、细胞质膜的透性改变及细胞质钙离子浓度变化做了研究。结果表明,不亲和锈菌CY25侵染可引起小麦叶片内2次ROS的爆发,第1次出现在接种后前期(接种后第2天),强度较小,第2次出现在接种后期(接种后第5天),强度较大;亲和锈菌CY29侵染只引起1次ROS爆发,出现在接种后期(接种后第5和第6天之间),但强度极大。过敏性坏死反应HR只出现在不亲和互作小麦叶片上前期ROS爆发之后,表明前期ROS爆发与HR的产生有关。伴随着后期小麦叶片中强度极高的ROS的爆发,叶片细胞原生质膜遭到了破坏,细胞内物质外渗,细胞不久便死亡,表明高强度的ROS爆发会导致细胞死亡。根据不同互作体系ROS爆发时期SOD、CAT和APX等保护酶的活性变化分析,不亲和互作体系前期强度较小的ROS爆发主要成分是H₂O₂,后期强度较高的ROS爆发主要成分是O₂^-_·和H₂O₂;亲和互作体系强度极高的ROS爆发主要成分是O₂^-_·和H₂O₂。由此说明H₂O₂是引起小麦抗病反应HR发生的因素,而O₂^-_·则是引起细胞死亡的因素。细胞质钙离子浓度变化研究表明,HR的发生与细胞质钙离子浓度增加相关。细胞质钙离子浓度的降低推迟了HR的发生,这说明小麦叶片细胞内细胞质钙离子浓度的增加是HR的必要条件,同时也说明Ca²⁺是植物HR的胞内第二信使参与植物抗病防卫反应。     

木霉分生孢子和厚垣孢子对黄瓜叶片抗氧化系统及枯萎病防效的影响

采用前期筛选出的对黄瓜枯萎病菌有较好拮抗作用的3株木霉菌,即哈茨木霉菌(Trichoderma harzianum)809、拟康氏木霉菌(Trichoderma pseudokoningii)886和棘孢木霉菌(Trichoderma asperellum)525,利用盆栽试验,测定了木霉菌分生孢子和厚垣孢子对黄瓜幼苗抗氧化能力及对枯萎病防效的影响。结果显示:3株木霉菌分生孢子和厚垣孢子对黄瓜枯萎病的盆栽防效均在66.81%以上,且以拟康氏木霉菌886厚垣孢子防效最高,达到81.46%;当黄瓜幼苗长至三叶一心时,与CK(即只接种病原菌)相比,经哈茨木霉菌809、拟康氏木霉菌886、棘孢木霉菌525分生孢子以及厚垣孢子处理后,黄瓜幼苗叶片相对电导率和丙二醛(MDA)含量均呈下降趋势,而保护性酶包括过氧化物酶(POD)、过氧化氢酶(CAT)、抗坏血酸过氧化物酶(APX)、超氧化物歧化酶(SOD)活性则呈上升趋势,其中以拟康氏木霉菌886厚垣孢子的变化幅度最显著;拟康氏木霉菌886厚垣孢子处理的黄瓜幼苗叶片相对电导率、MDA含量分别比CK下降了47.74%、41.40%;而叶片中的POD、CAT、APX、SOD活性则分别比CK增加了318.11%、155.36%、157.09%和300.34%。研究表明3株木霉菌分生孢子和厚垣孢子均能通过改善黄瓜幼苗叶片抗氧化能力,增加保护酶活性,提高了对黄瓜枯萎病的防治效果。     

虱螨脲亚致死浓度对小菜蛾保护酶系和解毒酶系活力的影响

为阐明虱螨脲对小菜蛾亚致死效应的生化机制,采用室内生物测定法测定了LC_(10)和LC_(25)亚致死浓度处理小菜蛾3龄幼虫1、6、12、24、48和72 h后其体内保护酶系和解毒酶系的活力变化。结果表明:虱螨脲LC_(10)处理对小菜蛾幼虫体内超氧化物歧化酶(SOD)活力表现为早期诱导、后期抑制的作用,而对过氧化物酶(POD)则表现为先抑制后诱导的作用,对过氧化氢酶(CAT)仅在72h时具有诱导作用;LC_(25)处理对SOD表现为诱导-抑制-诱导-抑制的作用,对POD具有抑制作用,而对CAT具有诱导作用。经虱螨脲亚致死浓度处理后,小菜蛾幼虫体内酯酶和多功能氧化酶活力被明显抑制,且基本表现为浓度越高抑制作用越强;而谷胱甘肽S-转移酶(GST)活力仅在处理后1 h被诱导,之后被显著抑制。表明虱螨脲进入小菜蛾幼虫体内后,初期可诱导SOD、CAT和GST活力升高,以适应外界毒害的影响;但随着时间的延长,3种解毒酶活力被抑制,使其难以代谢,从而增强了对小菜蛾的毒性。     

普通烟对烟粉虱取食及其与TYLCCNV共侵染的生理生化反应

为明确普通烟遭受烟粉虱Bemisia tabaci取食及与中国番茄黄化曲叶病毒(Tomato yellow leaf curl China virus,TYLCCNV)共侵染时的响应机制,采用常规生理生化指标测定法分析比较MED隐种烟粉虱取食及其与TYLCCNV共侵染处理对普通烟植株中营养组分、防御物质积累及主要氧化酶活性的影响。结果显示,与对照植株相比,烟粉虱取食及其与TYLCCNV共侵染均可导致普通烟植株中可溶性糖和可溶性蛋白含量明显降低,多酚含量显著升高,过氧化氢酶(CAT)活性显著升高,超氧化物歧化酶(SOD)和过氧化物酶(POD)活性呈现先升后降的变化趋势。与烟粉虱取食植株相比,烟粉虱与TYLCCNV共侵染后1 d时普通烟植株中可溶糖含量显著降低了21.05%,3、7和9 d时可溶性蛋白含量分别显著降低了27.40%、26.84%和20.90%;共侵染后的1、3、5和9 d时多酚含量分别显著降低了34.60%、14.30%、3.07%和15.19%;共侵染后的1、3、5和7 d时CAT活性分别提高了3.04倍、1.42倍、1.68倍和1.96倍;共侵染后7 d时POD活性显著降低,共侵染后9 d时SOD活性显著升高。表明普通烟可通过改变其营养组分及积累防御物质或提高氧化酶活性来防御烟粉虱取食及其与TYLCCNV的共侵染,而烟粉虱与TYLCCNV共侵染可诱导植株内产生较单独取食时更少的多酚类防御物质,以提高其在宿主植物上的适应性。     

Salicylic acid alleviates growth inhibition and oxidative stress caused by zucchini yellow mosaic virus infection in Cucurbita pepo leaves

The effects of zucchini yellow mosaic virus (ZYMV) infection and pretreatments with salicylic acid (SA) on biomass accumulation of pumpkin (Cucurbita pepo cv. Eskandarani) were investigated. The response of photosynthesis, transpiration and the activities of antioxidant enzymes in leaves was also considered. Significant reductions in growth parameters (i.e. leaf area, biomass and shoot height), photosynthesis and chlorophyll a and b content were detected in ZYMV-infected leaves in comparison to healthy controls. Antioxidant enzyme activities were increased up to 3-fold for peroxidase (POD), 2-fold for ascorbate peroxidase (APX) and catalase (CAT) activities and 1.3-fold for SOD activity by virus infection. ZYMV infection also caused increases in H₂O₂ and malondialdehyde (MDA) contents. These results suggest that ZYMV infection causes oxidative stress in pumpkin leaves leading to the development of epidemiological symptoms. Interestingly, spraying pumpkin leaves with SA led to recovery from the undesirable effects of ZYMV infection. Leaves treated with 100 μM SA three days before inoculation had the appearance of healthy leaves. No distinct disease symptoms were observed on the leaves treated with 100 μM SA followed by inoculation with ZYMV. In non-infected plants, SA application increased activities of POD and superoxide dismutase (SOD) and inhibited APX and CAT activities.In contrast, SA treatment followed by ZYMV inoculation stimulated SOD activity and inhibited activities of POD, APX and CAT. In addition, MDA displayed an inverse relation, indicating inhibition of lipid peroxidation in cells under SA treatment. It is suggested that the role of SA in inducing plant defense mechanisms against ZYMV infection might have occurred through the SA-antioxidant system. Such interference might occur through inhibition or activation of some antioxidant enzymes, reduction of lipid peroxidation and induction of H₂O₂ accumulation following SA application.