Oxidative stress response induced in an atrazine phytoremediating plant: Physiological responses of Pennisetum glaucum to high atrazine concentrations

This research presented here, for the first time, elucidates the responses of several antioxidants in Pennisetum leaves exposed to varying concentrations of atrazine (0–200 mgkg^?1). Pennisetum has been reported to be resistant to atrazine; however, its physiological response to high concentrations (≥50 mgkg^?1) of atrazine is not well documented. The contents of reduced (AsA) and oxidized (DHA) ascorbate increased significantly with increase in atrazine concentration and exposure time; but the increase was more evident under higher (50 and 100 mgkg^?1) atrazine concentrations. Increase in atrazine concentration to 200 mgkg^?1 significantly decreased AsA, but increased DHA content, throughout the experiment. Seedlings treated with 200 mgkg^?1 atrazine showed significantly lowest reduced glutathione (GSH) content, while oxidized glutathione (GSSG) was not significantly affected, after 68 d. Seedlings treated with 100 mgkg^?1 atrazine showed increased glutathione-S-transferase (GST) activity after 48 d and 68 d, while treatment with 200 mgkg^?1 atrazine significantly increased glutathione reductase (GR) after 58 d. This result suggests that Pennisetum may tolerate lower atrazine concentrations. However, higher concentrations (≥50 mg kg^?1), which could have longer residency period in the soil, could induce more physiological damage to the plant.     

水稻施用硅肥对稻纵卷叶螟幼虫保护酶和解毒酶活性的影响

【目的】研究施硅水稻对稻纵卷叶螟Cnaphalocrocis medinalis(Guenée)幼虫保护酶(过氧化氢酶CAT、过氧化物酶POD、超氧化物歧化酶SOD)和解毒酶(谷胱甘肽-S-转移酶GST、乙酰胆碱酯酶ACh E)活性的影响,为探明硅增强水稻抗稻纵卷叶螟的机理和稻纵卷叶螟的综合治理提供依据。【方法】采用感虫水稻品种TN1,设置2种施硅水平,即0.32 g Si/kg土壤(Si+)和不施硅(Si-),测定取食水稻24、48、72、96 h后3龄幼虫体内CAT、POD、SOD、GST、ACh E活性的动态变化。【结果】幼虫取食Si+水稻前期(24~48 h),CAT、POD、SOD活性呈上升趋势,并在48 h时达到最高值;取食后期(48~96 h),CAT、POD、SOD活性下降并在96 h时显著低于Si-处理。在Si-处理中,幼虫CAT和POD活性"先降低、再升高",SOD活性呈一直增大的趋势。取食Si+水稻幼虫GST活性始终显著高于Si-处理,而ACh E活性均低于Si-水稻,并在72 h和96 h时显著低于Si-水稻。处理间幼虫存活率存在显著差异,Si+水稻上1龄和3龄幼虫的存活率显著低于Si-水稻。这些结果说明,硅处理可能使稻纵卷叶螟幼虫产生应激反应引起保护酶活性在短时间内增大,但取食一段时间后保护酶活性下降。ACh E活性受到抑制,会引起昆虫神经传导的异常反应,造成昆虫死亡。【结论】硅可能通过参与植物的生理代谢过程,诱导植物增加次生代谢物的合成和积累,引起稻纵卷叶螟保护酶和解毒酶的活性变化,可能会影响稻纵卷叶螟的存活。     

Controlling and Defence‐related Mechanisms of Bacillus Strains Against Bacterial Leaf Blight of Rice

Bacillus strains are broadly studied for their beneficial role in plant growth and biological control of plant disease and pest; however, little is known about their underlying mechanisms. In this study, we assessed the controlling and defence‐related mechanisms of three Bacillus strains including rice seed‐associated strain B. subtilis A15, rhizobacterial strains B. amyloliquefaciens D29 and B. methylotrophicus H8, all of which are against bacterial leaf blight (BLB) caused by Xanthomonas oryzae pv. oryzae. Results indicated that all three strains showed strong biofilm formation ability. The culture filtrates of each strain significantly suppressed the growth and biofilm formation of X. oryzae, while changes in bacterial cell morphology such as cell swell and severe cell wall alterations were observed through the transmission electron microscopy images. PCR analysis revealed that all three strains harbour the antimicrobial‐associated genes that are responsible for biosynthesis of bacillomycin, fengycin, iturin and surfactin. Subsequent real‐time qPCR analysis revealed the upregulated expression of fenD and srfAA genes in D29 and H8, and fenD and ituC genes in A15 during their in vitro interaction with X. oryzae. It suggests that the antibacterial mechanisms of the three strains may be at least partially associated with their ability to secrete corresponding lipopeptides. Interestingly, the applications of the three strains in greenhouse conditions were found to be effective in controlling the BLB disease, which was achieved through the activation of inducing systemic resistance resulted from the enhanced activities of defence‐related enzymes. This is the first report of demonstration of the mode of antibacterial effect of Bacillus strains against X. oryzae. Overall, data from the current study provide valuable information for biological control of BLB disease in rice.     

Tinospora cordifolia induces enzymes of carcinogen/drug metabolism and antioxidant system, and inhibits lipid peroxidation in mice

The present study is an effort to identify a potent chemopreventive agent against various diseases (including cancer) in which oxidative stress plays an important causative role. Here, we investigated the effect of a hydroalcoholic (80% ethanol: 20% distilled water) extract of aerial roots of Tinospora cordifolia (50 and 100mg/kg body wt./day for 2 weeks) on carcinogen/drug metabolizing phase-I and phase-II enzymes, antioxidant enzymes, glutathione (GSH) content, lactate dehydrogenase and lipid peroxidation in liver of 8-week-old Swiss albino mice. The modulatory effect of the extract was also examined on extrahepatic organs, i.e., lung, kidney and forestomach, for the activities of GSH S-transferase (GST), DT-diaphorase (DTD), superoxide dismutase (SOD) and catalase. Significant increases in the levels of acid-soluble sulfhydryl (-SH) and cytochrome P(450) contents, and enzyme activities of cytochrome P(450) reductase, cytochrome b(5) reductase, GST, DTD, SOD, catalase, GSH peroxidase (GPX) and GSH reductase (GR) were observed in the liver. Both treated groups showed decreased malondialdehyde (MDA) formation. In lung SOD, catalase and GST; in kidney SOD and catalase; and in forestomach SOD, DTD and GST showed significant increase at both dose levels of treatment. BHA (0.75%, w/w in diet), a pure antioxidant compound, was used as a positive control. This group showed increase in hepatic levels of GSH content, cytochrome b(5), DTD, GST, GR and catalase, whereas MDA formation was inhibited significantly. In the BHA-treated group, the lung and kidney showed increased levels of catalase, DTD and GST, whereas SOD was significantly increased in the kidney and forestomach; the latter also showed an increase in the activities of DTD and GST. The enhanced GSH level and enzyme activities involved in xenobiotic metabolism and maintaining antioxidant status of cells are suggestive of a chemopreventive efficacy of T. cordifolia against chemotoxicity, including carcinogenicity, which warrants further investigation of active principle (s) present in the extract responsible for the observed effects employing various carcinogenesis models.     

Antioxidant defense system and cadmium uptake in barley genotypes differing in cadmium tolerance

Using two barley (Hordeum vulgare) cultivars (cvs. Tokak and Hamidiye) nutrient solution experiments were conducted in order to study the genotypic variation in tolerance to Cd toxicity based on (i) development of leaf symptoms, (ii) decreases in dry matter production, (iii) Cd concentration and (iv) changes in antioxidative defense system in leaves (i.e., superoxide dismutase, ascorbate peroxidase, glutathione reductase, catalase, ascorbic acid and non-protein SH-groups). Plants were grown in nutrient solution under controlled environmental conditions, and subjected to increasing concentrations of Cd (0, 15, 30, 60 and 120 micromol/L Cd) for different time periods. Of the barley cultivars Hamidiye was particularly sensitive to Cd as judged by the severity and earlier development of Cd toxicity symptoms on leaves. Within 48 h of Cd application Hamidiye rapidly developed severe leaf Cd toxicity symptoms whereas in Tokak the leaf symptoms appeared only slightly. Hamidiye also tended to show more decrease in growth caused by Cd supply when compared to Tokak. The differences in sensitivity to Cd between Tokak and Hamidiye were not related to Cd concentrations in roots and shoots or Cd accumulation per plant. With the exception of catalase, activities of the enzymes involved in detoxification of reactive oxygen species (ROS) were markedly enhanced in Hamidiye by increasing Cd supply. By contrast, in Tokak there was either only a slight increase or no change in the activities of the antioxidative enzymes. Similarly, levels of ascorbic acid and especially non-protein SH-groups were increased in Hamidiye by Cd supply, but not affected in Tokak. The results indicate the existence of a large genotypic variation between barley cultivars for Cd tolerance. The differential Cd tolerance found in the barley cultivars was not related to uptake or accumulation of Cd in plants, indicating importance of internal mechanisms in expression of differential Cd tolerance in barley. As a response to increasing Cd supply particular increases in antioxidative mechanisms in the Cd-sensitive barley cultivar Hamidiye suggest that the high Cd sensitivity of Hamidiye is related to enhanced production and oxidative damage of ROS.     

EPR and redox properties of periplasmic nitrate reductase from Desulfovibrio desulfuricans ATCC 27774

Nitrate reductases are enzymes that catalyze the conversion of nitrate to nitrite. We report here electron paramagnetic resonance (EPR) studies in the periplasmic nitrate reductase isolated from the sulfate-reducing bacteria Desulfovibrio desulfuricans ATCC 27774. This protein, belonging to the dimethyl sulfoxide reductase family of mononuclear Mo-containing enzymes, comprises a single 80-kDa subunit and contains a Mo bis(molybdopterin guanosine dinucleotide) cofactor and a [4Fe–4S] cluster. EPR-monitored redox titrations, carried out with and without nitrate in the potential range from 200 to −500 mV, and EPR studies of the enzyme, in both catalytic and inhibited conditions, reveal distinct types of Mo(V) EPR-active species, which indicates that the Mo site presents high coordination flexibility. These studies show that nitrate modulates the redox properties of the Mo active site, but not those of the [4Fe–4S] center. The possible structures and the role in catalysis of the distinct Mo(V) species detected by EPR are discussed.Electronic Supplementary Material Supplementary material is available for this article at and is accessible for authorized users.     

干旱胁迫对坡柳等抗旱树种幼苗膜脂过氧化及保护酶活性的影响

以金沙江干热河谷主要树种坡柳、银合欢、山毛豆实生幼苗为材料,通过盆栽苗自然干旱胁迫,同时以浇水处理为对照,研究了干旱胁迫对坡柳、银合欢、山毛豆3个树种丙二醛含量、膜相对透性及保护酶活性的影响。结果表明,干旱胁迫下3个树种幼苗的细胞膜透性、MDA及SOD, POD酶活性都发生了变化,只是变化的幅度和进程不同。干旱胁迫对银合欢膜系统损伤生成的主要降解产物不是MDA;山毛豆清除活性氧毒害作用主要不是通过SOD和POD的作用;通过叶片相对保水力测定及膜透性、MDA相对含量、酶活性变化情况的分析,3个树种中坡柳耐旱性最强,其次为银合欢,山毛豆居后。     

蚯蚓纤溶酶新基因EfP-0的电子克隆及其编码区序列RT-PCR验证

利用生物信息学手段,以期获得蚯蚓纤溶酶F-Ⅰ-0组分的基因。根据从粉正蚓(Lumbricusrubellus)中分离的F-Ⅰ-0组分的N端氨基酸序列VVGGSDTTIGQYPHQL,利用DNAMAN软件通过电子克隆方法,从Lumbricidae的dbEST中获得该组分的核酸序列信息,设计特异引物,经过RT-PCR,成功地从赤子爱胜蚓(Eiseniafoetida)中克隆到一条蚯蚓纤溶酶新基因,命名为EfP-0。EfP-0基因全长678bp,编码225个氨基酸的成熟肽,属丝氨酸蛋白酶,胰蛋白酶家族,与F-Ⅰ-0组分的氨基酸组成非常接近。BLAST证明,EfP-0与已报道的蚯蚓纤溶酶基因之间的相似性均低于40%,因此为蚯蚓纤溶酶中的一个新基因,GenBank登录号为DQ836917。构建的pMAL-c2x-EfP-0重组质粒,在大肠杆菌TB1中获得融合蛋白MBP-EfP-0的可溶性表达,表达产物有酪蛋白平板溶解活性。     

黄土高原长期培肥土壤团聚体中养分和酶的分布

土壤养分和生物酶是衡量土壤肥力的指标,土壤团聚体性质则决定了土壤物理结构的好坏,探讨不同培肥措施下土壤养分和生物酶在团聚体中的分布,对合理培肥和改善土壤性状具有重要的实际意义。对连续25a的长期有机培肥定位试验地进行测定,结果表明：培肥增加了大级别土壤团聚体、特征微团聚体的数量,显著地改善了土壤结构。培肥土壤团聚体平均重量直径、特征微团聚体C／F0.01较无肥对照土壤有显著增加。长期培肥增强了土壤化学、生物性质的“微域”变异性。相同培肥措施下土壤主要养分含量和酶活性均随土壤团聚体直径的增大而减小。除厩肥处理土壤碱性磷酸酶和蔗糖酶外,其它各培肥土壤的养分含量和酶活性均在〈0．01mm级别特征微团聚体中含量高、活性大。相同级别团聚体和特征微团聚体中,养分含量和酶活性在不同培肥土壤间基本表现出施厩肥处理〉施秸秆处理〉施化肥处理〉对照的规律性。将团聚体的含量和养分含量或酶活性在团聚体中的丰度结合考虑,发现〉5mm级别团聚体中养分含量或酶活性对土壤的贡献率最大。结果充分地显示着土壤性质在不同空间尺度下、不同管理措施下变异的复杂性和多样性。     

低温胁迫下钼对冬小麦抗氧化防御系统及膜脂过氧化的影响

研究了低温（0～5℃）胁迫下钼对冬小麦抗氧化系统和膜脂过氧化的影响。结果表明,低温胁迫下施钼植株电解质渗漏率和丙二醛含量显著降低。施用钼肥提高了冬小麦叶片中抗氧化酶类如超氧化物歧化酶（SOD,EC1．15．1．1）、过氧化物酶（POX,EC1．11．1．7）和过氧化氢酶（CAT,ECl．11．1．6）的活性。低温胁迫下施钼对抗氧化酶（SOD、POX和CAT）活性提高幅度比常温下高。不论常温还是低温下,施铝均提高了冬小麦叶片中抗坏血酸和脯氨酸含量,低温胁迫下提高幅度更大。常温下缺钼和施钼处理后,叶片中类胡萝卜素含量差异不显著;低温下施钼后,冬小麦叶片类胡萝卜素含量显著增加。因此,低温胁迫下施钼植株活性氧清除能力增强、细胞膜伤害减轻可能是冬小麦抗寒力提高的原因之一。