Cloning and Differential Gene Expression of Two Catalases in Suaeda salsa in Response to Salt Stress

Two different cDNA clones (Sscat1 and Sscat2) encoding catalase, the primary important H2O2-scavenging enzyme, were isolated from a AZap-cDNA library constructed from a 400 mmol/L NaCl-treated library of Suaeda salsa ( L.) Pall aerial tissue. Sscat1 (1.7 kb) contains a full open reading frame of 492 amino acids and Sscat2 (1.1 kb) is a partial clone. BLAST analysis indicates that the two clones share 71.9% identity in nucleotide sequence and 75% identity in deduced amino acid sequence within the last 287 amino acid residues of Sscat1. Southern blotting analysis showed that Sscat1 is multicopy in S. salsa genome, while Sscat2 is a single copy gene. Northern blotting analysis showed a rapid increase in the steady-level of both genes in roots after 48 It salt treatment, but only Sscat1 was induced in salinity treated leaves. Time-course analysis carried out in leaves confirmed that Sscat1 was induced by salt stress, in contrast to Sscat2. These implied that the expression of Sscat1 and Sscat2 genes are differentially regulated in S. salsa. The activity of total catalase is dramatically increased in response to salt stress.

A viral movement protein targets host catalases for 26S proteasome-mediated degradation to facilitate viral infection and aphid transmission in wheat

The infection of host plants by many different viruses causes reactive oxygen species(Ros)accumulation and yellowing symptoms,but the mechanisms through which plant viruses counteract RoS-mediated immunity to facilitate infection and symptom development have not been fully elucidated.Most plant viruses are transmitted by insect vectors in the field,but the molecular mechanisms underlying virus-host-insect interactions are unclear.In this study,we investigated the interactions among wheat,barley yellow dwarf virus(BYDV),and its aphid vector and found that the BYDV movement protein(MP)interacts with both wheat catalases(CATs)and the 26S proteasomeubiquitin receptor non-ATPase regulatorysubunit2homolog(PSMD2)to facilitate the 26S proteasome-mediateddegradation of CATs,promotingviral infection,disease symptom development,and aphid transmission.Overexpression of the BYDV MP gene in wheat enhanced the degradation of CATs,which leading to increased accumulation of ROS and thereby enhanced viral infection.Interestingly,transgenic wheat lines overexpressing BYDV MP showed significantly reduced proliferation of wingless aphids and an increased number of winged aphids.Consistent with this observation,silencing of CAT genes also enhanced viral accumulation and reduced the proliferation of wingless aphids but increased the occurrence of winged aphids.In contrast,transgenic wheat plants overexpressing TaCAT1 exhibited the opposite changes and showed increases in grain size and weight upon infection with BYDV.Biochemical assays demonstrated that BYDV MP interacts with PSMD2 and promotes 26S proteasome-mediated degradation of TaCAT1 likely in a ubiquitination-independent manner.Collectively,our study reveals a molecular mechanism by which a plant virus manipulates the Ros production system of host plants to facilitate viral infection and transmission,shedding new light on the sophisticated interactions among viruses,host plants,and insect vectors.

Remote Control of Bovine Catalase Activity with Specific Frequencies of Human and Bovine Catalase with and without Bound NADP+

Remote control enzyme technology is widely used today through resonance. In this study, we showed that the use of frequencies of the catalase enzyme itself to increase enzymatic rate is successful not only in test tubes but also remotely. The present study also suggests that, under optimal temperature, the use of bovine catalase frequency (the specific frequency of that enzyme) has a superior rate promoting vibration than the human catalase frequency, and so increases very significantly the chemical rate of bovine catalase (about 120% at 40˚C). It also suggests that bovine catalase subjected to bovine and human frequencies with catalase bound NADP+ experienced more resonance weight towards NADP+ and so were more slowly reduced back to catalase bound NADPH, increasing compound II formation rate, and slowing down the catalase activity rate.

细叶百合Catalase基因的克隆及表达分析

为探究细叶百合(Lilium pumilum)Catalase(CAT)基因与耐盐碱胁迫的关系,从细叶百合鳞茎中成功克隆出LpCat基因。该基因的ORF区长度为1479 bp,编码492个氨基酸,对其进行序列比对和系统进化树分析,发现细叶百合Catalase蛋白与通江百合(L.sargentiae)、菠萝(Ananas comosus)、油棕(Elaeis guineensis)等植物的Catalase蛋白亲缘关系较近。构建了pQE-LpCat原核表达载体,以1 mol·L^(-1)IPTG为诱导剂进行LpCat蛋白的体外诱导表达与纯化。在添加50 mmol·L^(-1)NaHCO_(3)胁迫下,携带PQE-LpCat蛋白的菌液生长浓度高于对照菌株。在1 mol·L^(-1)NaHCO_(3)、2.5 mol·L^(-1)H_(2)O_(2)胁迫下过表达LpCAT基因烟草(Nicotiana plumbaginifolia)植株与野生型相比,枯萎程度相对较低。通过净光合速率、气孔导度、胞间CO_(2)和蒸腾速率,H_(2)O_(2)含量,丙二醛(MDA)等生理指标检测说明过表达LpCat基因烟草植株比野生型烟草植株更耐盐碱。

Antioxidant status in patients with bladder cancer regarding cancer stage and grade

Objective:The imbalance of antioxidants and pro-oxidants plays a crucial role in the carcinogenesis of bladder cancer(BC).This study aimed to evaluate serum antioxidant status in patients with BC and determine its potential use in the diagnosis and progression potential considerations following histopathological assessment.Methods:A cross-sectional study included 90 patients with BC,divided into Ta,T1,and T2eT4 stage subgroups,and according to cancer progression potential,into low-grade(LG)and highgrade(HG)subgroups.The control group(CG)included 30 healthy volunteers.Antioxidant status was determined using the spectrophotometric method and standard laboratory tests.Results:Serum superoxide dismutase activity was significantly higher in BC patients regarding cancer stage in comparison to the CG(p<0.001).Catalase activity was highest in T2eT4 subgroup and was significantly higher compared to the Ta(p<0.01)and T1(p<0.05)subgroups.Serum albumin level was significantly lower in the BC group compared to the CG(p<0.001).In addition,it was significantly lower in T2eT4 subgroup compared to T1 and Ta subgroups(p<0.01).A significant negative correlation was found between tumor size and serum albumin level only(r=0.386,p<0.01).Catalase activity was higher in HG subgroup(p=0.009),while bilirubin level was higher in LG subgroup(p=0.035).The optimal cut-off value of catalase activity in differentiating patients with LG and HG BC subgroups was 11.96 IU/L,and the specificity and sensitivity were 51.1% and 82.2%,respectively.Bilirubin level,for a calculated optimal cut-off value of 11.95 mmol/L,had a specificity of 44.1%and sensitivity of 80.0%.Conclusion:More invasive stages of BC with greater progression potential are associated with an increase in enzymatic antioxidant activity and a decrease in non-enzymatic antioxidant capacity.It may suggest a possible role of antioxidants in the prediction and monitoring of illness trajectory.

Effects of Plant Extracts and Beauveria bassiana on the Activity of Defense-Related Enzymes in Solanum lycopersicum L.during Interaction with Fusarium oxysporum f.sp.lycopersici

The objective to this work was to evaluate the enzymatic activity in the culture of Solanum lycopersicum L.infected with Fusarium oxysporum after the combined application of Beauveria bassiana and plant extracts.Solanum lycopersicum plantlets were transplanted 15 days after the emergency.Five days after transplanting,Beauveria bassiana spores were applied at a concentration of 1×10^(7)spores mL^(−1)onto soil(along with A.indica(N)and P.auritum(H)leaf extracts)where S.lycopersicum plants were planted.Eight days after transplanting,spores of F.oxysporum strain were applied at a concentration of 1×10^(6)spores mL^(−1)to soil where S.lycopersicum plants were growing.The development of S.lycopersicum plants was monitored for 114 days,whereby a randomized complete block treatment design was used,the roots of the plants were crushed with liquid nitrogen,after which UV/VIS spectrophotometry was used to determine protein concentration,as well as the activity ofβ-1,3-glucanases,peroxidases(POX,EC1.11.1.7),catalase and chitinases.The treatments combining B.bassiana and A.indica and P.auritum extracts,had a significant difference in enzymatic activity levels,thus contributing to better defense mechanisms and a greater protection to S.lycopersicum plants in the presence of F.oxysporum.The application of B.bassiana and plant extracts to the ground mitigated damage caused by F.oxysporum on S.lycopersicum plants.

Functional Analysis of the Genotypic Differences in Response of Pea (Pisum sativum L.) to Calcareous-Induced Iron Deficiency

Lime-induced iron chlorosis is a major nutritional disorder causing severe plant growth and yield reduction in the calcareous soils of Tunisia.The understanding the behavior of key metabolic functions of peas on calcareous soils,the identification of useful traits of tolerance,and the exploration of the genotypic differences in response to this constraint remain the most efficient approaches due to their coast,environmental benefits,and sustainability.For this purpose,a greenhouse experiment was conducted on three pea genotypes(Alexandra:Alex,Douce de provence:DP,and Merveille de Kelvedon:MK)cultivated on calcareous soil(Fe-deficient)and fertile soil(control).Plant growth,SPAD index,iron nutrition and distribution,photosynthesis,and antioxidant enzymes were deeply analyzed to discriminate genotypic differences.Calcareous-induced iron deficiency reduced SPAD index,plant growth,net photosynthesis,and tissue Fe content against a significant stimulation of the oxidative stress indicators,H2O2 and Malondialdehyde(MDA).Moreover,we reported a significant induction of SOD and CAT activity in shoots and roots of the Alexandra genotype.Fe use efficiency increased on calcareous soil and clearly discriminated the studied genotypes.Alexandra genotype was found to be the most tolerant to lime-induced iron chlorosis.This genotype protects its tissues against oxidative stress by stimulating enzyme activities(SOD and CAT)and develops significant efficiency of Fe uptake,translocation to shoots and use when cultivated on calcareous soil.

Targeting biosignatures of hyperglycemia and oxidative stress in diabetes comorbid depressive rats: effectiveness of hydroethanolic extract of the whole plant of Ludwigia octovalvis

Background:The prime objective of the current research was to evaluate the whole plant hydroalcoholic extract of Ludwigia octovalvis(HLO)against hyperglycemia,and oxidative stress biomarkers in rats induced with diabetes comorbid depression,diabetes comorbid depression(streptozotocin-nicotinamide+electric footshocks).Methods:2,2-Diphenyl-1-picrylhydrazyl assay of HLO versus ascorbic acid was done.Effects of 200 and 400 mg/kg body weight/day HLO doses versus 25 mg/kg body weight/day metformin was studied through insulin,glucose,superoxide dismutase,lipid peroxidation,catalase,and behavioral assessment(forced swim and open field tests).Results:IC50 values of HLO and ascorbic acid were 33.52 and 27.86μg/mL respectively.Both the HLO doses showed intended results with respect to oxidative stress biomarkers in diabetes comorbid depression rats in comparison to metformin.Open field test showed better results for HLO in diabetes comorbid depression rats.However,hypoglycemic effects,and forced swim test performance of metformin was slightly higher than the 400 mg dose,followed by the 200 mg dose of HLO.Ethyl gallate,gallic acid,β-sitosterol,and quercetin in HLO might resulted in attenuating diabetic as well as depression biomarkers.Conclusion:Inhibition of glucosidase and lipase activity,and AMP-activated protein kinase phosphorylation might be the possible biochemical changes occurred in HLO treated rats.

Stress Biomarkers in the Giant Manta Mobula birostris Associated to Tourism in the Revillagigedo National Park, Mexico

A constant increase in dive tourism over the past years in the Revillagigedo National Park, Mexico, could result in a stressful scenario for giant mantas (Mobula birostris). The purpose of this study was to determine the degree of oxidative stress in terms of changes in catalase units (CAT) and muscle glycogen concentration in this species during two periods of different tourism intensity in this protected area. A total of 21 muscle biopsies were collected in March (peak tourism) and November (lower tourism), 2019. Stress biomarkers were analysed by commercial kits from the company Cayman Chemical. Oxidative stress (catalase activity) was significantly higher during the period with lower tourism (p = 0.002), compared to the period with more tourism, suggesting the presence of the general adaptation syndrome. In males, there was a significant difference (p = 0.0005) in oxidative stress between periods of different tourism intensity, suggesting that the reproductive season may be a stressor. Morphotypes showed different oxidative stress (p = 0.031);however, the reason is unknown. No statistical differences were detected in glycogen concentrations between the tourism periods (p = 0.123), probably because this polysaccharide is not a proper indicator of chronic stress in giant mantas. Based on these findings, giant mantas may have an adequate response in terms of oxidative stress due to an increase in tourism;however the observed increase in catalase suggests that it is within the tolerance range of these organisms.

Occurrences of Yttrium in Soil and Its Potential Impacts on Paddy Rice Triticum aestivum

[Objective] The effects of yttrium nitrate （YNO3） on biomass and antioxi- dant systems of paddy rice （Yttrium （Y）; Oxidative stress; Dismutases （SOD）; Per- oxidases （POD）, Catalases （CAT）, Paddy rice （Triticum aestivum）） together with the occurrences of Y in soils were investigated to assess its ecotoxicological effects on plant. [Method]Y solutions with various concentrations were sprinkled on soil sam- ples, which were well mixed and then put into culture dishes to culture paddy rice seeds for further evaluation. [Result] The results indicated that 25-100 mg/kg Y treatments significantly increased the biomass （total weight, root weight, shoot weight and leaf weight）, chlorophyll （CHL） content and protein content of paddy rice, whereas 200-800 mg/kg Y treatments had a converse effect. Similarly, biomarker for the antioxidant systems including superoxide dismutases （SOD）, peroxidases （POD） and catalases （CAT） all exhibited similar trends in both shoots and roots of paddy rice. At the same time, the malonaldehyde （MDA） content increased at from 25 to 100 mg/kg and decreased with concentrations of Y from 100 to 800 mg/kg in both shoots and roots of paddy rice. This indicated that Y could stimulate the growth of plant at low concentration, but inhibit the growth at relatively high concen- tration. [Conclusion] The levels of Y were 641＋49, 328_＋16 and 473_＋40 mg/kg in soils collected from mining area, farmland and navel orange orchard respectively. The levels of Y in the investigated area were higher than the benefit level （100 mg/kg）, which could cause low biomass as well as low activity of SOD, POD and CAT in paddy rice. Therefore, a more careful use of Y is necessary in crop management.

Effects of Hg^(2+) on Isozymes of Peroxidase,Catalase and Superoxide Dismutase in Wheat Seedlings

[Objective] This work was aimed to explore the mechanism of Hg2+ toxicity on plants.[Method]Activities of peroxidase(POD),catalase(CAT)and superoxide dismutase(SOD)were investigated in wheat(Triticum aestivum L.)seedlings under Hg2+ stress at different concentrations.[Result]① There were no obvious effects on the growth of seedlings when the concentration of Hg2+ was lower than 0.10 mmol/L.However,toxic effects on the growth of seedling were observed when the concentration of Hg2+ was higher than 0.10 mmol/L.② Different tissues showed different resistant ability in response to Hg2+ stress.The leaves and roots of wheat seedlings were more insensitive to Hg2+ toxicity.③ CAT was more sensitive to Hg2+ stress compared to POD and SOD.[Conclusion]The toxic effect was related to the concentration of Hg2+(0.10 mmol/L).The higher concentration of Hg2+ could affect the expression of POD,CAT,and SOD isozymes in the leaves,roots of wheat seedlings and germinated seeds,which further affect the normal metabolism of membrane lipid and inhibit the growth of wheat seedlings at last.

Effect of Chemical Gametocide EXP on Membrane Lipid Peroxidation and Protective Enzyme Activity in Rapeseed (Brassica napus)

[Objective] The aim of this study was to investigate the characteristics and mechanism of chemical emasculation in rapeseed and to provide the theoretical basis for development and utilization of new chemical gametocides.[Method] The activity of peroxidase,catalase and the content of hydrogen peroxide,malondialdehyde in leaves and flower buds of Brassica napus cultivars Qinyou No.3 and L89 induced by the chemical gametocide EXP in the course of male sterility were studied.[Result] Protective enzyme activity and the content of hydrogen peroxide,malondialdehyde in rapeseed treated with EXP changed significantly,which indicated that active oxygen metabolism was abnormal.Furthermore,there was a significant difference in the reaction degree of different cultivars and organs treated by EXP.[Conclusion] There was a correlation between the disturbance of active oxygen metabolism and the male sterility induced by chemical gametocide EXP.

幽门螺杆菌Catalase基因的克隆、表达及其抗原性鉴定

目的：构建含人幽门螺杆菌（Hpylori，Hp）过氧化氢酶（catalase，KatA）编码基因的重组质粒，测定、分析其核酸序列，并在E.coli中表达，研究其抗原性。方法：应用PCR技术从HpDNA染色体中扩增KatA编码基因片段，将其T-A克隆和测序，并与GenBank公布的其他Hp菌株的基因序列比较，再将目的基因插入至融合表达载体pGEX-4T-1中进行表达，用GST亲和层析对其进行纯化。纯化产物用于对29株小鼠抗Hp-全菌单克隆抗体（mAb）的鉴定及与Hp啊感染患者血清进行Western blot。结果：KatA基因全长为1 515 bp，并在GenBank上登录（No．DQ333889），与GenBank公布的其他Hp菌株的核酸的同源性为96％～97％，表达的KatA融合蛋白的相对分子质量（Mr）为85 000，29株小鼠抗Hp全菌mAb中有4株mAb是针对KatA的，表达产物可被Hp感染患者的血清特异性识别。结论：重组KatA具有较好的抗原性，为坳检测试剂和疫苗的研究奠定了基础。

Effects of Catalase and Ascorbate Peroxidase on the Root Growth of Rice under Cadmium Stress

[Objective] The research aimed to study the correlations between catalase（CAT） and ascorbate peroxidase（ASP） and the growth and development of rice roots under cadmium stress.[Method] Taking rice variety Zhonghua No.11 as materials,the changes of rice seedlings under the treatment conditions of Cd,Cd＋CAT inhibitor,Cd＋APX inhibitor were studied.[Result] Under Cd stress,inhibition of CAT activity caused the significant inhibition on the growth of aerial parts,decreased the number of adventitious roots and lateral roots,but it can significant promote the elongation growth of adventitious roots and lateral roots.Moreover,the length of the first lateral root from root tip on the primary roots and adventitious roots was also increased than control.When APX activity was inhibited,the growth changes of rice were similar with that treated by CAT inhibitor.[Conclusion] CAT and APX may play important roles in the regulation of rice root system growth in both non-stress and Cd-stressed rice

幽门杆菌Catalase/GST融合蛋白的表达、标签切除及鉴定

旨在利用GST融合基因表达系统表达幽门螺杆菌Catalase融合蛋白,并利用凝血酶切除GST标签。将重组质粒Catalase/pGEX-4T-1转化大肠杆菌BL21(DE3)感受态中,用IPTG进行诱导表达,菌体经反复冻融、溶菌酶裂解及超声破菌后,Catalase/GST融合蛋白以部分可溶性的形式表达在上清中。采用谷胱甘肽琼脂糖树脂Glutathione Sepharose 4B对其进行纯化,得到Catalase/GST融合蛋白,再用凝血酶进行GST标签的切除,所得产物进行Western blotting鉴定。高效表达出Catalase/GST融合蛋白的相对分子质量约85 kD,凝血酶成功地切除了GST标签,Western blotting证实Catalase蛋白能被鼠抗Catalase单克隆抗体识别。

Effects of Cadmium Pollution in Soil on Physiological and Biochemical Index of Allium sativum L.

[Objective] The experiment aimed to study the effects of cadmium pollution in soil on physiological and biochemical index of Allium sativum L. and provided reference for the recovery of cadmium pollution in soil. [Method]By setting eleven Cd concentrations from 0.21 to 500 mg/kg in soil and the pot test, ecological corresponding mechanism of plant height, chlorophyll (Chl) content, catalase (CAT) activity and malondialdehyde (MDA) of Allium sativum L. was analyzed. [Result] The plant height had a strong tolerance to cadmium pollution in soil, while the total chlorophyll content and chlorophyll a content had no significant difference compared with control treatment, except Cd concentration was 500 mg/kg. The high Cd concentration would increase the damage to membrane of Allium sativum L. however with the regulation of physiological mechanism, the damage was gradually decreased.[Conclusion] Allium sativum L. had strong eco-physiological adaptability to Cd contaminated soil and it had potential for recovering Cd contaminated soil.

Responses of soil enzymes to long-term CO_2 enrichment in forest ecosystems of Changbai Mountains

A study was conducted to determine the responses of soil enzymes （invertase, polyphenol oxidase, catalase, and dehydrogenase） to long-term CO2 enrichment at the Research Station of Changbai Mountain Forest Ecosystems, Chinese Academy of Sciences （42°24＇N, 128°28＇E; 738 m in elevation） in the northeast China during 1999-2006. Three treatments of the CO2 enrichment, designed as 500 μmol·mol-1 CO2 open-top chamber （OTC）, ambient control chamber and unchambered field （approx. 370 μmol·mol^-1CO2）, were conducted with Pinus koraiensis and Pinus sylvestriformis tree species. Soil sampling was made and analyzed separately in spring, summer and autumn in 2006 after the soil enzymes were exposed to elevated CO2 concentration （500 μmol·mol^-1） for eight growing seasons. Results showed that, at elevated CO2 concentration （500 μmol·mol^-1）, the activities of invertase （except for the summer samples of P. koraiensis） presented a remarkable decline in all growing seasons, while the activities of dehydrogenase had an increase but only part of the results was remarkable; the activities of polyphenol oxidase in P. sylvestriformis rhizosphere showed a remarkable decrease; the catalase activities increased in spring, while in turn were decline in other seasons. This study also revealed that the soil enzyme activities are significantly correlated with the tree species under the CO2 enhancement.

Studies on the Anti-lipid Peroxidative Actions of the Methanolic Extract of the Root of Aegle marmelos and Its Constituents In Vivo and In Vitro

The inhibitory effect of the methanolic extract of the root of Aegle marmelos (MERA) and its constituents on the lipid peroxidation in vivo and in vitro were studied. The results suggested that MERA increased the activities of superoxide dismutase (SOD) and GSH-peroxidase in the liver cytosol of mice, but showed no significant effect on the activity of catalase, and one of its major constituents, 4-methoxy-1-methyl-2-quinolone (MMQ) increased the activity of SOD in liver tissue of mice intoxicated with FeCl2-ascorbic acid (AA)-ADP in vivo. Various constituents isolated from the root of title plant inhibited the lipid peroxidation in rat liver homogenate, which was in vitro induced by FeCl2-ascorbic acid, CCl4-NADPH, or ADP- NADPH. Of the test compounds, MMQ and its derivatives integriquinolone were similar to (-tocopherol in inhibiting MDA production in rat liver microsomes induced by Fe2+-ascorbate, CCl4-NADPH, or ADP-NADPH.

Effects of Breviscapine on the Changes in Antioxidant Enzyme Activity Induced by Cerebral Ischemia-reperfusion in Rats

\ Effects of breviscapine, the active ingredient isolated from Erigeron breviscapus (Vant) Handmazz, on the changes in antioxidant enzyme activity induced by cerebral ischemiareperfusion in rats were explored. It was found that breviscapine improved the activities of superoxide dismutase (SOD), GSHperoxidase and catalase, while decreasing the malondialdehyde (MDA) content in the brain, which was benificial in reducing the damage from cerebral ischemiareperfusion.

烟草过氧化氢酶基因CAT2克隆与表达特征分析

基于已有的植物Catalase 2基因(CAT2)序列设计烟草Nicotiana tabacum CAT2的特异性引物,通过RT-PCR扩增克隆获得Nicotiana tabacum var. NC89 CAT2全长mRNA序列,包含1479 bp完整的读码框(ORF),可编码492个氨基酸残基。遗传进化分析显示,烟草NC89 CAT2与N. benthamiana CAT2基因亲缘关系最近。利用Quantitative Real-time PCR技术分析了CAT2基因在NC89烟草中的组织特异性表达和其应对生物和非生物胁迫的表达差异。结果表明,CAT2基因在烟草NC89的根、茎、叶、花瓣、花萼、种子中均有表达,其中在花中相对表达量最高,其次为叶、茎、种子和根。生物和非生物胁迫处理烟株,其CAT2诱导表达分析显示,机械损伤、渗透压、低温和高温、干旱和感染PVY时CAT2表达量均上调。上述研究表明烟草CAT2基因可能参与了应对非生物和生物胁迫的过程。