Cd胁迫下超富集植物圆锥南芥抗氧化机理

采用土培的方法研究了不同Cd2+添加量(0、10、20、40、80、160、240mg.kg-1)对多金属超富集植物圆锥南芥(Arabis paniculata Franch)抗氧化酶(超氧化物歧化酶SOD、过氧化氢酶POD、抗坏血酸过氧化物酶CAT、过氧化物酶APX)活性、非酶物质(巯基SH、谷胱甘肽GSH、植物络合素PCs)含量以及生物量、叶绿素、丙二醛(MDA)、半胱氨酸(Cysteine)、脯氨酸(Proline)含量的影响.结果表明,随着Cd添加量的增加,圆锥南芥的生物量、Cd含量、富集系数、转运系数呈增加趋势,SOD活性、叶绿素含量呈下降趋势.POD、CAT、APX活性以及MDA含量均呈先升高后降低的变化趋势.圆锥南芥叶片中SH、GSH、PCs含量随Cd添加量的增加而增加,当Cd添加量为240mg.kg-1时,SH、GSH、PCs含量分别比对照提高了92.7%、70.2%、143.4%,表明SH、GSH、PCs在解毒Cd毒害中起着重要作用.     

HPLC法测定镉、砷胁迫下水稻根系的植物螯合肽

利用monobromobimane衍生试剂,通过反相高效液相色谱-荧光检测体系对镉、砷胁迫下水稻根系内的植物螯合肽(PCs)等巯基化合物进行形态和含量的分析测定.结果表明,采用乙腈和0.1％三氟乙酸组成的两元梯度流动相,可以将标准样品中的谷胱甘肽(GSH)和PCs等巯基化合物很好地分离.利用上述方法,能较好地分离Cd、...     

Effects of nitrogen dioxide and its acid mist on reactive oxygen species production and antioxidant enzyme activity in Arabidopsis plants

Nitrogen dioxide (NO₂) is one of the most common and harmful air pollutants. To analyze the response of plants to NO₂ stress, we investigated the morphological change, reactive oxygen species (ROS) production and antioxidant enzyme activity in Arabidopsis thaliana (Col-0) exposed to 1.7, 4, 8.5, and 18.8 mg/m³ NO₂. The results indicate that NO₂ exposure affected plant growth and chlorophyll (Chl) content, and increased oxygen free radical (O₂⁻) production rate in Arabidopsis shoots. Furthermore, NO₂ elevated the levels of lipid peroxidation and protein oxidation, accompanied by the induction of antioxidant enzyme activities and change of ascorbate (AsA) and glutathione (GSH) contents. Following this, we mimicked nitric acid mist under experimental conditions, and confirmed the antioxidant mechanism of the plant to the stress. Our results imply that NO₂ and its acid mist caused pollution risk to plant systems. During the process, increased ROS acted as a signal to induce a defense response, and antioxidant status played an important role in plant protection against NO₂/nitric acid mist-caused oxidative damage.     

Assessing the bioremediation potential of arsenic tolerant bacterial strains in rice rhizosphere interface

The arsenic tolerant bacterial strains Staphylococcus arlettae(NBRIEAG-6), Staphylococcus sp.(NBRIEAG-8) and Brevibacillus sp.(NBRIEAG-9) were tested for their roles in enhancing plant growth and induction of stress-related enzymes in rice(Oryza sativa L. cv. NDR-359) plants at two different concentrations, 30 and 15 mg/kg of As(V) and As(III), respectively. An experiment was conducted to test the effect of these strains on plant growth promotion and arsenic uptake. We found 30%–40% reduction in total As uptake in bacteria-inoculated plants, with increased plant growth parameters compared to non-inoculated plants. Moreover, the bacteria-inoculated plants showed reduced activity of total glutathione(GSH) and glutathione reductase(GR) compared to their respective controls, which suggests the bacteria-mediated reduction of oxidative stress in plants. Thus, these strains were found to be beneficial in terms of the biochemical and physiological status of the plants under arsenic stress conditions.Furthermore, one-way ANOVA and principal component analysis(PCA) on enzymatic and non-enzymatic assays also revealed clear variations. The results support the distinction between control and treatments in both shoots and roots. Therefore, this study demonstrates the potential of rhizobacteria in alleviating arsenic stress in rice plants.     

纳米TiO2在拟南芥中的富集、转运及对其生长和生理的影响

为了探讨纳米材料对植物的生理效应,以拟南芥(Arabidopsis thaliana)作为受试生物,研究了纳米TiO_2在拟南芥中的富集、转运及对其生长和生理的影响.本研究将拟南芥分别暴露于浓度为0、0.05、0.1、0.5、1.0、1.5 mg·L-1的纳米TiO_2中,通过ICP-MS测定纳米TiO_2在根和叶中的生物积累量,并采用TEM技术观察纳米TiO_2转运至叶片后对叶绿体结构的影响.在处理0、7和14 d后,测定拟南芥生物量、根系活力、叶片相对电导率和超氧化物歧化酶(SOD)、丙二醛(MDA)、谷胱甘肽(GSH)、活性氧自由基(ROS)等生长生理指标的变化.结果显示:拟南芥能吸收纳米TiO_2并向地上部转运,但转运系数较低;TEM切片观察到纳米TiO_2能进入拟南芥体内并造成叶绿体内囊体中质体小球数目增多、体积变大.与对照组相比,幼苗地上部生物量、叶片相对电导率、叶肉原生质体活力呈下降的趋势,地下部生物量及根系活力呈上升趋势;纳米TiO_2还可引起叶片及根部活性氧(ROS)含量的变化,进而导致MDA、SOD、GSH等抗氧化体系酶活性的应激变化,同时与根部相比,叶片引起的毒性效应更强.说明纳米TiO_2在拟南芥中富集并转运至地上部后,对拟南芥叶片的生长具有显著的抑制作用,诱导其产生氧化应激.同时,一定浓度的纳米TiO_2胁迫下,拟南芥根部与叶片对纳米TiO_2的响应不同,敏感度存在差异,叶生长受到抑制的同时却对根有促进作用.     

Effect of nitrogen/phosphorus concentration on algal organic matter generation of the diatom Nitzschia palea: Total indicators and spectroscopic characterization

Critical algal blooms in great lakes increase the level of algal organic matters(AOMs),significantly altering the composition of natural organic matters(NOMs) in freshwater of lake.This study examined the AOM's characteristics of Nitzschia palea(N.palea),one kind of the predominant diatom and an important biomarker of water quality in the great lakes of China,to investigate the effect of AOMs on the variation of NOMs in lakes and the process of algal energy.Excitation–emission matrix fluorescence(EEM) spectroscopy,synchronous fluorescence(SF) spectroscopy and deconvolution UV–vis(D-UV) spectroscopy were utilized to characterize AOMs to study the effects of nutrient loading on the composition change of AOMs.From results,it was revealed that the phosphorus is the limiting factor for N.palea's growth and the generation of both total organic carbon and amino acids but the nitrogen is more important for the generation of carbohydrates and proteins.EEM spectra revealed differences in the composition of extracellular organic matter and intracellular organic matter.Regardless of the nitrogen and phosphorus concentrations,aromatic proteins and soluble microbial products were the main components,but the nitrogen concentration had a significant impact on their composition.The SF spectra were used to study the AOMs for the first time and identified that the protein-like substances were the major component of AOMs,creating as a result of aromatic group condensation.The D-UV spectra showed carboxylic acid and esters were the main functional groups in the EOMs,with –OCH_3,–SO_2NH_2,–CN,–NH_2,–O– and –COCH_3functional groups substituting into benzene rings.     

EDTA和BR对黑麦草铅积累与耐性的调控作用

通过盆栽试验研究了乙二胺四乙酸(EDTA)与植物生长调节剂芸苔素内酯(BR)单施或配施对Pb胁迫条件下黑麦草生长、生理特性及铅富集与转运特征的影响.结果表明,叶面喷施0.05~0.1 mg·L~(-1)的BR提高了黑麦草地上部Pb含量与积累量、植株干重、叶绿素a、叶绿素b、酸溶性巯基(SH)和络合素(PCs)含量.单施2.5 mmol·kg-1EDTA对黑麦草生长没有显著影响,但显著提高了植株Pb含量和积累量;EDTA与BR配施处理中,随着BR浓度的升高,地上部Pb含量与积累量、转移系数和富集系数随之提高,生物量则没有显著变化;叶面喷施0.1 mg·L~(-1)的BR配合根施2.5 mmol·kg~(-1)的EDTA,可使地上部Pb积累量提高8.79倍.BR单施在提高Pb积累的同时,通过提高SH和PCs含量来提高其对Pb的耐性;BR与EDTA配施处理则在进一步提高黑麦草对Pb的富集与转移能力的同时,通过提高As A含量来提高其对Pb胁迫的耐性.     

Arsenic methylation by an arsenite S-adenosylmethionine methyltransferase from Spirulina platensis

Arsenic-contaminated water is a serious hazard for human health. Plankton plays a critical role in the fate and toxicity of arsenic in water by accumulation and biotransformation.Spirulina platensis(S. platensis), a typical plankton, is often used as a supplement or feed for pharmacy and aquiculture, and may introduce arsenic into the food chain, resulting in a risk to human health. However, there are few studies about how S. platensis biotransforms arsenic. In this study, we investigated arsenic biotransformation by S. platensis. When exposed to arsenite(As(Ⅲ)), S. platensis accumulated arsenic up to 4.1 mg/kg dry weight.After exposure to As(Ⅲ), arsenate(As(Ⅴ)) was the predominant species making up 64% to86% of the total arsenic. Monomethylarsenate(MMA(Ⅴ)) and dimethylarsenate(DMA(Ⅴ))were also detected. An arsenite S-adenosylmethionine methyltransferase from S. platensis(Sp Ars M) was identified and characterized. Sp Ars M showed low identity with other reported Ars M enzymes. The Escherichia coli AW3110 bearing Spars M gene resulted in As(Ⅲ) methylation and conferring resistance to As(Ⅲ). The in vitro assay showed that Sp Ars M exhibited As(Ⅲ) methylation activity. DMA(Ⅴ) and a small amount of MMA(Ⅴ) were detected in the reaction system within 0.5 hr. A truncated Sp Ars M derivative lacking the last 34 residues still had the ability to methylate As(Ⅲ). The three single mutants of Sp Ars M(C59S, C186 S, and C238S) abolished the capability of As(Ⅲ) methylation, suggesting the three cysteine residues are involved in catalysis. We propose that Sp Ars M is responsible for As methylation and detoxification of As(Ⅲ) and may contribute to As biogeochemistry.     

Quantitative determination of AI-2 quorum-sensing signal of bacteria using high performance liquid chromatography–tandem mass spectrometry

Autoinducer 2(AI-2), an important bioactive by-product of the Lux S-catalyzed S-ribosylhomocysteine cleavage reaction in the activated-methyl-cycle, has been suggested to serve as a universal intra- and inter-species signaling molecule. The development of reliable and sensitive methods for quantitative determination of AI-2 is highly desired.However, the chemical properties of AI-2 cause difficulty in its quantitative analysis.Herein, we report a high performance liquid chromatography-tandem mass spectrometric method that enables reproducible and sensitive measurement of AI-2 concentrations in complex matrixes. 4,5-Dimethylbenzene-1,2-diamine(DMBDM), an easy-to-obtain commercial reagent, was used for the derivatization treatment. The assay was linear in the concentration range of 1.0–1000 ng/m L(R~2= 0.999) and had a lower limit of quantification of0.58 ng/m L. The method exhibited several advantages, e.g., high selectivity, wide linear response range, and good sensitivity. Furthermore, the effectiveness of the method was further validated through measuring AI-2 concentrations in the cell-free culture supernatant from Escherichia coli wild type.     

镉对不同生长期籽粒苋植物螯合肽的影响

植物螯合肽（PCs）由于其含有大量的巯基,能够螯合重金属离子,对重金属的积累和解毒起着重要作用.以镉（Cd）富集型植物籽粒苋K472为研究对象,采用土培法,设置6个Cd处理水平为：0（CK）、10（T1）、25（T2）、50（T3）、100（T4）和200 mg·kg^-1（T5）,分析Cd胁迫下营养生长初期、中期和后期的K472植物螯合肽的变化特征,探究植物螯合肽对Cd的螯合解毒机制.结果表明,各生长阶段的K472的株高、根长、生物量随着Cd胁迫含量的升高而下降显著,其降幅随着K472生长发育逐渐降低.而营养生长中期的K472对Cd的富集能力最强,最大Cd富集量为6695.35 mg,最大富集系数为6.3.此外,随着Cd胁迫含量的增加,不同生长阶段的K472根、茎和叶内Cd含量与PCs含量呈显著正相关.其中,根系及茎部中对Cd胁迫响应最强的植物螯合肽为PC3,叶片中对Cd胁迫响应最强的植物螯合肽为PC2.实际应用中,在营养生长中期采收籽粒苋K472可获得更高的修复效率.     

A combined process coupling phytoremediation and in situ flushing for removal of arsenic in contaminated soil

Phytoremediation and soil washing are both potentially useful for remediating arsenic(As)-contaminated soils.We evaluated the effectiveness of a combined process coupling phytoremediation and in situ soil flushing for removal of As in contaminated soil through a pilot study.The results showed that growing Pteris vittata L.(P.v.) accompanied by soil flushing of phosphate(P.v./Flushing treatment) could significantly decrease the total As concentration of soil over a 37 day flushing period compared with the single flushing(Flushing treatment).The P.v./Flushing treatment removed 54.04% of soil As from contaminated soil compared to 47.16% in Flushing treatment,suggesting that the growth of P.vittata was beneficial for promoting the removal efficiency.We analyzed the As fractionation in soil and As concentration in soil solution to reveal the mechanism behind this combined process.Results showed that comparing with the control treatment,the percent of labile arsenate fraction significantly increased by 17% under P.v./Flushing treatment.As concentration in soil solution remained a high lever during the middle and later periods(51.26–56.22 mg/L),which was significantly higher than the Flushing treatment.Although soil flushing of phosphate for more than a month,P.vittata still had good accumulation and transfer capacity of As of the soil.The results of the research revealed that combination of phytoremediation and in situ soil flushing is available to remediate As-contaminated soils.     

Effects of EDTA and plant growth-promoting rhizobacteria on plant growth and heavy metal uptake of hyperaccumulator Sedum alfredii Hance

Phytoremediation is a cost-effective and environment-friendly strategy for decontaminating heavy-metal-contaminated soil. However, the practical use of phytoremediation is constrained by the low biomass of plants and low bioavailability of heavy metals in soil. A pot experiment was conducted to investigate the effects of the metal chelator ethylenediaminetetraacetic acid (EDTA) and EDTA in combination with plant growth-promoting rhizobacteria (Burkholderia sp. D54 or Burkholderia sp. D416) on the growth and metal uptake of the hyperaccumulator Sedum alfredii Hance. According to the results, EDTA application decreased shoot and root biomass by 50% and 43%, respectively. The soil respiration and Cd, Pb, Zn uptake were depressed, while the photosynthetic rate, glutathione and phytochelatin (PC) contents were increased by EDTA application. Interestingly, Burkholderia sp. D54 and Burkholderia sp. D416 inoculation significantly relieved the inhibitory effects of EDTA on plant growth and soil respiration. Compared with the control, EDTA + D416 treatment increased the Cd concentration in shoots and decreased the Pb concentration in shoots and roots, but did not change the Zn concentration in S. alfredii plants. Furthermore, EDTA, EDTA + D54 and EDTA + D416 application increased the cysteine and PC contents in S. alfredii (p < 0.05); among all tested PCs, the most abundant species was PC2, and compared with the control, the PC2 content was increased by 371.0%, 1158.6% and 815.6%, respectively. These results will provide some insights into the practical use of EDTA and PGPR in the phytoremediation of heavy-metal-contaminated soil by S. alfredii.     

Syntrophic co-culture of Bacillus subtilis and Klebsiella pneumonia for degradation of kraft lignin discharged from rayon grade pulp industry

In order to search the degradability of kraft lignin, the potential bacterial strains Bacillus subtilis (GU193980) and Klebsiella pneumoniae (GU193981) were isolated, screened and applied in axenic and co-culture conditions. Results revealed that mixed culture showed better decolorization efficiency (80%) and reduction of pollution parameters (COD 73% and BOD 62%) than axenic culture. This indicated syntrophic growth of these two bacteria rather than any antagonistic effect. The HPLC analysis of degraded samples of kraft lignin has shown the reduction in peak area compared to control, suggesting that decrease in color intensity might be largely attributed to the degradation of lignin by isolated bacteria. Further, the GC–MS analysis showed that most of the compounds detected in control were diminished after bacterial treatment. Further, the seed germination test using Phaseolus aureus has supported the detoxification of bacterial decolorized kraft lignin for environmental safety. All these observations have revealed that the developed bacterial co-culture was capable for the effective degradation and decolorization of lignin containing rayon grade pulp mill wastewater for environmental safety.     

硫酸盐对两种硒形态处理下小麦硒吸收和转运的影响

为了揭示硫酸盐处理下小麦对亚硒酸盐和硒酸盐的吸收与转运规律,通过溶液培养,研究了不同浓度硫硒交互下小麦各部位的硒含量变化.结果表明,与无硫处理相比,0.1 mmol·L-1硫使小麦对硒酸盐的吸收潜力(Vmax值)和亲和力(1/Km值)分别降低了25.7%和90.8%,硫酸盐主要是通过降低小麦根系对硒酸盐的亲和力来缩小其与亚硒酸盐的吸收差异.小麦硒转移系数(TF值)随亚硒酸盐浓度的升高显著下降,降幅最高达35.3%,硫酸盐对其无显著影响;TF值随溶液中硒酸盐浓度的升高显著上升,最高增幅达53.8%,硫酸盐对其有显著促进作用.经亚硒酸盐处理后,无论施硫与否,小麦根部累积的硒均会随培养时间的延长显著向地上部运输;而经硒酸盐处理后,小麦根部累积的硒只在硫酸盐存在下才会随培养时间的延长显著运往地上部.富硒地区施用硫肥既可防止土壤硒的过度消耗,又可促进硒向可食部位的转移.     

NR途径参与调节SO2胁迫下拟南芥含硫抗氧化物水平

以拟南芥为材料,研究了硝酸还原酶(NR)途径在植株响应SO_2过程中对含硫抗氧化物水平的影响.结果发现,野生型拟南芥植株中谷胱甘肽过氧化物酶(GPX)和谷胱甘肽硫转移酶(GST)活性高于NR缺失突变体(nia1nia2);SO_2暴露后,野生型和突变体nia1nia2植株中半胱氨酸(Cys)和谷胱甘肽(GSH)含量显著增加,GPX和GST的活性诱导性增高,且野生型植株中上述4项指标的增幅高于突变体;拟南芥逆境响应基因GSTU24和GPX7在野生型和突变体nia1nia2中差异表达,且对SO_2的响应程度不同.研究表明,植株体内NR途径与拟南芥细胞中含硫抗氧化物水平有关,NR缺失影响逆境应答中含硫抗氧化物水平的提高及抗逆基因转录,说明NR途径参与调节了植株逆境响应中含硫抗氧化物的合成及相关氧化还原反应与解毒过程.     

Photocatalytic decomposition of acrylonitrile with N–F codoped TiO2/SiO2 under simulant solar light irradiation

The solid acid catalyst, N–F codoped TiO₂/SiO₂ composite oxide was prepared by a sol–gel method using NH₄F as nitrogen and fluorine source. The prepared materials were characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), X-ray photoelectron spectroscopy (XPS), UV–Visible diffuse reflectance spectroscopy (UV–Vis), ammonia adsorption and temperature-programmed desorption (NH₃-TPD), in situ Fourier transform infrared spectroscopy (FT-IR) and N₂ physical adsorption isotherm. The photocatalytic activity of the catalyst for acrylonitrile degradation was investigated under simulant solar irradiation. The results showed that strong Lewis and Brønsted acid sites appear on the surface of the sample after N–F doping. Systematic investigation showed that the highest photocatalytic activity for acrylonitrile degradation was obtained for samples calcined at 450°C with molar ratio (NH₄F to Ti) of 0.8. The degradation ratio of 71.5% was achieved with the prepared catalyst after 6-min irradiation, demonstrating the effectiveness of photocatalytic degradation of acrylonitrile with N–F codoped TiO₂/SiO₂ composite oxide. The photocatalyst is promising for application under solar light irradiation. Moreover, the intermediates generated after irradiation were verified by gas chromatography–mass spectrometry (GC–MS) analysis and UV–Vis spectroscopy to be simple organic acids with lower toxicity, and the degradation pathway was also proposed for acrylonitrile degradation with the prepared catalyst.     

“Arabidobrassica”: Plant-genome engineering by protoplast fusion

After fusion of somatic cells (isolated protoplasts) of Arabidopsis thaliana and Brassica campestris, plants could be regenerated in which genetic material from parents of taxonomically different tribes is combined. Between these regenerants asymmetric hybrids have also been obtained, in which one parental genome is represented by reduced chromosome numbers. Results concerning the analysis and genesis of these plants are used to discuss the question whether such monster plants are of any importance.     

TBT对大鼠肝脏ROS、抗氧化酶和解毒系统酶的影响

 研究了三丁基锡(TBT)对大鼠肝脏ROS、抗氧化酶和解毒系统酶的影响.SD大鼠随机分为6组,TBT染毒剂量各组分别为,0,0.1,0.3,1.0,3.0,10.0mg/kg(B·W),连续灌胃7d,第8d处死,取肝脏检测活性氧(ROS)水平、谷胱甘肽转硫酶(GST)活性、过氧化氢酶(CAT)活性、谷胱甘肽还原酶(GR)活性和谷胱甘肽(GSH)含量.结果表明,ROS随TBT浓度增加而升高,各浓度组均有显著性差异.在0.3,1.0mg/kg时,GST活性升高且具有显著性差异,在3.0,10.0mg/kg有下降趋势;GR活性也升高且具有显著性差异.CAT活性及GSH含量在各浓度组均无明显变化.研究结果提示,染毒各组ROS水平明显升高,而与ROS去除有直接关系的GSH水平和CAT活性没有发生相关的变化,因此TBT在代谢转化过程中产生的自由基不能及时被清除可能是导致ROS水平明显升高的重要原因.