Response of microbial activity and microbial community composition in soils to long-term arsenic and cadmium exposure

Arsenic (As) and cadmium (Cd) in soils can affect soil microbial function and community composition and, therefore, may have effects on soil ecosystem functioning. The aim of our study was to assess the effects of long-term As and Cd contamination on soil microbial community composition and soil enzyme activities. We analyzed soils that have been contaminated 25 years ago and at present still show enhanced levels of either As, 18 and 39 mg kg⁻¹, or Cd, 34 and 134 mg kg⁻¹. Soil without heavy metal addition served as control. Polymerase chain reaction (PCR) followed by denaturing gradient gel electrophoresis (DGGE) showed that bacterial community composition in As and Cd contaminated soils differed from that in the control soil. The same was true for the microbial community composition assessed by analysis of respiratory quinones. Soil fungi and Proteobacteria appeared to be tolerant towards As and Cd, while other groups of bacteria were reduced. The decline in alkaline phosphatase, arylsulphatase, protease and urease activities in the As- and Cd-contaminated soils was correlated with a decrease of respiratory quinones occuring in Actinobacteria and Firmicutes. Xylanase activity was unaffected or elevated in the contaminated soils which was correlated with a higher abundance of fungal quinones, and quinones found in Proteobacteria.     

不同芥菜品种(系)对镉胁迫的响应

为了解不同芥菜品种对Cd 的响应和富集特征,为芥菜安全生产和Cd 污染土壤修复提供理论依据,通过水培镉胁迫试验,在143 个芥菜品种(系)中筛选出9 种长势较好的芥菜进行土壤盆栽试验,分析不同Cd 浓度(CK、处理1、处理2、处理3 分别为0、5、25、50 mg·kg-1)对芥菜株高、鲜重、叶绿素的影响,总结Cd 在芥菜根和地上部的分布特征。研究发现,(1)品种5 在3 个处理均未存活,其他8 个品种在处理1 均可存活,在处理2 多数品种(品种6、9 除外)可以存活,在处理3 均未存活;与CK 相比,其株高、鲜重在处理1 无显著变化,在处理2 显著下降。8 种芥菜叶绿素对Cd 的响应规律不一致,但多数品种(1、3、7、8、9)叶绿素a 有下降趋势,且处理2 与CK 相比,下降显著。(2)土壤Cd 浓度在5 mg·kg-1 以上时,8 种芥菜均可富集Cd,不能作为低Cd 吸收品种种植;其地上部Cd 富集系数均>1、转运系数均<1,Cd 的主要富集部位在根部,品种3、7、8 在处理2 时地上部Cd 含量(干重)接近100 mg·kg-1,品种2、4 在处理1、处理2 地上部Cd 提取量高于140μg·株-1。(3)8 种芥菜均不属于Cd 超积累植物,但其在土壤中Cd 浓度为5 mg·kg-1 时生物量未受影响,富集系数均>1,具备富集Cd 的能力。可选种品种2、3、4、7、8[即高9、2017442297(血经芥菜)、2017442241、澳洲清甜香油芥菜、香港甜脆竹芥]修复5 mg·kg-1 以下的Cd 污染土壤。     

Soil pH influence on cadmium uptake by tobacco in high cadmium exposure

Cadmium (Cd) is a non‐essential toxic element, which is highly accumulated by tobacco leaves and is transferred to tobacco smoke thus contributing substantially to the permitted level of Cd intake by humans. Therefore, measures to reduce Cd accumulation by tobacco are of significant importance. The purpose of the present study was to investigate the influence of soil liming on Cd uptake by tobacco plants in high Cd exposure. A pot experiment was conducted with Nicotiana tabacum L. Samsun 53 on an acid Typic Haploxeralf amended with refuse sugarbeet lime to obtain a pH range from 5.3 to 7.0 and contaminated with 20 mg kg^‐1 Cd applied as CdCl₂. Tobacco was grown in pots for three months under natural conditions, harvested in four cuts and Cd uptake by leaves was estimated. In soil samples selected, pH, Cd extracted by DTPA method, and Cd fractions such as exchangeable, organically bound, carbonate and residual were determined. The results showed that Cd uptake influenced negatively tobacco yield. Soil liming decreased Cd uptake by tobacco plants. The DTPA method was not a good indicator for Cd availability in tobacco. A very strong relationship was recorded between exchangeable Cd and total Cd uptake showing that this Cd fraction is available to tobacco plants. Soil pH was correlated strongly in a negative way with this Cd fraction suggesting that this soil factor is very important in controlling Cd accumulation by tobacco.     

不同状态绿豆根边缘细胞对Cd2+的响应

以绿豆(Phaseolus radiatus)为材料,采用琼脂悬空培养法和培养皿滤纸培养法,以不同浓度的Cd2+处理洗脱根边缘细胞和未洗脱根边缘细胞的绿豆根尖,研究了离体状态和活体状态绿豆根边缘细胞对镉毒的响应.结果表明:在Cd2+的诱导下,离体状态的绿豆根边缘细胞凋亡和分泌黏液,同时随着Cd2+浓度的升高,其存活率逐渐递减;Cd2+具有诱导根尖产生根边缘细胞的作用,根尖释放的根边缘细胞数量随着Cd2+浓度的增加先增多后减少,其中,50 μmol L-1 Cd2+处理在洗脱组和未洗脱组均呈现最大诱导效应;Cd2+对根边缘细胞的活性具有影响,随着Cd2+浓度的增加,根边缘细胞的存活率在未洗脱组逐渐降低,而洗脱组则先升高后降低.与对照相比,25 μmol L-1,200 μmol L-1,400 μmol L-1 Cd2+在各处理时间段具有显著性差异(p＜0.05);根冠果胶甲基酯酶活性(PME)随着Cd2+浓度的增加呈现上升趋势,处理24 h高浓度Cd2+均会引起根冠PME活性的显著性升高(p＜0.05).以上结果表明,受到Cd2+胁迫时,绿豆根尖可释放更多根边缘细胞,并通过离体根边缘细胞形成黏液层、凋亡等来抵御Cd2+对根尖的毒害作用.     

Protective effect of selenium in Broccoli (Brassica oleracea) plants subjected to cadmium exposure

The protective effect of selenium against the cadmium-induced oxidative effect in broccoli ( Brassica oleracea) plants was studied. Plants grown in hydroponic culture were supplied with selenium [as Se(IV)] and cadmium [as Cd(II)], individually or simultaneously. Cadmium accumulation in roots was noticeably higher than in the aerial parts of the plants, and this effect was even more acute when selenium was simultaneously added. Cadmium phytotoxicity was evidenced by an increase in the malondialdehyde (MDA) concentration in the roots and a decrease of photosynthetic pigment and tocopherol concentration in the aerial parts of the plant. The simultaneous addition of selenium alleviated cadmium-induced stress in the roots after 40 days of exposition. In the leaves, a more remarkable decrease of tocopherol and chlorophyll concentration was observed in the cadmium-enriched plants after 10 days of exposure. The results provided evidence that selenium supplementation helps the plant to minimize the cadmium oxidant effect. Tocopherol concentration in broccoli fruit of cadmium-supplied plants was not affected in comparison to control. However, the proportion of alpha-tocopherol increases with the addition of selenium. This response is important not only for the protective effect against oxidative damage in the plant but also in terms of human nutrition.     

镉积累差异水稻品种生长发育对清除NO的响应

[目的]研究一氧化氮(NO)对镉(Cd)胁迫下水稻苗期生理生化及氮代谢响应的调节作用,探讨通过清除NO提高水稻耐Cd能力的措施.[方法]以Cd高积累(TN1)和Cd低积累(春江06)品种为材料,进行了Cd胁迫(40 nmol/L)水培试验.以Cd胁迫营养液为对照,在对照基础上设置添加硝普钠(Cd+SNP)、添加NO清除...     

高镉累积水稻对镉污染农田的修复潜力

为探究高镉累积水稻品种扬稻6号和玉珍香对镉污染农田土壤的修复潜力,通过大田小区试验,测定6个不同生育期(返青、分蘖、孕穗、齐穗、蜡熟、完熟)5个部位(根、茎0～10 cm、茎10～20 cm、茎20 cm以上和谷)稻草的镉含量,开展高镉累积水稻镉累积规律、移除时间和移除高度研究。结果表明,水稻扬稻6号和玉珍香各部位镉含量随生育期的延长而增加,孕穗期增幅最大,完熟期达到最大值,在同一时期不同部位镉含量分布随株高呈递减趋势;完熟期,水稻品种扬稻6号和玉珍香的根、茎0～10 cm、茎10～20 cm、茎20 cm以上部分和谷中镉含量分别为19.3、11.8、9.4、8.1和3.9mg/kg与19.5、16.3、14.3、9.7和3.7 mg/kg,其对应的镉富集系数均大于1,对镉表现出高积累特性;稻草镉的移除含量在全育期均表现从大到小依次为整株收割、地上部全收割、离地10 cm收割、离地20 cm收割,完熟期整株移除情况下,扬稻6号稻草镉累积移除含量达1 652.11μg/株,玉珍香稻草镉累积移除含量达1 547.70μg/株;一年种植一季水稻扬稻6号和玉珍香,整株移除情况下土壤镉移除效率分别为9.1%和8.5%,地上部全移除情况下土壤镉移除效率分别为7.2%和7.1%。因此为兼顾水稻移除修复效果和可操作性,建议稻草在完熟后按地上部全收割的方式移除。研究结果可为镉污染稻田的植物修复治理提供新的思路。     

Derivatization of phytochelatins from Silene vulgaris, induced upon exposure to arsenate and cadmium: comparison of derivatization with Ellman's reagent and monobromobimane

Phytochelatins (PCs) are a family of thiol-rich peptides, with the general structure (gamma-Glu-Cys)(n)()-Gly, with n = 2-11, induced in plants upon exposure to excessive amounts of heavy metals and some metalloids, such as arsenic. Two types of PC analyses are currently used, i.e., acid extraction and separation on HPLC with either precolumn derivatization (pH 8.2) with monobromobimane (mBBr) or postcolumn derivatization (pH 7.8) with Ellman's reagent [5, 5'-dithiobis(2-nitrobenzoic acid), DTNB]. Although both methods were satisfactory for analysis of Cd-induced PCs, formation of (RS)(3)-As complexes during extraction of As-induced PCs rendered the DTNB method useless. This paper shows that precolumn derivatization with mBBr, during which the (RS)(3)-As complexes are disrupted, provides a qualitative and quantitative analysis of both Cd- and As-induced PCs. In addition, derivatization efficiencies of both methods for the oligomers with n = 2-4 (PC(2)(-)(4)) are compared. Derivatization efficiency decreased from 71.8% and 81.4% for mBBr and DTNB derivatization, respectively, for PC(2) to 27.4% and 50.2% for PC(4). This decrease is most likely due to steric hindrance. Correction of measured thiol concentration is therefore advised for better quantification of PC concentrations in plant material.     

两种基因型番茄对镉胁迫响应差异

【目的】使用两个番茄品种YSL189高镉积累和HZ903低Cd积累进行苗期水培实验,旨在揭示番茄品种间Cd耐性差异的机理,为分子育种改良番茄重金属耐性提供新的信息。【方法】本研究以前期鉴定的番茄高Cd积累品种YSL189和低Cd积累品种HZ903为材料,采用苗期人工气候箱水培的方式,分析了在Cd 50和100μmol/L胁迫下,供试品种间形态学指标、抗氧化酶活性、Cd积累量及相关基因表达的差异,同时分析了供试品种苗期对离子态Cd的吸收速率差异。【结果】1)在Cd 50和100μmol/L胁迫下,根长对Cd胁迫最敏感,株高和生物量次之;与低Cd积累品种HZ903相比,高Cd积累品种YSL189表现出生物量大、根系短、株高较高的特点。2)在50和100μmol/L Cd胁迫下,植株Cd积累量以根茎叶的趋势递减;同等胁迫下,YSL189植株根、茎、叶部的Cd积累量都显著高于HZ903。3)当培养介质中Cd浓度大于20μmol/L时,YSL189根系对离子态Cd的吸收速率显著高于HZ903;定量PCR结果显示,在50和100μmol/L Cd胁迫下,品种间Nramp2、Nramp3和ZIP的表达量差异趋势与Cd积累的差异趋势一致,即YSL189根系Cd吸收相关基因Nramp2,Nramp3和ZIP的表达量高于HZ903。4)在Cd 50和100μmol/L胁迫下,根、茎部的SOD(超氧化物歧化酶)、POD(过氧化物酶)和CAT(过氧化氢酶)活性都是YSL189高于HZ903,同时YSL189叶部POD和SOD活性也显著高于HZ903,同时,TBARS(巴比妥酸反应底物)含量所指示的根、茎部膜质过氧化伤害程度则没有品种间差异。【结论】YSL189根系对离子态Cd的高吸收速率以及Nramp2、Nramp3和ZIP基因的高表达量,可能是其获得高镉积累量的原因之一;同等胁迫条件下,YSL189植株体内高Cd浓度诱导了抗氧化酶的高活性,这说明YSL189植株对活性氧的清除能力强于HZ903。     

Spatial patterns of cadmium contents related to soil heterogeneity

To investigate Cd behavior in a heterogeneous soil, soil samples were taken along a transect in a field. Total Cd content, Cd concentration in solution, and soil properties controlling Cd sorption were measured. The scale and degree of variability of these parameters were assessed using statistical and spectral analysis. No substantial correlation was found between total Cd and Cd in solution. However, with a scaled sorption model that accounts for effects of pH and organic matter content the variability of total Cd was predicted well as a function of measured concentrations and soil properties. Apparently, total Cd contents of this soil are primarily controlled by the organic matter content and pH.     

锌镉超富集植物滇苦菜中碳酸酐酶对镉的响应

A number of higher plants are able to hyperaccumulate cadmium(Cd). However, it is unknown whether cadmium(Cd) plays a biological functional role in the carbonic anhydrase(CA) of hyperaccumulators. A hydroponic experiment was conducted to explore the potentially physiological function of Cd in CA and the accumulation and tolerance of Cd in the Zn/Cd hyperaccumulator Picris divaricata Vant. P. divaricata was exposed to nutrient solutions with six Cd concentrations(0, 5, 10, 25, 50 and 75 μmol L~(-1)). After 12 d, plants were harvested for the analysis of plant biomass, Cd concentration and CA activity. The Cd concentrations in plant increased with the increasing Cd in nutrient solution, reaching 640 and 3 100 mg kg~(-1) in shoot and root, respectively, at the 75 μmol L~(-1) Cd treatment. Meanwhile, plant growth was enhanced by the Cd treatments at 5–25 μmol L~(-1), but it was significantly inhibited when the plants were exposed to solutions with higher Cd concerntrations(50 and 75 μmol L~(-1)). Exposure to Cd significantly increased the CA activity in P. divaricata, which reached a maximum value of 21.27 U mg~(-1) proteins at the 25 μmol L~(-1)Cd treatment, and the CA activity and shoot Cd concentration were positively correlated at solutions Cd of ≤ 25 μmol L~(-1). Moreover, two protein bands appeared on the denatured gel electrophoresis of purified CA, indicating that P. divaricata may have CA isomers with their respective molecular weights at around 60 and 55 k Da, at least one of which is Cd-bound. In addition, trace amounts of Cd in purified CA significantly increased with the supplied Cd concentration in nutrient solution(5–25 μmol L~(-1)). The results suggested that Cd may play a biological role by enhancing the activities and forming the active Cd-specific CA in the hyperaccumulator P. divaricata.     

Rapid isocratic HPLC quantification of metallothionein-like proteins as biomarkers for cadmium exposure in the earthworm Eisenia andrei

Metallothioneins (MTs) are regarded as sensitive biomarkers of cadmium (Cd) exposure in a number of organisms. An isocratic high-performance liquid chromatography (HPLC) method using the fluorophore ammonium-7-fluorobenz-2-oxa-1,3-diazole-4-sulfonate (SBD-F) and UV detection was developed for the quantification of MT-like proteins (MTLPs) in earthworm. This method was developed using a rabbit MT (MT-1) standard, and optimized concentrations of reagents including EDTA, SBD-F, and tris(2-carboxyethyl)phosphine (TCEP). The fluoro-tagged MT-1 (SBD-MT) and MTLP (SBD-MTLP) co-eluted at 7.2-7.3 min retention time on HPLC chromatograms. The optimized method was applied to quantify MTLPs in earthworms exposed to increasing concentrations of Cd (0, 2.7, 26.7, and 267.5 μg g⁻¹ of wet artificial soil) for 1-4 weeks. The amounts of MTLPs increased significantly in a dose- and time-dependent manner except in controls and in earthworms exposed to the lowest concentration of Cd. Since earthworm MTLPs are temporal- and dose-responsive to Cd exposure, they can be used as biomarkers to assess the impact of Cd contamination in soils. The method appeared to be rapid, sensitive, and suitable for direct quantification of total earthworm MTs.     

重金属铜、镉对蝌蚪毒性的研究

试验研究铜、镉对蝌蚪的急性毒性和联合毒性结果表明,Cu~(2 )对蝌蚪24h、48h、96h的半致死浓度LC_(50)分别为0.201mgg/kg、0.138mg/kg和0.118mg/kg,Cd~(2 )对蝌蚪24h、48h和96h的LC_(50)分别为32.1mg/kg、23.3mg/kg和18.9mg/kg。Cu~(2 )和Cd~(2 )共存对蝌蚪24h、48h和96h联合毒性的相加指数AI分别为1.03、1.2和1.20,均为协同作用。     

Characterization of cadmium-tolerant carrot cells in response to cadmium stress

Non-tolerant (CW) and Cd-tolerant (CCdl) carrot (Daucus carota L.) cells were compared for their response to cadmium (Cd). The CCdl cells were induced by selection from the CW cells on a B5 medium containing 100 µM Cd. The CCdl cells were more tolerant to Cd than the CW cells. In an 80 µM Cd treatment, the growth rate (8 d/O d) based on the fresh weight of the CCdl cells and CW cells was 69 and 24%, respectively, compared to the controls. Cd stress led to the decrease of the total glutathione content and to the increase of the contents of non-protein thiols (t-SH, phytochelatins) and Cd in both types of cells. Under Cd stress, the contents of t-SH and Cd of the CW cells were higher than those of the CCdl cells. The CW cells in the 80 and 160 µM Cd treatments accumulated Cd with the increase in time. In contrast, the Cd content in the CCdl cells in all the treatments and in the CW cells in the 40 µM Cd treatment gradually decreased after 1 d. In an additional experiment, both types of cells pretreated with 20–80 µM Cd for 1 d were transferred to a Cd-free medium. The CCdl cells in all the pretreatments continuously excluded Cd over a 4-d period, unlike the CW cells. Therefore, a larger depletion of Cd was found in the CCdl cells, which may be partially attributed to Cd tolerance.     

镉汞胁迫对花生种子的毒害效应

采用室内水培法,分别研究了镉(Cd2+)、汞(Hg2+)单一胁迫对花生种子萌发与幼苗的主要生理生化特性的影响。结果表明,当Hg2+浓度达到5 mg/L、Cd2+浓度达到或大于20 mg/L时出苗率下降;当Hg2+浓度达到25 mg/L、Cd2+浓度大于10 mg/L时不仅显著抑制生长,而且根系变黑腐烂;不同浓度Hg2+与叶绿素a的含量呈显著负相关,而与叶绿素b的含量呈显著正相关;不同浓度Hg2+与花生叶片过氧化氢酶活性呈极显著正相关。过氧化氢酶活性和叶绿素含量都与Cd2+浓度呈极显著负相关。     

Chemical forms of cadmium in a calcareous soil treated with different levels of phosphorus and cadmium and planted to spinach

The objective of the investigation was to evaluate the effect of applied phosphorus (P) and cadmium (Cd) on Cd chemical forms determined by sequential extraction and the relationship between these forms and plant responses, i.e., dry weights, concentration, and total uptake of Cd and P in a greenhouse experiment. Treatments consisted of five levels of Cd (5, 10, 20, 40 and 80 mg kg^?1 soil as cadmium sulfate) and four levels of P (0, 15, 30 and 60 mg kg^?1 soil as monocalcium phosphate), which were added to the soil and left to equilibrate for 1 month under greenhouse conditions. Spinach seeds (Spinacea oleracea L., cv Viroflay) were sown and then grown for 8 weeks. The chemical composition in the aerial part of the pant and soil was determined. Application of Cd decreased plant dry matter and increased Cd concentration in the plant, whereas at each level of applied Cd, P increased plant dry matter and decreased plant Cd concentration. All chemical forms of Cd in soil, as determined by a fractionation method, were increased following Cd application, the highest being the carbonatic form. Phosphorous application decreased exchangeable and carbonatic forms of Cd, whereas it increased other forms.     

Response of wheat to sulfur fertilization

Abstract

Coker 747³ wheat (Triticum aestivum L.) was grown on Keo silt loam (coarse‐silty, mixed, thermic Dystric Fluventic Entrochrepts) with four S sources applied at various rates for two years. The innate S level of this soil was not adequate for optimum grain yield; therefore, additional S significantly increased grain yield and S concentration and decreased N/S ratios in wheat tissue. Minimum S concentration and N/S ratios in plant tissue for maximum yield ranged from 1.3 to 2.73 g S/kg and 9.5 to 19.2, respectively.     

小麦施用硅钙肥效应的研究

适量施用硅钙肥能促进小麦对氮、磷、钾、钙、镁等营养的吸收并能改善小麦的生物学性状,具有显著的增产效应,但施用过量则会得到相反效果.硅钙肥对小麦锰、锌的吸收有明显的抑制作用,且随用量增加而增强.小麦对硅的吸收主要发生在拔节期以后,植株中硅的含量随处理量的增加而持续上升,直至出现毒害.该毒害主要表现在分蘖能力受抑制、茎蘖数下降、植株矮小、千粒重下降,最终导致产量降低.本试验条件下,施用硅钙肥750～1500kg/hm2能促进小麦的生长发育,提高产量,与此对应的植株SiO2含量,在初花期叶片中约小于0.8%,在成熟期秸秆中的硅含量约为2.0%.     

黄花菜施钾效果研究

宿豫县是黄花菜的传统生产基地 ,近年来 ,黄花菜生产上出现了落蕾增多、病害加重、产量锐减等现象 ,其原因是土壤缺钾 ,1 999年在黄花菜田进行追施钾肥试验。1　材料与方法试验设在宿豫县大兴镇农场村 ,供试土壤为黄泛冲积物发育的潮土类两合土 ,有机质含量为1 0 9% ,全氮 0