Cu和Cd胁迫下接种外生菌根真菌对油松根际耐热蛋白固持重金属能力的影响

铜和镉是我国土壤重金属常见种类,生物固定过量重金属以降低其生理毒害是修复土壤重金属污染的有效方法之一.外生菌根能够通过分泌大量有机物质来降低土壤中重金属的生物有效性.在Cu或Cd胁迫下,外生菌根真菌接种油松幼苗根际分泌的耐热蛋白具有固持重金属的潜力.结果表明,纯培养条件下接种并没有增加耐热蛋白的分泌,但是在Cu或Cd的胁迫培养下,接种幼苗耐热蛋白的分泌量增加.不同Cu浓度处理下,接种油松幼苗总提取耐热蛋白(total thermostable protein,TTP)和易提取耐热蛋白(easily extracted thermostable protein,EETP)的分泌量为未接种油松的2.64~11.79倍;Cd处理下接种油松TTP和EETP的分泌量,分别是未接种油松的1.49~7.56倍.虽然Cu处理下无论是接种还是未接种,根际耐热蛋白和根系细胞中固持的总Cu量显著大于空白对照,但接种显著增加了根际的重金属固持量,其Cu总相对含量是未接种的1.81~2.75倍,且绝大部分重金属固持在耐热蛋白中,使得根际耐热蛋白中的Cu相对含量是根系细胞含量的4.19~43.00倍.Cd处理下也得到了相似的结论.说明接种菌根真菌促进了植物分泌耐热蛋白量,加强了根系对重金属的固持能力,从而达到减少过量重金属的生物有效性,缓解土壤重金属污染的目的.     

外生菌根对欧洲赤松苗(Pinussylvestris)Cu、Zn积累和分配的影响

通过测定２种不同施锌和施铜水平下苗木和离体菌丝中铜锌含量研究了接种外生菌根（Ｓｕｉｌｌｕｓｂｏｖｉｎｕｓ）对欧洲赤松（Ｐｉ－ｎｕｓｓｙｌｖｅｓｔｒｉｓ）苗生长和微量元素积累和分配的影响． 结果表明菌丝的侵染增加了苗木生长同时也增加了植物体内重金属的含量．然而菌根植物中的重金属大部分分布在根部在铜处理中菌根植物地下部分的铜含量是无菌根植物的２．６倍在锌处理中锌是无菌根植物的１．３ 倍． 说明菌丝侵染使植物将过量的重金属滞留在根部从而增加了植物对过量重金属的抗性．通过进一步测定培养在过量重金属中的离体菌丝的重金属含量结果显示随着外界重金属浓度的增加菌丝分泌物内重金属含量比菌丝内重金属含量增加快说明滞留在根部的重金属可能并没有进入根系而是以某种形态滞留在菌丝分泌物和菌丝内．     

Soluble protein and acid phosphatase exuded by ectomycorrhizal fungi and seedlings in response to excessive Cu and Cd

Fungi and their symbionts can alleviate heavy metal stress by exuding soluble proteins and enzymes. This study examined the role of soluble protein and acid phosphatase (APase) exuded by Xerocomus chrysenteron, an ectomycorrhizal fungus, and the seedlings of its symbiont, Chinese pine (Pinus tabulaeformis), under conditions of excessive Cu and Cd. The growth type showed that this poorly studied ectomycorrhizal fungus was capable of tolerating high concentrations of Cu, and may be useful in phytoremediation. X. chrysenteron grew well at 80 mg/L Cu, and the EC50 for Cd was 17.82 mg/L. X. chrysenteron also showed enhanced exudation of soluble protein in both isolated and inoculated cultivations under the influence of Cu and Cd. Soluble protein exudation, however, di ered under Cu and Cd stress in isolates. In mediums containing Cu, soluble protein exudation increased with concentration, but in mediums containing Cd the content of soluble protein increased to a comparable level at all concentrations. This study demonstrated that soluble protein was related to heavy metal tolerance, although the di erent ions played di erent roles. While APase activity in exudates of fungi and seedlings decreased under Cu and Cd stress in comparison to the control, the APase activity in seedlings was maintained by inoculation. Thus, X. chrysenteron facilitated the ability of plant to maintain a normal nutrient uptake, and therefore to protect it from heavy metal toxicity.     

水稻幼苗对纳米氧化铜的吸收及根系形态生理特征响应

以纳米氧化铜nano-CuO(10, 100mg/L)为研究对象,以微米氧化铜micron-CuO(10, 100mg/L)及铜离子Cu2+(1.4, 2.3mg/L)为对照,通过水培实验,探讨水稻对nano-CuO的吸收积累及其根系形态和生理特征响应.结果表明,各浓度(10, 100mg/L)nano-CuO处理条件下,水稻根部及地上部铜含量(根351~1444mg/kg dw;地上部9~45mg/kg dw)总体高于micron-CuO处理(根248~817mg/kg dw;地上部1.57~1.60mg/kg dw)及Cu2+处理(根147~220mg/kg dw;地上部14~26mg/kg dw),且在水稻幼苗根及茎透射电镜图片中均观察到nano-CuO的存在,指示水稻可通过纳米颗粒的形式吸收、转运nano-CuO. Nano-CuO在水稻根细胞中主要存在于核内体中,指示内吞作用是其进入根细胞的主要方式. Nano-CuO对水稻幼苗根系有较强的毒性作用,对各根系形态指标的抑制率为28%~74%,其中总根长、根体积和比表面积为最敏感;nano-CuO的吸收累积及纳米效应是其水稻根系毒性的主要原因.低浓度nano-CuO (10mg/L)胁迫时,水稻幼苗根系活力显著提高,总吸收面积和活跃吸收面积无显著差异;高浓度nano-CuO (100mg/L)胁迫时,水稻幼苗根系活力和总吸收面积显著降低.     

Effects of arbuscular mycorrhizal fungi inoculation on arsenic accumulation by tobacco (Nicotiana tabacum L.)

A pot experiment was conducted to study the e ects of arbuscular mycorrhizal (AM) fungi (from contaminated or uncontaminated soils) on arsenic (As) uptake of tobacco (Nicotiana tabacum L.) in As-contaminated soil. Mycorrhizal colonization rate, dry weight, As and P uptake by plants, concentrations of water-extractable As and As fractions were determined. A low mycorrhizal colonization rate (< 25%) was detected. Our research indicated that AM fungi isolated from polluted soils were no more e ective than those from unpolluted soils when grown in symbiosis with tobacco. No significant di erences were observed in roots and stalks dry weights among all treatments. Leaves and total plant dry weights were much higher in Glomus versiforme treatment than that in control treatment. As contents in roots and stalks from mycorrhizal treatments were much lower than that from control treatment. Total plant As content exhibited the same trend. P concentrations in tobacco were not a ected by colonization, nor were stalks, leaves and total plant P contents. Roots P contents were remarkably lower in HN treatments than in other treatments. Meanwhile, decreased soil pH and lower water-extractable As concentrations and higher levels of As fraction bound to well-crystallized hydrous oxides of Fe and Al were found in mycorrhizal treatments than in controls. The protective e ect of mycorrhiza against plant As uptake may be associated with changes in As solubility mediated by changing soil pH. These results indicated that under As stress, proper mechanisms employed by AM fungi can protect tobacco against As uptake. Results confirmed that AM fungi can play an important role in food quality and safety.     

水培条件下秋华柳对重金属Cd的富集特性及光合响应

采用水培试验的方法,研究了不同Cd胁迫浓度〔ρ(Cd2+)分别为0、1、5、10和20 mg/L〕下秋华柳的生长、光合耐性及其对Cd的富集特性和转移能力.结果表明:①随着Cd胁迫浓度的升高,秋华柳生长所受到的抑制程度有所增加.除20 mg/L处理组外,其余各处理组的耐性指数(TI)均大于70,表现出较高的耐性.②低Cd胁迫浓度(1 mg/L)下秋华柳具有较好的光合耐性,当Cd胁迫浓度达到20 mg/L时,秋华柳的光合系统才受到明显影响.③秋华柳对外源Cd的富集能力较强,随着Cd胁迫浓度的升高,秋华柳所积累的Cd显著增加且主要积累于根部,表现为叶<茎<根.当Cd胁迫浓度小于5 mg/L时,秋华柳根部吸收的Cd向地上部分转移的效果较好.试验显示,秋华柳对于较低Cd胁迫浓度(≤10 mg/L)具有较好的耐受性和富集能力,可用于三峡库区消落带表层土壤Cd污染严重区域进行植物修复.      

Phytotoxicity of cadmium on protein, proline and antioxidant enzyme activities in growing Arachis hypogaea L. seedlings

Phytotoxicity of cadmium on growing Arachis hypogaea L. seedlings was studied. Seeds were exposed to 25, 50, and 100 μmol/L CdCl2 concentrations, for a period of 10, 15, 20 and 25 d. The extent of damage to chlorophyll, protein, proline, nitrate and nitrite reductase, antioxidant enzyme activity in leaves and roots were evaluated after 10 d of cadmium stress. The higher concentration of cadmium （100 μmol/L） resulted （leaves and roots） total chlorophyll 91.01%, protein 79.51%, 83.61%, nitrate reductase 79.39%, 80.72% and nitrite reductase 77.07%, 75.88% activity decreased with increase in cadmium concentrations and exposure periods. Cadmium caused significant changes in the activity of antioxidative enzymes. Contrastingly Cd treated plant tissues showed an increase in proline 159.87%, 239.6%, gluthion reductase （GR） 337.72%, 306.14%, superoxide disumutase （SOD） 688.56%, 381.72%, ascorbate peroxidase （APX） 226.47%, 252.14%, peroxidase （POD） 72.19%, 60.29% and catalase （CAT） 228.96%, 214.74% as compared to control. Cadmium stress caused a significant increase in the rate of SOD activity in leaves and roots of plant species. Results show the crop A. hypogaea is highly sensitive even at very low cadmium concentrations.     

4种外生菌根真菌对滴滴涕的耐受性及生理响应

在纯培养条件下,研究了外生菌根真菌--美味牛肝菌(Boletus edulis)、铆钉菇(Gomphidius viscidus)、双色蜡蘑(Laccaria bicolor)、褐疣柄牛肝菌(Leccinum scabrum)在不同质量浓度DDT处理下的生长效应、耐受性和生理响应. 结果表明:DDT没有影响外生菌根真菌的生长模式,只是改变了其生长速度和生物量积累. 在5.0 mg/L的DDT处理下,美味牛肝菌、铆钉菇和双色蜡蘑的生长没有受到显著影响. 在25.0 mg/L的DDT处理下,美味牛肝菌的生长没有受到显著影响,显示出了较好的耐受性. 在高ρ(DDT)处理下,铆钉菇的多酚氧化酶、漆酶和过氧化物酶活性显著增强,液体培养基Ph下降幅度较大. 美味牛肝菌和铆钉菇通过不同的方式来响应DDT的胁迫,它们均具有较好地降解DDT的潜力.      

Cd、Cu单一污染对小麦体内活性氧消除系统影响

采用溶液培养法,研究不同浓度Cd、Cu单一污染对小麦SOD、POD、CAT活性及MDA含量的影响。结果表明:(1)SOD和CAT活性对Cd胁迫反应较敏感,而POD活性变化不大,叶片中有效的活性氧自由基清除系统的平衡被打破,MDA含量增加;(2)Cu胁迫下,小麦叶片中MDA含量的变化趋势与根部的相反,在Cu处理浓度为25、50mg/L时,叶片中有效的活性氧自由基清除系统的平衡未被打破,MDA含量保持在较低的水平;而根部恰好相反,该系统出现了失衡现象,MDA含量偏高。     

Woody species Rhus chinensis Mill. seedlings tolerance to Pb: Physiological and biochemical response

Screening potential plant species is a crucial consideration in phytoremediation technology. Our previous study demonstrated that Rhus chinensis Mill. seedlings had potentials for phytoremediation of Pb contaminated soil. However, its bioaccumulation and tolerance characteristics remain unclear. Seedling growth, LMWOAs secreted by roots, Pb subcellular distribution and chemical forms, and mineral elements in R. chinensis tissues were evaluated under different Pb concentrations (0, 25, 50, 100, 200 and 400?mg/L) in culture solution at 14?days after planting. R. chinensis did not show visual symptoms of Pb toxicity under lower Pb treatments; however, Pb significantly declined the growth of seedlings under higher Pb treatments. Higher Pb stress also decreased the concentrations of nitrogen in leaves, but increased the concentrations of P and K in roots. Pb stress also decreased Mn concentrations in leaves. A great quantity of Pb was uptake and mostly retained in R. chinensis roots. Nonetheless, R. chinensis can still concentrate 459.3 and 1102.7?mg/kg Pb in leaves and stems, respectively. Most of Pb in R. chinensis tissues was stored in the cell wall with HAc-, HCl-, and NaCl-extractable form. LMWOAs secreted by R. chinensis roots showed a strong positive correlation with Pb concentrations in all plant tissues and with P in roots. Our results suggested that Pb deposited in the cell wall and integration with phosphate or oxalate might be responsible for the tolerance of R. chinensis under Pb stress in short period.     

Ability of Agrogyron elongatum to accumulate the single metal of cadmium, copper, nickel and lead and root exudation of organic acids

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施用沼渣和接种丛枝菌根真菌对甘草生长及矿质营养的影响

沼渣是厌氧发酵的残余物,可作为肥料施用,但因其含有一定量的重金属等有害物质可能导致环境污染风险.丛枝菌根(Arbuscular mycorrhiza,AM)真菌作为植物共生真菌,可以促进植物对矿质养分的吸收,同时能够通过不同途径减轻重金属对植物的毒害.本文采用甘草(Glycyrrhiza uralensis Fisch.)为供试植物开展盆栽试验,考察施用沼渣结合接种AM真菌对甘草生长和矿质营养的影响.试验结果表明,施用沼渣显著促进了植物生长,提高了植物生物量、磷含量和叶片叶绿素含量,与此同时提高了土壤有机质和磷、铬、铜、铅含量,并导致植物重金属含量显著升高.另一方面,AM真菌能够和甘草根系形成良好共生关系,但施用沼渣对菌根侵染表现出显著抑制作用.接种AM真菌促进了甘草生长、提高了根系磷含量及叶片叶绿素含量,同时显著降低了植株重金属含量至安全阈值以内.本试验表明,施用沼渣同时接种AM真菌可在促进甘草生长的同时阻控重金属污染风险,因而可作为沼渣安全利用的一种可行技术途径.     

Effect of inoculation with arbuscular mycorrhizal fungi on the degradation of DEHP in soil

The effect of inoculation with arbuscular mycorrhizal(AM) fungi( Acaulospora lavis) on the degradation of di(2-ethylhexyl) phthalate(DEHP) in soil was studies. Cowpea plants( Pigna sinensis) were used as host plants and grown in a specially designed rhizobox. The experimental results indicated that, both in sterile and non-sterile soil,mycorrhizal colonization rates were much higher in the mycorrhizal plants than in the non-mycorrhizal plants. Addition of 4 mg/kg DEHP slightly affected mycorrhizal colonization, but the addition of 100 mg/kg DEHP significantly decreased mycorrhizal colonization. DEHP degradation in the mycorrhizosphere (Ms) and hyphosphere (Hs),especially in the Hs, increased after inoculation with Acaulospora lavis. It is concluded that mycorrhizal hyphae play an important role in the plant uptake, degradation and translocation of DEHP. The mechanism might be attributed to increased numbers of bacteria and actinomycetes and activity of dehydrogenase, urease and acid phosphatase in the Ms and Hs by mycorrhizal fungi.     

丛枝菌根真菌对铜尾矿上植物生长和矿质营养的影响

通过温室盆栽试验研究了白三叶草(Trifolium repens Linn)和黑麦草(Lolium perenne L.)在单独和混合种植情况下单独或混合接种丛枝菌根真菌Glomus mosseae和Glomus versiforme对铜尾矿砂中植物生长和矿质营养的影响.试验结果表明,不同接种处理下均以单独种植白三叶草菌根侵染率较高,平均为25%.在单种和混种情况下,接种处理均显著提高了白三叶草地上部的干物重,但对黑麦草生长影响不明显同时,接种处理显著提高了三叶草植株中的磷含量,同时降低了植株铜含量,但对黑麦草植株体内元素含量影响不明显.两种植物混合种植情况下黑麦草显示了相对竞争优势,而菌根共生体能够提高三叶草的竞争能力.试验初步证明丛枝菌根真菌对于豆科植物适应铜尾矿复合逆境(养分贫瘠和重金属污染),以及在尾矿上重建具有物种多样性的植被具有潜在作用,但有必要进一步筛选耐性菌株,并验证自然条件下菌根真菌的作用潜力.     

纳米二氧化钛缓解镉胁迫下小麦幼苗生长及生理变化

研究了水培环境中不同浓度纳米二氧化钛（TiO₂-NPs）（0,25,50,100,200 mg/L）缓解Cd对小麦幼苗造成的生理毒害的作用。结果显示:在水培环境下TiO₂-NPs的施用,缓解了Cd对小麦带来的氧化胁迫,减轻了Cd对小麦的毒害作用,显著提高了小麦生物量、根长及株高,改善了小麦光合作用。中浓度（50,100 mg/L）TiO₂-NPs的使用显著提高了小麦幼苗净光合速率。对于所有浓度TiO₂-NPs的施用,小麦幼苗POD酶活均显著低于Cd处理组;SOD酶活在较低浓度（<200 mg/L）时显著低于Cd处理组。TiO₂-NPs通过减轻Cd氧化胁迫作用,有效缓解了Cd对小麦幼苗的毒害,提高了小麦幼苗的光合作用。     

Combined effects of cadmium and fluoranthene on germination, growth and photosynthesis of soybean seedlings

The single and combinational effects of cadmium (Cd) and fluoranthene (FLT) on germination, growth and photosynthesis of soybean seedlings were investigated. Exposure to 5, 10, or 15 mg Cd/L or 1, 5, or 10 mg FLT/L individually or in combination significantly decreased germination vigor (3 days) and final germination rate of soybean seeds, except at 1 and 5 mg FLT/L. The results of two-way ANOVA analysis and the Bliss independence model showed that at lower concentrations of FLT (1 mg/L), the interaction between Cd and FLT on germination was antagonistic, whereas the interaction was synergistic when the concentration of FLT was 5 or 10 mg/L and the concentration of Cd was 15 mg/L. Growth, expressed as dry weight, length of shoot and root, leaf area, and photosynthesis, expressed as net photosynthetic rate, intercellular CO2 concentration, chlorophyll contents and fluorescence of soybean seedlings were also reduced by exposure to 5 or 10 mg Cd/L or 1 or 5 mg FLT/L, singly or jointly. Significant antagonistic effects of exposure to 5 or 10 mg Cd/L or 1 or 5 mg FLT/L on shoot growth and photosynthesis were observed, whereas synergy and antagonism of Cd and FLT were both observed for root growth.     

铜胁迫对黄芪幼苗的生理学毒性与凹凸棒黏土的缓解作用

为探究黄芪幼苗对铜离子（Cu²⁺）胁迫的耐性机理以及凹凸棒黏土对Cu²⁺污染的缓解作用,研究了不同浓度CuSO₄（2~20mmol/L）胁迫对黄芪幼苗的生理学毒性与凹凸棒黏土的缓解作用.结果表明,2mmol/L CuSO₄胁迫使得根系Cu²⁺含量、H₂O₂（过氧化氢）含量、MDA（丙二醛）含量分别较对照显著上升1.82倍、1.04倍、2.14倍.CuSO₄胁迫浓度达8mmol/L时,根系SOD（超氧化物歧化酶）活性、根尖膜损伤程度和叶片Cu²⁺含量分别较对照显著上升1.13倍、1.12倍和2.62倍;同时,叶片PS Ⅱ（光系统II）实际光化学效率[Y（II）]、光化学淬灭系数（qP）、PSⅡ电子传递速率（ETR）和叶绿素含量较对照分别显著降低22.88%、24.44%、21.49%和28.31%,而NPQ和qN（非光化学荧光猝灭系数）则较对照分别显著上升2.35倍和1.58倍.根系POD（过氧化物酶）和CAT（过氧化氢酶）活性、可溶性糖和可溶性蛋白含量在8mmol/L CuSO₄处理下达到最高值后呈下降趋势.CuSO₄浓度为15~20mmol/L时,根系APX活性和叶片光适应下PS II潜在最大光化学效率（F_v'/F_m'）,以及幼苗全株鲜重、全株干重、地下部鲜重、地下部干重较对照显著下降.在非CuSO₄胁迫条件下,基质中凹凸棒黏土的存在使得幼苗根系MDA含量较对照显著降低15.93%,但未对其它所测生理学指标产生显著影响;在20mmol/L CuSO₄胁迫条件下,基质中凹凸棒黏土的存在使得幼苗根系和叶片中Cu²⁺含量分别显著下降30.78%和23.12%;同时显著缓解了20mmol/L CuSO₄胁迫对根系活性氧水平、抗氧化酶活性、膜脂质过氧化程度、根尖膜损伤程度、可溶性蛋白和可溶性糖含量,叶片PS II光化学活性和叶绿素含量的不良影响,以及对幼苗生长的抑制作用.研究结果表明,培养基质中凹凸棒黏土的存在能够显著降低幼苗组织中Cu²⁺的生物有效性,继而缓解CuSO₄胁迫对黄芪幼苗的生理学毒性作用.     

丛枝菌根菌丝对重金属元素Zn和Cd吸收的研究

丛枝菌根菌丝对重金属元素Ｚｎ和Ｃｄ吸收的研究陶红群李晓林张俊伶（中国农业大学资源与环境学院，北京１０００９４）关键词丛枝菌根；锌；镉．近年来，由于工业排放及一些农业措施（如污灌等）造成许多土壤中重金属元素的污染．其中锌和镉是两个较为重要的重金属污染...     

Uptake and accumulation of copper by roots and shoots of maize(Zea may L.)

The effects of different concentrations of copper sulfate on rootand shoot growth of maize(Zea mays L.) and the uptake and accumulation of Cu2+ by its roots and shoots were investigated in the present study. The concentrations of opper sulfate (CuSO4.5H2O) used were in the range of 10-5-10-3mol/L. Root growth decreased progressively with increasing concentration of Cu2+ in solution. The seedlings exposed to 10-3 mol/L Cu exhibited substantial growth reduction, yielding only 68% of the root length of the control. The shoot growth of the seedlings grown at 10-5-10-4 mol/L Cu2+ were more or less than the same as the control seedlings. The leaves treated with 10-3 mol/L Cu2+ were obviously inhibited in shoot growth. The fresh and dry weights both in roots and shots decreased progressively with increasing Cu2+ concentration.This fits well with the above mentioned effects of copper sulfate on root growth. Zea mays has considerable ability to remove Cu from solutions and accumulate it. The Cu content in roots of Z. Mays increased with increasing solution concentration of Cu2+. The amount of Cu in roots of plants treated with 10-3, 10-4 and 10-5 mol/L Cu2+ were 10, 8 and 1.5 fold, respectively, greater than that of roots of control plant. However, the plants transported and concentrated only a small amount of Cu in their shoots.     

铝对马尾松幼苗光合作用的若干影响

本工作在溶液培养条件下,研究了铝对马尾松针叶叶绿素含量、叶绿素吸收光谱、光合作用和叶绿体甘油醛-3-磷酸脱氢酶活性的影响。