水稻突变体对镉的吸收及其亚细胞分布和化学形态特点

Wild-type （Zhonghua 11） and mutant rice （Oryza sativa L.） plants were used to investigate the effect of cadmium （Cd） application on biomass production, to characterize the influx of Cd from roots to shoots, and to determine the form, content, and subcellular distribution of Cd in the roots, leaf sheaths, and leaves of the rice plants. Seedlings were cultivated in a nutrient solution and were treated with 0.5 mmol L^-1 of Cd^2＋ for 14 d. The sensitivity of rice plants to Cd toxicity was tested by studying the changes in biomass production and by observing the onset of toxicity symptoms in the plants. Both the wild-type and mutant rice plants developed symptoms of Cd stress. In addition, Cd application significantly （P ≤ 0.01） decreased dry matter production of roots, leaf sheaths, and leaves of both types, especially the mutant. The Cd content in roots of the mutant was significantly （P ≤0.05） higher than that of the wild-type rice. However, there was no significant difference in the Cd content of roots, leaf sheaths, and leaves between the wild-type and mutant rice. Most of the Cd was bound to the cell wall of the roots, leaf sheaths, and leaves, and the mutant had greater Cd content in cell organelles than the wild type. The uneven subcellular distribution could be responsible for the Cd sensitivity of the mutant rice. Furthermore, different chemical forms of Cd were found to occur in the roots, leaf sheaths, and leaves of both types of rice plants. Ethanol-, water-, and NaCl-extractable Cd had greater toxicity than the other forms of Cd and induced stunted growth and chlorosis in the plants. The high Cd content of the toxic forms of Cd in the cell organelles could seriously damage the cells and the metabolic processes in mutant rice plants.     

施用磷和锌对植株体中锌营养及代谢的影响

A solution culture experiment was conducted to investigate the growth,the accumulation and translocation of Zn,and the metabolic changes of 24 days old plants of corn and wheat with the varied suply of phosphorus(0,0.12,0.6 and 3.0mmol L^-1)and zinc (0.1 and 2.0umol L^-1) under controlled environmental conditions.The results showed the highest dry matter production of both corn and wheat under the moder ate combination of phosphorus(0.6mmol L^-1) and zinc(2.0 umolL^-1) as compared with other imbalance applications of phosphorus and zinc.Excessive P supply significantly inhibited the translocation of Zn from roots of corn to the aboveground part,thus decreasing the content of Zn in the shoots.Application of 3.0 mmolL^-1 P could also reduce the water-soluble Zn in plant tissues,leading to an increase in the cell plasma membrane permeability,a decrease in the dehydrogenase activity in roots and the activity of nitrate reductase in leaves,and a decline in the uptake of nitrate by plants.A similar decrease occurred in superoxide dismutase(SOD) and plasma membrane adenosine triphosphatase(ATPase)activity in Zn-deficient plants.But,with increasing P supply the activity of ATP ase in both corn and wheat increased and reached the maximum at the P-supplying level of 3.0 mmolL^-1.Similar to the effect of high P supply.no or low P(0.12mmolL^-1) supply could be detrimental to dry matter production and physiological functioning of the plants.Corn plants showed a more significant response to the imbalance supply of P and Zn than wheat plants.The possible physiological and biochemical mechanism of phosphorus-zinc antagonistic interaction in corn and wheat might be attributed to decrease in physiological availability and activation of Zn.     

铝和镉胁迫对两个大麦品种矿质营养和根系分泌物的影响

A hydroponic experiment was carried out to study the effect of aluminum （Al） and cadmium （Cd） on Al and mineral nutrient contents in plants and Al-induced organic acid exudation in two barley varieties with different Al tolerance. Al- sensitive cv. Shang 70-119 had significantly higher Al content and accumulation in plants than Al-tolerant cv. Gebeina, especially in roots, when subjected to low pH （4.0） and Al treatments （100 μmol L^-1 Al and 100 μmol L^-1 Al ＋1.0 μmol L^-1 Cd）. Cd addition increased Al content in plants exposed to Al stress. Both low pH and Al treatments caused marked reduction in Ca and Mg contents in all plant parts, P and K contents in the shoots and leaves, Fe, Zn and Mo contents in the leaves, Zn and B contents in the shoots, and Mn contents both in the roots and leaves. Moreover, changes in nutrient concentrations were greater in the plants exposed to both Al and Cd than in those exposed only to Al treatment. A dramatic enhancement of malate, citrate, and succinate was found in the plants exposed to 100 μmol L^-1 Al relative to the control, and the Al-tolerant cultivar had a considerable higher exudation of these organic acids than the Al-sensitive one, indicating that Al-induced enhancement of these organic acids is very likely to be associated with Al tolerance.     

Root cell walls and phytochelatins in low-cadmium cultivar of Brassica parachinensis

‘Lvbao-701’ is a cultivar of Chinese flowering cabbage(Brassica parachinensis) that exhibits low cadmium(Cd) accumulation and high Cd tolerance.In this study, this cultivar was compared with a high-Cd accumulating cultivar, ‘Chixin-4’, to characterize the mechanisms influencing Cd accumulation in B. parachinensis. Root cell walls were isolated by dissolving the cytoplasm with an organic solvent, and root Cd and phytochelatin(PC) contents were analyzed. In addition, a PC synthase gene fragment was cloned and expressed under Cd stress conditions. The proportions of Cd bound to root cell walls were higher in the ‘Lvbao-701’ plants(68.32%–76.80%) than in the ‘Chixin-4’ plants(35.36%–54.18%) after exposure to Cd stress. The proportions of Cd bound to root cell walls measured using cell walls isolated with a non-destructive method were higher than those obtained using a conventional method that required grinding and centrifugation. Exposure to Cd stress induced the PC production and resulted in higher PC contents in the ‘Lvbao-701’ plants than in the ‘Chixin-4’ plants. Cloning and expression analysis of a B. parachinensis PC synthase cDNA fragment indicated that PC synthase gene expression was induced by Cd and occurred mainly in the roots of both ‘Lvbao-701’ and ‘Chixin-4’ plants. However, the PC synthase gene expression level was higher in the‘Lvbao-701’ roots than in the ‘Chixin-4’ roots. Therefore, a higher abundance of Cd in the root cell walls of ‘Lvbao-701’ and up-regulated PC production in the roots are probably the main reasons why ‘Lvbao-701’ exhibits lower Cd translocation to the shoots and higher tolerance to Cd than ‘Chixin-4’.     

植物汁液化学组成及其与菜心抗性和Cd累积的关系

Sap mixtures of the xylem, phloem, and vacuoles from low and high Cd accumulator varieties of Brassica parachinensis L. H. Bailey were analyzed under Cd stress to understand the biochemical mechanisms of Cd accumulation in plants. Low Cd accumulator （‘Teqing-60＇） and high Cd accumulator （‘Chixin-2＇） plants were grown in Cd-treated soil in pots in a greenhouse. Percentage of cell wall-bound Cd was estimated, pH level and the concentrations of amino acids, organic acids, anions, and cations in both stem and root saps were determined for the calculation of Cd speciations using the computer program GEOCHEM. The results showed that ‘Teqing-60＇ had a significantly higher （P ≤ 0.05） percentage of Cd bound to cell walls in roots and a significantly higher （P ≤ 0.05） pH in the root sap. ‘Teqing-60＇ also contained a higher concentration of total amino acids in both roots and stems compared with the high Cd accumulator variety ‘Chixin- 2＇. However, between the two accumulators, for stems and for roots, there were no significant differences in non-amino organic acids. GEOCHEM calculations showed that Cd in the root sap of ‘Teqing-60＇ mainly combined with amino acids, especially alanine. Compared with ‘Chixin-2＇, in the root sap of ‘Teqing-60＇, much lower levels of Cd as free ions or bound to simple ligands were found, indicating that less ‘Teqing-60＇ is transferred to stems and leaves. Cadmium activity in the shoot sap of ‘Teqing-60＇ was much lower than that in ‘Chixin-2＇; therefore, ‘Teqing-60＇ exhibited higher Cd resistance. However, direct determination of the Cd complexes from xylem and phloem sap is needed to verify these results.     

Effects of Exogenous Nitric Oxide and 24-Epibrassinolide on the Physiological Characteristics of Peanut Seedlings Under Cadmium Stress

Cadmium(Cd) is highly toxic to plants, animals, and humans. Limited information is available on the role of nitric oxide(NO)and/or 24-epibrassinolide(EBR) in response of plants to Cd stress. In this study, a hydroponic experiment was performed to investigate the effects of NO and/or EBR on peanut plants subjected to Cd stress(200 μmol L~(-1)) with sodium nitroprusside(SNP, an exogenous NO donor)(250 μmol L~(-1)) and/or EBR(0.1 μmol L~(-1)) addition. The results showed that Cd exposure inhibited plant growth, and this stress was alleviated by exogenous NO or EBR, and especially the combination of the two. Treatment with Cd inhibited the growth of peanut seedlings, decreased chlorophyll content, and significantly increased the Cd concentration in plants. Furthermore, the concentration of reactive oxygen species(ROS) markedly increased in peanut seedlings under Cd stress, resulting in the accumulation of malondialdehyde(MDA) and proline in leaves and roots. Under Cd stress, applications of SNP, EBR, and especially the two in combination significantly reduced the translocation of Cd from roots to leaves, increased the chlorophyll content, decreased the concentrations of ROS, MDA, and proline, and significantly enhanced the activities of superoxide dismutase(SOD), peroxidase(POD), and catalase(CAT) in peanut seedlings. Exogenous NO and/or EBR also stimulated the activities of nitrate reductase(NR)and nitric oxide synthase(NOS) and increased the contents of antioxidants, such as ascorbic acid(AsA) and reduced glutathione(GSH). Furthermore, exogenous NO and/or EBR enhanced Cd accumulation in the cell wall and thus decreased Cd distribution in the organelles in the roots. The concentrations of calcium(Ca), iron(Fe), magnesium(Mg), and zinc(Zn) were also regulated by exogenous NO or EBR, and especially by the two in combination. These results indicated that SNP and EBR, alone and particularly in combination, can mitigate the negative effects of Cd stress in peanut plants.     

土壤－水分－植物体系中铅和镉的交互作用及其机制: Ⅱ. 根际中Pb-Cd的交互作用

The interaction of Pb-Cd can be observed not only in the uptake process of elements by plants and in their influence on the growth,but also in rhizosphere.The changes in extractable Cd and Pb concentrations in the rhizosphere soil of rice plants ,root exudates from wheat and wheat plant and their complexing capacity,with Pa and Cd were investigated under different Pb and Cd treatments.Results showed that the concentration of extractable Cd in the rhizosphere of rice in red soil was markedly increased by Pb-Cd interaction,It increased by 56% in the treatment with Pb and Cd added against that in the treatment with only Cd added in soil . The considerable differences in both composition and amount of root exudate from wheat and rice were found among different treatments.Pb and Cd might be complexed by root exudates ,The concentrations of free Pb and Cd in the solution were increased markedly by adding root exudate from wheat and decreased by that from rice due to Pd-Cd interaction.The distribution patterns of Pb and Cd in roots were affected by Pb-Cd interaction,which accelerated transport of Pb into internal tissue and retarded accumulation of Cd in external tissue.     

盐胁迫下两种草坪草膜脂过氧化和抗氧化酶比较

Salinity stress is a major factor limiting the growth of turfgrass irrigated with recycled wastewater. The change in lipid peroxidation in terms of malondialdehyde (MDA) content and the activities of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxide (APX) and glutathione reductase (GR) in the shoots and roots of Kentucky bluegrass and tall fescue were investigated under salinity stress. Plants were subjected to 0, 50, 100, 150 and 200 mmol L 1 NaCl for 40 d. The MDA content under salinity stress was lower in tall fescue than in Kentucky bluegrass in both shoots and roots. Activities of SOD in the shoots of both species increased with salinity stress. The activities of CAT and APX decreased in Kentucky bluegrass, but no significant difference in the activities of CAT and APX was observed in tall fescue. The activities of SOD, CAT and APX in the shoots of tall fescue were higher than those in Kentucky bluegrass. In the roots of Kentucky bluegrass, SOD and GR activities increased and CAT and APX activities decreased in comparison with the control. In the roots of tall fescue, salinity increased the activities of SOD, CAT, and APX. These results suggested that tall fescue exhibited a more effective protection mechanism and mitigated oxidative stress and lipid peroxidation by maintaining higher SOD, CAT and APX activities than Kentucky bluegrass.     

厌氧去除活性污泥中痕量有机污染物:15年实验研究

Knowledge of cellular metal homeostasis will provide a better understanding of the mechanisms involved in metal tolerance and hyperaccumulation in metal-hyperaccumulating plants. Energy dispersive X-ray spectrometry (EDS) was used to determine the localization of cadmium (Cd) in leaves of the Zn/Cd hyperaccumulator Picris divaricata which had a shoot Cd concentration of 565 mg kg 1 after 2 weeks of growth in solution culture supplying 10 μmol L-1 CdCl2 . The results indicated that Cd was distributed mainly in the trichomes, upper and lower epidermis and bundle sheath cells, with a relatively low level of Cd in mesophyll cells. Mesophyll protoplasts isolated from leaves remained viable after 24 h exposure to CdCl2 at a concentration up to 1 mmol L-1 , indicating their high tolerance to Cd. The intracellular Cd was visualized by staining with Leadmium Green dye, a cellular permeable Cd fluorescence probe. The results showed that the majority of protoplasts (> 82%) did not accumulate Cd, with only a minority (< 18%) showing Cd accumulation. In the Cd-accumulating protoplasts, Cd accumulation was depressed by the addition of Fe2+ , Mn2+ and the metabolic inhibitor carbonyl cyanide m-chlorophenylhydrazone (CCCP), but not by Ca 2+ or Zn2+ . Furthermore, the entire process of Cd uptake from external solution into the cytoplasm and subsequent sequestration into vacuoles was successfully recorded by confocal images. These results suggested that reduced cellular Cd accumulation and efficient Cd vacuolar sequestration in mesophyll cells might be responsible for cellular Cd tolerance and distribution in the leaves of P. divaricata.     

玉米、小麦细胞磷、锌营养及交互作用的研究

采用溶液培养研究玉米、小麦在不同磷、锌浓度下细胞磷、锌营养及交互作用。结果表明,随着外界磷浓度（０,０．１２,０．６,３．０ｍｍｏｌ／Ｌ）的提高,玉米、小麦根系和叶片细胞壁、细胞质和液泡中磷含量也随之增高,但细胞壁和液泡中磷的含量高于细胞质中的含量。介质中磷浓度的提高使根细胞壁中锌的含量增加,但高磷（３．０ｍｍｏｌ／Ｌ）处理使细胞质、液泡中锌的含量下降。高磷处理抑制了两种作物的锌由根系向地上部的转运,同时叶片中大部分锌被结合在细胞壁中,因而叶细胞中的细胞质和液泡处于相对的低锌状态。与足量锌（２．０ｍｏｌ／Ｌ）供应比较,低锌（０．１ｍｏｌ／Ｌ）处理使玉米、小麦根系和叶细胞壁、细胞质和液泡中磷的浓度增高。低锌和过量磷的供给抑制了植株的生长,最终使干物质的积累下降。     

EFFECT OF CADMIUM ON PHYSIOLOGICAL RESPONSES OF WHEAT AND CORN TO IRON DEFICIENCY

This work evaluated the effect of cadmium (Cd) on the physiological responses of corn (Zea Mays L.) and wheat (Triticum aestivum L.) to iron (Fe) deficiency. For this purpose, seedlings of corn and wheat were cultivated under controlled conditions, plants were grown in different strength Hoagland's solutions for one month. In the fifth week, some seedlings were still in full strength Hoagland's solution (+Fe) and others were in full strength Hoagland's solutions without iron (?Fe). The plants were exposed to different cadmium (Cd) concentrations for four days. The plant chlorophyll content of young leaves, Fe and Cd content in shoots and roots, biomass production, and phytosiderophores (PS) release by roots were assessed. Results showed that Cd decreased the chlorophyll content of young leaves, accompanied by a significant shoot and root biomass reduction for Fe-deficient and Fe-sufficient wheat and corn across all Cd treatments. However, chlorophyll content and shoot and root biomass of Fe-deficient wheat and corn were lower than Fe-sufficient plants at different Cd concentrations. Iron-deficiency induced Cd accumulation compared to Fe-sufficient in wheat and corn; however, a depressive effect of Cd on iron acquisition in shoots and roots of Fe-deficient and Fe-sufficient wheat and corn across all Cd treatments was observed. Cadmium also inhibited PS release in Fe-deficient and Fe-sufficient wheat and corn. Iron-deficient PS release was higher than Fe-sufficient corn and wheat across all Cd treatments. These results suggested that Cd might reduce capacity of plants to acquire iron from solution by inhibiting PS release.     

不同蔬菜对镉的吸收累积及亚细胞分布

采用营养液培养,基于叶绿体分离方法,研究了不同种类蔬菜对镉吸收、运输和亚细胞的分布规律。结果表明,蔬菜在不同浓度镉的营养液中培养1周后,蔬菜生物量没有产生显著差异,而不同种类的蔬菜生物量差异较大。镉在蔬菜叶片中大部分存在于细胞壁中,占总量的62％～85％,少量存在于原生质（不含叶绿体）和叶绿体中;随着营养液中镉浓度的增加,各组分中镉的含量明显增加,但分配比例变化不大。各种蔬菜根中镉的含量高于地上部镉含量;随着镉浓度的增加,根中镉分配比例从44％-59％降低至27％～38％;不同蔬菜根部对镉的吸收能力及镉向地上部转移的能力有显著性差异。     

硫对超积累东南景天镉累积、亚细胞分布和化学形态的影响

采用差速离心技术和化学试剂逐步提取法研究了硫对超积累东南景天镉累积、亚细胞分布和化学形态的影响。结果表明,增硫处理（S2和S3）显著提高超积累东南景天根、茎和叶的镉含量、累积量及整株累积总量。镉在超积累东南景天根、茎和叶中的含量和分配比例为F1（细胞壁）F3（可溶性部分）F2（细胞器与膜）,细胞壁（F1）是Cd在超积累东南景天细胞内的主要结合位点;超积累东南景天根、茎和叶F1、F2、F3组分中的镉含量随着硫水平的增加而增加,但分配比例变化不一致;超积累东南景天植物体内镉形态以氯化钠提取态（FNacl）、醋酸提取态（FHAc）和水提取态（FW）占优势。增施硫处理,提高超积累东南景天根FNacl、FHAc和FW提取态镉含量和分配比例,降低FE和FHCl提取态镉含量和分配比例;茎FNacl和FHAc提取态镉含量和分配比例增加,FW、FE和FHCl提取态镉分配比例降低;叶FNacl、FHAc和FW提取态镉的含量增加,但对其分配比例影响不大。     

镉胁迫下钙对镉在玉米细胞中分布及对叶绿体结构与酶活性的影响

采用溶液培养试验、亚细胞组分分级分离技术、透射电子显微镜观察及生化分析等方法研究钙对镉胁迫下玉米根、叶组织镉的亚细胞分布、叶绿体超微结构及叶片RuBP 羧化酶和PEP 羧化酶活性的影响。结果表明,与缺钙加镉处理比较,镉毒害下添加钙,玉米根、叶细胞器和细胞质Cd 组分均显著下降,相应Ca 组分则显著增加,而细胞壁Cd 、Ca 含量各处理间均十分接近;镉毒害下添加钙,维管束鞘细胞基质片层及叶肉细胞基粒和基粒片层排列较好,维管束鞘细胞嗜锇粒大为减少,叶片RuBP和PEP羧化酶活性显著增加。说明Ca 对于Cd 胁迫下玉米叶片正常结构与功能的保持具有十分重要的意义。     

镉对水稻幼苗根系细胞膜电位和膜透性的影响

镉（Cd）对膜透性、跨膜电势差等膜性质所产生的直接影响是造成Cd胁迫下植物对营养元素吸收异常的重要原因。以对Cd2＋敏感性不同的两个水稻品种（淮稻11号和扬稻6号）作为试验材料,采用玻璃微电极技术原位考察了水稻幼苗根细胞膜电位对Cd2＋胁迫的响应,同时分析了Cd2＋对水稻根系细胞膜透性的影响。结果表明,Cd2＋可使根表皮细胞膜电位在短时间内发生明显的去极化,去极化程度随Cd2＋浓度提高而增加;相同浓度Cd2＋所诱导的膜电位去极化程度与水稻品种有关,扬稻6号幼苗根系膜电位的去极化程度明显高于淮稻11号;在迅速的去极化之后,随后的30 min内是否发生复极化与水稻品种和Cd2＋浓度有关;就更长时间（0-12 h）来看,膜电位在经过了去极化阶段后仍有缓慢的恢复,淮稻11号在1.0 mmo1·L-1 Cd2＋处理6 h后已接近初始的膜电位值,而扬稻6号在处理12 h后仍不能恢复到处理前的水平;Cd2＋处理增大了水稻根细胞膜透性,且扬稻6号比淮稻11号膜透性增加的程度要大。     

Silicon induced cadmium tolerance of rice seedlings

Cadmium (Cd²⁺) toxicity and effects of silicon (Si) applications on the cellular and intracellular accumulations and distributions of Cd were investigated by conventional electron microscopy and EDX analysis. The Si‐deprived rice (Oryza sativa L.) plants (‐Si) differ greatly from Si‐replete ones in cell walls and vacuoles distributions of Ca in their leaves and roots. Energy dispersive x‐ray microanalysis revealed that considerable amounts of Cd could be detected in the cytoplasm, vacuole or cellular organelles in ‐Si rice plants, while very little to be found in +Si ones. From the nanochemical and nanobiological points of views, cell wall templates mediated the formation of colloidal silica with the high specific adsorption property to prevent the uptake of Cd into the cell.     

水稻镉安全材料分蘖期根部镉积累分布特征

【目的】镉 (Cd) 低积累作物的培育和应用是减少土壤中Cd通过食物链危害人体健康的重要途径。通过研究Cd处理下水稻分蘖期根部Cd的积累分布特征,揭示水稻Cd安全材料根部Cd的固持机理,为水稻Cd安全品种的培育提供理论依据。【方法】以水稻Cd安全材料D62B为供试材料,普通材料Luhui17为对照,进行水培试验。水稻秧苗于三叶一心时移栽至盆中 (40 cm × 60 cm × 15 cm),以CdCl₂·2.5H₂O加入营养液,设0 (CK)、0.5 (Cd0.5)、1.0 (Cd1)、2.0 (Cd2) mg/L 4个Cd浓度处理,30天后收获,分为根和地上部测定其Cd全量。采用化学试剂逐步提取法和差速离心法分别测定根部Cd化学形态和亚细胞分布特征,并进一步结合细胞壁多糖,研究其对Cd的响应特征。【结果】1) Cd处理下D62B各部位Cd含量显著低于Luhui17,转移系数较小,其根部Cd向地上部转移较少。2) 随Cd处理浓度升高D62B根部水提取态Cd分配比例降低,盐酸提取态Cd、残渣态Cd分配比例升高,Cd移动性减弱。D62B根部Cd以氯化钠提取态为主 (48.9%～52.1%),高浓度Cd处理 (2.0 mg/L) 下其分配比例是Luhui17的1.11倍,水提取态是Luhui17的82.3%,其根中Cd的移动性弱于普通材料。3) D62B根部Cd主要分布于可溶部分和细胞壁,其中细胞壁Cd分配比例为38.6%～41.8%,高于Luhui17。随Cd处理浓度升高,D62B根细胞壁Cd分配比例降幅小于Luhui17,其细胞壁对Cd的固持作用有限但强于普通材料。4) D62B根细胞壁半纤维素1的Cd含量是果胶的7.74～8.40倍,Cd主要与细胞壁中半纤维素1结合。半纤维素1 Cd含量随Cd处理浓度升高而显著增加,2.0 mg/L Cd处理下D62B和Luhui17半纤维素1单位总糖Cd结合量较1.0 mg/L Cd处理分别增加32.6%、11.2%,D62B根细胞壁半纤维素1的Cd结合能力强于Luhui17。【结论】水稻Cd安全材料D62B各部位Cd含量低于Luhui17,其转移系数较小。D62B根中Cd主要为氯化钠提取态,随Cd处理浓度升高,根部Cd向移动性较弱的化学形态转化。D62B根细胞壁中Cd主要与半纤维素1结合,由于其Cd结合能力较强,D62B根细胞壁对Cd的固持作用强于普通材料。因此,D62B对Cd的转移能力弱于普通材料,是其籽粒Cd安全的重要原因。