Subcellular distribution,chemical forms,and physiological response to cadmium stress in Hydrilla verticillata

This study investigated the subcellular distribution and chemical forms of cadmium (Cd) in Hydrilla verticillata and the physiological mechanism underlying H. verticillata responses to Cd stress. Hydrilla verticillata was grown in a hydroponic system and was treated with various Cd concentrations (0, 10, 50, 100, 125, and 150?µM) for 7?days. Cadmium analysis of the leaves at the subcellular level showed that Cd was mainly stored in the soluble fraction (77.98–83.62%) and in smaller quantities in the cell wall fraction (11.99–17.30%) and the cell organelles (4.30–4.88%). The Cd taken up by H. verticillata was in different chemical forms. In the leaves and stems, the Cd was mostly extracted using 1?M NaCl and smaller amounts of Cd were extracted using 2% acetic acid. The malondialdehyde content significantly increased at all Cd concentrations, which indicated oxidative stress. The superoxide dismutase, guaiacol peroxidase, and catalase activities were enhanced. The proline, ascorbate, and glutathione contents increased at lower Cd concentrations, but decreased consistently as the Cd concentration rose. These results suggest that H. verticillata can be successfully used in the phytoremediation of Cd-contaminated water.     

Subcellular distribution,chemical forms and thiol synthesis involved in cadmium tolerance and detoxification in Siegesbeckia orientalis L.

Siegesbeckia orientalis L. is a promising species for cadmium (Cd) phytoextraction with large biomass and fast growth rate, while little information about their intracellular mechanisms involved in Cd tolerance and detoxification has been explored. A soil pot experiment with total target Cd concentrations of 0, 10, 50, 100, and 150 mg kg^?1 were designed to investigate the subcellular distribution, chemical forms and thiol synthesis characteristics of Cd in S. orientalis. More than 90% of Cd was bound to the soluble fractions (48.4–76.5%) and cell walls (19.9–46.3%). Increasing soil Cd concentrations enhanced Cd sequestration into the cell walls. Most of the Cd (69.8–82.7%) in the plant organ was mainly in the forms of pectate and protein integrated Cd and undissolved Cd phosphate, while a minor portion (6.8–20.9%) was in the forms of the inorganic Cd and the water soluble Cd. Nonprotein thiols and phytochelatins significantly increased with increasing soil Cd treatment levels, while glutathione concentrations had no obvious change trends. Therefore, intracellular detoxification mechanisms of Cd in S. orientalis mainly rely on formation of less toxic Cd chemical forms, store of a large amount of Cd in cell wall and synthesis of thiol compounds.     

Effects of extended growth periods on subcellular distribution,chemical forms,and the translocation of cadmium in Impatiens walleriana

Impatiens walleriana plants accumulate sufficiently high concentrations of cadmium (Cd) for this species to be considered a potential Cd hyperaccumulator. Rooted cuttings were grown hydroponically for 25 and 50 days in solutions spiked with various Cd concentrations. The subcellular distribution and chemical forms of Cd in different organs were analyzed, and its upward translocation was also assessed. The plants accumulated large amounts of Cd; the Cd concentration in the roots and shoots reached 120–1900 and 60–1600 mg/kg, respectively. Regardless of the growth period, the Cd accumulated in the roots was primarily compartmentalized in the soluble fraction or ethanol and deionized water extractable chemical forms with high migration abilities. Translocation to the shoots was followed by an association of Cd mainly in the cell wall or with pectate and protein. The roots’ Cd showed a high migration capacity for predicting the shoots’ Cd concentrations. Different exposure periods significantly affected the subcellular distribution of Cd in the stems, and thus the upward translocation.     

霞多丽苗木中镉的积累、亚细胞分布及化学存在形态

以一年生盆栽葡萄品种霞多丽(Vitis vinifera L.cv.Chardonnay)自根苗及SO4砧嫁接苗为试材,采用CdCl2和CaCl2根部灌入的方法,研究了镉在霞多丽植株内的亚细胞分布和存在形态,以及外源添加氯化钙对植株镉吸收的影响.结果表明:大部分镉积累在霞多丽自根苗及嫁接苗的地下部器官;4 mmol·L-1浓度的CdCl2处理下,镉在自根苗根及根颈部分的积累量占77.1％,在叶片中的积累量占1.4％,而嫁接苗中镉在嫁接口以下部分的积累量高达93.9％,叶片中的积累量仅占0.1％;5 mmol·L-1外源钙缓解了植株对镉的吸收积累,而10mmol ·L-1外源钙则显著增加了植株对镉的吸收积累.镉在根系和叶片中的亚细胞分布规律为细胞壁＞可溶性部分＞细胞器,且在细胞壁中积累50％以上;镉在根系中主要以氯化钠提取态存在,其次为乙酸提取态,去离子水提取态含量最少.随着镉处理浓度的增加,各提取态含量有所变化.     

Subcellular distribution and chemical forms of antimony in Ficus tikoua

Ficus tikoua (F. tikoua) was a potential species for antimony (Sb) phytoremediation due to its wide growth in the mining area. However, little was known about its tolerance mechanisms toward Sb. The determination of the distribution and chemical speciation of Sb in F. tikoua is essential for understanding the mechanisms involved in Sb accumulation, transportation, and detoxification. The present study investigated the subcellular distribution and chemical forms of Sb in F. tikoua. The plant was exposed to different Sb concentrations (0, 30, 90, and 180 μmol/L) for 30 days. The results showed that F. tikoua possessed a marked ability to tolerate and accumulate Sb. The proportional Sb increased with increasing Sb concentration in the solution, and the highest Sb concentration occurred in roots (1274.5–1580.9 mg/kg), followed by stems (133.5–498.9 mg/kg) and leaves (4.1–15.7 mg/kg). In the subcellular sequestration of Sb in F. tikoua, the largest accumulation of Sb occurred in cell walls (72.4–87.5%) followed by cytoplasmic organelles (8.2–18.6%) and cytoplasmic supernatant. The results suggested that cell walls act as important protective barriers against Sb toxicity in F. tikoua. Although Sb in all plant tissues found primarily in the fractions extracted by ethanol and distilled water, the current study found that the Sb amounts in the HAc-extractable fraction, HCl-extractable fraction, and residue fraction increased at the highest Sb level (180 μmol/L) compared to that under lower Sb levels. These results indicate that excessive Sb accumulated in F. tikoua under Sb stress is bound to non-dissolved or low-bioavailable compounds, a biochemical mechanism that benefits F. tikoua because it helps alleviate Sb toxicity.     

镉在旱柳中亚细胞分布及存在的化学形态

以2个旱柳无性系幼苗为材料,通过营养液培养并结合差速离心与化学试剂提取法,分析了不同浓度Cd2+胁迫下旱柳叶和根中Cd的亚细胞分布及其存在的化学形态.结果显示,(1)随着培养介质Cd2+浓度升高,旱柳无性系幼苗叶和根中各亚细胞组分Cd含量随之增加.叶片的Cd主要富集于细胞壁、叶绿体和可溶性部分,它们的含量分别占65%～69%、14%～22%、6.8%～7.7%,仅少量Cd发现于膜部分;而根中Cd主要积累于细胞壁和可溶性部分,其中含量分别占59%～66%和14%～25%,Cd在根亚细胞组分中积累量依次为细胞壁>可溶性部分>质体>膜部分.(2)旱柳体内Cd以不同的化学形态存在,大部分为HCl(FHCl)、NaCl(FNaCl)、醋酸(HAC,FHAC)提取态,极少部分为乙醇(EtOH,FEtOH)和水提取态(Fwater),叶和根中5种Cd提取态含量依次为FHCl>FNaCl>FHAC>Fwater>FEtOH,而叶和根中HCl和NaCl提取态Cd占有比例大于30%以上.研究表明,旱柳无性系中Cd主要与蛋白质和有机酸螯合或以金属磷酸盐沉淀的形态存在,其根、叶的细胞壁和液泡在Cd忍耐与解毒中起到重要作用.     

铅在茶树体内的分布及化学形态特征

采用水培试验,结合亚细胞组分分离和化学形态提取技术,研究了茶树品种龙井43和迎霜不同器官的Pb含量,及其在根系亚细胞中的分布和化学结合形态.结果表明： 在Pb胁迫下,两个品种茶树根系的形态特征不同;龙井43新叶中Pb含量随营养液Pb浓度的提高显著增加,而迎霜的变化不显著.两个品种茶树根系亚细胞中Pb的含量分布及结合形态均存在差异,低浓度Pb处理下, 龙井43根亚细胞组分（除可溶物质外）中Pb含量均低于迎霜;而高浓度Pb处理下,龙井43根亚细胞组分（除细胞壁外）中Pb含量均高于迎霜;龙井43根中醋酸(HAc)提取态的Pb含量比例最高,其他化学形态的高低顺序为NaCl提取态>HCl提取态/H₂O提取态>乙醇提取态;而迎霜根中NaCl提取态Pb成分比例最高,其他化学形态的高低顺序为HAc提取态>HCl提取态/H₂O提取态>乙醇提取态.迎霜对Pb毒害的耐受能力强于龙井43.     

不同铅水平下紫茎泽兰细胞内铅的分布和化学形态的分析

该研究运用差速离心法和化学试剂逐步提取法,分析了重金属铅在紫茎泽兰亚细胞内的分布和主要化学形态。结果表明:随着Pb浓度的升高,紫茎泽兰的叶、根、茎中各亚细胞组分Pb含量逐渐增加;紫茎泽兰中的Pb在叶片分布于可溶性部分和细胞壁中,两者占总量的75.34%~84.63%;茎也主要分布于可溶性部分和细胞壁中,占总量的36.10%~57.14%和20.07%~36.52%;而在根中则富集于细胞壁和可溶性部分,分别占39.2%~49.78%和28.27%~37.62%,其他细胞器中的Pb含量均较少。紫茎泽兰叶中的Pb以盐酸提取态和水提取态为主,两者占总量的58.74%~73.04%;茎中的Pb以醋酸提取态和氯化钠提取态为主;而根中的Pb则以醋酸提取态和盐酸提取态占优势,两者占总量的39.15%~52.91%。     

Subcellular distribution and chemical forms of cadmium in Bechmeria nivea (L.) Gaud.

Bechmeria nivea (L.) Gaud. (Ramie) is a promising species for Cd phytoextraction with large biomass and fast growth rate. Nevertheless, little information is available on its tolerance mechanisms towards Cd. Determination of Cd distribution and chemical speciation in ramie is essential for understanding the mechanisms involved in Cd accumulation, transportation and detoxification. In the present study, ramie plants were grown in hydroponics with increasing Cd concentrations (0, 1, 3, 7 mg l^?1). The subcellular distribution and chemical forms of Cd in different tissues were determined after 20 days exposure to this metal. To assess the effect of Cd uptake on plant performance, nitrate reductase activity in leaves and root activity were analyzed during the entire experimental period. Increased Cd level in the medium caused a proportional increase in Cd uptake, and the highest Cd concentration occurred in roots, followed by stems and leaves. Subcellular fractionation of Cd-containing tissues indicated that about 48.2–61.9% of the element was localized in cell walls and 30.2–38.1% in soluble fraction, and the lowest in cellular organelles. Cd taken up by ramie rapidly equilibrated among different chemical forms. Results showed that the greatest amount of Cd was found in the extraction of 1 M NaCl and 2% HAC, and the least in residues in all test tissues. In roots, the subdominant amount of Cd was extracted by d-H₂O and 80% ethanol, followed by 0.6 M HCl. While in stems and leaves, the amount of 0.6 M HCl-extractable Cd was comparable with that extracted by 80% ethanol or d-H₂O. 1 mg l^?1 Cd stimulated nitrate reductase activity in leaves and root activity, while a concentration-dependent inhibitory effect was observed with increasing Cd concentration, particularly at 7 mg l^?1 Cd. It could be suggested that the protective mechanisms evolved by ramie play an important role in Cd detoxification at relatively low Cd concentrations (below 3 mg l^?1 Cd) but become restricted to maintain internal homeostasis with higher Cd stress.     

Lead bioaccumulation,subcellular distribution and chemical form in sugarcane and its potential for phytoremediation of lead-contaminated soil

Abstract

Sugarcane is a promising species for lead (Pb) phytoextraction due to its large biomass and high tolerance toward Pb content. To understand the mechanisms involved in Pb tolerance and detoxification and its potential for phytoremediation in sugarcane, the bioaccumulation, subcellular distribution, and chemical forms of Pb in different tissues were investigated through pot cultivation sugarcane with increasing Pb concentrations in the present study. Results showed that sugarcane could tolerate high concentrations of Pb (up to 1250.0–1750.0?mg/kg); the Pb content in roots and shoots increased with increasing Pb concentration. A large amount of Pb content was stored in roots. The subcellular distribution of Pb in sugarcane revealed that the majority of Pb was bound to the cell wall. Meanwhile, the greatest amount of Pb was extracted by 2.0% acetic acid and 0.6?mol/L HCl, which indicated that most of Pb was combined with undissolved phosphate and oxalate. These results implied that the Pb formation of undissolved salts and compartmentalization in the cell wall may be a key strategy for Pb detoxicity and tolerance in sugarcane.     

锌在3种乔木中的积累及其亚细胞分布和化学形态

为了探索园林木本植物对重金属锌(Zn)的积累和耐性机制,本研究以栾树(Koelreu-teria paniculata)、臭椿(Ailanthus altissima)和银杏(Ginkgo biloba)3种北京常见乔木为试验树种,通过盆栽污染模拟试验,研究不同浓度Zn处理(O、250、500、1000、2000 mg...     

Subcellular distribution and chemical forms of Cd in Bougainvillea spectabilis Willd. as an ornamental phytostabilizer: An integrated consideration

The effects of Cd stress on the growth and Cd accumulation of Bougainvillea spectabilis Willd. as an ornamental plant and the related mechanisms were investigated in the study. We studied the impact of Cd on the plant ultrastructure, examined the cellular distribution of Cd, explored the Cd chemical forms and transformation, and determined the organic acid secretion in the plants. The results showed that B. spectabilis could grow well in the Cd treatment groups, and the roots could accumulate high concentration of Cd. The soluble fraction (primarily in the vacuole) as the form of citrate in leaves of B. spectabilis was the major compartment for Cd storage. The citric acid secreted by B. spectabilis played an important role in the detoxification of Cd, as well as the growth of plants and Cd accumulation. As an ornamental plant, B. spectabilis has the potential to be used in the phytostabilization of Cd-contaminated soils and can beautify the environment at the same time.     

镉在水花生叶片中的亚细胞分布及其毒理学

以水花生为试验材料,研究了不同浓度镉(Cd)(0、0.05、0.1、0.2、0.4 mmol·L-1)胁迫下水花生叶中Cd及矿质元素的亚细胞分布、抗氧化能力、叶绿素、可溶性蛋白和植物络合素(PCs)含量的变化.结果表明:随着培养液中Cd浓度的增加,叶片内各亚细胞组分中Cd含量显著增加,但分布不均,主要积累在细胞壁,其次为可溶成分,线粒体和叶绿体中分布最少.Cd处理浓度大于0.2 mmol·L-1时,呈现Cd向水花生叶的可溶成分中分配增加、细胞壁中分配减少的趋势.矿质元素的吸收明显失衡,表现为各亚细胞组分中Ca含量显著增加且主要分布在细胞壁中,细胞壁和可溶成分中P和K含量不断降低,叶绿体中Mg和Fe的含量逐渐减少.叶绿素和可溶性蛋白含量持续下降.谷胱甘肽和抗坏血酸含量先升后降,总抗氧化能力(T-AOC)不断提高,PCs大量累积.水花生对水体中的Cd有一定耐性,且Cd在亚细胞结构中的富集量与其毒性间具有明显的剂量-效应关系;亚细胞中矿质元素的紊乱以及叶绿素和可溶性蛋白含量的显著降低,Cd在亚细胞中的积累对植物体造成了明显的毒害作用;Ca在细胞壁中的累积以及PCs和T-AOC水平的提高说明水花生具有较强的抗性;PCs的合成量与Cd对水花生的毒性变化有一定的相关性,可以作为Cd胁迫的生物标记物.     

Alleviation of Cadmium Toxicity to <i>Medicago Truncatula</i> by AMF Involves the Changes of Cd Speciation in Rhizosphere Soil and Subcellular Distribution

Arbuscular mycorrhizal fungi (AMF) can improve plant tolerance to several abiotic stresses, including heavy metals, drought or salinity exposure. However, the role of AMF in alleviation of soil cadmium (Cd)-induced toxicity to plants is still largely unknown. In this study, Cd speciation in soil and subcellular distribution of Cd were used to characterize the roles of application AM fungi in the alleviation of Cd toxicity in alfalfa plants. Our results showed that the addition of Glomus mosseae in Cd contaminated soil (10 mg/Kg) significantly increased soil pH, cation exchange capacity (CEC) and organic matter in rhizosphere soil with Medicago truncatula L., and then account for significantly decreased contents of exchangeable and carbonate-bounded Cd speciation in rhizosphere soil, indicating alleviation of plant toxicity by reduction of bioavailable fractions of Cd. Although there is no significant difference found in Cd accumulation by roots and shoots respectively between Cd and AM-Cd treatments, more portion of Cd was recorded compartmentalization in cell wall fraction of both root and shoot in treatment of Cd with AM application, indicating alleviation of Cd toxicity to plant cell. Herein, application of AM fungi in Cd treatments performed to inhibit the appearance of Cd toxicity symptoms, including the improvement of leaf electrolyte leakage, root elongation, seedling growth and biomass. This information provides a clearer understanding of detoxification strategy of AM fungi on Cd behavior with development and stabilization of soil structure and subcellular distribution of plant.     

镉在水生植物中的富集与亚细胞分布及其化学形态特征

以7种水生植物(长苞香蒲、水生美人蕉、黑藻、粉绿狐尾藻、花叶芦竹、香根鸢尾、水葫芦)为研究材料,构建生物塘系统,通过差速离心法和五步提取法,提取植物各亚细胞组分和不同化学形态的镉,并用原子吸收分光光度法测定镉含量,分析镉在水生植物体中富集特征,揭示植物对镉的耐性机制。结果表明:(1)水生植物根中镉富集量大于茎和叶;3种类型水生植物富集镉能力表现为沉水植物(粉绿狐尾藻、黑藻)浮水植物(水葫芦)挺水植物(长苞香蒲、水生美人蕉、花叶芦竹、香根鸢尾)。(2)镉在黑藻、水葫芦和花叶芦竹中的亚细胞分布量均呈现为细胞壁可溶组分原生质体线粒体,所占比例分别为37.16%~50.86%、20.69%~31.21%、10.81%~23.83%、8.15%~19.83%。(3)赋存化学形态表现为以氯化钠提取态、醋酸提取态为主,所占比例分别为29.37%~56.27%和15.06%~36.19%。研究发现,粉绿狐尾藻、水葫芦、水生美人蕉和花叶芦竹4种植物为富集镉较好的植物,而黑藻、香根鸢尾和长苞香蒲3种植物的镉富集能力相对较弱;镉主要以果胶酸盐、蛋白质结合态或吸附态存在于植物根的细胞壁和液泡中,以减弱镉对根细胞器和植物地上部分的毒害。     

Agrobacterium-mediated transient MaFT expression in mulberry (Morus alba L.) leaves

To optimize Agrobacterium-mediated transient transformation assay in mulberry (Morus alba L.), various infiltration methods, Agrobacterium tumefaciens (A. tumefaciens) strains, and bacterial concentrations were tested in mulberry seedlings. Compared with LBA4404, GV3101 harboring pBE2133 plasmids presented stronger GUS signals at 3 days post infiltration using syringe. Recombinant plasmids pBE2133:GFP and pBE2133:GFP:MaFT were successfully constructed. Transient expression of MaFT:GFP protein was found in leaves, petiole (cross section), and shoot apical meristem (SAM) of mulberry according to the GFP signal. Moreover, MaFT:GFP mRNA was also detected in leaves and SAM via RT-PCR and qRT-PCR. An efficient transient transformation system could be achieved in mulberry seedlings by syringe using A. tumefaciens GV3101 at the OD₆₀₀ of 0.5. The movement of MaFT expression from leaves to SAM might trigger the precocious flowering of mulberry.     

Exogenous spermidine elevating cadmium tolerance in Salix matsudana involves cadmium detoxification and antioxidant defense

In this study, exogenous spermidine role on Salix matsudana tolerance to cadmium was evaluated. Spermidine and cadmium presented antagonistic effects on the biomass, copper and zinc concentrations in S. matsudana. cadmium mainly distributed in the cell wall of subcellular fraction; 46.97%–60.43% of cadmium existed in a sodium chloride-extracted form. Cadmium contents in roots, leaves, and twigs ranged from 2002.67 to 3961.00, 111.59 to 229.72, and 102.56 to 221.27?mg/kg, respectively. Spermidine application elevated cadmium concentrations in the roots, cuttings, and cell wall and the ratio of deionized water-extracted cadmium, but decreased cadmium levels in the twigs and leaves and the fractions of cadmium extracted by ethanol and sodium chloride, respectively. Putrescine and malondialdehyde were important indicators of cadmium-induced oxidative damage. Exogenous spermidine alleviated the accumulation of superoxide anion, hydrogen peroxide, malondialdehyde via promoting the levels of spermidine, soluble protein, superoxide dismutase, reductive ascorbate, glutathione reductase, and glutathione peroxidase in S. matsudana leaves under the corresponding cadmium stress. The results indicated that S. matsudana was a candidate for cadmium rhizoremediation and extraction in leaves; the spermidine application enhanced the cadmium tolerance of S. matsudana through promoting cadmium accumulation in roots, cell wall, and less bioactive chemical forms and the antioxidative ability.     

镉和锌在皖景天细胞内的分布及化学形态

运用差速离心法和化学试剂逐步提取法,分析了Cd和Zn在皖景天根、茎和叶的亚细胞分布及其化学形态.结果表明:10 μmol·L^-1 Cd处理下,Cd在皖景天细胞内的主要分布位点是其可溶部分;在100 μmol·L^-1 Cd处理下,Cd在根中主要分布在细胞壁、茎中主要分布在细胞壁和可溶部分、叶中超过90%的Cd分布在可溶部分.高Cd浓度处理时,皖景天根、茎和叶的细胞壁中Cd分布比例增加,而可溶部分Cd分布比例相对减少.在1和800 μmol·L^-1 Zn处理条件下,Zn在皖景天根、茎和叶的主要分布位点是可溶部分;高Zn浓度处理时,皖景天叶、茎和根的可溶部分和细胞壁中Zn的分布比例无明显变化.细胞器中Zn和Cd分布都很少.Cd在皖景天根、茎和叶内主要以氯化钠提取态和水提取态存在,Zn在皖景天根、茎和叶内以多种化学形态存在.     

镉在东南景天中的亚细胞分配

采用差速离心技术,比较研究了Cd在矿山生态型东南景天(Sedum alfredii Hance)根、茎、叶中的亚细胞分布.结果表明:矿山生态型东南景天对Cd具有很好的忍受耐和累积能力,非矿山生态型东南景天则不具有这种能力.Cd在矿山生态型东南景天根、茎、叶各部分的亚细胞分配满足F1(细胞壁部分)>F3(可溶部分)>F2(细胞器及膜)规律,且Cd在细胞壁部分的分配占绝对优势;同时矿山生态型东南景天地上部有很好的Cd累积能力.与众多超累积植物相似的Cd亚细胞分配规律和地上部的良好的Cd累积能力.对此植物的Cd亚细胞分布完全不同于非矿山生态型东南景天的情况作了比较分析,结果显示矿山生态型东南景天很可能是一种新的镉超累积种质资源.     

Kinetics of the early subcellular distribution of cadmium in rat hepatocytes

The kinetics of the early subcellular distribution of cadmium (Cd) was characterized in primary cultures of rat hepatocytes exposed to 10, 50 and 100 M Cd in a serum-free WME medium for 10, 30 or 60 min. Our results demonstrate a time- and concentration-dependent increase in Cd content with the highest metal concentration measured in the cytosol, whereas the lowest was observed in the mitochondria. With the exception of early localization in the plasma membrane, Cd concentrations in fractions were characterized by the following decreasing order of magnitude: cytosol > low density molecules nuclei > lysosomes mitochondria. We also found evidence for: (i) a two-step process for Cd distribution in the nuclei and mitochondria; and (ii) a time-dependent slow process of transfer from the plasma membrane to the cytosol. Saturation in Cd uptake was observed at 50 M in most cell fractions at 10 and 30 min, except for the plasma membrane. The lack of apparent saturation for Cd accumulation at 60 min was not related to an increase in metallothionein synthesis. Altogether, our data provide insights into the dynamics of transfer between intracellular compartments, and allow a better identification of the organelles that are the most subjected to Cd toxicity for early exposure conditions.Deceased March 25, 2004.