砂培条件下施加钙、砷对蜈蚣草吸收砷、磷和钙的影响

在砂培条件下 ,研究施加钙、砷对蜈蚣草生长和砷、磷和钙的吸收及转运的影响。添加砷对蜈蚣草的生物量 (根、叶柄和羽叶的干物重 )虽未达到显著影响 (p<0 .0 5) ,但添加 0 .1 mmol/L砷时 ,表现出刺激生长效应。提高介质中钙浓度明显抑制蜈蚣草根系生长 ,钙浓度过高还会显著限制地上部生长。供应 0 .0 3mmol/L钙时 ,蜈蚣草羽片砷浓度为 42 1 8mg/kg,明显高于 2 .5和 5 mmol/L钙处理下相应的砷浓度。砷的转运系数 (羽片 /根 )随着介质中砷浓度的升高而增大 ,随着介质中钙浓度的升高而减少。这说明一定范围内提高介质中砷浓度促进砷向地上部运输 ,而钙却明显抑制砷向地上部转运。钙和砷浓度过高时 ,植株均会出现中毒症状。钙中毒表现为叶脉变褐和叶肉坏死 ;而砷中毒现象表现在叶尖和叶缘变褐。介质中砷限制蜈蚣草根部对磷的吸收 ,但对地上部磷浓度无显著影响。介质中添加砷 ,植物体内钙浓度升高 ,可能起缓解砷毒的作用。钙、砷对蜈蚣草羽片砷累积量和总累积量均有极显著的交互作用 ,钙是负交互效应 ,砷是正交互效应。添加 2 .5和 5.0 mmol/L钙时 ,相对于 0 .0 3 mmol/L钙处理分别减少地上部砷累积量 2 0 .8%和73.1 %。这表明在应用蜈蚣草进行植物修复时 ,介质中出现过高浓度的钙是不利于提高土壤修复效率     

蜈蚣草砷超富集机制及其在砷污染修复中的应用

蕨类植物蜈蚣草能够从土壤中吸收砷,并储存于地上部分羽叶的液泡中。蜈蚣草具有高效的抗氧化系统,以降低砷的毒害;其砷酸还原系统和液泡区隔化是蜈蚣草进行砷解毒和砷超富集的重要机制。本文综述了目前蜈蚣草砷超富集机制研究的主要进展,并对其在修复砷污染环境的应用中进行了讨论。     

不同生态型摩西球囊霉菌株对蜈蚣草砷吸收的影响

砷超富集植物——蜈蚣草无论是在野外或是在室内均能被丛枝菌根真菌(AM真菌)侵染,但其对蜈蚣草砷吸收及转运的机理尚不清晰.本研究将分离于湖南省郴州市金川塘某铅锌尾矿蜈蚣草根际土壤(Glomus mosseae BGC GD01,简称污染菌株)和云南省未污染土壤(G.mosseae BGC YN05,简称非污染菌株)的2种摩西球囊霉菌株分别接种于非污染生态型和污染生态型蜈蚣草根际,8周后利用菌根化蜈蚣草幼苗在浓度为100 μmol·L-1砷(Na2HAsO4·7H2O)营养液中进行为期24 h的水培试验.结果表明,2种生态型摩西球囊霉菌株分别与蜈蚣草形成中等程度侵染,侵染率为25.2％ ～31.3％.无论是接种污染菌株或是非污染菌株,均明显促进了蜈蚣草根部对磷的吸收.在24 h水培试验期间,接种非污染菌株显著促进了蜈蚣草根部砷的吸收,但接种污染菌株对蜈蚣草根部砷吸收的促进作用有限,说明AM真菌对蜈蚣草砷吸收存在种内差异.     

AM真菌对蜈蚣草根围土壤砷形态及其砷吸收的影响

为探明AM真菌对蜈蚣草Pteris vittata根围土壤砷形态及其吸收砷的效应,采用盆栽实验,接种摩西管柄囊霉Funneliformis mosseae(Fm)、幼套近明球囊霉Claroideoglomus etunicatum(Ce)和变形球囊霉Glomus versiforme(Gv)。实验结果表明:接种Ce处理对蜈蚣草根围p H影响不显著,但提高了根围土壤中非专性吸附态砷、结晶水合铁铝氧化物结合态砷比例,分别达35%和13%,同时降低了无定形和弱结晶水合铁铝氧化物结合态砷、残渣态砷比例,分别达3%和11%。蜈蚣草生物量及其体内砷浓度分别提高了111%和15%。研究表明接种Fm或Ce处理相比接种Gv处理对提高根围土壤中弱吸附态砷比例或降低较强吸附态砷比例的效果更好。而与接种Fm和Gv处理相比,接种Ce处理对提高蜈蚣草生物量及砷浓度、砷累积量的效果更显著。接种Ce可显著提高蜈蚣草对砷的提取效率,研究结果为蜈蚣草-AM真菌联合修复As污染土壤提供了技术指导。     

不同比例盐生小球藻-枯草芽孢杆菌共生体对砷酸盐耐性及富集差异

采用盐生小球藻与枯草芽孢杆菌为供试材料,构建不同藻菌比(1∶0、1∶1、1∶2、1∶3、1∶4)的共生体,在不同浓度砷酸盐[As(Ⅴ)]处理7 d后,测定藻菌共生体生长及其对砷的富集、吸附、吸收和形态转化。结果表明: 在As(Ⅴ)处理下,随着枯草芽孢杆菌比例的增加,藻菌共生体叶绿素含量、干重、比增长率显著提高,在750 μg·L^-1As(Ⅴ)处理时,1∶4的藻菌共生体分别达到1.81 mg·L^-1、125.0 mg、0.28 mg·L^-1·d^-1。藻菌比例由1∶0变为1∶4时,藻菌共生体对砷的富集和吸收呈下降趋势;砷的富集方式随砷浓度的增加而变化,在75～150 μg·L^-1As(Ⅴ)处理下以吸收为主,在300～750 μg·L^-1 As(Ⅴ)处理下以吸附为主。藻菌共生体内存在As(Ⅴ)和As(Ⅲ)两种形态,且随枯草芽孢杆菌比例的上升,As(Ⅴ)还原率增大(最高达到12.6%)。综上,枯草芽孢杆菌的添加提升了藻菌共生体对As(Ⅴ)的耐性和还原,但减少了对As(Ⅴ)的富集。     

苦草对砷的富集作用

为了探求合适的水体砷污染修复植物及砷在食物链中传递、累积的特点,以常见的沉水植物-苦草为研究对象,对受砷污染的水体进行修复,结果表明:苦草对水环境中砷的富集能在较短的时间内(3 d)达到一个较大值,到第14天,不同砷水平(2 mg/L)处理下的苦草对砷富集系数均超过200;苦草中砷浓度随处理时间及外源砷浓度的增加而增加,且与外源砷浓度之间存在极显著地正相关;苦草在不同浓度砷处理下都生长良好,对砷胁迫表现出较强的耐受性。因此,苦草对于水体的砷污染有着很好的去除效果,同时也能很好地反映出一个地区的砷污染水平。     

不同采收方式对富养化河道浮床空心菜生物产出的影响

以浮床空心菜为研究对象,在过水河道条件下,研究了不同采收周期(14 d采收1次、21 d采收1次、28 d采收1次)和留茬高度(15 cm、25 cm、35 cm)组合对浮床植物生物产出的影响.结果表明:刈割后的植物仍能够适应过水河道环境,并在其生长周期内持续生长.经110 d的生长,分枝数提高了近5倍,最长根长27.55 cm,平均根长15 cm,根系直径达11cm.单株根鲜重可达146 g,每平方米浮床根部可吸附颗粒物3.36 kg.新芽生长速率随采收次数增多,处理组均呈现先不断提高然后减小的趋势,变化幅度为0.54～3.7cm·d-1,对照组为1.63 cm·d-1.适当刈割能提高新芽再生速率.单次收获生物量同新芽变化规律相似,即先逐渐增多然后减小,生物量增长速率变化幅度3.83～37.9 g·m-2·d-1.从总生物量来看,28 d采收1次留茬25 cm、35 cm组的生物量(干重)产出最高,达2112 g·m-2;从茎叶比看,每14 d采收1次3个留茬高度,及每21d采收1次留茬15 cm和25 cm方式收获生物量茎叶比较佳.综合考虑产量、质量及浮床便捷管理,每21 d采收1次留茬15 cm的采收方式效果最佳,此时新芽平均生长速率1.88 cm·d-1,平均茎叶比＜1,总生物量1966 g·m-2.     

砷对植物衰老的影响

供试植物有：油菜（Brassicacampestris）品种“上海青”、空心菜（Alternantheraphiloxeroides）品种“湘早”、大豆（Glycinemax）品种“中黄4号”、豌豆（Pisumsativum）品种“绿珠”和蚕豆（Viciafaba）品种“浙抗60”。采用Hoagland营养液（pH5．5）作为基础培养液，根据不同试验目的，设置不同砷处理水平，每个处理3次重复。各种植物分别于3～5月份播种。所用砷盐为Na3AsO4·12H2O。培养器皿容积为IL，每瓶6～7株苗，每周更换一次培养液，用加氧泵和0．3％HZO。联合供氧。取30d龄植株不同部位叶，测定生理指标。叶绿素测定用改…     

中亚热带天然林土壤CH_4吸收速率对模拟N沉降的响应

陆地森林土壤是重要的大气甲烷(CH4)汇,大气氮(N)沉降增加对森林土壤CH4吸收速率影响突出。运用静态箱-气相色谱法对中亚热带天然林土壤CH4吸收速率对模拟N沉降的响应进行连续3a的观测;试验作3种N处理,分别为对照(CK,0 kg N·hm-2·a-1)、低氮(LN,50 kg N·hm-2·a-1)和高氮(HN,100 kg N·hm-2·a-1),每种处理重复3次,每个月采集气体1次,同时测定0—5 cm土壤温度和0—12 cm土壤含水量;分析不同N沉降水平土壤CH4吸收速率的差异、动态变化以及对土壤含水量和土壤温度响应,并探讨N沉降对土壤理化性质的影响。结果显示:天然林土壤(CK)平均CH4吸收速率为(-62.78±14.39)μg·m-2·h-1,LN和HN土壤平均CH4吸收速率分别下降了30.21%、7.24%,CK、LN和HN处理土壤CH4吸收速率季节变化趋势相似;观测期间土壤CH4吸收速率对LN响应达到显著水平(P0.05),对HN响应则不显著(P0.05);LN、HN处理前两年对土壤CH4吸收速率抑制作用均不显著(P0.05),但在第3年LN极显著降低了土壤CH4吸收速率(P0.01),HN处理对土壤CH4吸收速率的影响则在第3年表现为显著抑制作用(P0.05),表明土壤CH4吸收速率对N沉降的响应随着N沉降时间的持续呈抑制效应加剧的趋势。相关分析表明:CK与HN土壤CH4吸收速率与土壤温度和土壤含水量均有显著相关性(P0.05),但LN土壤CH4吸收速率仅与土壤含水量显著相关(P0.05),表明土壤含水量是控制各N沉降处理土壤CH4吸收速率动态的主要环境因子。此外,LN、HN处理下土壤pH均极显著降低(P0.01),但LN土壤pH极显著低于HN(P0.01);LN处理极显著提高了土壤C/N比(P0.01),HN处理则相反;LN和HN处理对土壤NH+4-N、NO-3-N、可溶性总N(TDN)、可溶性有机碳(DOC)、地面凋落物量、地下0—10 cm细根生物量影响均不显著(P0.05),表明一定时期内N沉降首先引起了土壤pH和土壤C/N比的显著变化。     

长白山三种主要林地土壤甲烷通量

森林土壤甲烷(CH4)通量及主要影响因素的研究对于降低全球温室气体收支评估的不确定性具有重要价值.本研究通过室内培养实验,分析了土壤湿度、温度和氮添加对长白山3种主要林型(白桦林、山杨林和阔叶红松林)土壤甲烷通量的影响.结果表明:3种林型土壤均为甲烷汇,15 d平均吸收速率分别为2.27 μg·kg-1·h-1(山杨林)、1.54μg·kg-1·h-1(阔叶红松林)和1.46 μg·kg-1·h-1(白桦林).重复测量多元方差分析结果显示:林型、温度、土壤湿度及氮素处理对甲烷通量均有极显著影响(P＜0.0l),林型与其他因子交互作用显著;3种林型土壤甲烷吸收的最佳含水量为45％ ～ 60％;在10 ～20℃条件下,甲烷吸收速率随温度增加而增加;氮对甲烷吸收有明显抑制作用.     

蜈蚣草中砷超富集的分子机制研究进展

砷是一种毒性很强的类金属元素,土壤砷污染可引发一系列食品安全问题,进而威胁人类健康。蜈蚣草具有极强的富集砷的能力,在砷污染土壤的植物修复中具有重要的应用价值。深入阐释蜈蚣草超富集砷的分子机制是植物修复技术的核心理论基础。文中综述了蜈蚣草超富集砷的组学研究进展,以及目前鉴定到的砷富集过程中的重要分子元件,并对未来的研究方向和趋势进行了展望。     

Arsenic hyperaccumulation in gametophytes of Pteris vittata. A new model system for analysis of arsenic hyperaccumulation

The sporophyte of the fern Pteris vittata is known to hyperaccumulate arsenic (As) in its fronds to >1% of its dry weight. Hyperaccumulation of As by plants has been identified as a valuable trait for the development of a practical phytoremediation processes for removal of this potentially toxic trace element from the environment. However, because the sporophyte of P. vittata is a slow growing perennial plant, with a large genome and no developed genetics tools, it is not ideal for investigations into the basic mechanisms underlying As hyperaccumulation in plants. However, like other homosporous ferns, P. vittata produces and releases abundant haploid spores from the parent sporophyte plant which upon germination develop as free-living, autotrophic haploid gametophyte consisting of a small (<1 mm) single-layered sheet of cells. Its small size, rapid growth rate, ease of culture, and haploid genome make the gametophyte a potentially ideal system for the application of both forward and reverse genetics for the study of As hyperaccumulation. Here we report that gametophytes of P. vittata hyperaccumulate As in a similar manner to that previously observed in the sporophyte. Gametophytes are able to grow normally in medium containing 20 mm arsenate and accumulate >2.5% of their dry weight as As. This contrasts with gametophytes of the related nonaccumulating fern Ceratopteris richardii, which die at even low (0.1 mm) As concentrations. Interestingly, gametophytes of the related As accumulator Pityrogramma calomelanos appear to tolerate and accumulate As to intermediate levels compared to P. vittata and C. richardii. Analysis of gametophyte populations from 40 different P. vittata sporophyte plants collected at different sites in Florida also revealed the existence of natural variability in As tolerance but not accumulation. Such observations should open the door to the application of new and powerful genetic tools for the dissection of the molecular mechanisms involved in As hyperaccumulation in P. vittata using gametophytes as an easily manipulated model system.     

Comparison of root absorption, translocation and tolerance of arsenic in the hyperaccumulator Pteris vittata and the nonhyperaccumulator Pteris tremula

* Several fern species can hyperaccumulate arsenic, although the mechanisms are not fully understood. Here we investigate the roles of root absorption, translocation and tolerance in As hyperaccumulation by comparing the hyperaccumulator Pteris vittata and the nonhyperaccumulator Pteris tremula. * The two species were grown in a pot experiment with 0-500 mg As kg-1 added as arsenate, and in a short-term (8 h) uptake experiment with 5 microM arsenate under phosphorus-sufficient conditions. * In the pot experiment, P. vittata accumulated up to 2500 mg As kg-1 frond d. wt and suffered no phytotoxicity. P. tremula accumulated<100 mg As kg-1 frond d. wt and suffered severe phytotoxicity with additions of >or=25 mg As kg-1. In the short-term uptake experiment, P. vittata had a 2.2-fold higher rate of arsenate uptake than P. tremula, and distributed more As taken up to the fronds (76%) than did P. tremula (9%). * Our results show that enhanced root uptake, efficient root-to-shoot translocation, and a much elevated tolerance through internal detoxification all contribute to As hyperaccumulation in P. vittata.     

Effects of arsenic species and concentrations on arsenic accumulation by different fern species in a hydroponic system

Two hydroponic experiments were conducted to evaluate factors affecting plant arsenic (As) hyperaccumulation. In the first experiment; two As hyperaccumulators (Pteris vittata and P. cretica mayii) were exposed to 1 and 10 mg L(-1) arsenite (AsIII) and monomethyl arsenic acid (MMA) for 4 wk. Total As concentrations in plants (fronds and roots) and solution were determined In the second experiment P. vittata and Nephrolepis exaltata (a non-As hyperaccumulator) were exposed to 5 mgL(-1) arsenate (AsV) and 20 mgL(-1) AsIIIfor 1 and 15 d. Total As and AsIII concentrations in plants were determined Compared to P. cretica mayii, P. vittata was more efficient in arsenic accumulation (1075-1666 vs. 249-627mg kg(-1) As in the fronds) partially because it is more efficient in As translocation. As translocation factor (As concentration ratio in fronds to roots) was 3.0-5.6 for P. vittata compared to 0.1 to 4.8 for P. cretica. Compared to N. exaltata, P. vittata was significantly more efficient in arsenic accumulation (38-542 vs. 4.8-71 mg kg(-1) As in thefronds) as well asAs translocation (1.3-5.6 vs. 0.2-0.5). In addition, P. vittata was much more efficient in As reduction from AsV to AsIII (83-84 vs. 13-24% AsIII in the fronds). Little As reduction occurred after 1-d exposure to AsV in both species indicates that As reduction was not instantaneous even in an As hyperaccumulator. Our data were consistent with the hypothesis that both As translocation and As reduction are important for plant As hyperaccumulation.     

Effectiveness of applying arsenate reducing bacteria to enhance arsenic removal from polluted soils by Pteris vittata L

Arsenic is a common contaminant in soils and water. It is well established that the fern Pteris vittata L. is an As hyperaccumulator and therefore has potential to phyroremediate As-polluted soils. Also, it is accepted that rhizosphere microflora play an enhancing role in plant uptake of metallic elements from soils. Studies showed that hydroponiclly grown P. Vittata accumulated arsenite more than the arsenate form of As apparently because arsenate and phosphate are analogues and therefore its absorption is inhibited by phosphate. The objective of this study was to determine whether addition of five different arsenate-reducing bacteria would enhance arsenic uptake by P. vittata grown in arsenic polluted soils in afield experiment. Results showed that addition of the As reducing bacteria promoted the growth of P. vittata, increased As accumulation, activated soil insoluble As, and reduced As leaching compared to the untreated control. Plant biomass increased by 53% and As uptake by 44%. As leaching was reduced by 29% to 71% depending on the As reducing bacterium. The results in their entirety permitted some insight into the mechanisms by which the arsenate reducing bacteria enhanced the effectiveness of P. vittata to remove As from the polluted soil.     

Mechanisms of arsenic hyperaccumulation in Pteris vittata. Uptake kinetics,interactions with phosphate,and arsenic speciation

The mechanisms of arsenic (As) hyperaccumulation in Pteris vittata, the first identified As hyperaccumulator, are unknown. We investigated the interactions of arsenate and phosphate on the uptake and distribution of As and phosphorus (P), and As speciation in P. vittata. In an 18-d hydroponic experiment with varying concentrations of arsenate and phosphate, P. vittata accumulated As in the fronds up to 27,000 mg As kg(-1) dry weight, and the frond As to root As concentration ratio varied between 1.3 and 6.7. Increasing phosphate supply decreased As uptake markedly, with the effect being greater on root As concentration than on shoot As concentration. Increasing arsenate supply decreased the P concentration in the roots, but not in the fronds. Presence of phosphate in the uptake solution decreased arsenate influx markedly, whereas P starvation for 8 d increased the maximum net influx by 2.5-fold. The rate of arsenite uptake was 10% of that for arsenate in the absence of phosphate. Neither P starvation nor the presence of phosphate affected arsenite uptake. Within 8 h, 50% to 78% of the As taken up was distributed to the fronds, with a higher translocation efficiency for arsenite than for arsenate. In fronds, 49% to 94% of the As was extracted with a phosphate buffer (pH 5.6). Speciation analysis using high-performance liquid chromatography-inductively coupled plasma mass spectroscopy showed that >85% of the extracted As was in the form of arsenite, and the remaining mostly as arsenate. We conclude that arsenate is taken up by P. vittata via the phosphate transporters, reduced to arsenite, and sequestered in the fronds primarily as As(III).     

A novel arsenate reductase from the arsenic hyperaccumulating fern Pteris vittata

Pteris vittata sporophytes hyperaccumulate arsenic to 1% to 2% of their dry weight. Like the sporophyte, the gametophyte was found to reduce arsenate [As(V)] to arsenite [As(III)] and store arsenic as free As(III). Here, we report the isolation of an arsenate reductase gene (PvACR2) from gametophytes that can suppress the arsenate sensitivity and arsenic hyperaccumulation phenotypes of yeast (Saccharomyces cerevisiae) lacking the arsenate reductase gene ScACR2. Recombinant PvACR2 protein has in vitro arsenate reductase activity similar to ScACR2. While PvACR2 and ScACR2 have sequence similarities to the CDC25 protein tyrosine phosphatases, they lack phosphatase activity. In contrast, Arath;CDC25, an Arabidopsis (Arabidopsis thaliana) homolog of PvACR2 was found to have both arsenate reductase and phosphatase activities. To our knowledge, PvACR2 is the first reported plant arsenate reductase that lacks phosphatase activity. CDC25 protein tyrosine phosphatases and arsenate reductases have a conserved HCX5R motif that defines the active site. PvACR2 is unique in that the arginine of this motif, previously shown to be essential for phosphatase and reductase activity, is replaced with a serine. Steady-state levels of PvACR2 expression in gametophytes were found to be similar in the absence and presence of arsenate, while total arsenate reductase activity in P. vittata gametophytes was found to be constitutive and unaffected by arsenate, consistent with other known metal hyperaccumulation mechanisms in plants. The unusual active site of PvACR2 and the arsenate reductase activities of cell-free extracts correlate with the ability of P. vittata to hyperaccumulate arsenite, suggesting that PvACR2 may play an important role in this process.     

Arsenic in the rhizosphere soil solution of ferns

The aim of this study was to explore the evidence of arsenic hyperaccumulation in plant rhizosphere solutions. Six common fern plants were selected and grown in three types of substrate: arsenic (As) -tailings, As-spiked soil, and soil-As-tailing composites. A rhizobox was designed with an in-situ collection of soil solutions to analyze changes in the As concentration and valence as well as the pH, dissolved organic carbon (DOC) and total nitrogen (TN). Arsenite composed less than 20% of the total As, and As depletion was consistent with N depletion in the rhizosphere solutions of the various treatments. The As concentrations in the rhizosphere and non-rhizosphere solutions in the presence of plants were lower than in the respective controls without plants, except for in the As-spiked soils. The DOC concentrations were invariably higher in the rhizosphere versus non-rhizosphere solutions from the various plants; however, no significant increase in the DOC content was observed in Pteris vittata, in which only a slight decrease in pH appeared in the rhizosphere compared to non-rhizosphere solutions. The results showed that As reduction by plant roots was limited, acidification-induced solubilization was not the mechanism for As hyperaccumulation.     

Molecular cloning and characterization of a phytochelatin synthase gene, PvPCS1, from Pteris vittata L.

Pteris vittata L. is a staggeringly efficient arsenic hyperaccumulator that has been shown to be capable of accumulating up to 23,000 microg arsenic g(-1), and thus represents a species that may fully exploit the adaptive potential of plants to toxic metals. However, the molecular mechanisms of adaptation to toxic metal tolerance and hyperaccumulation remain unknown, and P. vittata genes related to metal detoxification have not yet been identified. Here, we report the isolation of a full-length cDNA sequence encoding a phytochelatin synthase (PCS) from P. vittata. The cDNA, designated PvPCS1, predicts a protein of 512 amino acids with a molecular weight of 56.9 kDa. Homology analysis of the PvPCS1 nucleotide sequence revealed that it has low identity with most known plant PCS genes except AyPCS1, and the homology is largely confined to two highly conserved regions near the 5'-end, where the similarity is as high as 85-95%. The amino acid sequence of PvPCS1 contains two Cys-Cys motifs and 12 single Cys, only 4 of which (Cys-56, Cys-90/91, and Cys-109) in the N-terminal half of the protein are conserved in other known PCS polypeptides. When expressed in Saccharomyces cerevisae, PvPCS1 mediated increased Cd tolerance. Cloning of the PCS gene from an arsenic hyperaccumulator may provide information that will help further our understanding of the genetic basis underlying toxic metal tolerance and hyperaccumulation.