大叶井边草——一种新发现的富集砷的植物

自1999年以来对位于湖南省一些高砷区的植物和土壤进行了一系列的野外调查,以着力寻找砷的超富集植物。结果表明,与砷超富集植物蜈蚣草同属的另一种植物－－大叶井口边草,对砷也具有显著的富集特征。这一发现为研究揭示砷在植物中的超富集机理提供了一种新的材料,建议深入开展蜈蚣草与大叶井口边草这两种砷富集植物的对比研究。     

砷胁迫下不同砷富集能力植物内源生长素与抗氧化酶的关系

采用室内土培法,研究了砷胁迫(0,50,100和200 mg/kg)对凤尾蕨属中砷超富集植物大叶井口边草(Pteris cretica var.nervosa)和非砷超富集植物剑叶凤尾蕨(Pteris ensiformis)生物量、株高、叶片内源3-吲哚乙酸(IAA)含量、IAA氧化酶(IAAO)活性、砷含量、抗氧化酶(超氧化物歧化酶SOD、过氧化物酶POD、过氧化氢酶CAT)活性以及细胞膜脂过氧化产物丙二醛(MDA)含量的影响,并用逐步回归法分析了IAA含量与砷含量、抗氧化酶活性、MDA含量之间的关系。此外,研究了100 mg/kg砷处理下IAA、IAAO、3种抗氧化酶和MDA的时间动态。结果表明,与对照(不加砷)相比,加砷处理下大叶井口边草的株高和生物量未出现显著变化,但剑叶凤尾蕨在中、高浓度砷胁迫下则显著降低;中、高浓度砷胁迫均使2种植物叶片含砷量、IAA含量显著增加和IAAO活性显著下降,但这种改变在大叶井口边草中更为显著;加砷处理使大叶井口边草叶片3种抗氧化酶活性维持或增加,剑叶凤尾蕨中SOD和CAT活性虽能维持,但POD活性则显著下降,说明砷胁迫下大叶井口边草具有更强的抗氧化能力。逐步回归分析结果显示,2种植物叶片IAA含量均与砷含量成显著正相关,但在大叶井口边草中,IAA含量还与CAT活性成显著负相关。时间动态研究表明,第13天,大叶井口边草具有最高的IAA含量、最低的IAAO活性以及最低的CAT活性,而剑叶凤尾蕨中的变化规律则不明显。因此,叶片保持较高的IAA含量、较低的IAAO和CAT活性有助于大叶井口边草超量富集砷。     

砷超富集植物中砷化学形态及其转化的EXAFS研究

通过同步辐射扩展X射线吸收精细结构(SR EXAFS)研究砷超富集植物大叶井口边草(Pteris nervosa)中砷的化学形态及其在植物体中的转化。结果表明,在大叶井口边草中砷主要与O配位,根部存在与谷胱苷肽(GSH)结合的砷,但是在羽叶中没有发现与GSH结合的砷,在NaAsO_2和Na_2HAsO_4处理中,植物根系的砷分别以As(Ⅲ)和As(Ⅴ)为主,但是在叶柄和羽叶中砷都以As(Ⅲ)的形态为主,植物根系吸收的As(Ⅴ)在向上转运的过程中具有向As(Ⅲ)转化的趋势,其转化过程主要发生在根部。实验证明,与GSH结合并不是大叶井口边草中砷解毒的主要机理,超富集植物可能具有与一般耐性植物不同的重金属解毒机制。     

砷超富集植物中砷化学形态其转化的EXAFS研究

通过同步辐射扩展X射线吸收精细结构(SR EXAFS)研究砷超富集植物大叶井口边草(Pteris nervosa)中砷的化学形态及其在植物体中的转化. 结果表明, 在大叶井口边草中砷主要与O配位, 根部存在与谷胱苷肽(GSH)结合的砷, 但是在羽叶中没有发现与GSH结合的砷. 在NaAsO2和Na2HAsO4处理中, 植物根系的砷分别以As(Ⅲ)和 As(Ⅴ)为主, 但是在叶柄和羽叶中砷都以As(Ⅲ)的形态为主. 植物根系吸收的As(Ⅴ)在向上转运的过程中具有向As(Ⅲ)转化的趋势, 其转化过程主要发生在根部. 实验证明, 与GSH结合并不是大叶井口边草中砷解毒的主要机理, 超富集植物可能具有与一般耐性植物不同的重金属解毒机制.     

砷超富集植物中砷化学形态及其转化的EXAFS研究

通过同步辐射扩展X射线吸收精细结构(SR EXAFS)研究砷超富集植物大叶井口边草(Pteris nervosa)中砷的化学形态及其在植物体中的转化. 结果表明, 在大叶井口边草中砷主要与O配位, 根部存在与谷胱苷肽(GSH)结合的砷, 但是在羽叶中没有发现与GSH结合的砷. 在NaAsO₂和Na₂HAsO₄处理中, 植物根系的砷分别以As(Ⅲ)和As(Ⅴ)为主, 但是在叶柄和羽叶中砷都以As(Ⅲ)的形态为主. 植物根系吸收的As(Ⅴ)在向上转运的过程中具有向As(Ⅲ)转化的趋势, 其转化过程主要发生在根部. 实验证明, 与GSH结合并不是大叶井口边草中砷解毒的主要机理, 超富集植物可能具有与一般耐性植物不同的重金属解毒机制.     

砷超富集植物蜈蚣草原生质体的分离及其抗砷性分析

蜈蚣草(Pteris vittata)是一种砷超富集植物, 能够通过根从土壤中吸收砷, 并将其输送至羽叶中富集。为了探索蜈蚣草单个细胞在砷积累和砷抗性中的特性, 本文首次通过酶解方法获得了这一砷超富集蕨类植物的原生质体, 并研究了原生质体在不同浓度砷胁迫下的生活力。结果显示, 蜈蚣草原生质体的抗砷性远高于烟草原生质体的抗砷性, 与其整体植株的抗性一致。这为探索砷抗性和超富集机理提供了一个新的研究体系。     

砷超富集植物蜈蚣草原生质体的分离及其抗砷性分析

蜈蚣草（Pteris vittata）是一种砷超富集植物,能够通过根从土壤中吸收砷,并将其输送至羽叶中富集.为了探索蜈蚣草单个细胞在砷积累和砷抗性中的特性,本文首次通过酶解方法获得了这一砷超富集蕨类植物的原生质体,并研究了原生质体在不同浓度砷胁迫下的生活力.结果显示,蜈蚣草原生质体的抗砷性远高于烟草原生质体的抗砷性,与其整体植株的抗性一致.这为探索砷抗性和超富集机理提供了一个新的研究体系.     

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。     

基于蕨素聚类分析的凤尾蕨属药用植物化学分类研究

目的:研究凤尾蕨属5种药用植物蕨素类活性成分,为其质量评价、鉴定及亲缘关系研究提供依据,为凤尾蕨属植物化学分类深入研究奠定基础。方法:采用HPLC-DAD测定5种药用植物的石油醚、乙酸乙酯、正丁醇和水部位的蕨素类成分。比较蕨素种类和含量差异,并以蕨素类特征成分及其含量为分类性状,采用聚类分析对凤尾蕨属药用植物进行化学分类研究。结果:从大叶井口边草、半边旗、全缘凤尾蕨、蜈蚣草和凤尾草中分别检测出蕨素类成分59、39、3、7和22个,总体来看大叶井口边中蕨素类成分含量最高,而全缘凤尾蕨中蕨素类成分含量最低。运用系统聚类将5种药用植物聚成3类:第Ⅰ类为全缘凤尾蕨、蜈蚣草和凤尾草;第Ⅱ类为半边旗;第Ⅲ类为大叶井口边草。结论:半边旗与全缘凤尾蕨、蜈蚣草和凤尾草之间的化学亲缘关系较远。蕨素聚类结果与传统分类结果有较好的一致性。蕨素类成分HPLC-DAD图谱可用于凤尾蕨属植物的鉴别和分类研究,并可为其质量评价提供重要参考。     

改良FIASCO方法筛选砷超富集植物蜈蚣草SSR分子标记

蜈蚣草（Pteris vittata L.）是目前用于砷污染土壤修复最好的超富集植物,但其分子水平上的研究数据较少。为了开发蜈蚣草特异性SSR遗传标记,本文采用改良的FIASCO方法从蜈蚣草AG和AC微卫星富集文库中随机挑选100个克隆,分离得到51个微卫星位点,其中60%为完美型（Perfect）SSR。根据这些位点设计、合成了25对引物,并对江西庐山及湖北恩施两地蜈蚣草种群各20个个体进行了遗传多样性检测,结果发现：其中8个完美型及1个间断型（intermittent）SSR位点的引物能够扩增出清晰、稳定且具有多态性的条带。9对引物共扩增出41个等位基因,各位点等位基因数在2~7之间,平均等位基因数为4.56个;期望杂合度在0.0494~0.8169之间;没有连锁不平衡现象发生。采用大叶井栏边草（Pteris multifida Poir.）进行跨种扩增,结果发现其中6对引物能够进行种间扩增。这些SSR分子标记的开发有助于蜈蚣草生态适应性进化分析、揭示蜈蚣草地理分布格局以及探讨蜈蚣草遗传多样性,还可用于品种鉴定及选育等。     

EXAFS study on arsenic species and transformation in arsenic hyperaccumulator

Synchrotron radiation extended X-ray absorption fine structure (SR EXAFS) was employed to study the transformation of coordination environment and the redox speciation of arsenic in a newly discovered arsenic hyperaccumulator, Cretan brake (Pteris cretica L. var nervosa Thunb). It showed that the arsenic in the plant mainly coordinated with oxygen, except that some arsenic coordinated with S as As-GSH in root. The complexation of arsenic with GSH might not be the predominant detoxification mechanism in Cretan brake. Although some arsenic in root presented as As(V) in Na₂HAsO₄ treatments, most of arsenic in plant presented as As(III)-O in both treatments, indicating that As(V) tended to be reduced to As(III) after it was taken up into the root, and arsenic was kept as As(III) when it was transported to the above-ground tissues. The reduction of As(V) primarily proceeded in the root.     

EXAFS study on arsenic species and transformation in arsenic hyperaccumulator

As a cost-effective, efficient and environmental friendly method for the remediation of contaminated soils and waters, phytoremediation of arsenic-con- taminated soils has drawn more and more attention[1]. The plants with the special ability to accumulate arse-nic (hyperaccumulators) are a prerequisite for phy-toremediation. Cretan brake (Pteris cretica L. var nervosa Thunb) has been shown to accumulate arsenic as much as 694 mg/kg in pinna in field investigation[2], and such elevated arsenic…     

Phytoremediation of arsenic contaminated soil by Pteris vittata L. II. Effect on arsenic uptake and rice yield

A greenhouse experiment evaluated the effect of phytoextraction of arsenic from a contaminated soil by Chinese Brake Fern (Pteris vittata L.) and its subsequent effects on growth and uptake of arsenic by rice (Oryza sativa L.) crop. Pteris vittata was grown for one or two growing cycles of four months each with two phosphate sources, using single super phosphate (SSP) and di-ammonium phosphate (DAP). Rice was grown on phytoextracted soils followed by measurements of biomass yield (grain, straw, and root), arsenic concentration and, uptake by individual plant parts. The biomass yield (grain, straw and rice) of rice was highest in soil phytoextracted with Pteris vittata grown for two cycles and fertilized with diammonium phosphate (DAP). Total arsenic uptake in contaminated soil ranged from 8.2 to 16.9 mg pot(-1) in first growing cycle and 5.5 to 12.0 mg pot(-1) in second growing cycle of Pteris vittata. There was thus a mean reduction of 52% in arsenic content of rice grain after two growing cycle of Pteris vittata and 29% after the one growing cycle. The phytoextraction of arsenic contaminated soil by Pteris vittata was beneficial for growing rice resulted in decreased arsenic content in rice grain of <1 ppm. There was a mean improvement in rice grain yield 14% after two growing cycle and 8% after the one growing cycle of brake fern.     

Phytoextraction: simulating uptake and translocation of arsenic in a soil-plant system

The uptake, transport, and accumulation of metals by plants are functions central to successful phytoextraction. This study investigates the uptake and translocation of arsenic from a contaminated sandy soil by a mature Chinese brake fern (Pteris vittata L.). An existing mathematical model for the coupled transport of water, heat, and solutes in the soil-plant-atmosphere continuum (CTSPAC) was modified to examine the flow of water as well as the uptake and translocation of total arsenic in the xylem of the fern. This model was calibrated using greenhouse measurements before its application. Simulation results showed that about 20% of the soil arsenic was removed by the fern in 10 d, of which about 90% of the arsenic was stored in the fronds and 10% in the roots. Although arsenic mass in the plant tissues increased consecutively with time, arsenic concentration in the xylem sap of the root tips has a typical diurnal distribution pattern: increasing during the day and decreasing at night, resulting from daily variations of frond surface water transpiration. The largest difference in simulated arsenic concentration in the root tips between the day and night was about 5%. This study also suggests that the use of transpiration stream concentration factor (TSCF), which is defined as the ratio of chemical concentration in the xylem sap to that in the external solution, to evaluate the translocation efficiency of arsenic for the hyperaccumulator Chinese brake fern (Pteris vittata L.) could be limited.     

Phytoremediation of an arsenic-contaminated site using Pteris vittata L.: a two-year study

A field study was conducted to determine the efficiency of Chinese brake fern (Pteris vittata L.), an arsenic hyperaccumulator, on removal of arsenic from soil at an arsenic-contaminated site. Chinese brake ferns were planted on a site previously used to treat wood with chromated copper arsenate (CCA). Arsenic concentrations in surface and profile soil samples were determined for 2000, 2001, and 2002. In both 2001 and 2002, senesced and senescing fronds only, as well as all fronds, were harvested. Frond arsenic concentrations were not significantly different between the three harvests. Compared to senesced fronds, live fronds resulted in the greatest amount of arsenic removal. There were no significant differences in soil arsenic concentrations between 2000, 2001, and 2002, primarily due to the extreme variability in soil arsenic concentrations. However, the mean surface soil arsenic was reduced from 190 to 140 mg kg(-1). Approximately 19.3 g of arsenic were removed from the soil by Chinese brake fern. Therefore, this fern is capable of accumulating arsenic from the CCA -contaminated site and may be competitive, in terms of cost, to conventional remediation systems. However, better agronomic practices are needed to enhance plant growth and arsenic uptake to obtain maximum soil arsenic removal and to minimize remediation time.     

Expression of a Pteris vittata glutaredoxin PvGRX5 in transgenic Arabidopsis thaliana increases plant arsenic tolerance and decreases arsenic accumulation in the leaves

Chinese brake fern Pteris vittata hyperaccumulates arsenic in its fronds. In a study to identify brake fern cDNAs in arsenic resistance, we implicated a glutaredoxin, PvGRX5, because when expressed in Escherichia coli , it improved arsenic tolerance in recombinant bacteria. Here, we asked whether PvGRX5 transgenic expression would alter plant arsenic tolerance and metabolism. Two lines of Arabidopsis thaliana constitutively expressing PvGrx5 cDNA were compared with vector control and wild-type lines. PvGRX5-expressors were significantly more tolerant to arsenic compared with control lines based on germination, root growth and whole plant growth under imposed arsenic stress. PvGRX5-expressors contained significantly lower total arsenic compared with control lines following treatment with arsenate. Additionally, PvGRX5-expressors were significantly more efficient in their arsenate reduction in vivo . Together, our results indicate that PvGRX5 has a role in arsenic tolerance via improving arsenate reduction and regulating cellular arsenic levels. Paradoxically, our results suggest that PvGRX5 from the arsenic hyperaccumulator fern can be used in a novel biotechnological solution to decrease arsenic in crops.     

砷对土壤-蜈蚣草系统中磷生物有效性的影响

磷是植物必需的大量营养元素,而其同族元素砷却不是植物生长发育所必需的。通过等温吸附平衡实验发现土壤中存在的砷可以降低褐土对磷的吸附,褐土对砷的吸附率大于等于褐土对磷的吸附率。对砷超富集植物蜈蚣草而言,土壤中砷的添加量不超过800mg／kg时,蜈蚣草地上部和地下部磷含量显著提高,结果初步表明,砷可以提高土壤中磷的生物有效性。     

A Critical Review of the Arsenic Uptake Mechanisms and Phytoremediation Potential of Pteris vittata

The discovery of the arsenic hyperaccumulator, Pteris vittata (Chinese brake fern), has contributed to the promotion of its application as a means of phytoremediation for arsenic removal from contaminated soils and water. Understanding the mechanisms involved in arsenic tolerance and accumulation of this plant provides valuable tools to improve the phytoremediation efficiency. In this review, the current knowledge about the physiological and molecular mechanisms of arsenic tolerance and accumulation in P. vittata is summarized, and an attempt has been made to clarify some of the unresolved questions related to these mechanisms. In addition, the capacity of P. vittata for remediation of arsenic-contaminated soils is evaluated under field conditions for the first time, and possible solutions to improve the remediation capacity of Pteris vittata are also discussed.     

Antioxidant responses of hyper-accumulator and sensitive fern species to arsenic

Plant species capable of hyper-accumulating heavy metals are of considerable interest for phytoremediation, and differ in their ability to accumulate metals from the environment. This work aims to examine (i) arsenic accumulation in three fern species [Chinese brake fern (Pteris vittata L.), slender brake fern (Pteris ensiformis Burm. f.), and Boston fern (Nephrolepis exaltata L.)], which were exposed to 0, 150, or 300 muM of arsenic (Na(2)HAsO(4).7H(2)O), and (ii) the role of anti-oxidative metabolism in arsenic tolerance in these fern species. Arsenic accumulation increased with an increase in arsenic concentration in the growth medium, the most being found in P. vittata fronds showing no toxicity symptoms. In addition, accumulation was highest in the fronds, followed by the rhizome, and finally the roots, in all three fern species. Thiobarbituric acid-reacting substances, indicators of stress in plants, were found to be lowest in P. vittata, which corresponds with its observed tolerance to arsenic. All three ferns responded differentially to arsenic exposure in terms of anti-oxidative defence. Higher levels of superoxide dismutase, catalase, and ascorbate peroxidase were observed in P. vittata than in P. ensiformis and N. exaltata, showing their active involvement in the arsenic detoxification mechanism. However, no significant increase was observed in either guaiacol peroxides or glutathione reductase in arsenic-treated P. vittata. Higher activity of anti-oxidative enzymes and lower thiobarbituric acid-reacting substances in arsenic-treated P. vittata correspond with its arsenic hyper-accumulation and no symptoms of toxicity.     

Solubility and accumulation of metals in Chinese brake fern, vetiver and rostrate sesbania using chelating agents

Greenhouse experiments were conducted to study the effects of chelating agents on the growth and metal accumulation of Chinese brake fern (Pteris vittata L.), vetiver (Vetiveria zizanioides L.), and rostrate sesbania (Sesbania rostrata L.) in soil contaminated with arsenic (As), Cu, Pb, and Zn. Among the five chelating agents used [ethylenediaminetriacetic acid (EDTA), hydroxyethylenediaminetriacetic acid (HEDTA), nitrilotriacetic acid (NTA), oxalic acid (OA), and phytic acid (PA)], OA was the best to mobilize As, EDTA to mobilize Cu and Pb, and HEDTA to mobilize Zn from soil, respectively. The biomass of vetiver was the highest, followed by rostrate sesbania. All chelating agents inhibited the growth of Chinese brake fern and rostrate sesbania, but HEDTA significantly increased the aboveground biomass of vetiver. Dry weights of both Chinese brake fern and rostrate sesbania decreased with increasing EDTA concentrations amended in the soil, especially in treatments with high EDTA concentrations. EDTA and HEDTA enhanced Cu, Zn, and Pb, but lowered As accumulation in all three plant species, except for As in vetiver, while OA significantly enhanced As accumulation in the aboveground part of vetiver. Concentrations of Cu, Zn, and Pb in the aboveground parts of plants increased significantly with the increase of EDTA concentrations and treatment time. In addition to As, Chinese brake fern also accumulated the highest Cu, Pb, and Zn in its aboveground parts among the three plant species grown in metal-contaminated soil with EDTA/HEDTA treatments. This species, therefore, can be used to simultaneously clean up As, Cu, Pb, and Zn from contaminated soils with the aid of EDTA or HEDTA.