Role of trichome ofPteris vittata L. in arsenic hyperaccumulation

Environmental scanning electron microscope (ESEM) fitted with an energy dispersive X-ray microanalyzer (EDX) was used to investigate the surface micromorphology and arsenic (As) micro-distribution in Chinese brake (Pteris vittata L.). It was found that amounts of trichome, which possessed multicellular structure with the average length of 160 μm and with an average diameter of 28 μm, existed in the frond ofP. vittata, and the density of trichome on the pinnate axial surface was higher than that on the petiole. Visible X-ray peak of As was recorded in the epidermal cell and trichome. The relative weight of As in the pinnate trichome, which contained the highest concentration of As among all tissues of the plant, was 2.4 and 3.9 times as much as that in the epidermal and mesophyllous cells, respectively. The As concentrations in the basal and stalk cells of the same trichome were higher than that in its cap cell. This is the first time to report that the trichome ofP. vittata plays an important role in arsenic hyperaccumulation. The finding from the present study implies that much attention should be paid to the role of the trichome in understanding the hyperaccumulation and detoxicity of As in the hyperaccumulator and improving the ability of As accumulation.     

Role of trichome of Pteris vittata L. in arsenic hyperaccu-mulation

Heavy metal pollution of soils, caused by various anthropogenic sources, is a major environmental problem. Due to its cost-effectiveness and environ-mental friendliness, phytoremediation of arsenic-con- taminated soils has attracted more and more attention. An arsenic (As) hyperaccumulator, Chinese brake (Pteris vittata L.) was discovered by Chen et al. in China[1]. The field phytoremediation in Chenzhou City, Hunan Province has been successfully carried out by Chen et al. since 2000[2,3].…     

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。     

Elemental allelopathy by an arsenic hyperaccumulating fern,Pteris vittata L.

Aims The functional advantages of arsenic (As) hyperaccumulation by plants are poorly understood. One proposed benefit, termed elemental allelopathy, occurs when hyperaccumulated As is cycled from the plant back into the top layer of soil, allowing As hyperaccumulators to gain an advantage over intolerant species by increasing soil As concentrations ([ As]) underneath their canopy. To date, there are no studies that detail the presence of increased soil [ As] associated with As hyperaccumulators. In this study, we documented variation in the soil [ As] associated with the Chinese brake fern, Pteris vittata L. and also compared the effects of environmentally relevant soil and solution [ As] on competitor plant growth.Methods Four populations of P. vittata were identified in central Florida, USA. P. vittata tissue samples and soil samples were collected at the base of and at 3 m away from ferns in each population (n = 36). Five sample locations were randomly selected from each site, and soils from the base and 3 m away from each fern were collected to examine the effects of naturally occurring soil [ As] on the germination and growth of a potential competitor plant (Oxalis stricta). Solutions with increasing [ As] were also used to examine the threshold for negative effects of [ As] on O. stricta growth. [ As] were measured using inductively coupled plasma mass spectrometry (ICP-MS).Important findings Overall, soil [ As] from the base of ferns was nearly twice that of soil 3 m away indicating that ferns hyperaccumulate As. However, ferns and their associated soil, contained different [ As] depending on their collection site, indicating that these populations accumulate and use [ As] differently. O. stricta growth decreased and germination was delayed as solution and soil [ As] increased. However, the relative distance from the fern that the soil was collected from did not affect growth, which would be expected with elemental allelopathy. Our results show that P. vittata is associated with higher soil [ As] and these concentrations are sufficient to inhibit growth of competitors. However, the absence of a strong inhibitory relationship associated with proximity to the fern across all locations suggests that the possible functional advantages of elemental allelopathy may depend on site specific characteristics.     

Distributions of arsenic and essential elements in pinna of arsenic hyperaccumulator Pteris vittata L.

The distributions of arsenic and 6 essential elements in the pinna of As hyperaccumulator, Pteris vittata L., were studied using synchrotron radiation X-ray fluorescence (SRXRF). Significant correlation between the distribution and mobility of the elements revealed that SRXRF study on the elemental distribution was feasible to inspect the transportations of elements in plants. The distribution of As in the pinna showed that As had great abilities to be transported in xylem vessels and from xylem to mesophyll. The distribution of K, one of the most mobile elements in plants, was similar to that of As, whereas the distributions of Fe and Ca with less mobility in plants were almost opposite to that of As in the pinna.     

Metabolic adaptation of Pteris vittata L. gametophyte to arsenic induced oxidative stress

The sporophyte and gametophyte of Pteris vittata are arsenic hyperaccumulators, however, little is known about the mechanism by which the gametophyte deals with this toxic element. An in vitro system (spores grown in arsenic amended nutrient media) was used to investigate the impact of arsenic on growth of the gametophyte and the role of antioxidative systems in combating As-stress. When mature spores of P. vittata were grown in medium amended with 0-50 mg kg(-1) of arsenic (as arsenate), the arsenic concentration in the gametophyte increased, with increasing arsenate in the media, but did not inhibit the spore germination and biomass development. Increases in the level of antioxidant enzymes, superoxide dismutase, catalase, ascorbate peroxidase, glutathione reductase, and glutathione-Stransferase) and of ascorbic acid and glutathione probably enabled the gametophyte to withstand the oxidative stress caused by arsenate.     

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.     

Effects of arsenate and phosphate on their accumulation by an arsenic-hyperaccumulator Pteris vittata L.

Arsenate and phosphate interactions are important for better understanding their uptake and accumulation by plant due to their similarities in chemical behaviors. The present study examined the effects of arsenate and phosphate on plant biomass and uptake of arsenate and phosphate by Chinese brake (Pteris vittata L.), a newly-discovered arsenic hyperaccumulator. The plants were grown for 20 weeks in a soil, which received the combinations of 670, 2670, or 5340 mol kg^–1 arsenate and 800, 1600, or 3200 mol kg^–1 phosphate, respectively. Interactions between arsenate and phosphate influenced their availability in the soil, and thus plant growth and uptake of arsenate and phosphate. At low and medium arsenate levels (670 and 2670 mol kg^–1), phosphate had slight effects on arsenate uptake by and growth of Chinese brake. However, phosphate substantially increased plant biomass and arsenate accumulation by alleviating arsenate phytotoxicity at high arsenate levels (5340 mol kg^–1). Moderate doses of arsenate increased plant phosphate uptake, but decreased phosphate concentrations at high doses because of its phytotoxicity. Based on our results, the minimum P/As molar ratios should be at least 1.2 in soil solution or 1.0 in fern fronds for the growth of Chinese brake. Our findings suggest that phosphate application may be an important strategy for efficient use of Chinese brake to phytoremediate arsenic contaminated soils. Further study is needed on the mechanisms of interactive effects of arsenate and phosphate on Chinese brake in hydroponic systems.     

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。     

Behavior of chloroplasts and chloroplast nuclei during spermatogenesis in the fern,Pteris vittata L.

Summary The fate of the chloroplasts and chloroplast nuclei (cp-nuclei) was followed during spermatogenesis in the fernPteris vittata L. by epifluorescence microscopy after staining with 4-6-diamidino-2-phenylindole (DAPI) and by quantitation of chloroplast DNA (cp-DNA) by fluorimetry using a video intensified microscope photon counting system (VIMPICS). The spores were grown on solid medium that contained antheridiogen (An_ptd), and formed an antheridium initial on the protonema cell. The antheridium initial divided and produced 16 spermatocytes and 3 surrounding cells. The chloroplasts in the spermatocytes decreased in volume as cell division was repeated, until finally the volume of each chloroplast was 1/15 of that of the primary chloroplasts. The DNA content of the chloroplasts was also reduced to 1/5 of the original value and when the sperm matured, the fluorescence of cp-DNA disappeared. In the 16-cell spermatocyte, the recognition of the fluorescence of chlorophyll in the chloroplasts with a green excitation filter became difficult. But, the plastids could be observed until the final stage of the sperm. From these observations, it appears that there are two steps in the metamorphosis of chloroplasts during spermatogenesis in the fern. The first step involves the decrease in the volume of chloroplasts, accompanied by reduction of the DNA content, and the second step involves the change of the physical state of chloroplasts to amyloplasts and the disappearance of the cp-DNA from the amyloplasts.     

Arsenic-resistant bacteria solubilized arsenic in the growth media and increased growth of arsenic hyperaccumulator Pteris vittata L

The role of arsenic-resistant bacteria (ARB) in arsenic solubilization from growth media and growth enhancement of arsenic-hyperaccumulator Pteris vittata L. was examined. Seven ARB (tolerant to 10 mM arsenate) were isolated from the P. vittata rhizosphere and identified by 16S rRNA sequencing as Pseudomonas sp., Comamonas sp. and Stenotrophomonas sp. During 7-d hydroponic experiments, these bacteria effectively solubilized arsenic from the growth media spiked with insoluble FeAsO? and AlAsO? minerals (from < 5 μg L?1 to 5.04-7.37 mg L?1 As) and enhanced plant arsenic uptake (from 18.1-21.9 to 35.3-236 mg kg?1 As in the fronds). Production of (1) pyochelin-type siderophores by ARB (fluorescent under ultraviolet illumination and characterized with thin layer chromatography) and (2) root exudate (dissolved organic C) by P. vittata may be responsible for As solubilization. Increase in P. vittata root biomass from 1.5-2.2 to 3.4-4.2 g/plant dw by ARB and by arsenic was associated with arsenic-induced plant P uptake. Arsenic resistant bacteria may have potential to enhance phytoremediation of arsenic-contaminated soils by P. vittata.     

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.     

Hydathode structure and localization in Pteris vittata fronds and evidence for their involvement in arsenic leaching

Pteris vittata, a fern able to hyperaccumulate arsenic (As) in its fronds, has been object of a number of studies aimed to understand the mechanisms involved in As absorption and tolerance. This study has focused on a new mechanism, As leaching, already observed in P. vittata, but not explained, based on the possible involvement of hydathodes, not yet described in this fern; moreover, the results contained in this article will provide information on a more detailed frond anatomy of P. vittata. A combination of light and electron microscopic techniques (transmission electron microscopy and environmental scanning electron microscopy with energy dispersive X-ray) and chemical analyses (inductively coupled plasma atomic emission spectroscopy) was used. The results suggest that in phytoremediation processes under field conditions there could be the need to know the atmospheric conditions before harvesting plants, to avoid the loss of As in the environment.     

Arsenic hyperaccumulation and localization in the pinnule and stipe tissues of the gold-dust fern (Pityrogramma calomelanos (L.) Link var. austroamericana (Domin) Farw.) using quantitative micro-PIXE spectroscopy

Spatial distribution patterns of arsenic (As) in the tissues of a lesser-known As hyperaccumulating fern Pityrogramma calomelanos (L.) Link var. austroamericana (Domin) Farw. (Pteridaceae) have been studied. Quantitative micro-proton-induced X-ray emission (micro-PIXE) spectroscopy was employed to examine As localization in pinnule and stipe cross-sections of this species. In addition, As hyperaccumulation status of P. calomelanos var. austroamericana was compared with the well-known As hyperaccumulating fern Pteris vittata L. Both species were grown in pots under controlled conditions and exposed to four levels of As (0–500 mg As kg⁻¹) for 20 weeks. Pityrogramma calomelanos var. austroamericana accumulated up to 16 415 mg As kg⁻¹ dry weight (DW), however, phytotoxicity symptoms such as necrotic pinnule tips and margins, appeared in fronds with concentrations >3,008 mg As kg⁻¹ DW. Arsenic was readily translocated to fronds, with concentrations up to 75 times greater in fronds than in roots. Quantitative elemental maps of As generated using micro-PIXE analysis revealed that As concentrations in pinnule cross-sections were higher than in stipe cross-sections with concentrations of 3.7 × 10³ and 1.6 × 10³ mg As kg⁻¹ DW, respectively (as determined by region selection analysis; RSA). In pinnules, RSA revealed variable concentrations of As, however did not resolve a clear pattern of compartmentalization across different anatomical regions. In stipe tissues, As concentrations followed the order vascular bundle > cortex > epidermis (as determined by RSA). Our results show that P. calomelanos var. austroamericana is an As hyperaccumulator and has the potential for use in phytoremediation of soils with low levels (up to 50 mg kg⁻¹) of As contamination.     

砷、钙对蜈蚣草中金属元素吸收和转运的影响

蜈蚣草是砷的超富集植物和钙质土壤的指示植物。本试验在砂培条件下,研究砷、钙对蜈蚣草吸收和转运必需金属元素K、Mg、Mn、Fe、Zn和Cu的影响。结果表明。提高营养液中的砷浓度显著降低根部Mg和Zn的吸收。但对根部其它元素的浓度没有明显影响;叶柄中的Mn和地上部的Fe浓度因介质中添加砷而显著减少。其它元素在地上部的分布不受抑制。添加砷限制Fe从地下部向地上部转运,但促进其从叶柄向羽叶中运输;另外,还显著促进Mn由叶柄向羽叶和Zn由根向羽叶的转运。提高钙处理浓度对蜈蚣草吸收Fe、Zn、Cu无显著影响,但显著限制K、Mg和Mn的吸收。Mn是研究的6种金属元素中惟一一种明显向地上部转运富集的元素。从根部到羽叶中。金属元素间的相关性增强,在根部Ca与各种金属元素都无相关性;叶柄中Ca和Fe浓度呈极显著正相关;在羽叶中,Ca与K、Mg、Mn和Zn浓度呈显著负相关。     

An arsenic hyperaccumulating fern,Pteris vittata L. (Pteridaceae) broadly affects terrestrial invertebrate abundance

1. The Chinese brake fern (Pteris vittata L.; Pteridaceae) can accumulate up to 27 000 mg kg^?1 dry wt. of arsenic (As) from the soil into its above‐ground biomass. They may use this As to deter invertebrate threats. 2. This study explored how As concentrations [As] in the fern, and in soil associated with the fern, influenced the abundance and composition of various invertebrates. 3. Populations of P. vittata were identified in the field. Soils from the base of the fern and from 3 m away of each plant were collected and pitfall traps were installed. Soil and fern arsenic concentrations ([As]) were measured via inductively coupled plasma mass spectrometry and invertebrates were identified to order and classified by feeding guild. 4. Increased [As] did not affect all feeding guilds and orders equally. For example, individual herbivore abundance did not decrease as [As] increased, but predator abundance did. In many cases, the impact of soil [As] on invertebrates depended on the distance from the fern. Fern [As] also influenced components of the community, but only at 3 m away from the fern. Furthermore, the abundances of many invertebrate groups were higher beneath the fern, where [As] was higher. 5. These results suggest that hyperaccumulated As can impact the invertebrate community, but the defensive benefits of hyperaccumulation are more complex than have been previously described. The authors advocate that future studies examining the potential defensive benefits of hyperaccumulation should do so in a natural setting that incorporates this complexity and invertebrate richness.     

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.     

Phytoremediation of arsenic contaminated soil by Pteris vittata L. I. Influence of phosphatic fertilizers and repeated harvests

A greenhouse experiment was conducted to evaluate the effectiveness of diammonium phosphate (DAP), single superphosphate (SSP) and two growing cycles on arsenic removal by Chinese Brake Fern (Pteris vittata L.) from an arsenic contaminated Typic Haplustept of the Indian state of West Bengal. After harvest of Pteris vittata the total, Olsen's extractable and other five soil arsenic fractions were determined. The total biomass yield of P. vittata ranged from 10.7 to 16.2 g pot(-1) in first growing cycle and from 7.53 to 11.57 g pot(-1) in second growing cycle. The frond arsenic concentrations ranged from 990 to 1374 mg kg(-1) in first growing cycle and from 875 to 1371 mg kg(-1) in second growing cycle. DAP was most efficient in enhancing biomass yield, frond and root arsenic concentrations and total arsenic removal from soil. After first growing cycle, P. vittata reduced soil arsenic by 10 to 20%, while after two growing cycles Pteris reduced it by 18 to 34%. Among the different arsenic fractions, Fe-bound arsenic dominated over other fractions. Two successive harvests with DAP as the phosphate fertilizer emerged as the promising management strategy for amelioration of arsenic contaminated soil of West Bengal through phyotoextraction by P. vittata.     

耐砷促生菌对超富集植物蜈蚣草砷吸收及根际微生物群落的调控作用

【目的】分析添加外源促植物生长微生物(plant growth-promoting bacteria,PGPB)对植物生长、砷富集和根际微生物的影响,为植物-微生物联合修复砷污染土壤提供参考。【方法】通过添加外源PGPB,研究蜈蚣草生物量和砷富集量与外源微生物促植物生长特性的关系,采用高通量测序技术分析蜈蚣草根际微生物群落在外源PGPB干预下的变化规律。【结果】2株根际菌(假单胞菌PG12、芽孢杆菌R19)和1株内生菌(恶臭假单胞菌S6)具备典型的促植物生长特性,对蜈蚣草的促生作用顺序为：PG12>S6>R19,与对照组相比,生物量分别提高了234%(P<0.01)、136%(P<0.01)和67%;添加外源PGPB后,蜈蚣草砷含量从对照的18.50 mg提高到了31.25-46.95 mg,增幅高达153%(PG12)和139%(S6),对应的蜈蚣草砷浓度从2616.34mg/kg降至1348.04-2 156.23 mg/kg,呈现出典型的砷“稀释效应”;α多样性指数Sobs、Chao和Ace显示,仅R19处理显著提高了根际微生物的群落多样性,而β多样性指数...