The Use of DNA Fingerprinting in Ecological Studies of Phragmites australis (Cav.) Trin. ex Steudel

A number of reed populations (Phragmites australis) from natural locations on the shore of Lake Ammersee in the south of Bavaria as well as artificially grown reed plants were analyzed for restriction fragment length polymorphisms (RFLPs) in order to evaluate for further ecological studies the dynamics of the restriction pattern within the genome of plants which propagate themselves asexually. The artificially grown reed plants were raised in four ponds supplemented with different phosphate concentrations (8 μg, 16 μg, 50 μg, and 200 μg/l) from rhizomes taken from a neighbouring fish pond. The genomic DNA of 25 of these plants and the DNA from 37 plants out of 18 stands from the lakeshore was isolated and restricted with the enzyme EcoRI, EcoRV, XbaI and HindIII and hybridized to three probes from a shot-gun cloned reed plasmid library. The probes gave between 15 and 26 bands (DNA fingerprints) of high polymorphic frequency (68% ? 100%) after hybridization with repetitive sequences of restricted DNA. A cluster analysis based on the banding pattern as a measure of genetic similarity (GS) was performed with the two sets of reed plants. The plants from the lakeshore exhibited a clear formation of clusters with GS up to 100%; the 20 plants grown from rhizomes, in contrast, did not show any clear correlation among themselves, neither to the phosphate concentration they grew in, nor to the five plants tested from the pond their rhizomes came from. The value of genetical investigations of the plant genome in context with environmental impact on plants is being discussed.     

Nitrogen and carbohydrate pools of two rhizome types of Phragmites australis (Cav.) Trin. ex Steudel

Carbohydrate and total nitrogen contents as well as free amino acid (FAA) concentrations were determined in distinct types of rhizomes of two genetically homogeneous stands of reed (Phragmites australis (Cav.) Trin. ex Steudel) differing in morphology, productivity and nutrient supply in order to evaluate the storage capacity of vertical rhizomes and expansion rhizomes. The expansion rhizomes possess significantly higher amounts of FAA and of total nitrogen but similar carbohydrate concentrations in comparison to the vertical rhizomes. However, no significant differences were found for total nitrogen, FAA and total carbohydrates between both investigated stands indicating a comparable storage capacity of rhizomes independent of nutrient availability. Only the composition of the FAA pool varied in the alanine/asparagine ratio probably influenced by the oxygen supply of the rhizome/root system.     

The speciation and distribution characteristics of Cu in Phragmites australis (Cav.) Trin ex. Steudel

• Heavy metal allocation and the mechanism(s) of metal sequestration in different clonal organs, micro‐domains and subcellular structures has not been systematically studied for rhizomatous perennial plants. It is thus pertinent to investigate knowledge of the speciation and distribution characteristics of Cu in Phragmites australis to elucidating the mobility of metals in wetland plants after their uptake via root systems so as to facilitate development of strategies to enhance Cu tolerance.
• This study investigated the distributions of Cu in P. australis root, stem and leaf using ICP‐MS, synchrotron‐based X‐ray micro‐fluorescence and X‐ray absorption spectroscopy, then evaluated the effects of Cu on cellular structure and ultrastructure via transmission electron microscopy.
• The results indicate a clear preferential localisation of Cu in the roots as compared with the shoots (stems and leaves). The intensity of Cu in the vascular bundles was higher than that in the surrounding epidermis and the endodermis and parenchyma outside the medullary cavity. The dominant chemical form of Cu in P. australis was similar to Cu citrate.
• The results suggest that although Cu can be easily transported into the vascular tissues in roots and stems via Cu citrate, most of the metal absorbed by plants is retained in the roots because if its high binding to the cell wall, thus preventing metal translocation to aerial parts of the plants. Therefore, P. australis showed a high capacity to accumulate Cu in roots, being therefore a suitable species for phytostabilisation interventions.

     

Factors affecting iron plaque on the roots ofPhragmites australis (Cav.) Trin. ex Steudel

Phragmites australis (the common reed) was collected at six sites in southern Québec and Ontario, Canada, in order to study the accumulation of iron plaque on the roots. The deposition of iron oxides on roots ofP. australis did not correlate directly with soil measurements; however, the amounts of iron-bound-to-carbonates fraction of the soil/sediment, responsible for the iron plaque accumulation, correlated with the % of water, % of organic matter, % of clay and pH of the substrate. Plants located very near flowing water accumulated more iron plaque on the roots than plants in other habitats through the summer; it is hypothesized that carbonates associated with iron come from the flowing water. In wetlands or sites near flowing water, most root iron was found on the surface, as iron plaque, while there was more iron inside the root in dry environments. Radial oxygen loss from the roots is probably the most important source of oxygen for the oxidation of iron.     

Effects of external iron concentration upon seedling growth and uptake of Fe and phosphate by the common reed, Phragmites australis (Cav.) Trin ex. Steudel

The objectives of this study were to determine whether, and to what degree, the aqueous iron concentration in the growing medium affects the growth of, and Fe uptake by, Phragmites australis, and whether the presence of iron in the growing environment affects the uptake of the essential element phosphate. The wetland macrophyte P. australis was grown under laboratory conditions in nutrient solution (0.31 mg L(-1) phosphate) containing a range of iron concentrations (0-50 mg L(-1) Fe). A threshold of iron concentration (1 mg L(-1)) was found, above which growth of P. australis was significantly inhibited. No direct causal relationship between iron content in aerial tissues and growth inhibition was found, which strongly suggests that iron toxicity cannot explain these results. Phosphate concentrations in aerial tissues were consistently sufficient for growth and development (2-3 % d. wt) despite significant variation in concentration of phosphate associated with roots. External Fe concentration had a significant effect on the growth of P. australis and on both Fe and phosphate concentrations associated with roots. However, neither direct toxicity nor phosphate deficiency could explain the reduction in growth above 1 mg L(-1) external Fe concentration     

Arbuscular mycorrhizal fungi effect growth and photosynthesis of Phragmites australis (Cav.) Trin ex. Steudel under copper stress

• Arbuscular mycorrhizal fungi (AMF) is an effective way to remove heavy metals’ inhibition on plants, however, few relevant research attempts have been made to determine the contribution of AMF to the physiological and biochemical changes related to the enhanced copper tolerance of Phragmites australis under metal‐stressed conditions.
• In this study, the effects of AMF inoculation on P. australis under different concentrations of copper stress were investigated according to the changes in the parameters related to growth and development, and photosynthetic charateristics. Then, differentially expressed proteins (DEPs) were evaluated by the Isobaric Tag for Relative and Absolute Quantification (iTRAQ) system, which could accurately quantify the DEPs by measuring peak intensities of reporter ions in tandem mass spectrometry (MS/MS) spectra.
• It was found that AMF inoculation may relieve the photosynthesis inhibition caused by copper stress on P. australis and thus promote growth. Proteomic analysis results showed that under copper stress, the inoculation of R. irregularis resulted in a total of 459 differently‐expressed proteins (200 up‐regulated and 259 down‐regulated) in root buds. In addition, the photosynthetic changes caused by AMF inoculation mainly involve the up‐regulated expression of transmembrane protein–pigment complexes CP43 (photosystem II) and FNR (ferredoxin‐NADP+ oxidoreductase related to photosynthetic electron transport).
• These results indicate that AMF could effectively improve the growth and physiological activity of P. australis under copper stress, and thus provides a new direction and instructive evidence for determining the mechanisms by which AMF inoculation enhances the copper tolerance of plants.

     

Desodification from calcareous saline sodic soil through phytoremediation with Phragmites australis (Cav.) Trin. ex Steud. and gypsum

The reclamation of saline sodic soils requires sodium removal and the phytoremediation is one of the proven low-cost, low-risk technologies for reclaiming such soils. However, the role of Phragmites australis in reclaiming saline sodic soils has not been evaluated extensively. The comparative reclaiming role of P. australis and gypsum was evaluated in a column experiment on a sandy clay saline sodic soil with EC_e 74.7 dS m^?1, sodium adsorption ratio (SAR) 63.2, Na⁺ 361 g kg^?1, and pH 8.46. The gypsum at 100% soil requirement, planting common reed (P. australis) alone, P. australis + gypsum at 50% soil gypsum requirements, and leaching (control without plant and gypsum) were four treatments applied. After 11 weeks of incubation, the results showed that all treatments including the control significantly reduced pH, EC, exchangeable Na⁺, and SAR from the initial values, the control being with least results. The gypsum and P. australis + gypsum were highly effective in salinity (EC_e) reduction, while sodicity (SAR) and Na⁺ reductions were significantly higher in P. australis + gypsum treatment. The reclamation efficiency in terms of Na⁺ (83.4%) and SAR (86.8%) reduction was the highest in P. australis + gypsum. It is concluded that phytoremediation is an effective tool to reclaim saline sodic soil.     

Plant regeneration from embryogenic cultures of Phragmites australis (Cav.) Trin. Ex Steud.

Embryogenic cultures of the common reed [Phragmites australis (Cav.) Trin. Ex. Steud.] were induced on Murashige and Skoog (1962)-based medium with 2% (w/v) sucrose, B5 vitamins and 4.5 μM 2,4-dichlorphenoxyacetic acid. Four independent culture lines, two initiated from stem nodes and two from roots, were established. These cultures underwent somatic embryogenesis. In one line of stem node origin, the somatic embryos germinated and developed into plants, following transfer of embryogenic cultures to Murashige and Skoog (1962)-based medium lacking growth regulators, with 108 ± 17 plants being recovered per 100 mg fresh weight of culture. In other lines, the somatic embryos developed roots, but not shoots. Shoot regeneration via somatic embryogenesis offers potential as anin vitro system for physiological studies, including assessments of the response of common reed to environmental pollutants. This revised version was published online in June 2006 with corrections to the Cover Date.     

用分子标记技术分析不同生态型芦苇的遗传多样性

用分子标记技术对河西走廊4种不同生态型芦苇进行了遗传多态性分子标记分析。从30条ISSR引物和45条RAPD引物中分别筛选出适合4种不同生态型芦苇分析的9条和13条引物,前者共扩增出99条带,多态性位点数为51;后者共扩增出195条带,多态性位点数为87。两种分子标记分析所得的遗传相似系数呈显著正相关(r=0.845,P<0.05)。4种生态型芦苇表现出由水生芦苇经盐渍芦苇向沙丘芦苇逐渐演化的趋势。     

Tissue culture and long-term regeneration of Phragmites australis (Cav.) Trin. ex Steud.

Phragmites australis tissue cultures were initiated from mature seeds on MS medium supplemented with 1 mgl^-1 each of 2,4-D and IAA. Cultures displayed typical embryogenic callus that was compact and bright yellow. Selection for embryogenic callus established long-term regenerable cultures. Removal of auxin from the basal medium allowed numerous complete plants to be recovered from the cultures. Histological study indicated both the presence of embryogenic-type cells and the bipolar development of regenerated plants.     

Zinc, Lead and Cadmium Tolerance, Uptake and Accumulation by the Common Reed,Phragmites australis(Cav.) Trin. ex Steudel

Zinc (Zn), lead (Pb) and cadmium (Cd) tolerance in populationsof seedlings ofPhragmites australisraised from seeds collectedfrom a mine site (Plombières, Belgium) contaminated withZn, Pb and Cd and three ‘clean’ sites (Felixstowe,UK; Wisbech, UK; and Mai Po, Hong Kong) were studied under glasshouseconditions. Small differences were found between the metal-contaminatedpopulation and the three ‘clean’ populations whenseedlings were grown in 1.0 µg  ml^-1Zn and 10.0 µgml^-1Pb treatment solutions. In general, however, different populationsof seedlings showed similar growth responses, metal uptake andindices of Zn, Pb and Cd tolerance when cultured in the samemetal-contaminated media for 89 d or in the same metal treatmentsolutions (ZnSO₄:1.0 and 4.0 µg ml^-1Zn; Pb(NO₃)₂: 10.0and 25.0 µg ml^-1Pb; CdSO₄: 0.5 and 1.0 µg ml^-1Cd)for 3 weeks. There was insufficient evidence to support thehypothesis that the metal-contaminated population has evolvedto a Zn-, Pb- or Cd-tolerant ecotype but the results indicatedsome differentiation between the populations with that fromHong Kong being the least productive under the experimentalconditions used. The implications of the findings on selectionof provenances for use in constructed wetlands for wastewatertreatment are discussed. Metal accumulation; heavy metal tolerance; Phragmites australis; population differentiation     

The effect of sewage sludge flooding on growth and morphometric parameters of Phragmites australis (Cav.) Trin. ex Steudel

Phragmites australis is the most frequently used species in constructed wetlands (CW) for dewatering and mineralization of sludge. Such CW create a very hostile environment for plants because their roots are exposed to permanently anaerobic conditions. Our working hypothesis assumed a simple relationship between the success of reed in colonising sludge flood stands and maximalisation of shoot oxygen production and transport to rhizomes and roots. Morphometric parameters and density of common reed shoots growing in reed bed systems inundated with sewage sludge were investigated. It was proved that flooding with sewage sludge significantly influenced characteristics of common reed populations. In comparison with natural stands, on stands treated with sewage sludge, maximal density of shoots was higher, biomass was twice as high, shoot diameter was significantly greater, and leaf weight ratio of an individual shoot was also significantly higher. Tendency to shorten shoots was noted. Growing period was longer for reed shoots treated with sewage sludge than on natural stands.     

The Effect of pH and Plaque on the Uptake of Cu and Mn in Phragmites australis(Cav.) Trin ex. Steudel

The physico-chemical properties of iron oxyhydroxide plaquesformed on the roots of Phragmites australis under field andlaboratory conditions were determined using electron microscopyand energy dispersive spectrometry (EDS) analysis. Plaques werepresent as an amorphous coating on roots with an uneven distribution.They were shown to be composed of iron in both the field andlaboratory, and phosphorus was found to be adsorbed onto thesurface of plaques formed in the laboratory. The uptake of copperand manganese in the presence and absence of plaque was investigatedunder two different pH regimes. Concentrations of Cu were lowerin the shoots of P. australis in the presence of plaque (565mg kg^-1) than when it was absent (1400 mg kg^-1), under growthconditions of higher pH (6.0). The adsorption of Cu and Mn ontothe plaque surface was not the mechanism by which plaque reducedthe uptake of other metals. Alternatively, the plaque may simplyact as a physical barrier. Under conditions of lower pH (3.5)the activity of hydrogen ions at the root surface interferedwith the movement of metals into the root and masked any potentialeffect of iron plaque. Copyright 2000 Annals of Botany Company Phragmites australis, iron oxyhydroxide plaque, pH, manganese, copper, EDS, SEM, wetlands     

北京地区芦苇资源状态及其多样性

为探明北京地区芦苇(Phragmites australis)的资源状态和多样性, 实地考察北京主要河流、湿地和水库, 发现北京地区芦苇总生长面积已超过600 hm²。芦苇染色体倍性以八倍体为主, 四倍体次之。在面积较大的湿地内, 八倍体单一芦苇群落占据优势地位; 而在城市的浅河内有形态和遗传性多样的混合种群。研究表明, 植物性状和倍性水平之间无显著相关性。在小清河发现了6种形态各异的芦苇克隆, 均属于叶绿体DNA片段的P单倍型; 其单倍体基因组大小为(0.499±0.019) pg, 变异系数为3.8%。这表明表型与单倍型之间也不具相关性。此外, 发现1个具有变叶特性的芦苇, 将其命名为金条芦苇。北京地区芦苇形态和遗传多样性为研究芦苇基因型与环境适应性之间的关系提供了珍贵的资源。     

Interaction of cadmium with glutathione and photosynthesis in developing leaves and chloroplasts of Phragmites australis (Cav.) Trin. ex Steudel

We investigated how the presence of cadmium (Cd) at the emergence of Phragmites australis Trin. (Cav.) ex Steudel plants from rhizomes interacted with leaf and chloroplast physiological and biochemical processes. About 8.5 nmol Cd mg-1 chlorophyll was found in leaves, and 0.83 nmol Cd mg-1 chlorophyll was found in chloroplasts of plants treated with 50 microm Cd. As a result, a 30% loss of chlorophyll was measured concomitantly with a comparable percentage reduction in light-saturated photosynthesis. Rubisco content and activity were lowered by 10% and 60%, respectively. Antioxidant activity was stimulated by Cd treatment and was associated with an increase in the glutathione and pyridine pools, and with a larger pool of reduced glutathione. It is suggested that the glutathione pool and its predominance in the reduced state protected the activity of many key photosynthetic enzymes against the thiophilic binding of Cd. Chloroplast ultrastructure was not significantly altered with 50 microm treatment and the efficiency of photosystem II, measured as the fluorescence ratio Fv/Fm, remained high because F0 and Fm were proportionally decreased. In plants treated with 100 microm Cd, all effects were exacerbated, but Fv/Fm remained close to that of control leaves and the glutathione and pyridine nucleotides pools were lowered. The results suggest that glutathione exerted a direct important protective role on photosynthesis in the presence of Cd.     

Morphology and Genetic Diversity of Phragmites australis in Beijing

为探明北京地区芦苇(Phragmites australis)的资源状态和多样性, 实地考察北京主要河流、湿地和水库, 发现北京地区芦苇总生长面积已超过600 hm²。芦苇染色体倍性以八倍体为主, 四倍体次之。在面积较大的湿地内, 八倍体单一芦苇群落占据优势地位; 而在城市的浅河内有形态和遗传性多样的混合种群。研究表明, 植物性状和倍性水平之间无显著相关性。在小清河发现了6种形态各异的芦苇克隆, 均属于叶绿体DNA片段的P单倍型; 其单倍体基因组大小为(0.499±0.019) pg, 变异系数为3.8%。这表明表型与单倍型之间也不具相关性。此外, 发现1个具有变叶特性的芦苇, 将其命名为金条芦苇。北京地区芦苇形态和遗传多样性为研究芦苇基因型与环境适应性之间的关系提供了珍贵的资源。