Effect of pH,Temperature, and Lead Concentration on the Bioremoval of Lead from Water Using Lemna Minor

This study examined the ability of the aquatic plant Lemna minor (duckweed) to remove soluble lead under various laboratory conditions. In a batch process L. minor was exposed to different pH values (4.5–8.0) and temperature (15–35°C) in presence of different lead concentrations (0.1–10.0 mg L^?1) for 168 h. The amount of biomass obtained in the study period on a dry weight basis, the concentrations of lead in tissue and in medium and net uptake of lead by Lemna all have been determined in each condition. The percentages of lead uptake ratios (PMU) and bioconcentration factors (BCF) were also calculated for these conditions. Bioaccumulated lead concentrations and the PMU were obtained at lowest pH of 4.5, and at 30°C. The highest accumulated lead concentration was found at pH 4.5 as 3.599 mg Pb g^?1 in 10.0 mg L^?1. It decreased to pH 6.0, but it did not change at pH 6.0–8.0 range. The maximum lead accumulation was obtained at 30°C as 8.622 mg Pb g^?1 in 10 mg L^?1 at pH 5.0, and the minimum was at 15°C as 0.291 mg g^?1 in 0.1 mg L^?1. Lead accumulation gradually increased with increasing lead in medium, but the opposite trend was observed for PMU. Lead accumulation increased up to 50 mg L^?1, but did not change significantly in the 50.0–100.0 mg L^?1 range. The lead uptake from water was modeled and the equation fit the experimental data very well.     

Callus Induction and Plant Regeneration in Lemna Minor L.

Callus induction was obtained on Murashige and Skogg agar medium with 45 M 2,4-dichlorophenoxyacetic acid under dark at 25°C. Among the four explant types investigated, the best callus induction was obtained from two-week old fronds to which a surgical incision was applied in the basal (meristematic) region. This treatment resulted in 89.11% of fronds producing callus which continued to proliferate for another 24 months. To obtain plant regeneration pieces of calluses were transferred onto Murashige and Skoog agar medium containing 22 M indole-3-acetic acid and 4.6 M kinetin and maintained under 16-h photoperiod (irradiance of 30 mol m^–2 s^–1) at 23°C. Green fronds formed on all callus pieces. The regenerated fronds were later transferred onto Wang medium where they formed roots. The regenerated Lemna minor L. plants obtained through indirect organogenesis did not differ morphologically from individuals forming the stock collection.     

Correlations in metal release profiles following sorption by Lemna minor

Following the rapid uptake of contaminants in the first few hours of exposure, plants typically attempt to cope with the toxic burden by releasing part of the sorbed material back into the environment. The present study investigates the general trends in the release profiles of different metal(loid)s in the aquatic macrophyte Lemna minor and details the correlations that exist between the release of metal(loid) species. Water samples with distinct contamination profiles were taken from Nilüfer River (Bursa, Turkey), Yeniça?a Lake (Bolu, Turkey), and Bey?ehir Lake (Konya, Turkey) and used for release studies; 36 samples were tested in total. Accumulation and release profiles were monitored over five days for 11 metals and a metalloid (²⁰⁸Pb, ¹¹¹Cd, ⁵²Cr,⁵³Cr,⁶⁰Ni,⁶³Cu,⁶⁵Cu,⁷⁵As,⁵⁵Mn, ¹³⁷Ba, ²⁷Al, ⁵⁷Fe, ⁶⁶Zn,⁶⁸Zn) and correlation, cluster and principal component analyses were employed to determine the factors that affect the release of these elements. Release profiles of the tested metal(loid)s were largely observed to be distinct; however, strong correlations have been observed between certain metal pairs (Cr/Ni, Cr/Cu, Zn/Ni) and principal component analysis was able to separate the metal(loid)s into three well-resolved groups based on their release.     

Aqueous two-phase systems

Biphasic systems formed by mixing of two polymers or a polymer and a salt in water can be used for separation of cells, membranes, viruses, proteins, nucleic acids, and other biomolecules. The partitioning between the two phases is dependent on the surface properties and conformation of the materials, and also on the composition of the two-phase system. The mechanism of partitioning is, however, complex and not easily predicted. Aqueous two-phase systems (ATPS) have proven to be a useful tool for analysis of biomolecular and cellular surfaces and their interactions, fractionation of cell populations, product recovery in biotechnology, and so forth. Potential for environmental remediation has also been suggested. Because ATPS are easily scalable and are also able to hold high biomass load in comparison with other separation techniques, the application that has attracted most interest so far has been the large-scale recovery of proteins from crude feedstocks. As chemicals constitute the major cost factor for large-scale systems, use of easily recyclable phase components and the phase systems generated by a single-phase chemical in water are being studied.     

The distribution of lead in duckweed (Lemna minor L.) root tip

While considerable information on lead distribution in the cells of terrestrial plants has been collected, little is known about lead localization in the cells of the aquatic plant. Lemna minor L. (duckweed) roots were examined using X-ray microanalysis. After 1-h treatment with lead, its concentration was the highest in small vacuoles. After 6 and 12 h, the lead content of cell walls gradually increased. The changes of lead level between vacuoles and cell walls may result from redistribution of this metal from symplast to cell walls or it may reflect increased apoplastic transport. Lead was not found in the ground cytoplasm of any variants of the experiments. This fact and presence of lead in small vesicles suggests that endocytosis may play the role in lead uptake in Lemna.     

Genetic transformation of duckweed Lemna gibba and Lemna minor

Summary We developed efficient genetic transformation protocols for two species of duckweed, Lemna gibba (G3) and Lemna minor (8627 and 8744), using Agrobacterium-mediated gene transfer. Partially differentiated nodules were co-cultivated with Agrobacterium tumefaciens harboring a binary vector containing β-glucuronidase and nptII expression cassettes. Transformed cells were selected and allowed to grow into nodules in the presence of kanamycin. Transgenic duckweed fronds were regenerated from selected nodules. We demonstrated that transgenic duckweed could be regenerated within 3 mo. after Agrobacterium-mediated transformation of nodules. Furthermore, we developed a method for transforming L. minor 8627 in 6 wk. These transformation protocols will facilitate genetic engineering of duckweed, ideal plants for bioremediation and large-scale industrial production of biomass and recombinant proteins.     

Phytoremediation Potential of Duckweed (Lemna minor L.) On Steel Wastewater

An eco-friendly and cost effective technique- phytoremediation was used to remediate contaminants from waste water. This study demonstrated that phytoremediation ability of duckweed (Lemna minor L.) to remove chloride, sulphate from Biological Oxygen Treatment (BOT) waste water of coke oven plant. The BOT water quality was assessed by analyzing physico-biochemical characters – pH, Biological oxygen demand (BOD), Chemical oxygen demand (COD), total dissolved solids (TDS) and elemental concentration. It was observed that an increase in pH value indicated an improvement of water quality. The experimental results showed that, duckweed effectively removed 30% chloride, 16% sulphate and 14% TDS from BOT waste water, which suggested its ability in phytoremediation for removal of chloride and sulphate from BOT waste water. A maximum increase of 30% relative growth rate of duckweed was achieved after 21 days of experiment. Thus, it was concluded that duckweed, an aquatic plant, can be considered for treatment of the effluent discharged from the coke oven plant.     

Applications of the aquatic higher plant Lemna gibba for ecotoxicological assessment

Although higher plants represent a significant portion of the total biomass in some aquatic environments, their use in ecosystem evaluation has lagged behind that of other organisms. This is partly due to a lack of convenient aquatic higher plant systems that can be employed for ecotoxicological assessment. However, the aquatic C-3 monocot Lemna gibba has many attributes that makes it useful for ecosystem health assessment. In this report, using examples from the literature and our research, some of the applications Lemna has for environmental research are discussed. Toxicant impacts on Lemna can be readily assessed in terms of growth; the plants multiply quickly and changes in biomass (which doubles approximately every 2 days) can be accurately measured by counting leaves. The plants are small, allowing for simultaneous multiple replication. The small size also makes the lighting conditions easy to control; sunlight can be accurately simulated and specific spectral regions can be enhanced or deleted. Lemna is amenable to in vitro chlorophyll and photosynthesis assays, which make excellent companion endpoints for growth. The plants assimilate chemicals directly from the growth medium, facilitating controlled toxicant application. Furthermore, Lemna has a high bioconcentration capacity, indicating a potential for use in bioremediation technologies.     

Engineering triacylglycerol accumulation in duckweed (Lemna japonica)

Duckweeds are amongst the fastest growing of higher plants, making them attractive high-biomass targets for biofuel feedstock production. Their fronds have high rates of fatty acid synthesis to meet the demand for new membranes, but triacylglycerols (TAG) only accumulate to very low levels. Here we report on the engineering of Lemna japonica for the synthesis and accumulation of TAG in its fronds. This was achieved by expression of an estradiol-inducible cyan fluorescent protein-Arabidopsis WRINKLED1 fusion protein (CFP-AtWRI1), strong constitutive expression of a mouse diacylglycerol:acyl-CoA acyltransferase2 (MmDGAT), and a sesame oleosin variant (SiOLE(*)). Individual expression of each gene increased TAG accumulation by 1- to 7-fold relative to controls, while expression of pairs of these genes increased TAG by 7- to 45-fold. In uninduced transgenics containing all three genes, TAG accumulation increased by 45-fold to 3.6% of dry weight (DW) without severely impacting growth, and by 108-fold to 8.7% of DW after incubation on medium containing 100 μm estradiol for 4 days. TAG accumulation was accompanied by an increase in total fatty acids of up to three-fold to approximately 15% of DW. Lipid droplets from fronds of all transgenic lines were visible by confocal microscopy of BODIPY-stained fronds. At a conservative 12 tonnes (dry matter) per acre and 10% (DW) TAG, duckweed could produce 350 gallons of oil/acre/year, approximately seven-fold the yield of soybean, and similar to that of oil palm. These findings provide the foundation for optimizing TAG accumulation in duckweed and present a new opportunity for producing biofuels and lipidic bioproducts.     

Photosynthesis in ozone-exposed duckweed (Lemna gibba)

The photosynthetic light saturation curve in duckweed was lowered by 20–25% after ozone exposure (300 nmol mol⁻¹, 1 h). The light flux and oxygen concentration during ozone-exposure had no effect on reduction of net photosynthesis. Net photosynthesis and photorespiration were both depressed by about 40% after exposure for 1 h to 360 nmol mol⁻¹ ozone. We could not find any change in dark respiration after ozone exposure below 300 nmol mol⁻¹. When the concentration of ozone was doubled from 150 nmol mol⁻¹ to 300 nmol mol⁻¹, the uptake of ozone in duckweed changed from 100 nmol m⁻² s⁻¹ to 170 nmol m⁻² s⁻¹. We found no differences in fluorescence (pattern) between ozone treated plants and the control plants during a period of 150 min after ozone treatment, but there was an increase in synthesis of the Dl-protein and a significant reduction in degradation after ozone treatment (300 nmol mol⁻¹, 1 h). These results, together with fluorescence measurements, indicate that photochemical electron transport was not responsible for the ozone-induced reduction in net photosynthesis.     

Enhanced Removal of Lead from Soil Using Biosurfactant Derived from Edible Oils

ABSTRACT

Lead contamination in soil due to anthropogenic activities has amplified and therefore, remediation is of prime significance due to its nonbiodegradability and toxicity effects. This study focuses on lead removal from the soil collected from a rifle range using biosurfactants produced from native microorganisms and edible oils. Native microorganisms in contaminated soil served as a source for biosurfactant production aided by edible vegetable oils such as palm oil and gingelly oil. Preliminary isolation and characterization studies indicated the presence of Pseudomonas aeruginosa that produced biosurfactant and removed lead simultaneously. Batch adsorption experiments showed 96%–99.6% of lead adsorption following Langmuir isotherm model. Lead desorption of 23.6% occurred without biosurfactant. Whereas in the presence of biosurfactants, enhanced desorption of 62.3% was observed. Of both palm oil and gingelly oil derived biosurfactants, the former reached a lead removal efficiency of 93.6% indicating the feasibility and effectiveness of the biosurfactants for contaminated site remediation.     

Phytoremediation potential of Lemna minor L. for heavy metals

Phytoremediation potential of L. minor for cadmium (Cd), copper (Cu), lead (Pb), and nickel (Ni) from two different types of effluent in raw form was evaluated in a glass house experiment using hydroponic studies for a period of 31 days. Heavy metals concentration in water and plant sample was analyzed at 3, 10, 17, 24, and 31 day. Removal efficiency, metal uptake and bio-concentration factor were also calculated. Effluents were initially analyzed for physical, chemical and microbiological parameters and results indicated that municipal effluent (ME) was highly contaminated in terms of nutrient and organic load than sewage mixed industrial effluent (SMIE). Results confirmed the accumulation of heavy metals within plant and subsequent decrease in the effluents. Removal efficiency was greater than 80% for all metals and maximum removal was observed for nickel (99%) from SMIE. Accumulation and uptake of lead in dry biomass was significantly higher than other metals. Bio-concentration factors were less than 1000 and maximum BCFs were found for copper (558) and lead (523.1) indicated that plant is a moderate accumulator of both metals. Overall, L. minor showed better performance from SMIE and was more effective in extracting lead than other metals.     

A Novel Microbial Biofilm for Bioremoval of Nickel from Aqueous Media

This study evaluated the efficacy of a microbial biofilm in removing Ni ions in aqueous media. The biofilm was developed incorporating a garden soil fungus with a bacterium isolated from Ni-rich serpentinite soil. The biofilm was characterized using microscopy, scanning electron microscopy, Fourier transform infrared (FTIR) investigations, and Boehm and potentiometric titrations. Ni removal was determined using batch experiments as a function of pH, Ni concentration, and time. The adsorption isotherm assay was conducted with varying Ni concentrations from 25 to 500 mg/L for 4 days. Isotherm and kinetic modeling were applied to the experimental data to understand the mechanisms of Ni removal. The zero point charge at pH 4.5 indicated the pH values greater than 4.5 is favorable for Ni adsorption. Acidic nature of the biofilm was reflected from Boehm titration data showing higher number of acidic groups than basic groups. With the increase in initial Ni concentration, the uptake increased from 3.43 to 38.16 mg/g. Hill, the best-fitted isotherm model, indicated a maximum adsorption capacity of 165.37 mg/g. After 4 days, the adsorption rate reached an equilibrium with a maximum sorption of ～30 mg/g for an initial concentration of 100 mg/L. Kinetic model fitting with Power function further demonstrated the chemisorptive interaction of Ni with the biofilm surface. A clear involvement of functional groups of the biofilm in Ni bonding was observed from the attenuated total reflection (ATR)-FTIR spectrum. The microbial biofilm showed an efficient but slow removal of Ni from aqueous media.     

Diatom ecology in the phyllosphere of the common duckweed (Lemna minor L.)

Epiphyton associated with thick, floating mats of the common duckweed (Lemna minor L.) was studied at four sites in western Canada between 1985 and 1988. Maximum epiphyton abundance generally occurred in spring as biomass of the duckweed mat was increasing. Epiphytic biomass was low during summer and increased at some sites in autumn with mat decomposition. The community was composed mostly of diatoms and, during summer, photosynthetic bacteria. Species richness of the diatom flora was low, suggesting that duckweed mats are environments to which few species are adapted. Photosynthesis - irradiance curves indicated that duckweed epiphyton was not adapted to low light levels that occurred in the mat (< 1 % of ambient), suggesting they may survive via other means of nutrition. The mat phyllosphere was also characterized by wide spatial and temporal variation in temperature, and sharp vertical profiles of dissolved oxygen and nutrients.     

双水相电泳分离蛋白质的研究

近几年来,随着生物技术的迅速发展,制备型电泳技术的研究得到了重视。然而由于技术上的原因,大规模的制备型电泳技术的研究还未能取得突破。阻碍电泳放大的一个主要问题是由于电加热作用而导致的热对流对电泳分离的破坏。为解决这一问题,人们提出了许多方法。例如,在太空的微重力环境下进行电泳,应力稳定自由流动电泳,循环等电聚焦和区带电泳,色谱电泳和等电膜等电聚焦等。这些方法在电泳放大上都取得了一定的进展,但各有其局限性。最近,Clark提出利用双水相的液液界面阻止热对流的设想,为开发大规模的制备型电泳技术开辟了一条新途径、Raghava Rao等在两种双水相体系上施加电场后成倍地缩短了分相时间。Levine和Bier采用U型管电泳装置研究了双水相体系中血红蛋白的电泳迁移率,观测到界面有阻滞作用。Clark在柱型电泳装置中进行了一组双水相萃取肌红蛋白的简单实验。在10mA的恒电流下电泳40min之后,肌红蛋白的分配系数为7.5,而当电场反向后,分配系数变为0.04,界面阻力并不显著,两者结论并不一致。     

pH及无机氮化合物对细脉浮萍生长的影响

研究以细脉浮萍 ( Lemna aequinoctialis)为研究对象 ,在完全培养液的基础上模拟生活污水的氨氮、硝酸氮浓度和 p H范围 ,研究了培养条件对其生长的影响。结果表明 ,这个浮萍种对 p H耐受的低限在 4～ 5之间 ,最适 p H范围为 5～ 6。非离子态的氨会对其生长产生抑制作用 ,在氨氮为唯一氮源的条件下 ,非离子态氨的浓度大于 0 .1 mg/L时会对此种的生长产生明显的抑制作用 ,接近 2 mg/L时 ,基本上致死。在研究的氨氮和硝酸氮浓度范围内 ( <40 mg N /L) ,铵根离子和硝酸根离子浓度增加可促进细脉浮萍的生长。氨氮中生长的植株过氧化物酶活性高于硝酸氮中的活性。     

Influence of nutrient addition on growth and accumulation of cadmium and copper in Lemna gibba

Abstract

Aquatic plants have been identified as potentially useful for accumulating and bioconcentrating heavy metals. This study was developed to test the hypothesis that nutrient enrichment enhances the metal tolerance of floating macrophytes. Relative growth rates (RGR), photosynthetic pigments (chlorophyll a, b and carotenoid), malondialdehyde (MDA) content, and electrical conductivity (EC) were measured in Lemna gibba exposed to different cadmium and copper concentrations in laboratory conditions. Relative growth rates were negatively correlated with metal exposure, but nutrient addition suppressed this effect. Photosynthetic pigment levels were negatively correlated with metal exposures, and nutrient addition attenuated chlorophyll decrease in response to metal exposures. MDA content and EC also showed sharp increases at higher concentrations, indicating oxidative stress. This study indicates that nutrient enrichment increases the tolerance of Lemna gibba to metals, and that Lemna gibba is a suitable candidate for the phytoremediation of low-level copper and cadmium pollution.     

Strong growth limitation of a floating plant (Lemna gibba) by the submerged macrophyte (Elodea nuttallii) under laboratory conditions

1. The asymmetric competition for light and nutrients between floating and submerged aquatic plants is thought to be key in explaining why dominance by either of these groups can be stable and difficult to change. 2. Although the shading effect of floating plants on submerged plants has been well documented, the impact of submerged plants on floating plants has been poorly explored hitherto. 3. Here, we used laboratory experiments to examine how submerged plant (Elodea nuttallii) alter nutrient conditions in the water column and how this affects the growth of floating plants (Lemna gibba). 4. We demonstrate that, at higher nutrient concentrations, Lemna is increasingly likely to outcompete Elodea. 5. Under low nutrient concentrations (0.1–2 mg N L^?1) Elodea can strongly reduce the growth of Lemna. Growth of floating plants virtually stopped in some of the experiments with Elodea. 6. Extremely reduced tissue N, Mn, chlorophyll and elongated roots indicated that the growth inhibition of Lemna by Elodea was predominantly caused by the latter’s impact on the nutrient conditions for floating plants. 7. These results strengthen the hypothesis that submerged plants can prevent colonization of a lake by floating plants.     

Effects of artificial ultraviolet-B radiation on growth and fatty acid composition of duckweed (Lemna minor)

1. Duckweed (Lemna minor), collected either in summer or early fall was exposed under laboratory conditions to control (photosynthetically active and UV‐A radiation) or experimental (control plus UV‐B radiation) conditions. 2. Growth and survival were determined by counting the number of green, and brown/white fronds following 1–5 or 11 days of irradiation. Growth of duckweed was impaired by exposure to UV‐B radiation in the fall experiment but not in the summer. 3. Fatty acid compositions were analysed following 5 or 11 days of irradiation and a recovery period of 0, 5, 29 or 40 h. Concentrations of the major fatty acids, palmitic, linoleic (LA) and α‐linolenic (ALA) acids were similar in the summer and fall duckweed collections, but the summer samples had higher concentrations of the desaturation products of LA and ALA. 4. UV‐B exposure had small, but significant, and contrasting effects on duckweed fatty acid concentrations. In the summer experiment, duckweed exposed to UV‐B had slightly lower concentrations of major fatty acids than control duckweed, while the reverse was true in the fall experiments. 5. These minor effects of UV‐B on concentrations of LA and ALA would be unlikely to have a major impact on the supply of these essential fatty acids from duckweed to freshwater food webs.