Stimulatory role of smoke–water and karrikinolide on the photosynthetic pigment and phenolic contents of micropropagated ‘Williams’ bananas

At low concentrations, smoke–water (SW) and smoke-derived karrikinolide (KAR₁) are compounds with potential cytokinin and auxin-like activity. Their roles on the growth, photosynthetic pigment and phenolic contents of micropropagated ‘Williams’ bananas were investigated in comparison with meta-topolin (mT). Explants were cultured on modified Murashige and Skoog basal media supplemented with either SW (1:125; 1:250; 1:500; 1:1,000; 1:2,000 dilutions) or KAR₁ at four concentrations ranging from 4.8?×?10^?22 to 3.3?×?10^?12?M. After 42?days, growth parameters were measured while the photosynthetic pigments and phenolic contents were quantified using spectrophotometric methods. Chlorophyll a, b and total carotenoid contents were significantly enhanced by KAR₁ (4.8?×?10^?22?M) and SW (1:125 and 1:1,000 dilutions). The pigments in KAR₁-treated plantlets were approximately two-fold to three-fold higher than those in the control and mT-treated plants, respectively. Total phenolic content was highest with KAR₁ at 1.0?×?10^?19?M in the leaves and 7.8?×?10^?17?M in the roots. Furthermore, KAR₁-treated plants at 1.0?×?10^?19?M yielded the highest level of total phenolics (leaves) and proanthocyanidins (roots). At 1:500 dilutions, SW stimulated the highest total flavonoid content in the leaves across all the treatments. Combining mT with either SW (1:500) or KAR₁ (4.8?×?10^?22?M) significantly increased the quantity of secondary metabolites. However, the growth parameters and pigment contents were not improved. Based on the significant role of photosynthetic pigments and phenolic compounds on the defense and survival strategies of plants, current findings will have practical significance for important processes such as acclimatization and survival of micropropagated plants. These results are also demonstrating the potential of SW and KAR₁ as an eliciting agent for secondary metabolite production.     

Phytoextraction of Risk Elements by Willow and Poplar Trees

To characterize the phytoextraction efficiency of two clones of willow trees (Salix x smithiana Willd., Salix rubens) and two clones of poplar trees (Populus nigra x maximowiczii, Populus nigra Wolterson) were planted in contaminated soil (0.4–2.0 mg Cd.kg^?1, 78–313 mg Zn.kg^?1, 21.3–118 mg Cu.kg^?1). Field experiment was carried out in Czech Republic. The study investigated their ability to accumulate heavy metals (Cd, Zn, and Cu) in harvestable plant parts. The poplars produced higher amount of biomass than willows. Both Salix clones accumulated higher amount of Cd, Zn and Cu in their biomass (maximum 6.8 mg Cd.kg^?1, 909 mg Zn.kg^?1, and 17.7 mg Cu.kg^?1) compared to Populus clones (maximum 2.06 mg Cd.kg^?1, 463 mg Zn.kg^?1, and 11.8 mg Cu.kg^?1). There were no significant differences between clones of individual species. BCs for Cd and Zn were greater than 1 (the highest in willow leaves). BCs values of Cu were very low. These results indicate that Salix is more suitable plant for phytoextraction of Cd and Zn than Populus. The Cu phytoextraction potential of Salix and Populus trees was not confirmed in this experiment due to low soil availability of this element.     

Screening of Indigenous Ornamental Species from Different Plant Families for Pb Accumulation Potential Exposed to Metal Gradient in Spiked Soils

Contamination of surface soils with lead (Pb) is a global concern due to the release of hazardous materials containing the metal element. In order to explore ways to remediate contaminated soils with less impact on environment and costs, this study aimed at screening ornamental plant species exposed to Pb gradient in spiked soils for Pb phytoextraction. Twenty-one ornamental plant species that currently grow in Pakistan, were selected to assess their potential for Pb accumulation. Pot experiments were conducted to evaluate the accumulative properties of the different plant species in unspiked control (Pb = 0) and spiked soils with different levels of Pb at 500, 1000, 1500 and 2000 mg Pb kg^?1 of soil. Biotranslocation factor (TF), Enrichment factor (EF) and Bioconcentration factor (CF) were calculated to assess the phytoremediation potential of tested plant species after seven weeks of exposure. Out of 21 plant species, Pelargonium hortorum and Mesembryanthemum criniflorum performed better and accumulated more than 1000 mg Pb kg^?1 of shoot dry biomass when they were grown in 500, 1000 and 1500 mg Pb kg^?1 contaminated soils. Both plants had no significant (P < 0.05) variation in the total dry biomass with increasing soil Pb concentration indicating a high tolerance to Pb. Considering the capacity of Pb accumulation, total dry biomass, TF, EF &; CF indices, Pelargonium hortorum and Mesembryanthemum criniflorum could be considered as Pb hyperaccumulators and could have the potential to be used in phytoremediation.     

Phytoextraction potential of water fern (Azolla pinnata) in the removal of a hazardous dye,methyl violet 2B: Artificial neural network modelling

This study investigated the potential of Azolla pinnata (AP) in the removal of toxic methyl violet 2B (MV) dye wastewater using the phytoextraction approach with the inclusion of an Artificial Neural Network (ANN) modelling. Parameters examined included the effects of dye concentration, pH and plant dosage. The highest removal efficiency was 93% which was achieved at a plant dosage of 0.8 g (dye volume = 200 mL, initial pH = 6.0, initial dye concentration = 10 mg L^?1). A significant decrease in relative frond number (RFN), a growth rate estimator, observed at a dye concentration of 20 mg L^?1 MV indicated some toxicity, which coincided with the plant pigments studies where the chlorophyll a content was lower than the control. There were little differences in the plant pigment contents between the control and those in the presence of dye (5 to 15 mg L^?1) indicating the tolerance of AP to MV at lower concentrations. A three-layer ANN model was optimized (6 neurons in the hidden layer) and successfully predicted the phytoextraction of MV (R = 0.9989, RMSE = 0.0098). In conclusion, AP proved to be a suitable plant that could be used for the phytoextraction of MV while the ANN modelling has shown to be a reliable method for the modelling of phytoextraction of MV using AP.     

Enhanced Phytoremediation of Cadmium-Contaminated Soil by Parthenium hysterophorus Plant: Effect of Gibberellic Acid (GA3) and Synthetic Chelator,Alone and in Combinations

The roles of gibberellic acid (GA₃) and ethylenediaminetetraacetic acid (EDTA) in phytoremediation of cadmium (Cd)-contaminated soil by Parthenium hysterophorus plant was investigated. GA₃ (10^?9, 10^?7, and 10^?5M) was applied as a foliar spray. EDTA was added to soil in a single dose (160 mg/kg soil) and split doses (40 mg/kg soil, four split doses). GA₃ and EDTA were used separately and in various combinations. P. hysterophorus was selected due to its fast growth and unpalatable nature to herbivores to reduce the entrance of metal into the food chain. The Cd phytoextraction potential of the P. hysterophorus plant was evaluated for the first time. Cd significantly reduced plant growth and dry biomass (DBM). GA₃ alone increased the plant growth and biomass in Cd-contaminated soil, whereas EDTA reduced it. GA₃ in combination with EDTA significantly increased the growth and biomass. The highest significant DBM was found in treatment T3 (10^?5M GA₃). All treatments of GA₃ or EDTA significantly enhanced the plant Cd uptake and accumulation compared with control (C1). The highest significant root and stem Cd concentrations were found in the combination treatment T11 (GA₃ 10^?5M + EDTA split doses), whereas in leaves it was found in the EDTA treatments. Cd concentration in plant parts increased in the order of stem < leaves < roots. The combination treatment T9 (GA₃ 10^?7M + EDTA split doses) showed the significantly highest total Cd accumulation (8 times greater than control C1, i.e., only Cd used). The GA₃ treatments accumulated more than 50% of the total Cd in the roots, whereas the EDTA treatments showed more than 50% in the leaves. Root dry biomass showed a positive and significant correlation with Cd accumulation. GA₃ is environment friendly as compared with EDTA. Therefore, further investigation of GA₃ is recommended for phytoremediation research for the remediation of metal-contaminated soil.     

Potential of Mauritius Hemp (Furcraea gigantea Vent.) for the Remediation of Chromium Contaminated Soils

The present study was conducted to evaluate the ability of a high biomass producing, drought tolerant succulent plant Mauritius hemp (Furcraea gigantea Vent.) for its tolerance to different levels of Cr (0, 25, 50, 100 and 200 mg Cr kg soil^?1) and its potential for phytoremediation purposes. Based on the data on inhibition of the growth of plants with Cr, tolerance index and grade of growth inhibition, it was observed that the plant could tolerate up to 50 mg Cr kg ^?1 soil. Absorption of Cr from soil to plant and its translocation into plant tissues were discussed in terms of bio concentration factor (BCF), transfer factor (TF), and translocation efficiency (TE%). Cr was mainly accumulated in the roots and exclusion of Cr was found to be the principal physiological tolerance mechanism followed by a marked increase in proline, ascorbic acid, total free amino acids in the leaf tissue and malic acid in the rhizosphere samples to counter Cr stress. Based on the tissue concentration of Cr (< 300 μg g^?1 in the leaves and TF<1), it was concluded that, Furcraea gigantea could not be considered a hyperaccumulator and therefore unsuitable for phytoextraction of Cr. Nevertheless, Furcraea gigantea could be a suitable candidate for phytostablization of Cr contaminated soils.     

Hyperaccumulator Oilcake Manure as an Alternative for Chelate-Induced Phytoremediation of Heavy Metals Contaminated Alluvial Soils

The ability of hyperaccumulator oilcake manure as compared to chelates was investigated by growing Calendula officinalis L for phytoremediation of cadmium and lead contaminated alluvial soil. The combinatorial treatment T₆ [2.5 g kg^?1oilcake manure + 5 mmol kg^?1 EDDS] caused maximum cadmium accumulation in root, shoot and flower up to 5.46, 4.74 and 1.37 mg kg^?1and lead accumulation up to 16.11, 13.44 and 3.17 mg kg^?1, respectively at Naini dump site, Allahabad (S₃). The treatment showed maximum remediation efficiency for Cd (RR = 0.676%) and Pb (RR = 0.202%) at Mumfordganj contaminated site (S₂). However, the above parameters were also observed at par with the treatment T₅ [2.5 g kg^?1oilcake manure +2 g kg^?1 humic acid]. Applied EDDS altered chlorophyll–a, chlorophyll–b, and carotene contents of plants while application of oilcake manure enhanced their contents in plant by 3.73–8.65%, 5.81–17.65%, and 7.04–17.19%, respectively. The authors conclude that Calendula officinalis L has potential to be safely grown in moderately Cd and Pb-contaminated soils and application of hyperaccumulator oilcake manure boosts the photosynthetic pigments of the plant, leading to enhanced clean-up of the cadmium and lead-contaminated soils. Hence, the hyperaccumulator oilcake manure should be preferred over chelates for sustainable phytoremediation through soil-plant rhizospheric process.     

A two-year field study of phytoremediation using Solanum nigrum L. in China

A two-year in-situ phytoremediation trial was launched in Shenyang Zhangshi (Sewage) Irrigation Area (SZIA). The phytoremediation efficiency of Solanum nigrum L. was determined, by both monitoring the change of soil Cadmium level in the upper 20 cm of soil, and calculating the plant uptake of soil Cd. After two years experimental, by monitoring the soil Cd concentrations, The Cd concentrations decreased on average from 2.75 mg kg^?1to 2.45 mg kg^?1 in the first year and from 2.33 mg kg^?1 to 1.53 mg kg^?1 in the second year, amounting to a decrease by a factor of 10.6% in the first year and 12% in the second year. After two years phytoremediation by S. nigrum, Cd concentrations of the seven experimental plots with S. nigrum growth decreased from 2.75 mg kg^?1 to 1.53 mg kg^?1, a decrease by a factor of 24.9%. And the soil Cd concentration decreased only 2.1% and 1.7% in the bared experimental plot. And the calculating of Cd uptake by S. nigrum shown that, the plants uptake 4.46% and 5.18% of the total soil Cd in 2008 and 2009, while the soil Cd concentrations decreased by a factor of 10.6% in 2008 and 12.1% in 2009.     

Phytoremediation potential of some halophytic species for soil salinity

Phytoremediation potential of six halophytic species i.e. Suaeda nudiflora, Suaeda fruticosa, Portulaca oleracea, Atriplex lentiformis, Parkinsonia aculeata and Xanthium strumarium was assessed under screen house conditions. Plants were raised at 8.0, 12.0, 16.0, and 20.0 dSm^?1 of chloride-dominated salinity. The control plants were irrigated with canal water. Sampling was done at vegetative stage (60–75 DAS). About 95 percent seed germination occurred up to 12 dSm^?1 and thereafter declined slightly. Mean plant height and dry weight plant^?1 were significantly decreased from 48.71 to 32.44 cm and from 1.73 to 0.61g plant^?1 respectively upon salinization. Na⁺/K⁺ ratio (0.87 to 2.72), Na⁺/ Ca²⁺ + Mg²⁺ (0.48 to 1.54) and Cl^?/SO₄^2– (0.94 to 5.04) ratio showed increasing trend. Salinity susceptibility index was found minimum in Suaeda fruticosa (0.72) and maximum in Parkinsonia aculeata (1.17). Total ionic content also declined and magnitude of decline varied from 8.51 to 18.91% at 8 dSm^?1 and 1.85 to 7.12% at 20 dSm^?1 of salinity. On the basis of phytoremediation potential Suaeda fruticosa (1170.02 mg plant^?1), Atriplex lentiformis (777.87 mg plant^?1) were the best salt hyperaccumulator plants whereas Xanthium strumarium (349.61 mg plant^?1) and Parkinsonia aculeata (310.59 mg plant^?1) were the least hyperaccumulator plants.     

A Study on Cadmium Phytoremediation Potential of Indian Mustard,Brassica juncea

The objective of this study was to investigate Cd phytoremediation ability of Indian mustard, Brassica juncea. The study was conducted with 25, 50, 100, 200 and 400 mg Kg^?1 CdCl₂ in laboratory for 21 days and Cd concentrations in the root, shoot and leaf tissues were estimated by atomic absorption spectroscopy. The plant showed high Cd tolerance of up to 400 mg Kg^?1 but there was a general trend of decline in the root and shoot length, tissue biomass, leaf chlorophyll and carotenoid contents. The tolerance index (TI) of plants were calculated taking both root and shoot lengths as variables. The maximum tolerance (TI _shoot = 87.4 % and TI _root = 89.6 %) to Cd toxicity was observed at 25 mg Kg^?1, which progressively decreased with increase in dose. The highest shoot (10791 μg g^?1 dry wt) and root (9602 μg g^?1 dry wt) Cd accumulation was achieved at 200 mg kg^?1 Cd treatment and the maximum leaf Cd accumulation was 10071.6 μg g^?1 dry wt achieved at 100 mg Kg^?1 Cd, after 21 days of treatment. The enrichment coefficient and root to shoot translocation factor were calculated, which, pointed towards the suitability of Indian mustard for removing Cd from soil.     

Brassica juncea tested on urban soils moderately contaminated by lead: Origin of contamination and effect of chelates

Urban garden soils are a potential repository of heavy metal pollution, resulting from either anthropogenic or geogenic origin. The efficiency of phytoextraction was compared on two garden soils with the same texture and topsoil Pb concentration (170 mg kg^?1) but not the same origin: one geogenic, the other anthropogenic. Two varieties of Brassica juncea were tested with citric acid (25 mmol kg^?1) or ethylenediaminetetraacetic acid (EDTA, 2.5 mmol kg^?1). Geogenic Pb was shown to be two times less available than anthropogenic Pb, as a result of which the phytoextraction efficiency was reduced by 59%. Pb mobility in the soil was solely enhanced with EDTA, which increased the Pb concentration in shoots of B. juncea by between 14 and 26 times in comparison with the control. The highest Pb concentration in shoots still remained low, however (i.e., 45 mg kg^?1 dry weight). Regardless of the chelates introduced, B. juncea 426308 accumulated roughly twice as much lead as B. juncea 211000, but only for the anthropogenic contaminated soil. Under these conditions, the amount of Pb accumulated by B. juncea (even when assisted by EDTA) was not high enough to envision achieving soil clean-up within a reasonable time frame.     

Enhanced Phytoremediation Capacity of Chenopodium album L. Grown on Pb-Contaminated Soils Using EDTA and Reduction of Leaching Risk

Leaching of metals due to enhanced mobility during ethylenediaminetetraacetic acid (EDTA)-assisted phytoextraction has been demonstrated as one of the potential hazards associated with this technology. This study was conducted to determine phytoextraction efficiency of Chenopodium album L. for Pb and EDTA-assisted (1.5, 3, and 9 mmol kg^?1) phytoextraction and potential for leaching of Pb. The results demonstrated that BCF_shoot (bioconcentration factor) was relatively higher than the BCF_root. Translocation factor in the shoot was higher than the roots. Thus, plant species would be applicable for Pb phytoextraction. EDTA enhanced translocation of Pb from roots to shoots. Lead content in the plant parts was maximum in the shoot and root of 9EDTA and 3EDTA, respectively. However, there was no significant difference between 3EDTA and 9EDTA. Lead concentration in the plant parts increased significantly from vegetative stage into flowering stage. Lead content taken up by the plant was lowest when EDTA was applied in a single dose. Therefore, application of EDTA in several increments rather than a single split reduced the leaching risk. Totally, optimum phytoextraction was observed when 3 mmol kg^?1 EDTA was added in triple dosage 60 days after the plant cultivation under triple application mode. The results indicated the plant has the potential for Pb phytoextraction, but it should not be used unless the biomass containing such accumulated metal is removed for disposal. Significant improvement over current ETDA-assisted phytoextraction of Pb may be possible but should be implemented cautiously because of environmental risk.     

Impact of chelator-induced phytoextraction of cadmium on yield and ionic uptake of maize

Enhanced phytoextraction uses soil chelators to increase the bioavailability of heavy metals. This study tested the effectiveness of ethylenediaminetetraacetic acid (EDTA) and citric acid in enhancing cadmium (Cd) phytoextraction and their effects on the growth, yield, and ionic uptake of maize (Zea mays). Maize seeds of two cultivars were sown in pots treated with 15 (Cd₁₅) or 30 mg Cd kg^?1 soil (Cd₃₀). EDTA and citric acid at 0.5 g kg^?1 each were applied 2 weeks after germination. Results demonstrated that the growth, yield per plant, and total grain weight were reduced by exposure to Cd. EDTA increased the uptake of Cd in shoots, roots, and grains of both maize varieties. Citric acid did not enhance the uptake of Cd, rather it ameliorated the toxicity of Cd, as shown by increased shoot and root length and biomass. Cadmium toxicity reduced the number of grains, rather than the grain size. The maize cultivar Sahiwal-2002 extracted 1.6% and 3.6% of Cd from soil in both Cd+ EDTA treatments. Hence, our study implies that maize can be used to successfully phytoremediate Cd from soil using EDTA, without reducing plant biomass or yield.     

Effect of bacterial inoculation of strains of pseudomonas aeruginosa,alcaligenes feacalis and bacillus subtilis on germination,growth and heavy metal (cd,cr, and ni) uptake of brassica juncea

Bacterial inoculation may influence Brassica juncea growth and heavy metal (Ni, Cr, and Cd) accumulation. Three metal tolerant bacterial isolates (BCr3, BCd33, and BNi11) recovered from mine tailings, identified as Pseudomonas aeruginosa KP717554, Alcaligenes feacalis KP717561, and Bacillus subtilis KP717559 were used. The isolates exhibited multiple plant growth beneficial characteristics including the production of indole-3-acetic acid, hydrogen cyanide, ammonia, insoluble phosphate solubilization together with the potential to protect plants against fungal pathogens. Bacterial inoculation improved seeds germination of B. juncea plant in the presence of 0.1 mM Cr, Cd, and Ni, as compared to the control treatment. Compared with control treatment, soil inoculation with bacterial isolates significantly increased the amount of soluble heavy metals in soil by 51% (Cr), 50% (Cd), and 44% (Ni) respectively. Pot experiment of B. juncea grown in soil spiked with 100 mg kg^?1 of NiCl₂, 100 mg kg^?1 of CdCl₂, and 150 mg kg^?1 of K₂Cr₂O₇, revealed that inoculation with metal tolerant bacteria not only protected plants against the toxic effects of heavy metals, but also increased growth and metal accumulation of plants significantly. These findings suggest that such metal tolerant, plant growth promoting bacteria are valuable tools which could be used to develop bio-inoculants for enhancing the efficiency of phytoextraction.     

A microcosm investigation of fe (iron) removal using macrophytes of ramsar lake: A phytoremediation approach

The present study deals with the microcosm study of Fe (Iron) phytoremediation using Eichhornia crassipes, Lemna minor, Pistia stratiotes and Salvinia cucullata species collected from the Loktak Lake, a Ramsar Site which exists in north-eastern India (an Indo-Burma hotspot region). Efficiency of these four macrophytes was compared using different Fe concentrations of 1 mg L^?1, 3 mg L^?1 and 5 mg L^?1 for 4 days, 8 days and 12 days, respectively. E. crassipes was the most efficient macrophyte whereas L. minor was the least efficient. E. crassipes removed the highest percentage of Fe, i.e. 89% from 1 mg L^?1, 81.3% from 3 mg L^?1 and 73.2% from 5 mg L^?1 in 12-day experiment.     

Can the use of natural biostimulants be a potential means of phytoremediating contaminated soils from goldmines in South Africa?

Biostimulants offer great potential in improving phytoremediation of contaminated soils. In the current greenhouse-based study, Brassica juncea seedlings grown on soils collected from Krugersdorp Goldmine and the adjourning areas (a Game Reserve and private farmland) were supplemented with different biostimulants (Kelpak® = KEL, vermicompost leachate = VCL, smoke-water = SW). Indole-3-butyric acid (IBA) was included in the study for comparative purposes because these biostimulants are known to enhance rooting. Prior to the pot trial, concentrations of elements in the three soil types were determined using Inductively Coupled Plasma-Optical Emission Spectroscopy. Plants were harvested after 105 days and the growth and concentrations of elements in the various plant organs were determined. TheB. juncea seedlings with and without biostimulants did not survive when growing in soil from the Krugersdorp Goldmine. The Game Reserve and private farmland soils supplemented with KEL produced the highest plant biomass and the lowest accumulation of metals in the organs of B. juncea. High concentrations (>13 000 mg kg^?1) of zinc and aluminium were quantified in the roots of IBA-supplemented soils from the Game Reserve. Generally, IBA and SW enhanced the phytoremediation of B. juncea due to elevated levels of elements that accumulated in their different organs.     

Assessment of plant growth attributes,bioaccumulation, enrichment,and translocation of heavy metals in water lettuce (Pistia stratiotes L.) grown in sugar mill effluent

This study was conducted to assess the pollutant uptake capability of water lettuce (Pistia stratiotes L.) in terms of bioaccumulation, enrichment, and translocation of heavy metals grown in sugar mill effluent. Results showed that the maximum fresh weight (328.48 ± 2.04 gm kg^?1), total chlorophyll content (2.13 ± 2.03 mg g^?1 fwt), and relative growth rate, RGR (11.89 gg^?1 d^?1) of P. stratiotes were observed at 75% concentration of the sugar mill effluent after 60 days of phytoremediation experiment. The bioaccumulation factor (B_F) of different heavy metals was greater than 1 with 50% and 75% concentrations of sugar mill effluent and this indicated that P. stratiotes was hyperaccumulator or phytoremediator of these metals. The enrichment factor (E_F < 2 for Cu, Fe, Cr, Pb, Zn, and Mn) and (E_F > 2 for Cd) indicated that P. stratiotes mineral enrichment deficient and it moderately enriched the different heavy metals. Moreover, translocation factor (T_F) was less than 1 which indicated the low mobility of metals in different parts (root and leaves) of P. stratiotes after phytoremediation. Therefore, P. stratiotes can be used for phytotreatment of sugar mill effluent up to 50% to 75% concentrations and considered as hyperaccumulator aquatic plant for different heavy metals and other pollutants from the contaminated effluents.     

Phytoremediation of wastewater toxicity using water hyacinth (Eichhornia crassipes) and water lettuce (Pistia stratiotes)

This paper elucidates the phytoremediation potential of water hyacinth and water lettuce on the reduction of wastewater toxicity. Acute toxicity tests were performed in an aquarium with a population of Sarotherodon melanotheron, contaminated by different concentrations of wastewaters before and after phytoremediation with Eichhornia crassipes and Pistia stratiotes. Lethal concentrations (LC₅₀) of the fish's population obtained during 24 hours of exposures were determined. COD, BOD, ammonium, TKN and PO₄^3? concentrations in wastewaters were of 1850.29, 973.33, 38.34, 61.49 and 39.23 mg L^?1, respectively, for each plant. Phytoremediation reduced 58.87% of ammonium content, 50.04% of PO₄^3?, 82.45% of COD and 84.91% of BOD. After 15 days of the experiment, metal contents in treated wastewaters decreased from 6.65 to 97.56% for water hyacinth and 3.51 to 93.51% for water lettuce tanks. Toxicity tests showed that the mortality of fish exposed increased with increase in concentration of pollutants in wastewaters and the time of exposure. Therefore, the highest value of LC₅₀ was recorded for fish subjected to 3 hours of exposure (16.37%). The lowest rate was obtained after an exposure of 20 to 24 hours (5.85%). After phytoremediation, the effluents purified by Eichhornia crassipes can maintain the fish life beyond 24 hours of exposure.     

Effect of Different Amendments on Growing of Canna indica L. Inoculated with AMF on Mining Substrate

Canna indica L. (CiL) was used here in phytoremediation of mining soils. Our work evaluated the effect of AMF (i) on the growth and (ii) on the uptake of heavy metals (HM). The tests were conducted in the greenhouse on mining substrates collected from the Kettara mine (Morocco). The mine soil was amended by different proportions of agricultural soil and compost and then inoculated with two isolates of AMF (IN1) and (IN2) of different origins. After six months of culture, the results show that on mining soils (100%) only AMF (IN2) was able to colonize the roots of CiL with a frequency of 40 ± 7% and an intensity of 6.5 ± 1.5%. Also, the lowest values of shoot and root dry biomass are obtained on these mining soils with respectively 0.30 g and 0.27 g. In contrast, the accumulation of HM was higher and reached more than 50% of that contained in the mining soils, the highest values with 138 mg kg^?1 Cu²⁺, Zn²⁺ 270 mg kg^?1 and 1.38 mg kg^?1 Cd was recorded. These results indicate that the colonization of CiL roots by AMF (IN2) could significantly improve its potential to be used in phytoremediation of polluted soil.     

Enhanced accumulation of cadmium in Linum usitatissimum L. plants due to overproduction of metallothionein α-domain as a fusion to β-glucuronidase protein

As most cultivars of flax and linseed (Linum usitatissimum L.) are capable of accumulating cadmium (Cd), they are suitable candidates for phytoextraction of the metal from contaminated soils. In an attempt to enhance the phytoextraction capacity of L. usitatissimum through overproduction of an efficient heterologous Cd-binding peptide, we engineered linseed breeding line AGT 917 to constitutively express genetic fusion of α-domain of mammalian metallothionein 1a (αMT1a) and β-glucuronidase gus gene under the control of CaMV 35S promoter. An improved transformation protocol was developed, which involved co-cultivation of AGT 917 hypocotyl segments with partially removed epidermis with Agrobacterium suspension for 10 min in the presence of 200 mg l^?1 cellulase. The enzyme treatment increased the transformation efficiency (TE) 1.6-fold as compared to agroinfection without cellulase. Less pronounced impact on TE exerted 100 mg l^?1 acetosyringone, increasing TE 1.3-fold. When tested in soils amended with Cd at 20 and 360 mg kg^?1, the mature αMT1a::gus plants accumulated more Cd than parental AGT 917: the stem Cd concentrations in the best performing αMT1/2 line were 3.3- and 1.9-fold higher, respectively. Moreover, hypocotyl explants of αMT1/2 line showed 1.7-fold higher biomass than those of AGT 917 on media containing 15 mg Cd l^?1, indicating that αMT1a::gus did confer higher Cd tolerance to engineered plant. Overproduction of metal-binding peptides thus appears to be a viable strategy for the production of L. usitatissimum with improved phytoremediation capacity.