Phytoextraction of trace elements and physiological changes in Indian mustard plants (Brassica nigra L.) grown in post methanated distillery effluent (PMDE) irrigated soil

The metal accumulation potential and its physiological effects in Indian mustard plants (Brassica nigra L.) grown in soil irrigated with post methanated distillery effluent (25%, 50%, 75%, 100%, v/v) were studied after 30, 60 and 90 days after sowing. An increase in the chlorophyll and protein contents was recorded at the lower concentrations of post methanated distillery effluent (PMDE) at initial exposure periods followed by a decrease at higher concentrations of PMDE compared to their respective controls. An enhanced lipid peroxidation in tested plants was observed, which was evidenced by the increased malondialdehyde content in shoot, leaves and seeds at all the concentrations of PMDE and exposure periods compared to their respective controls. This study revealed that Indian mustard plants (B. nigra L.) are well adopted to tolerate and accumulate high quantities of trace elements due to increased level of antioxidants (cysteine and ascorbic acid) in root, shoot and leaves of the treated plants at all the concentrations and exposure periods except at 90 days, whereas a decrease was observed at 100% PMDE as compared to their respective controls.     

Enhanced uptake of soil Pb and Zn by Indian mustard and winter wheat following combined soil application of elemental sulphur and EDTA

Enhancement of Pb and Zn uptake by Indian mustard (Brassica juncea (L.) Czern.) and winter wheat (Triticum aestivumL.) grown for 50 days in pots of contaminated soil was studied with application of elemental sulphur (S) and EDTA. Sulphur was added to the soil at 5 rates (0–160 mmol kg^?1) before planting, and EDTA was added in solution at 4 rates (0–8 mmol kg^?1) after 40 days of plant growth. Additional pots were established with the same rates of S and EDTA but without plants to monitor soil pH and CaCl₂-extractable heavy metals. The highest application rate of S acidified the soil from pH 7.1 to 6.0. Soil extractable Pb and Zn and shoot uptake of Pb and Zn increased as soil pH decreased. Both S and EDTA increased soil extractable Pb and Zn and shoot Pb and Zn uptake. EDTA was more effective than S in increasing soil extractable Pb and Zn, and the two amendments combined had a synergistic effect, raising extractable Pb to ¿1000 and Zn to ¿6 times their concentrations in unamended control soil. Wheat had higher shoot yields than Indian mustard and increasing application rates of both S and EDTA reduced the shoot dry matter yields of both plant species to as low as about half those of unamended controls. However, Indian mustard hyperaccumulated Pb in all EDTA treatments tested except the treatment with no S applied, and the maximum shoot Pb concentration was 7100 mg kg^?1 under the highest application rates of S and EDTA combined. Wheat showed similar trends, but hyperaccumulation (1095 mg kg^?1) occurred only at the highest rates of S and EDTA combined. Similar trends in shoot Zn were found, but with lower concentrations than Pb and far below hyperaccumulation, with maxima of 777 and 480 mg kg^?1 in Indian mustard and wheat. Despite their lower yields, Indian mustard shoots extracted more Pb and Zn from the soil (up to 4.1 and 0.45 mg pot^?1) than did winter wheat (up to 0.72 and 0.28 mg pot^?1), indicating that the effects of S and EDTA on shoot metal concentration were more important than yield effects in determining rates of metal removal over the growth period of 50 days. Phytoextraction of Pb from this highly contaminated soil would require the growth of Indian mustard for nearly 100 years and is therefore impractical.     

Comparison of tolerance of <Emphasis Type="Italic">Brassica juncea</Emphasis> and <Emphasis Type="Italic">Vigna radiata</Emphasis> to cadmium

The effect of different cadmium concentrations (6–120 μM) on Hill reaction activity (HRA) of isolated chloroplasts, contents of chlorophylls (Chls) and carotenoids (Cars), and Cd uptake and accumulation in plant organs of Indian mustard (Brassica juncea L. cv. Vitasso) and mung bean [Vigna radiata (L.) Wilczek] were determined. The Cd stress inhibited photochemical activity of isolated chloroplasts of both species and in both tested developmental stages. On the basis of EC₅₀ values, the mung bean showed a higher sensitivity to Cd treatment than Indian mustard. The higher sensitivity of both species was determined in the earlier than in the older developmental stage. The leaves of Cd-treated plants possessed lower contents of Chls and Cars in both species and the negative effect increased with Cd concentration. A difference between species was also found in Cd uptake and accumulation. In both species, Cd was accumulated more in roots than in shoots, with higher accumulation in Indian mustard than in mung bean.     

Alleviation of Cadmium Toxicity in Brassica juncea L. (Czern. & Coss.) by Calcium Application Involves Various Physiological and Biochemical Strategies

Calcium (Ca) plays important role in plant development and response to various environmental stresses. However, its involvement in mitigation of heavy metal stress in plants remains elusive. In this study, we examined the effect of Ca (50 mM) in controlling cadmium (Cd) uptake in mustard (Brassica juncea L.) plants exposed to toxic levels of Cd (200 mg L⁻¹ and 300 mg L⁻¹). The Cd treatment showed substantial decrease in plant height, root length, dry weight, pigments and protein content. Application of Ca improved the growth and biomass yield of the Cd-stressed mustard seedlings. More importantly, the oil content of mustard seeds of Cd-stressed plants was also enhanced with Ca treatment. Proline was significantly increased in mustard plants under Cd stress, and exogenously sprayed Ca was found to have a positive impact on proline content in Cd-stressed plants. Different concentrations of Cd increased lipid peroxidation but the application of Ca minimized it to appreciable level in Cd-treated plants. Excessive Cd treatment enhanced the activities of antioxidant enzymes superoxide dismutase, ascorbate peroxidase and glutathione reductase, which were further enhanced by the addition of Ca. Additionally, Cd stress caused reduced uptake of essential elements and increased Cd accumulation in roots and shoots. However, application of Ca enhanced the concentration of essential elements and decreased Cd accumulation in Cd-stressed plants. Our results indicated that application of Ca enables mustard plant to withstand the deleterious effect of Cd, resulting in improved growth and seed quality of mustard plants.     

Overexpression of cystathionine-γ-synthase enhances selenium volatilization in<Emphasis Type="Italic"> Brassica juncea</Emphasis>

Selenium (Se) can be assimilated and volatilized via the sulfate assimilation pathway. Cystathionine--synthase (CGS) is thought to catalyze the synthesis of Se-cystathionine from Se-cysteine, the first step in the conversion of Se-cysteine to volatile dimethylselenide. Here the hypothesis was tested that CGS is a rate-limiting enzyme for Se volatilization. Cystathionine--synthase from Arabidopsis thaliana (L.) Heynh. was overexpressed in Indian mustard [Brassica juncea (L.) Czern & Coss], and five transgenic CGS lines with up to 10-fold enhanced CGS levels were compared with wild-type Indian mustard with respect to Se volatilization, tolerance and accumulation. The CGS transgenics showed 2- to 3-fold higher Se volatilization rates than wild-type plants when supplied with selenate or selenite. Transgenic CGS plants contained 20–40% lower shoot Se levels and 50–70% lower root Se levels than the wild type when supplied with selenite. Furthermore, CGS seedlings were more tolerant to selenite than the wild type. There were no differences in Se accumulation or tolerance from selenate, in agreement with the earlier finding that selenate-to-selenite reduction is rate-limiting for selenate tolerance and accumulation. In conclusion, CGS appears to be a rate-limiting enzyme for Se volatilization. Overexpression of CGS offers a promising approach for the creation of plants with enhanced capacity to remove Se from contaminated sites in the form of low-toxic volatile dimethylselenide.Abbreviations CGS cystathionine--synthase - DMSe dimethylselenide - SeCys selenocysteine - WT wild type     

The isochorismate pathway is negatively regulated by salicylic acid signaling in O<Subscript>3</Subscript>-exposed <Emphasis Type="Italic">Arabidopsis</Emphasis>

Phytochelatins (PCs) are heavy metal binding peptides that play an important role in sequestration and detoxification of heavy metals in plants. In this study, our goal was to develop transgenic plants with increased tolerance for and accumulation of heavy metals from soil by expressing an Arabidopsis thaliana AtPCS1 gene, encoding phytochelatin synthase (PCS), in Indian mustard (Brassica juncea L.). A 35S promoter fused to a FLAG–tagged AtPCS1 cDNA was expressed in Indian mustard, and transgenic lines, designated pc lines, were evaluated for tolerance to and accumulation of Cd and Zn. Transgenic plants with moderate AtPCS1 expression levels showed significantly higher tolerance to Cd and Zn stress, but accumulated significantly less Cd and Zn than wild type plants in both shoot and root tissues. However, transgenic plants with highest expression of the transgene did not exhibit enhanced Cd and Zn tolerance. Shoots of Cd-treated pc plants had significantly higher levels of phytochelatins and thiols than wild-type plants. Significantly lower concentrations of gluthatione in Cd-treated shoot and root tissues of transgenic plants were observed. Moderate expression levels of phytochelatin synthase improved the ability of Indian mustard to tolerate certain levels of heavy metals, but at the same time did not increase the accumulation potential for Cd and Zn.     

Exploring the selenium phytoremediation potential of transgenic Indian mustard overexpressing ATP sulfurylase or cystathionine-gamma-synthase

Indian mustard (Brassica juncea) plants overexpressing ATP sulfurylase (APS transgenics) were previously shown to have higher shoot selenium (Se) levels and enhanced Se tolerance compared to wild type when supplied with selenate in a hydroponic system. Other transgenic Indian mustard overexpressing cystathionine-gamma-synthase (CGS) showed a higher Se volatilization rate, lower shoot Se levels, and higher Se tolerance than wild type, also in hydroponic studies. In the present study, these APS and CGS transgenics were evaluated for their capacity to accumulate Se from soil that is naturally rich in Se. Wild-type Indian mustard and the Se hyperaccumulator Stanleya pinnata were included for comparison. After 10 weeks on Se soil, the APS transgenics contained 2.5-fold higher shoot Se levels than wild type Indian mustard, similar to those of S. pinnata. The CGS transgenics contained 40% lower shoot Se levels than wild type. Shoot biomass was comparable for all Indian mustard types and higher than that of S. pinnata. These results obtained with these transgenics on soil are in agreement with those obtained earlier using hydroponics. The significance of these findings is that they are the first report on the performance of transgenic plants on Se in soil and show the potential of genetic engineering for phytoremediation.     

Experimental complexities in evaluating the comparative phytotoxicity of chemicals with different modes of action

Careful attention should be paid to bioassay experiments to examine the comparative phytotoxicity of chemicals with different modes of actions. Experimental complexities in examining comparative phytotoxicity of chemicals with differing modes of action are determined and discussed to appreciate the importance of relevant secondary effects that can be quickly measured. The chemicals selected were: benzoic acid, isoxaflutole and rimsulfuron. Data on shoot and root length of 7-day-old mustard (Brassica napus L.) seedlings, shoot height of 4-week-old mustard plants and total leaf chlorophyll concentrations were determined when plants were grown in soil treated with different concentrations of the three chemicals. Scanning electron microscope (SEM) studies were conducted to determine any damage to mustard root hairs after treatment. Root growth of 7-day-old mustard seedlings was reduced when treated with different concentrations of benzoic acid or rimsulfuron. Root growth of mustard seedlings, however, largely remained unaffected when plants were grown in soil treated with isoxaflutole. While no significant reduction in either chlorophyll concentration or shoot height of 4-week-old mustard plants was observed when treated with soil-applied benzoic acid, both parameters were inhibited when mustard plants were treated with isoxaflutole or rimsulfuron. SEM studies revealed significant damage to root hairs in a 7-day-old mustard seedlings when plants were grown in soil treated with 500, 1000 and 2000 mg/L benzoic acid, and 0.5 mg/L rimsulfuron. No such damage was observed when mustard was grown in soil treated with isoxaflutole.     

Cadmium tolerance and accumulation in Indian mustard is enhanced by overexpressing gamma-glutamylcysteine synthetase

To investigate rate-limiting factors for glutathione and phytochelatin (PC) production and the importance of these compounds for heavy metal tolerance, Indian mustard (Brassica juncea) was genetically engineered to overexpress the Escherichia coli gshI gene encoding gamma-glutamylcysteine synthetase (gamma-ECS), targeted to the plastids. The gamma-ECS transgenic seedlings showed increased tolerance to Cd and had higher concentrations of PCs, gamma-GluCys, glutathione, and total non-protein thiols compared with wild-type (WT) seedlings. When tested in a hydroponic system, gamma-ECS mature plants accumulated more Cd than WT plants: shoot Cd concentrations were 40% to 90% higher. In spite of their higher tissue Cd concentration, the gamma-ECS plants grew better in the presence of Cd than WT. We conclude that overexpression of gamma-ECS increases biosynthesis of glutathione and PCs, which in turn enhances Cd tolerance and accumulation. Thus, overexpression of gamma-ECS appears to be a promising strategy for the production of plants with superior heavy metal phytoremediation capacity.     

Cadmium and zinc induced chlorosis in Indian mustard [<Emphasis Type="Italic">Brassica juncea</Emphasis> (L.) Czern] involves preferential loss of chlorophyll <Emphasis Type="Italic">b</Emphasis>

Plants of Indian mustard (Brassica juncea) were treated with either 50 μM Cd, 250 μM Zn, or 25 μM Cd+125 μM Zn and the progression of chlorosis in the mature leaves monitored. As relative chlorophyll (Chl) contents in the mature leaves decreased to 75, 50, and 25 % relative to controls, both mature and young leaves were harvested and the Chl pools extracted. The metal treatments caused a greater loss of Chl b than Chl a. As mature leaves underwent progressive chlorosis, the young leaves displayed a characteristic over-greening, due largely to increased content of Chl b. However, as the young leaves began to experience chlorosis, a greater loss of Chl b was also observed. Thus during metal induced chlorosis, there is a preferential turnover of the Chl b pool in mature and young leaves.     

Transport of Cd and Zn to seeds of Indian mustard (Brassica juncea) during specific stages of plant growth and development

The accumulation of excess Cd in the seeds of cereal and other crops compromises their commercial value and presents a potential risk to human health. Indian mustard [ Brassica juncea (L.) Czern.] is a moderate accumulator of heavy metals such as Cd and Zn, and the seeds are consumed throughout the world, particularly in the Indian subcontinent. The study here examined the transport of Cd into Indian mustard plants and to seeds as a function of external Cd and the stage of the life cycle (vegetative growth, flowering and seed set) to identify critical developmental windows where transport from roots to seeds was the greatest. Plants were also treated simultaneously with Zn to determine if Zn fertilization mitigated the transport of Cd to seeds. Plants treated with Cd during the seed set accumulated the highest concentrations of Cd, exceeding 8 mg kg⁻¹ dry weight in some instances. Cadmium accumulated during vegetative growth was not highly redistributed to seeds. No effects of Zn were observed with regard to Cd redistribution to seeds. This may be because of the relatively small Zn : Cd ratios tested. However, the results suggest that if Zn fertilization is to be used to reduce the Cd accumulation in seeds of this species, that plants should be treated during the seed set stage. As the seeds of Indian mustard consistently accumulated Cd to concentrations that exceed acceptable limits for food crops, additional study of Cd redistribution in this species is warranted.     

The role of root damage in the chelate-enhanced accumulation of lead by Indian mustard plants

In the present study, increasing ethylenediaminetetraacetic acid (EDTA) concentration from 0 to 0.5 mmol L(-1) resulted in progressive increases in root elongation and in shoot and root dry matter (DM) of Indian mustard seedlings (Brassica juncea. L.) exposed at 0.5 mmol L(-1) of lead (Pb). The highest concentration of Pb in the shoots of Indian mustard reached 1140 mg kg(-1) dry weight (DW) in the treatment with 0.5 mmol L(-1) of Pb + 0.25 mmol L(-1) of EDTA. A significantly positive correlation was found between the concentrations of Pb and EDTA in the shoots of mustard. Roots were pretreated with an MC (methanol:trichloromethane) solution, 0.1 mol L(-1) of HCl, and 65 degrees C hot water. The plants were then exposed to 0.5 mmol L(-1) of Pb + 3 mmol L(-1) of EDTA in solution for 2 d. The pretreatments with MC, HCl, and hot water all increased the concentration of Pb in shoots by 14-, 7-, and 15-fold, respectively, compared with the shoots that had not been pretreated. Therefore, some physiological damage to roots would be useful to enhance the uptake of metal by plants and to minimize the application of doses of chelates in the practical operation of chelate-assisted phytoremediation.     

Effect of salicylic acid on salinity-induced changes in Brassica juncea

Seeds of Indian mustard (Brassica juncea (L.) Czern.et Coss.) were exposed to 0,50,100 and 150 mmol/L NaCl for 8h and seeds were sown in an earthen pot.These stressed seedlings were subsequently sprayed with 10 μrnol/L salicylic acid (SA) at 30 d and were sampled at 60 d to assess the changes in growth,photosynthesis and antioxidant enzymes.The seedlings raised from the seeds treated with NaCl had significantly reduced growth and the activities of carbonic anhydrase,nitrate reductase and photosynthesis,and the decrease was proportional to the increase in NaCl concentration.However,the antioxidant enzymes (catalase,peroxidase and superoxide dismutase) and proline content was enhanced in response to NaCl and/or SA treatment,where their interaction had an additive effect.Moreover,the toxic effects generated by the lower concentration of NaCl (50 mmol/L) were completely overcome by the application of SA.It was,therefore,concluded that SA ameliorated the stress generated by NaCl through the alleviated antioxidant system.     

Turgor Maintenance by Osmoregulation inBrassica napusandB. junceaunder Field Conditions

Indian mustard (Brassica juncea(L) Czernjacw) maintains higherleaf turgor than canola (B. napusL.) under water deficits andthis is related to the greater yield of mustard under theseconditions. The work reported in this paper was designed tostudy the way mustard maintains this turgor advantage. It wasbased on three field experiments that each used at least twocultivars or lines of each species. The leaf water potentialat which leaves reached zero turgor was consistently lower inmustard than in canola (up to 1.1 MPa lower). This differencearose from a greater rate of decline in leaf osmotic potentialwith declining water potential in mustard rather than from anydifference in the osmotic potential at full turgor. Calculationsof solute accumulation showed that mustard had a greater capacityto osmoregulate than canola, with this capacity being the basisfor its advantage in turgor maintenance. Other differences inplant water relations were consistent with the differences inosmoregulation, with the predicted relative water content ofleaves at an osmotic potential of -2.5 MPa being 0.43 for canolaand 0.61 for mustard. Mustard's greater capacity to accumulatesolutes is concluded to be a major factor in its greater yieldunder water deficits. Brassica napusL.; Brassica juncea(L) Czernjacw; Indian mustard; canola; water deficit; plant water relations; osmoregulation; osmotic adjustment; turgor     

The Role of EDTA in Lead Transport and Accumulation by Indian Mustard

Indian mustard (Brassica juncea) plants exposed to Pb and EDTA in hydroponic solution were able to accumulate up to 55 mmol kg⁻¹ Pb in dry shoot tissue (1.1% [w/w]). This represents a 75-fold concentration of Pb in shoot tissue over that in solution. A threshold concentration of EDTA (0.25 mm) was found to be required to stimulate this dramatic accumulation of both Pb and EDTA in shoots. Below this threshold concentration, EDTA also accumulated in shoots but at a reduced rate. Direct measurement of a complex of Pb and EDTA (Pb-EDTA) in xylem exudate of Indian mustard confirmed that the majority of Pb in these plants is transported in coordination with EDTA. The accumulation of EDTA in shoot tissue was also observed to be directly correlated with the accumulation of Pb. Exposure of Indian mustard to high concentrations of Pb and EDTA caused reductions in both the transpiration rate and the shoot water content. The onset of these symptoms was correlated with the presence of free protonated EDTA (H-EDTA) in the hydroponic solution, suggesting that free H-EDTA is more phytotoxic than Pb-EDTA. These studies clearly demonstrate that coordination of Pb transport by EDTA enhances the mobility within the plants of this otherwise insoluble metal ion, allowing plants to accumulate high concentrations of Pb in shoots. The finding that both H-EDTA and Pb-EDTA are mobile within plants also has important implications for the use of metal chelates in plant nutritional research.The synthetic chelate EDTA forms a soluble complex with many metals, including Pb (Kroschwitz, 1995), and can solubilize Pb from soil particles (Means and Crerar, 1978). Recently, application of EDTA to Pb-contaminated soils has been shown to induce the uptake of Pb by plants (Jøgensen, 1993; Huang and Cunningham, 1996; Blaylock et al., 1997; Huang et al., 1997), causing Pb to accumulate to more than 1% (w/w) of shoot dry biomass (Huang and Cunningham, 1996; Blaylock et al., 1997; Huang et al., 1997). For the in situ remediation of Pb-contaminated soils it appears that this chelate-assisted phytoextraction strategy (Salt et al., 1998) may be more effective than a strategy based on the natural ability of certain wild plant species for metal hyperaccumulation (Chaney, 1983; Baker et al., 1988).For more than 40 years, synthetic chelates have been used to supply plants with micronutrients in both soil and hydroponics. Yet the mechanisms by which chelates enhance metal accumulation are still not well characterized (Wallace and Wallace, 1992), and what is known appears contradictory. For example, some evidence suggests that the Fe-chelate EDTA can be absorbed by plants and translocated to shoots (Weinstein et al., 1954; Hill-Cottingham and Llyod-Jones, 1961, 1965). However, Tiffin et al. (1960) concluded that Fe-chelates are excluded from root tissue, and this was supported by Chaney et al. (1972), who demonstrated that Fe is taken up by plants only after first being split from the Fe-chelate complex by the action of a specific plasma membrane-bound Fe-chelate reductase.To optimize the process of chelate-assisted phytoextraction, it is important to understand the biological mechanisms responsible for this process. Because of the stimulatory role of chelate application in the uptake of Pb and other metals by plants, we have investigated the role of EDTA in Pb accumulation in plants. In this study we have demonstrated that the previously described EDTA-enhanced Pb accumulation in Indian mustard (Brassica juncea) is based on the ability of EDTA to chelate and transport Pb from soil into shoot tissue.     

Root, shoot tissues of Brassica juncea and Cereal secale promote potato health

Brassica species are increasingly being used as cover crops to suppress soil-borne diseases in potato cropping systems. Experiments were conducted in controlled environments and in the field to evaluate the effects of cover crop root or shoot or a combination of root and shoot tissues on potato root and tuber health. In a lab assay we examined the extent to which volatile compounds released from tissues of two cover crop species, rye (Cereale secale L.) and oriental mustard (Brassica juncea L.), could inhibit mycelium growth of two important potato diseases, Rhizoctonia solani and Pythium ultimum. Twenty-four hours into the lab assay, volatile compounds from all residues suppressed fungal growth. After 48 h, marked suppression of hyphal growth continued in the presence of mustard residues but not in the presence of rye tissues or the control without tissues. A 75 L volume container experiment evaluated the effect of incorporating different quantities of mustard shoot and root tissues (none, comparable to field level and fourfold field level) into R. solani and P. ultimum infested soil on potato growth, root health and tuber disease. In the container study, incorporating mustard shoots at the highest dose increased potato yield by 54% and reduced disease rating to 2.3 compared to a severe rating of 4.4 in the control. In the field trial, potato growth, root health and tuber disease levels were evaluated in plots where disease management involved either incorporation of mustard or rye cover crop roots, shoots and whole plants (roots plus shoots) or standard farmer practice of a fumigated fallow as a control. White root tissue was used as a health indicator, and averaged 58 and 78% in the fumigated control and mustard cover crop treatments, respectively. The highest healthy root tissue status (91%) was recorded where whole plants of mustard were incorporated. In contrast to the visual assessment of root and tuber health, tuber yield in the field was not influenced by cover crop treatment. Across experiments, the incorporation of or exposure to whole mustard plants was consistently effective at suppressing soil-borne fungi and promoting healthy roots and tubers, especially at higher rates of biomass. Mustard should be managed so as to maximize incorporated biomass for effective biofumigation. Multipurpose management requiring removal of mustard shoots is incompatible with promoting potato rhizosphere health.     

Selenium assimilation and volatilization from selenocyanate-treated Indian mustard and muskgrass

Selenocyanate (SeCN(-)) is a major contaminant in the effluents from some oil refineries, power plants, and in mine drainage water. In this study, we determined the potential of Indian mustard (Brassica juncea) and muskgrass (a macroalga, Chara canescens) for SeCN(-) phytoremediation in upland and wetland situations, respectively. The tolerance of Indian mustard to toxic levels of SeCN(-) was similar to or higher than other toxic forms of Se. Indian mustard treated with 20 microM SeCN(-) removed 30% (w/v) of the Se supplied in 5 d, accumulating 554 and 86 microg of Se g(-1) dry weight in roots and shoots, respectively. Under similar conditions, muskgrass removed approximately 9% (w/v) of the Se supplied as SeCN(-) and accumulated 27 microg of Se g(-1) dry weight. A biochemical pathway for SeCN(-) degradation was proposed for Indian mustard. Indian mustard and muskgrass efficiently degraded SeCN(-) as none of the Se accumulated by either organism remained in this form. Indian mustard accumulated predominantly organic Se, whereas muskgrass contained Se mainly as selenite and organic Se forms. Indian mustard produced volatile Se from SeCN(-) in the form of less toxic dimethylselenide. Se volatilization by Indian mustard accounted for only 0.7% (w/v) of the SeCN(-) removed, likely because the biochemical steps in the production of dimethylselenide from organic Se were rate limiting. Indian mustard is promising for the phytoremediation of SeCN(-) -contaminated soil and water because of its remarkable abilities to phytoextract SeCN(-) and degrade all the accumulated SeCN(-) to other Se forms.     

Effects of below- and above-ground herbivores on plant growth,flower visitation and seed set

Separate and combined effects of root and leaf herbivores on plant growth, flower visitation and seed set were tested in a factorial experiment using potted mustard, Sinapis arvensis, at an old fallow field. A 50% leaf removal by cabbageworms (Pieris rapae) when the seedlings had their first four leaves reduced plant height and shoot mass, and delayed the onset of flowering. Root herbivory by two wireworms (Agriotes sp.) over the whole experiment changed flower visitation; the number of flower visitors per plant was higher in plants with root herbivores than in plants without root herbivores. Combined leaf and root herbivory affected flowering period, number of fruits per plant and number of seeds per fruit. Plants attacked by leaf and root herbivores had a shorter flowering period and produced fewer fruits per plant than plants with root herbivores only. Although the experimental plants faced major herbivore-induced growth changes, plant reproduction (seed set and weight per plant) was similar in all treatments, documenting their ability to effectively compensate for leaf and root herbivory.     

Overexpression of Glutathione Synthetase in Indian Mustard Enhances Cadmium Accumulation and Tolerance

An important pathway by which plants detoxify heavy metals is through sequestration with heavy-metal-binding peptides called phytochelatins or their precursor, glutathione. To identify limiting factors for heavy-metal accumulation and tolerance, and to develop transgenic plants with an increased capacity to accumulate and/or tolerate heavy metals, the Escherichia coli gshII gene encoding glutathione synthetase (GS) was overexpressed in the cytosol of Indian mustard (Brassica juncea). The transgenic GS plants accumulated significantly more Cd than the wild type: shoot Cd concentrations were up to 25% higher and total Cd accumulation per shoot was up to 3-fold higher. Moreover, the GS plants showed enhanced tolerance to Cd at both the seedling and mature-plant stages. Cd accumulation and tolerance were correlated with the gshII expression level. Cd-treated GS plants had higher concentrations of glutathione, phytochelatin, thiol, S, and Ca than wild-type plants. We conclude that in the presence of Cd, the GS enzyme is rate limiting for the biosynthesis of glutathione and phytochelatins, and that overexpression of GS offers a promising strategy for the production of plants with superior heavy-metal phytoremediation capacity.