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Absorption and degradation of metalaxyl in mustard plant (Brassica juncea).

Absorption and degradation of metalaxyl were studied in mustard (Brassica juncea) plants after application as a seed dresser, a foliar spray, and a combination of both under subtropical conditions in India. Results indicated that absorption of metalaxyl increased up to 30 days when it was applied as a seed dresser; thereafter, it started declining and was not detectable after 60 days of sowing. The maximum residues (average, 9.03 ppm) of metalaxyl were found after 1 day of spraying. The dissipation of metalaxyl after initial deposits on mustard plants was almost complete after 15 days of spraying. The safe waiting period of metalaxyl was calculated to be 62 and 8 days for seed dresser and foliar application, respectively. The seeds raised through treatments under study were completely free from any detectable amount of metalaxyl residues.

Absorption↗

Analysis of transgenic Indian mustard plants for phytoremediation of metal-contaminated mine tailings.

Transgenic Indian mustard [Brassica juncea (L.) Czern.] plants overproducing the enzymes gamma-glutamylcysteine synthetase (ECS) or glutathione synthetase (GS) were shown previously to have increased levels of the metal-binding thiol peptides phytochelatins and glutathione, and enhanced Cd tolerance and accumulation. Furthermore, transgenic Indian mustard plants overexpressing adenosine triphosphate sulfurylase (APS) were shown to have higher levels of glutathione and total thiols. These results were obtained with a solution culture. To better examine the phytoremediation potential of these transgenics, a greenhouse experiment was performed in which the transgenics were grown on metal-contaminated soil collected from a USEPA Superfund site near Leadville, Colorado. A grass mixture used for revegetation of the site was included for comparison. The ECS and GS transgenics accumulated significantly (P < 0.05) more metal in their shoot than wild-type (WT) Indian mustard, while the APS plants did not. Of the six metals tested, the ECS and GS transgenics accumulated 1.5-fold more Cd, and 1.5- to 2-fold more Zn, compared with wild-type Indian mustard. Furthermore, the ECS transgenics accumulated 2.4- to 3-fold more Cr, Cu, and Pb, relative to WT. The grass mixture accumulated significantly less metal than Indian mustard: approximately 2-fold less Cd, Cu, Mn, and Zn, and 5.7-fold less Pb than WT Indian mustard. All transgenics removed significantly more metal from the soil compared with WT Indian mustard or an unplanted control. While WT did not remove more metal than the unplanted control for any of the metals tested, all three types of transgenics significantly reduced the soil metal concentration, and removed between 6% (Zn) and 25% (Cd) of the soil metal. This study is the first to demonstrate enhanced phytoextraction potential of transgenic plants using polluted environmental soil. The results confirm the importance of metal-binding peptides for plant metal accumulation and show that results from hydroponic systems have value as an indicator for phytoremediation potential.

Biodegradation, Environmental↗

Field trial of transgenic Indian mustard plants shows enhanced phytoremediation of selenium-contaminated sediment.

Three transgenic Indian mustard [Brassica juncea (L.) Czern.] lines were tested under field conditions for their ability to remove selenium (Se) from Se- and boron-contaminated saline sediment. The transgenic lines overexpressed genes encoding the enzymes adenosine triphosphate sulfurylase (APS), gamma-glutamyl-cysteine synthetase (ECS), and glutathione synthetase (GS), respectively. The APS, ECS, and GS transgenic plants accumulated 4.3, 2.8, and 2.3-fold more Se in their leaves than wild type, respectively (P < 0.05). GS plants significantly tolerated the contaminated soil better than wild type, attaining an aboveground biomass/area almost 80% of that of GS plants grown on clean soil, compared to 50% for wild type plants. This is the first report showing that plants genetically engineered for phytoremediation can perform successfully under field conditions.

Biodegradation, Environmental↗

Effects of cadmium on nutrient uptake and translocation by Indian Mustard.

Plants that hyperaccumulate metals are ideal subjects for studying the mechanisms of metal and mineral nutrient uptake in the plant kingdom. Indian Mustard (Brassica juncea) has been shown to accumulate moderate levels of Cd, Pb, Cr, Ni, Zn, and Cu. In this experiment, 10 levels of Cd concentration treatments were imposed by adding 10-190 mg Cd kg(-1) to the soils as cadmium nitrate [Cd(NO3)2]. The effect of Cd on phosphorus (P), potassium (K), calcium (Ca), magnesium (Mg), and the micronutrients iron (Fe), manganese (Mn), copper (Cu), and zinc (Zn) in B. juncea was studied. Plant growth was affected negatively by Cd, root biomass decreased significantly at 170 mg Cd kg(-1) dry weight soils treatment. Cadmium accumulation both in shoots and roots increased with increasing soil Cd treatments. The highest concentration of Cd was up to 300 mg kg(-1) d.w. in the roots and 160 mg kg(-1) d.w. in the shoots. The nutrients mainly affected by Cd were P, K, Ca, Fe, and Zn in the roots, and P, K, Ca, and Cu in the shoots. K and P concentrations in roots increased significantly when Cd was added at 170 mg kg(-1), and this was almost the same level at which root growth was inhibited. Zn concentrations in roots decreased significantly when added Cd concentration was increased from 50 to 110 mg kg(-1), then remained constant with Cd treatments from 110 to 190 mg kg(-1). However, Zn concentrations in the shoots seemed less affected by Cd. It is possible that Zn uptake was affected by the Cd but not the translocation of Zn within the plant. Ca and Mg accumulation in roots and shoots showed similar trends. This result indicates that Ca and Mg uptake is a non-specific process.

Biodegradation, Environmental↗

Effects of mercury on visible/near-infrared reflectance spectra of mustard spinach plants (Brassica rapa P.).

Mustard spinach plants were grown in mercury-spiked and contaminated soils collected in the field under controlled laboratory conditions over a full growth cycle to test if vegetation grown in these soils has discernible characteristics in visible/near-infrared (VNIR) spectra. Foliar Hg concentrations (0.174-3.993ppm) of the Mustard spinach plants were positively correlated with Hg concentration of soils and varied throughout the growing season. Equations relating foliar Hg concentration to spectral reflectance, its first derivative, and selected vegetation indices were generated using stepwise multiple linear regression. Significant correlations are found for limited wavelengths for specific treatments and dates. Ratio Vegetation Index (RVI) and Red Edge Position (REP) values of plants in Hg-spiked and field-contaminated soils are significantly lower relative to control plants during the early and middle portions of the growth cycle which may be related to lower chlorophyll abundance or functioning in Hg-contaminated plants.

Brassica rapa↗

Systemic inhibition of arbuscular mycorrhiza development by root exudates of cucumber plants colonized by Glomus mosseae.

The arbuscular mycorrhizal (AM) non-host plants mustard, sugar beet, lupin and the AM host plant cucumber were used as test plants. Cucumber plants were grown either in the absence of the AM fungus (AMF) Glomus mosseae or in a split-root system, with one side mycorrhizal and one side non-mycorrhizal. Root exudates of the AM non-host plants, the non-mycorrhizal cucumber plants and the mycorrhizal and the non-mycorrhizal side of the split-root system of mycorrhizal cucumber plants were collected and applied to cucumber plants inoculated with the AMF. Root exudates of non-mycorrhizal cucumber plants showed a significant stimulatory effect on root colonization, whereas root exudates from the mycorrhizal and the non-mycorrhizal sides of a split-root system of a mycorrhizal cucumber plant did not show this stimulatory effect and were even slightly inhibitory. Root exudates of the two AM non-host plants mustard and sugar beet significantly reduced root colonization in cucumber plants, whereas no such effect was observed when root exudates of the AM non-host plant lupin were applied.

Beta vulgaris↗

Phytoremediation of arsenic and lead in contaminated soil using Chinese brake ferns (Pteris vittata) and Indian mustard (Brassica juncea).

Field and greenhouse experiments were performed to assess the performance of phytoremediation of arsenic and lead from contaminated soil at an EPA Superfund site (Barber Orchard). Chinese Brake ferns (Pteris vittata) were used to extract arsenic. On average, fern shoot arsenic concentrations were as high as 20 times the soil arsenic concentrations under field conditions. It was estimated that 8 years would be required to reduce the acid-extractable portion of soil arsenic to safe levels (40 mg/kg). The effect of soil pH on arsenic extraction was also investigated. Results indicate that increasing soil pH may improve arsenic removal. Indian mustard plants (Brassica juncea) were used under greenhouse conditions to phytoextract soil lead. EDTA was applied to soil and was found to improve lead extraction. When the EDTA concentration was 10 mmol EDTA/kg soil in soil containing 338 mg Pb/kg soil, mustard plants extracted approximately 32 mg of lead. In conclusion, phytoremediation would be a suitable alternative to conventional remediation techniques, especially for soils that do not require immediate remediation.

Arsenic↗

How can we deliver the large plant genomes? Strategies and perspectives.

The first sequenced plant genome, from the small mustard plant Arabidopsis thaliana, was published at the end of 2000. The sequencing of the rice genome is well under way. The sizes of plant genomes vary by a factor of up to 1000, and many important crop plants have genomes that are several times larger than the human genome. To gain insight into the gene toolbox of plant species, numerous large-scale EST sequencing projects have been launched successfully, and analysis procedures are constantly being refined to add maximum value to the sequence data. In addition, an alternative approach to exclude repetitive noncoding DNA and to enrich sequence libraries for gene-containing genomic regions has been developed. This strategy has the potential to deliver information about both genes and regulatory regions outside the transcribed regions.

Arabidopsis↗

Mercury volatilisation and phytoextraction from base-metal mine tailings.

Experiments were carried out in plant growth chambers and in the field to investigate plant-mercury accumulation and volatilisation in the presence of thiosulphate (S2O3)-containing solutions. Brassica juncea (Indian mustard) plants grown in Hg-contaminated Tui mine tailings (New Zealand) were enclosed in gastight volatilisation chambers to investigate the effect of ammonium thiosulphate ([NH4]2 S2O3) on the plant-Hg volatilisation process. Application of (NH4)2 S2O3 to substrates increased up to 6 times the Hg concentration in shoots and roots of B. juncea relative to controls. Volatilisation rates were significantly higher in plants irrigated only with water (control) when compared to plants treated with (NH4)2 S2O3. Volatilisation from barren pots (without plants) indicated that Hg in tailings is subject to biological and photochemical reactions. Addition of sodium thiosulphate (Na2S2O3) at 5 g/kg of substrate to B. juncea plants grown at the Tui mine site confirmed the plant growth chambers studies showing the effectiveness of thio-solutions at enhancing shoot Hg concentrations. Mercury extraction from the field plots yielded a maximum value of 25 g/ha. Mass balance studies revealed that volatilisation is a dominant pathway for Hg removal from the Tui mine site. A preliminary assessment of the risks of volatilisation indicated that enhanced Hg emissions by plants would not harm the local population and the regional environment.

Biodegradation, Environmental↗

The effect of illumination of the malic acid content and anion/cation balance of mustard leaves (Sinapis alba).

1. Mustard plants have been grown under conditions in which the length of artificial day could be controlled. 2. Leaf samples were analysed for malic acid and citric acid, and for a number of inorganic anions and cations. A simple method is described by which sap was obtained from 0.5g. samples of leaves. 3. In days of 16hr. or more, malic acid was accumulated; the chief cation accumulated was calcium. 4. When the day-length was reduced the malic acid content decreased considerably but the calcium content remained the same. There was little change in the pH value of the sap, the balance of anions and cations having been maintained mainly by increases in citrate and nitrate contents. Analyses of the whole leaf still showed some deficiency in anion after sodium, potassium, calcium, magnesium, nitrate, sulphate, inorganic phosphate, chloride, malate and citrate had been accounted for. 5. Analyses at shorter intervals revealed a large diurnal variation in malic acid content, which increased during the first 5-6hr. of the light period, and fell during darkness. 6. The significance of these findings is discussed, and it is suggested that malic acid accumulation is a by-product of photosynthesis, calcium being taken up irreversibly to maintain anion/cation balance, and hence creating a continuing need for anions to balance it.

Calcium↗

Study of calcium-dependent lead-tolerance on plants differing in their level of Ca-deficiency tolerance.

The main aim of the study was to determine the role of calcium in the amelioration of lead toxic effects in plants with accordingly high/low level of Pb-tolerance and high/low Ca-deficiency tolerance. The study was performed on maize, rye, tomato and mustard. Plants were cultivated in modified Knop's solution. They were subjected to Ca-deficiency, and to lead nitrate administered in the presence of four calcium nitrate concentrations 3.0, 2.4, 1.2, 0.3mM. Lead-tolerance and tolerance to Ca-deficiency were determined, as were concentration of the studied elements in plant tissues, and the Pb deposition pattern at the ultrastructural level (electron microscopy study, X-ray microanalysis). In all studied plants, lead toxicity increased as medium calcium content decreased, however, only in the Ca-deficiency sensitive mustard with low Pb-tolerance was it accompanied by a rise in tissue lead concentration. In contrast, lead root and shoot levels did not increase in the highly Ca-deficiency tolerant tomato, mustard and rye with high Pb-tolerance irrespective of the Ca(2+) regimens applied. Thus, in these plants, lead's unfavourable effects resulted only from the higher toxicity of the same amount of lead in tissues at low calcium in the medium. Of particular relevance is the finding by electron microscopy and X-ray microanalysis, that under low calcium in both highly Ca-deficiency tolerant and Ca-deficiency sensitive plants, less efficient Pb(2+) detoxification was accompanied by the restriction of the formation of large lead deposits in cell walls. Obtained results are novel in demonstrating calcium involvement in the lead deposition in the cell wall, thus in the regulation of the internal lead detoxification.

Air Pollutants↗

DNA fiber replication during a morphogenetic switch in the shoot meristematic cells of a higher plant.

Chromosomal DNA fiber replication was investigated in the shoot meristem of mustard plants during the morphogenetic transition from the leaf-forming (vegetative) to the flower-forming (evoked) condition. The replicon size, determined using the modal class, was 15 micron in the vegetative meristem and shifted to 7.5 micron in the evoked meristem. The average fork rate was 1.05 micron.h-1 in the vegetative meristem and only slightly increased to 1.55 micron.h-1 during the morphogenetic switch. Replicon activation was asynchronous but the pattern of activation of replicons was the same in both kinds of meristems. Thus the shortening of the S phase at the floral transition in mustard was essentially achieved by an increase of the number of replicon origins per unit length of DNA. These results are similar to those obtained in amphibian and Drosophila embryogenesis.

DNA Replication↗

Isolation and cloning of Bacillus thuringiensis var Kurstaki HD73 toxin gene and construction of a chimaeric gene for expression in plants.

Necessity for the production of transgenic crop plants of India, capable of expression of insecticidal Bt protein in plant to combat lepidopteran pest damage has been strongly felt. Bacillus thuringiensis Kurstaki HD73 crystal protein coded by CryIA(c)73 gene has been found to be sufficiently effective against the major pests of jute and chickpea. An attempt to isolate the gene and make its use in a chimaeric gene construct for expression in plant was carried out. The plasmid CryIA(c)73 gene was cloned and tailored at the 3' end. The expression of the truncated gene was monitored in the minicell systems of E. coli. The entomocidal property was found to be fully retained by the gene product. Deletion of the nucleotides at the 5' end was carried out and chimaeric gene construct of cryIA(c)73 was made in such a way that it was fused in frame with GUS gene under the control of the caMV 35S promoter with Nos polyadenylated terminus. Such a chimaeric gene construct was used as the passenger of a Ti plasmid derived plant vector with kanamycin gene (NPTII) as the additional plant marker. Transformation through infection of tobacco and mustard plant cells in culture was carried out. Plants regenerated from the transformed cells showed the presence of GUS gene indicating the expression of the cloned fused gene. Also, Northern analysis established the presence of cryIA(c)73 gene transcripts in the transgenic plants.

Bacillus thuringiensis↗