Phytoremediation of aspirin and tetracycline by Brassica juncea

With the increasing release of pharmaceutical drugs in the environment, research is in progress for investigating alternative methods for their remediation. Various studies have shown the phytoremediation potential of Brassica juncea for metals. The current study was aimed at evaluating the phytoremediation potential of B. juncea for two different pharmaceutical drugs i.e. aspirin and tetracycline in in-vitro conditions. The seeds of B. juncea were germinated and grown for a period of 28 and 24 days for aspirin and tetracycline, respectively. The study analyzed the remediation rate of B. juncea for the selected drugs in three different sets of varying concentration along with any phytotoxic effects exerted by the drugs on the seeds. Preliminary results showed that the average remediation rate of aspirin and tetracycline at the end of experiment was approximately 90% and 71%, respectively. As initial drug concentrations were increased in the media, the remediation rate also improved. However, at higher concentrations, the plants showed phytotoxicity as depicted by the decrease in shoot length of the germinated seeds. These preliminary results indicated that B. juncea could tolerate and remediate pharmaceutical drugs such as analgesics and antibiotics.     

Lithium,Vanadium and Chromium Uptake Ability of Brassica juncea from Lithium Mine Tailings

The potential for phytoremediation and phytostabilization of lithium in lieu with vanadium and chromium on a formulated acidic heterogeneous growth media engineered around lithium mine tailings, was investigated in four phases: (1) overall efficiency of the removal of the three metals, (2) bioaccumulation ratios of the three metals, (3) overall relative growth rate, and (4) translocation index of the three metals in the physiology of the hyperaccumulator plant. A pot study was conducted to assess the suitability of Brassica juncea (Indian mustard) in a phytoremediation process whereby it was lingered for eighty-six days under homogeneous growth conditions and irrigated bidaily with organic fertilizer amended with LiCl. A post harvest data analysis was achieved through ashing and the implementation of cold digestion procedure in a concentrated hydrochloric acidic matrix. In physiological efficiency parameters, the hyperaccumulator plant was twice as able to phytostabilize chromium and four times was able to phytostabilize vanadium in comparison to lithium. Moreover, it was extremely efficient in translocating and accumulating lithium inside its upper physiological sites, more so than chromium and vanadium, thereby demonstrating Indian mustard, as a hyperaccumulator plant, for phytoextraction and phytostabilization in an acidic heterogeneous rhizosphere, with an extremely low relative growth rate.     

Phytoremediation efficiency of Brassica juncea cultivars at vegetative and reproductive growth stages under individual and combined treatment of fluoride and aluminium

The objective of this study was to investigate phytoremediation ability of Brassica juncea cultivars for aluminium (Al) and fluoride (F) independently and in combination (Al + F). Out of 8 cultivars which were treated with Al, F, and (Al + F), 4 cultivars (Bio-902, Pusa-Tarak, CS-14, and Laxmi) were selected for further studies on the basis of growth parameters. These cultivars were exposed to soil (pH 5) supplemented with F (0, 25, 50, and 75 mg kg^?1) and Al (0, 50, 100, and 150 mg kg^?1) independently and in combination (Al + F) (0 + 0, 50 + 25, 100 + 50, and 150 + 75 mg kg^?1). We found that the accumulation of F, Al, and (Al + F) was highest in the roots followed by grains, shoots, and leaves. When the plants were treated with Al or F separately, the accumulation of Al or F were less as compared to when treated in combination (Al + F). Conclusively, the results also showed that maximum tolerance index, uptake, and translocation factor for F were highest in CS-14, Al in Bio-902, and (Al+ F) in Pusa-Tarak, and were found to be the lowest in Laxmi.     

Effect of Salinity on Zinc uptake by Brassica juncea

Salinity is a major worldwide problem that affects agricultural soils and limits the reclamation of contaminated sites. Despite the large number of research papers published about salt tolerance in Brassica juncea L., there are very few accounts concerning the influence of salinity on the uptake of trace metals. In this study, B. juncea plants divided through soil sets comprising 0, 900 and 1800 mg Zn kg^?1, were treated with solutions containing 0, 60 and 120 mmol L^?1 of NaCl, with the purpose of observing the effect of salt on Zn uptake, and some physiological responses throughout the 90 days experiment. Increasing concentrations of NaCl and Zn produced a decline in the ecophysiological and biochemical properties of the plants, with observable synergistic effects on parameters like shoot dry weight, leaf area, or photochemical efficiency. Nevertheless, plants treated with 60 mmol L^?1 of NaCl accumulated striking harvestable amounts of Zn per plant that largely exceed those reported for Thlaspi caerulescens. It was concluded that salinity could play an important role on the uptake of Zn by B. juncea. The potential mechanisms behind these results are discussed, as well as the implications for phytoremediation of Zn on saline and non-saline soils.     

不同品种芥菜对Cu胁迫响应的差异

采用水培法研究了不同品种芥菜对Cu胁迫响应的差异。结果表明:4μmol·L-1和8μmol·L-1Cu处理均抑制根和地上部生物量的积累,根受到的抑制程度大于地上部,敏感品种受到的抑制程度大于抗性品种。根中Cu积累量大于地上部,敏感品种根中Cu积累量大于抗性品种。8μmol·L-1Cu处理能明显诱导芥菜根中的MDA、Pro含量的增加及AsA含量的减少,APX、POD活性上升和GR活性下降,而不改变SOD活性。     

Callus Formation and Morphogenesis of Cultured Mustard (Brassica juncea) Protoplasts

Protoplasts from mustard seedling cotyledons were suspended in Nitsch medium and cultured at 26℃ under low intensity illumination. When colonies were observed in the cultures, fresh medium (3% sucrose replaced mannitol) was added by equalvolume. When protoplasts developed into small callus the cultures were transfered to modified MS agar medium, in which some of them produced roots while an others gave rise to green spots. Shoot-differentiation experiment is in progress.     

A RFLP-based linkage map of mustard [Brassica juncea (L.) Czern. and Coss.]

 A genetic linkage map of Brassica juncea was constructed based on restriction fragment length polymorphism (RFLP) detected by anonymous cDNA markers from B. napus, using a segregating F₁-derived doubled haploid (DH) progeny from a cross between a canola-quality mustard line (J90-4317) and a high-oil-content mustard line (J90-2733). The RFLP probes consisted of 229 cDNA probes from B. napus and a B. napus tandem repeat sequence, RDA2. The map consisted of 343 marker loci arranged in 18 major linkage groups plus five small segments with two to five marker loci, covering a total map distance of 2073 cM. Twenty-four percent of the markers were dominant in nature. Sixty-two percent of the marker loci were duplicated, and the majority were involved in inter-linkage group duplications, illustrating that complex duplications and subsequent rearrangements occurred after allopolyploidy. Deviation from the Mendelian segregation ratio for a DH population was observed for 27% of the markers. Two-thirds of these markers with a skewed segregation were clustered in 6 linkage groups and two unassigned segments. The overall average marker interval of the B. juncea map reported here was 6.6 cM, which would provide a marker density satisfactory for efficient use of the map in breeding applications, such as tagging of important agronomic traits and marker-assisted selection. Received: 14 May 1996 / Accepted: 11 October 1996     

Loss of C-genome-specific markers during transgene introgression from Brassica napus to wild Brassica juncea

Transgene flow from engineered Brassica napus to wild weed relatives could potentially have an environmental effect. To evaluate the introgression of transgenic B. napus into wild Brassica juncea, the hybrid F₁ and backcross progenies derived from B. juncea (genome constitution AABB) and transgenic B. napus (AACC) crosses were investigated. C-genome-specific simple sequence repeat (SSR) markers corresponding to linkage groups N11–N19 in B. napus were screened and used to estimate the marker frequency in hybrid F₁ and backcross progenies. C-genome-specific markers could be stably detected in hybrid F₁ and backcross BC₁ plants, but were only rarely found in the BC₂–BC₅ generations. For example, a specific SSR marker for linkage group N12 segregated in BC₂ generation but were completely lost in BC₃–BC₅, while a specific SSR marker of linkage group N15 segregated in BC₁, BC₂ and BC₃ generations and was absent in more advanced backcrossed generations (BC₄ and BC₅). The results indicate that a certain gene regions in Brassica napus plants are transmitted at a relatively lower frequency to wild relatives, and more rapidly disappeared in subsequent backcross generations. We propose that a foreign gene or transgene that is integrated in the C-chromosome of Brassica napus could reduce the risk of introgression in nature.     

Chlorophyllase and peroxidase activity during degreening of maturing canola (Brassica napus) and mustard (Brassica juncea) seed

Chlorophyllase and peroxidase activities were measured in relation to seed maturation and degreening in canola ( Brassica napus cvs Westar and Alto) and mustard ( Brassica juncea cvs Cutlass and Lethbridge 22A). Samples of seed collected at the same moisture content were pooled, then divided and used for each assay. During maturation the green pigment (chlorophyll and related pigments) content of canola seed decreased linearly and was lower than that measured in mustard at all moisture contents studied, except for the highest and lowest moisture contents. Chlorophyllides and pheophorbides were detected in canola and were essentially absent in mustard. This difference in accumulation of dephytylated pigments infers differences in the pigment degradation pathways in Brassica species. Interspecific differences in the enzymology of degreening were found. Green pigment degradation was associated with increased chlorophyllase activity and low peroxidase activity in canola and low Chlorophyllase and high perosidase activity in mustard. The possible role of ethylene in seed degreening is discussed.     

Transport and Partitioning of Lead in Indian Mustard (Brassica juncea) and Wheat (Triticum aestivum)

Lead (Pb) contamination in soils is a serious concern because it can be taken up by crops and then transferred through the food chain, posing a potential risk to human health. Indian mustard (Brassica juncea) and wheat (Triticum aestivum) are important crop species known to accumulate heavy metals in their tissues. This study aimed at understanding the transport and accumulation of Pb in these two species and the risk associated with consumption of these foods, which would help us in mitigating accumulation of Pb in edible tissues. The plants were grown at different Pb concentrations for the entire life cycle, and the partitioning of the metal to different tissues was examined. The results showed that plant species differ widely in their ability to transport and accumulate Pb in different tissues. In B. juncea, there was significant accumulation of Pb in both siliques and seeds, whereas most of the Pb in wheat was concentrated in the vegetative tissues and less to the flag leaf and reproductive tissues. In both species, although seed Pb concentrations exceeded acceptable limits, dietary intake did not exceed acceptable limits in most treatments, indicating that more studies on Pb transport and redistribution in crop species is necessary.     

Development of Plantlets from Leaf Protoplast Culture in Brassica juncea var. tsatsi

Protoplast of two mustard cultivars: Brassica juncea var. tsatsi cv. “Quxian Jiaoercai” and “Bangbangcai”, were isolated by enzymolysis from leaf grown in vitro. Protoplasts were suspended in liquid medium and semi-solidified medium with 0.35% low melting point agarose which formed a thin layer floating on the surface of the liquid medium. The first division appeared after 48h in the culture. One week after the original culture, a diluted medium with gradual dicrease of mannitol concentrations (6%→4%→zero) was then added to the culture three times respectively at one week's interval. In this culture method cell division and formation of microcalli were achieved. During the liquid culture of protoplasts, shaking at 20 rpm from time to time was beneficial in the formation of cell colonies and microcalli. Cell colonies developed into calli of approx 0.5—1mm in diameter one month after culture. The plating efficiency, which defined as the percentage of microcatli to numbers of protoplasts, was 0.2%—1%. Shoot regeneration occured when leaf protoplast-derived calli of “Quxian Jiaoercai” were transferred onto the modified MS medium supplemented with BAP 2.0mg/L, KT 1.0mg/L and NAA 0.2mg/L, and those of -'Bangbangcai" were transferred onto the modified MS medium supplemented with BAP 2.0mg/L. Individual shoot was rooted on a rooting medium supplemented with NAA 0.2 or 0.4 mg/L.     

Remediation of Nickel-Contaminated Soil by Brassica juncea L. cv. T-59 and Effect of the Metal on Some Metabolic Aspects of the Plant

Among four cultivars of Brassica juncea L., viz., TM-4, TM-2, RH-30, and T-59, cv. T-59 was relatively more tolerant to nickel (Ni) toxicity based on the growth parameters, seedling vigor index, and metal tolerance index. Nickel application inhibited the activity of the nitrate-assimilating enzyme nitrate reductase in the roots, stem, and leaves, whereas the total organic nitrogen, proline, and activity of a polyamine-metabolizing enzyme, diamine oxidase, increased in this tolerant cultivar (T-59). It accumulated a good amount of Ni from the soil in its root and shoot (i.e., 6.0–6.51 μg Ni g^?1 dry weight) during 2 months of cultivation with an 8.0 mM Ni supply in the soil. The data presented in this paper indicate that Ni tolerance and its removal by Indian mustard from subtropical Indian soil is cultivar dependent, possibly due to different genetic and physiological adaptations of the cultivars.     

Improved microspore culture and doubled-haploid plant regeneration in the brown condiment mustard (Brassica juncea)

 The availability of doubled haploids could greatly contribute to improving seed quality in condiment mustard (Brassica juncea). We have developed an efficient and reliable protocol of microspore culture, modified from that of Baillie et al. (1992), based on a modification of the sucrose concentration of culture media. A comparison of microspore culture media differing in their sucrose content showed that a decrease from 17% (w/v) sucrose during the first 48 h to 10% (w/v) thereafter favoured an increase in the production of embryos whatever the responding genotype tested. Thus, out of the 13 B. juncea genotypes studied, 12 gave rise to embryos, and seven of these embryos could be converted into plants. Doubled-haploid plants were produced after treatment with colchicine. Received: 16 January 2000 / Revision received: 8 August 2000 / Accepted: 20 September 2000     

Assessment of the root system of Brassica juncea (L.) czern. and Bidens pilosa L. exposed to lead polluted soils using rhizobox systems

The purpose of this study was to compare the behavior of the root system of one of the most frequently cited species in phytoremediation Indian mustard [Brassica juncea (L.) Czern.] and a representative perennial herb (Bidens pilosa L.) native of Argentina, for different concentrations of lead in soils through chemical and visualization techniques of the rhizosphere. Lead polluted soils from the vicinity of a lead recycling plant in the locality of Bouwer, were used in juxtaposed rhizobox systems planted with seedlings of B. juncea and B. pilosa with homogeneous and heterogeneous soil treatments. Root development, pH changes in the rhizosphere, dry weight biomass, lead content of root and aerial parts and potential extraction of lead by rhizosphere exudates were determined. In both species lead was mainly accumulated in roots. However, although B. juncea accumulated more lead than B. pilosa at elevated concentrations in soils, the latter achieved greater root and aerial development. No changes in the pH of the rhizosphere associated to lead were observed, despite different extractive potentials of lead in the exudates of the species analyzed. Our results indicated that Indian mustard did not behave as a hyperaccumulator in the conditions of the present study.     

提高榨菜离体培养植株再生频率

采用榨菜“浙桐1号”品种为材料,以MS为基本培养基,通过对不同植物生长调节剂的组合和不同外植体等主要因素的筛选,大幅度提高了榨菜离体培养植株再生频率。结果表明,2mg／L6．BA 0．2mg／L2,4-D的组合较为适宜,其不定芽再生频率可达50％,且外植体以下胚轴为好：而CPPU和2,4-D的适宜组合为1．5mg／L 0．2mg／L,其不定芽再生频率高达66．67％,最适外植体为带柄子叶。同时,研究结果显示,添加0．25～1mg／L的GA,对榨菜已分化的不定芽的伸长有抑制作用;子叶柄和下胚轴外植体的分化具有极性现象。     

Accumulation, detoxification, and genotoxicity of heavy metals in Indian mustard (Brassica juncea L.)

Plants of Indian mustard (Brassica juncea L.) were exposed to different concentrations (15, 30, 60, 120 microM) of (Cd, Cr, Cu, Pb) for 28 and 56 d for accumulation and detoxification studies. Metal accumulation in roots and shoots were analyzed and it was observed that roots accumulated a significant amount of Cd (1980 microg g(-1) dry weight), Cr (1540 microg g(-1) dry weight), Cu (1995 microg g(-1) dry weight), and Pb (2040 microg g(-1) dry weight) after 56 d of exposure, though in shoot this was 1110, 618, 795, and 409 microg g(-1) dry weight of Cd, Cr, Cu, and Pb, respectively. In order to assess detoxification mechanisms, non-protein thiols (NP-SH), glutathione (GSH) and phytochelatins (PCs) were analyzed in plants. An increase in the quantity of NP-SH (9.55), GSH (8.30), and PCs (1.25) micromol g(-1) FW were found at 15 microM of Cd, however, a gradual decline in quantity was observed from 15 microM of Cd onwards, after 56 d of exposure. For genotoxicity in plants, cytogenetic end-points such as mitotic index (MI), micronucleus formation (MN), mitotic aberrations (MA) and chromosome aberrations (CA) were examined in root meristem cells of B. juncea. Exposure of Cd revealed a significant (P < 0.05) inhibition of MI, induction of MA, CA, and MN in the root tips for 24 h. However, cells examined at 24 h post-exposure showed concentration-wise recovery in all the endpoints. The data revealed that Indian mustard could be used as a potential accumulator of Cd, Cr, Cu, and Pb due to a good tolerance mechanisms provided by combined/concerted action of NP-SH, GSH, and PCs. Also, exposure of Cd can cause genotoxic effects in B. juncea L. through chromosomal mutations, MA, and MN formation.     

Overexpression of enzymes involved in glutathione synthesis enhances tolerance to organic pollutants in Brassica juncea

Transgenic Indian mustard (Brassica juncea) overexpressing y-glutamylcysteine synthetase (ECS) or glutathione synthetase (GS) were shown previously to have two-fold higher levels of glutathione and total nonprotein thiols, as well as enhanced cadmium tolerance and accumulation. Here, the hypothesis was tested that these transgenics have enhanced tolerance to organic pollutants, based on the reasoning that many organic xenobiotics are detoxified via conjugation to glutathione. Both the ECS and GS transgenics showed enhanced tolerance to atrazine: while root growth of wildtype seedlings was inhibited 50% by 100 mg L(-1) atrazine, ECS and GS root growth was inhibited 20-30% (P < 0.05). The tolerance of the transgenics to CDNB (1-chloro-2,4-dinitrobenzene). metolachlor, and phenanthrene was also somewhat higher than wild type, but these differences were not as pronounced. Each of the organics treatments significantly enhanced total nonprotein thiol levels in all plant types (2 to 12-fold). Overall, these results suggest that GSH biosynthesis is limiting for atrazine detoxification in Indian mustard and that overexpression of enzymes involved in GSH biosynthesis offers a promising approach to create plants with the enhanced capacity to tolerate not only heavy metals, but also certain organics.     

雪里蕻非特异脂质转运蛋白基因的克隆与分析

非特异脂质转运蛋白是植物生命活动中一类重要的活性蛋白,这类蛋白在植物的抗性和防御中行使着重要的功能。近年来研究发现这类蛋白还与植物的有性生殖密切相关。通过已得到的普通白菜的脂质转运蛋白基因BcMF15的核苷酸序列,在其基因全长两侧设计引物,从雪里蕻中克隆得到该类活性蛋白基因,命名为BjLTP (登陆号: EU082009)。该基因全长650bp,不含内含子。不同组织的表达特征分析发现,BjLTP在雪里蕻的花蕾、开放的花中特异表达,推测BjLTP可能与花粉的发育有关。蛋白质特征预测及蛋白序列结构分析发现BjLTP是一个跨膜蛋白,具有显著的疏水区。序列同源比对表明该基因与白花芥蓝、拟南芥等的脂质转运蛋白基因有很高的相似性,证明BjLTP是LTP家族的成员之一。     

Differential response of Brassica juncea cultivars to Al; consequences for chlorophyll a fluorescence,antioxidants and psb A gene

The present study was undertaken to identify the aluminum (Al) tolerant cultivar of Brassica juncea. We examined the changes in antioxidant enzymes, proline level, chlorophyll a fluorescence and psb A gene expression at vegetative and reproductive growth stages of B. juncea cultivars (Bio-902, CS-14, Pusa-Tarak and Laxmi). The selected cultivars were exposed to soil (pH 5.2) supplemented with Al (0, 50, 100 and 150?mg?kg^?1). We observed the lowest decline in photosynthetic efficiency (ΔF/Fm′), ETR, PPFD values and psb A expression to Al stress in the cultivar Bio-902 (tolerant cultivar) followed by CS-14 and Pusa-Tarak whereas the highest decline was observed in the cultivar Laxmi (sensitive cultivar). The improved performance of the cultivar Bio-902 under Al stress was accompanied by an increase in proline level, CAT and APX activities but without any increase in SOD activity. However, significant increase in SOD activity was observed in Laxmi.