Overexpression of ATP sulfurylase in Indian mustard: effects on tolerance and accumulation of twelve metals

Indian mustard [Brassica juncea (L.) Czern.] transgenics overexpressing ATP sulfurylase (APS plants) were shown previously to have higher levels of total thiols, S, and Se. The present study explores the effect of ATP sulfurylase overexpression on tolerance and accumulation of other metals, both oxyanions and cations, reasoning that some anions may react directly with ATP sulfurylase, while other ions may be bound by its thiol end products. The APS transgenics were compared with wild-type plants with respect to tolerance and accumulation of As, Cd, Cr, Cu, Hg, Mn, Mo, Ni, Pb, V, W, and Zn, supplied individually in agar medium (seedlings) or in hydroponics (mature plants). At the seedling stage, APS transgenics were more tolerant than wild type to As(III), As(V), Cd, Cu, Hg, and Zn, but less tolerant to Mo and V. The APS seedlings had up to 2.5-fold higher shoot concentrations of As(III), As(V), Hg, Mo, Pb, and V, and somewhat lower Cr levels. Mature APS plants contained up to 2.5-fold higher shoot concentrations of Cd, Cr, Cu, Mo, V, and W than wild type. They also contained 1.5- to 2-fold higher levels of the essential elements Fe, Mo, and S in most of the treatments. Mature APS plants showed no differences in metal tolerance compared with the wild type. Overexpression of ATP sulfurylase may be a promising approach to create plants with enhanced phytoextraction capacity for mixtures of metals.     

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.