Summary• Here, Thlaspi caerulescens populations from contrasting soil types (serpentine, calamine and nonmetalliferous) were characterized with regard to tolerance, uptake and translocation of zinc (Zn), cadmium (Cd) and nickel (Ni) in hydroponic culture.• Results showed that high-level tolerances were apparently metal-specific and confined to the metals that were enriched at toxic levels in the soil at the population site.• With regard to metal accumulation, results suggested that, unlike Zn hyperaccumulation, Cd and Ni hyperaccumulation were not constitutive at the species level in T. caerulescens .• In general, the populations under study exhibited a pronounced uncorrelated and metal-specific variation in uptake, root to shoot translocation, and tolerance of Zn, Cd and Ni. The distinct intraspecific variation of these characters provides excellent opportunities for further genetic and physiological dissection of the hyperaccumulation trait.

Heavy metal hyperaccumulation in plants is an intriguing and poorly understood phenomenon. Transmembrane metal transporters are assumed to play a key role in this process. We describe the cloning and isolation of three zinc transporter cDNAs from the Zn hyperaccumulator Thlaspi caerulescens. The ZTP1 gene is highly similar to the Arabidopsis ZAT gene. Of the other two, one is most probably an allele of the recently cloned ZNT1 gene from T. caerulescens (Pence et al; Proceedings of the National Academy of Science USA 97, 4956–4960, 2000). The second, called ZNT2, is a close homologue of ZNT1. All three zinc transporter genes show increased expression in T. caerulescens compared with the non‐hyperaccumulator congener T. arvense, suggesting an important role in heavy metal hyperaccumulation. ZNT1 and ZNT2 are predominantly expressed in roots and ZTP1 is mainly expressed in leaves but also in roots. In T. arvense, ZNT1 and ZNT2 are exclusively expressed under conditions of Zn deficiency. Their expression in T. caerulescens is barely Zn‐responsive, suggesting that Zn hyperaccumulation might rely on a decreased Zn‐induced transcriptional downregulation of these genes. ZTP1 expression was higher in plants from calamine soil than in plants from serpentine or normal soil. The calamine plants were also the most Zn tolerant, suggesting that high ZTP1 expression might contribute to Zn tolerance.

The relation between loss of glutathione due to metal-induced phytochelatin synthesis and oxidative stress was studied in the roots of copper-sensitive and tolerant SiIene cucubalus (L.) Wib., resistant to 1 and 40 micromolar Cu, respectively. The amount of nonprotein sulfhydryl compounds other than glutathione was taken as a measure of phytochelatins. At a supply of 20 micromolar Cu, which is toxic for sensitive plants only, phytochelatin synthesis and loss of total glutathione were observed only in sensitive plants within 6 h of exposure. When the plants were exposed to a range of copper concentrations for 3 d, a marked production of phytochelatins in sensitive plants was already observed at 0.5 micromolar Cu, whereas the production in tolerant plants was negligible at 40 micromolar or lower. The highest production in tolerant plants was only 40% of that in sensitive plants. In both varieties, the synthesis of phytochelatins was coupled to a loss of glutathione. Copper at toxic concentrations caused oxidative stress, as was evidenced by both the accumulation of lipid peroxidation products and a shift in the glutathione redox couple to a more oxidized state. Depletion of glutathione by pretreatment with buthionine sulfoximine significantly increased the oxidative damage by copper. At a comparably low glutathione level, cadmium had no effect on either lipid peroxidation or the glutathione redox couple in buthionine sulfoximinetreated plants. These results indicate that copper may specifically cause oxidative stress by depletion of the antioxidant glutathione due to phytochelatin synthesis. We conclude that copper tolerance in S. cucubalus does not depend on the production of phytochelatins but is related to the plant's ability to prevent glutathione depletion resulting from copper-induced phytochelatin production, e.g. by restricting its copper uptake.In plants, both essential and nonessential heavy metals induce the formation of thiol-rich peptides, (-y-glutamylcysteinyl),-glycines with n = 2 to 1 1, also known as metal-binding compounds or phytochelatins (8,26). Experiments with BSO,2 an inhibitor of y-glutamylcysteine synthetase, showed that glutathione serves as a precursor in the phytochelatin biosynthesis and that phytochelatins are involved in the detoxification of heavy metals in vivo (17,23,25 2Abbreviations: BSO, buthionine sulfoximine; TBA-rm: 2-thiobarbituric acid-reactive material; SH, sulfhydryl.lyzed by a specific y-glutamylcysteine dipeptidyl transpeptidase, called phytochelatin synthase, which is activated in the presence of metal ions and uses GSH as a substrate (9). Phytochelatins are the major if not the only thiol-rich compounds induced in metal-exposed plants (8,26), although it has been reported that copper induces metallothionein-like compounds as well (27). Phytochelatins probably play a central role in the homeostatic control of metal ions in plants (26). They may also be involved in the physiological mechanism of metal tolerance of selected cell lines and intact plants (4,12,22...

(A.A.M.) Arsenate tolerance is conferred by suppression of the high-affinity phosphate/arsenate uptake system, which greatly reduces arsenate influx in a number of higher plant species. Despite this suppressed uptake, arsenate-tolerant plants can still accumulate high levels of As over their lifetime, suggesting that constitutive detoxification mechanisms may be required. Phytochelatins are thiol-rich peptides, whose production is induced by a range of metals and metalloids including arsenate. This study provides evidence for the role of phytochelatins in the detoxification of arsenate in arsenate-tolerant Holcus lanatus. Elevated levels of phytochelatin were measured in plants with a range of tolerance to arsenate at equivalent levels of arsenate stress, measured as inhibition of root growth. The results suggest that arsenate tolerance in H. lanatus requires both adaptive suppression of the high-affinity phosphate uptake system and constitutive phytochelatin production.