In its natural habitat, Astragalus bisulcatus can accumulate up to 0.65% (w/w) selenium (Se) in its shoot dry weight. X-ray absorption spectroscopy has been used to examine the selenium biochemistry of A. bisulcatus. High concentrations of the nonprotein amino acid Se-methylseleno-cysteine (Cys) are present in young leaves of A. bisulcatus, but in more mature leaves, the Se-methylseleno-Cys concentration is lower, and selenate predominates. Seleno-Cys methyltransferase is the enzyme responsible for the biosynthesis of Se-methylseleno-Cys from seleno-Cys and S-methyl-methionine. Seleno-Cys methyltransferase is found to be expressed in A. bisulcatus leaves of all ages, and thus the biosynthesis of Se-methylselenoCys in older leaves is limited earlier in the metabolic pathway, probably by an inability to chemically reduce selenate. A comparative study of sulfur (S) and Se in A. bisulcatus using x-ray absorption spectroscopy indicates similar trends for oxidized and reduced Se and S species, but also indicates that the proportions of these differ significantly. These results also indicate that sulfate and selenate reduction are developmentally correlated, and they suggest important differences between S and Se biochemistries.Many selenium (Se) compounds are toxic to mammals at high concentrations, but Se is also an essential micronutrient, and low doses have been implicated in cancer prevention (Clark et al., 1996; Combs et al., 1997). Not all diets provide adequate Se, and an obvious and inexpensive way to provide Se may be to engineer food plants to accumulate higher levels of the element (Ip et al., 1994). The Se hyperaccumulator Astragalus species, such as Astragalus bisulcatus, may be an excellent source of genetic material from which to isolate genes to develop such plants. In the wild, A. bisulcatus can accumulate Se levels of up to 0.65% (w/w) dry weight in the shoots (Byers, 1936), predominantly as Se-methylseleno-Cys (Trelease et al., 1960), and similar results are readily obtained in plants grown hydroponically in the laboratory (Orser et al., 1999). Understanding Se uptake in A. bisulcatus might also allow the development of highly effective cultivars for phytoremediation (Salt et al., 1998).A critical step in the biotransformation of selenate is the initial two-electron reduction to selenite. Hyperaccumulating plants might achieve this in at least three different ways: by substituting selenate into the sulfate reduction pathway (reduction by ATP sulfurylase/adenyl sulfate (APS) reductase; Shrift, 1969;Setya et al., 1996), by substituting selenate into the nitrate uptake pathway (microbial nitrate reductases can reduce selenate; Sabaty et al., 2001), or by a specific selenate reductase. For nonhyperaccumulating plants, there is good evidence that selenate reduction occurs via substitution for sulfate in the ATP sulfurylase/APS reductase system, and that this is the ratelimiting step in selenate transformation (Shrift, 1969;Shaw and Anderson, 1974; Burnell, 1981; Pilon-Smits et al., 1999). In these spe...