Selenium (Se) phytovolatilization, the process by which plants metabolize various inorganic or organic species of Se (e.g. selenate, selenite, and Se-methionine [Met]) into gaseous Se forms (e.g. dimethylselenide), is a potentially important means of removing Se from contaminated environments. Before attempting to genetically enhance the efficiency of Se phytovolatilization, it is essential to elucidate the enzymatic pathway involved and to identify its rate-limiting steps. The present research tested the hypothesis that S-adenosyl-l-Met:l-Met S-methyltransferase (MMT) is the enzyme responsible for the methylation of Se-Met to Se-methyl Se-Met (SeMM). To this end, we identified and characterized an Arabidopsis T-DNA mutant knockout for MMT. The lack of MMT in the Arabidopsis T-DNA mutant plant resulted in an almost complete loss in its capacity for Se volatilization. Using chemical complementation with SeMM, the presumed enzymatic product of MMT, we restored the capacity of the MMT mutant to produce volatile Se. Overexpressing MMT from Arabidopsis in Escherichia coli, which is not known to have MMT activity, produced up to 10 times more volatile Se than the untransformed strain when both were supplied with Se-Met. Thus, our results provide in vivo evidence that MMT is the key enzyme catalyzing the methylation of Se-Met to SeMM.The trace element selenium (Se) is an essential micronutrient with important benefits for animal and human nutrition; however, at high doses, Se is toxic (Wilber, 1980; Van Vleet and Ferrans, 1992;Lemly, 1997). Major sources of Se pollution are agricultural drainage from seleniferous soils and industrial wastewater. Se pollution is a worldwide problem and there is a tremendous demand for the cleanup of Se-contaminated soil and water. Phytoremediation, the use of plants to remove, stabilize, or detoxify pollutants, is a highly promising solution to counter the Se problem (Bañ uelos et al., 1995;Salt et al., 1998;Terry and Zayed, 1998).Se volatilization, the process by which gaseous Se forms are produced from inorganic or organic Se compounds (Lewis et al., 1966;Zieve and Peterson, 1984;Duckart et al., 1992;Terry et al., 1992Terry et al., , 2000, is particularly attractive for the phytoremediation of Se-contaminated environments because it completely removes Se from the local food chain (Atkinson et al., 1990;Terry and Zayed, 1998). The major volatile Se form produced by plants and microbes is dimethylselenide (DMSe; Lewis et al., 1974). DMSe is 600 to 700 times less toxic than selenate or selenite, two Se species that are commonly present in polluted areas (McConnell and Portman, 1952;Ganther et al., 1966;Wilber, 1980).The formation of DMSe in many plants is thought to proceed via the sulfur (S) assimilation pathway . To enhance the efficiency of Se volatilization by plants, it is essential that we fully elucidate the biochemical pathway involved in Se assimilation and volatilization. Once we have determined the rate-limiting steps in the pathway, it should be possible to enhance the effic...