The research objectives for this report are: (1) Transform poplar and other tree species to extend and optimize chlorinated hydrocarbon (CHC) oxidative activities. (2) Determine the mechanisms of CHC oxidation in plants. (3) Isolate the genes responsible for CHC oxidation in plants. We have made significant progress toward an understanding of the biochemical mechanism of CHC transformation native to wild-type poplar. We have identified chloral, trichloroethanol, trichloroacetic acid, and dichloroacetic acid as products of TCE metabolism in poplar plants and in tissue cultures of poplar cells.(Newman et al. 1997; Newman et al. 1999) Use of radioactively labeled TCE showed that once taken up and transformed, most of the TCE was incorporated into plant tissue as a non-volatile, unextractable residue.(Shang et al. 2001; Shang and Gordon 2002) An assay for this transformation was developed and validated using TCE transformation by poplar suspension cells. Using this assay, it was shown that two different activities contribute to the fixation of TCE by poplar cells: one associated with cell walls and insoluble residues, the other associated with a high molecular weight, heat labile fraction of the cell extract, a fixation that was apparently catalyzed by plant enzymes.
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