Improvement of Cadystins [(.GAMMA.-EC)nG] Synthesis Activity and Cd Tolerance in Tobacco Cells by Transfer of the Rice Type I Carboxypeptidase Gene (CPase Gene).

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Plants and certain fungi produce metal-binding peptides after uptaking heavy metals that have the general structure (6γ-Glutamyl-cysteinyl) n-Glycine, (gamma;-EC) nG, with n = 2 to 11. A heavy metal, cadmium, incorporated into the cell is detoxified by (γ-EC) nG and Cd tolerant plant accumulate much Cd in the roots and leaves. In order to improve (γ-EC) nG synthesis activity in tobacco cells, the structural gene of the rice carboxypeptidase gene (CPase gene) was ligated downstream of the CaMV 35 S promoter and the fused gene was transformed into Nicotiana tabacum L.cv. Bright Yellow 2 (BY-2). The (γ-Glu-Cys) n-Gly, (γ-EC) nG, synthesis activity and Cd tolerance ability were increased in transformed tobacco cells and that Cd concentration was increased in respect to the concentration of synthesized (γ-EC) ZG in Cd-exposed transformed tobacco cells. These results indicated that the CPase gene, when generated in transformed tobacco cells, causes an increase in (γ-EC) nG synthetic activity and tolerance to Cd. These results further indicated that a production of the heavy metal tolerance plants are possible by introducing the gene of the enzyme which has (γ-EC) nG synthesis ability.
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