Yeast metallothionein in transgenic tobacco promotes copper uptake from contaminated soils.

Abstract

Metallothioneins (MTs) are metal-binding proteins that confer heavy metal tolerance and accumulation in yeast. To augment higher plant metal sequestration, the yeast metallothionein (CUP 1) was introduced into tobacco plants. The CUP 1 gene expression and copper and cadmium phytoextraction were determined. To confirm transformation, selfed and kanamycin-resistant third generation plants were subjected to DNA blot and polymerase chain reaction (PCR) analysis. A 4 mM CuSO(4) stress for 7 days resulted in a decline in CUP 1 transcripts versus nonstress conditions. Despite low mRNA levels, CUP 1 transformants accumulated up to seven times more copper in older versus younger leaves during copper stress. Pooled leaves of transgenic plants grown in soils from copper stamp-sands contained two to three times the copper content as that of the control plants. Unlike some previous reports featuring MT overexpression in plants, CUP 1 seedlings did not significantly sequester or demonstrate tolerance to CdCl(2). Using this transgenic approach, yeast CUP 1 expression under nonstressed conditions contributed to copper metal phytoextraction during a subsequent copper challenge. This strategy could be incorporated into plants designed for enhanced phytoremediation of metal contaminants.