Abstract

Phytochelatins (PCs) are heavy metal binding peptides that play important roles in sequestration and detoxification of heavy metals in plants. To develop transgenic plants with increased tolerance and/or accumulation of heavy metals from soil, an Arabidopsis thaliana FLAG–tagged AtPCS1 cDNA encoding phytochelatin synthase (PCS) under the control of a 35S promoter was expressed in Indian mustard (Brassica juncea). Four transgenic Indian mustard lines, designated pc lines, with different levels of AtPCS1 mRNA accumulation and correspondent AtPCS1 protein levels were selected and analyzed for tolerance to cadmium (Cd) and zinc (Zn). Heavy metal tolerance was assessed by measuring root length of 10-day-old seedlings grown on agar medium supplemented with different concentrations of Cd (0, 100, 150, and 200 μm CdCl2) and Zn (200, 400, 600, and 800 μm ZnCl2). All transgenic lines showed significantly longer roots when grown on a medium supplemented with 100 μm CdCl2. No significant differences were observed between transgenic lines and wild type when plants were grown on higher levels of Cd. This indicated that only partial tolerance to Cd was observed in these transgenic lines. Similarly, partial tolerance for Zn was also observed in these transgenic lines, but up to levels of 400 μm ZnCl2. Expression levels of AtPCS1 protein were not related to tolerance responses for either Cd or Zn stresses in transgenic lines.

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