Figure 5.

Functional complementation of cad1-3 by AtPCS1construct. A, Cad1-3 mutants were transformed with C-terminal FLAG-tagged genomic AtPCS1 under the control of a 2.0-kb AtPCS1 promoter. After screening of transgenic (cad1pcs1) lines on kanamycin, selected T₂ seeds were used for testing the recovery of Cd hypersensitivity in cad1-3 plant. Approximately 20 seeds were germinated and grown in a vertical orientation for 10 d on Murashige and Skoog agar medium containing 0 μm (top), 50 μm (middle), or 85 μm (bottom) CdCl₂, and root lengths were then measured. Values correspond to means ± se. B, Western-blot analysis was performed on various cad1pcs lines to evaluate the amount of the expressed C-terminal FLAG-tagged AtPCS1. Proteins were extracted from 10-d-old seedlings and were analyzed as described in Figure 1. The pcs1 and pcs3 lines were used as standards.