Figure 3.

Expression of V. aconitifolia P5CSF129A in transgenic tobacco plants. A, Northern blot of total RNA from seedlings of 14-d-old wild-type (lane 1), P5CSF129A-3 (lane 2), and P5CS line 136 (lane 3). RNA blot was probed with ³²P-labeled V. aconitifolia P5CS cDNA (top panel) or ribosomal DNA as internal loading control (bottom panel). Molecular masses of RNA markers are indicated (in kb). B, Western blot of total soluble proteins from 14-d-old wild-type (W.T.), P5CSF129A-3 (F129A), and P5CS line 136 (P5CS) seedlings. Seeds were germinated and maintained on MS medium containing 0 or 200 mm NaCl for 2 weeks. The nitrocellulose membrane was reacted with antibodies to purified V. aconitifolia P5CS protein. The top band corresponds to the expected size of P5CS protein, whereas the bottom band appears to be a degradation product. Molecular sizes of protein markers (Bio-Rad, Hercules, CA) are indicated in kD.