Figure 7.

sec24B and sec24C Fail to Complement sec24A^R693K.

(A) Confocal images of T1 G92 cotyledon epidermal cells alone or transformed with 35S:sec24B or 35S:Sec24C constructs show the presence of globular structures (arrowheads) similar to those in G92 (arrowheads). This is in marked contrast to G92 plants transformed with 35S:Sec24A, which complements the phenotype. Bar = 40 μm.

(B) Expression levels of the sec24 transgenes were analyzed by RT-PCR from two independent T1 G92/35S:Sec24A, Sec24B, and Sec24C lines. Although the transcription levels of sec24B and sec24C in the respective transgenic lines are comparable to sec24A in complemented transgenic plants, sec24B and sec24C failed to complement the sec24A^R693K phenotype (A). Amplification was performed on 25 ng of cDNA. Amplification of UBQ10 was used as control. The number of PCR cycles is indicated below the RT-PCR bands. These data are representative of two biological replicates and two technical replicates. Additional evidence for plant transformation is provided in Supplemental Figure 7 online.