Figure 3.

RT-PCR analysis of hormone response of Pbf expression in rehydrated aleurone layers. Total RNA was isolated from aleurone layers after different hormone treatments and was reverse transcribed in the presence of random hexamers. The first strand cDNA was then amplified by PCR using gene-specific primers for the Pbf (BPBF) or the cathepsin B-like thiol protease (thiol protease) transcripts. Amplification of a region of the 18S RNA was used as the internal control (18S). The PCR products were fractionated in agarose gels and were visualized by ethidium bromide staining. A, Comparative Pbf expression pattern in aleurone layers incubated in the presence of 1 μm GA₃ for 0, 4, 8, 16, or 24 h. B, Expression analysis of Pbf and of the cathepsin B-like thiol protease transcripts in aleurone layers incubated for 4 h with (+) or without (−) 1 μm GA₃, 10 μm ABA, or both, as indicated above.