Figure 3.
Complementation of aao3-2 and aao3-3 mutants by AAO3 gene. A, Seed germination and early seedling growth in the presence of NaCl or mannitol. Wild-type (Col), aao3-2, aao3-3, and T3 homozygous progeny of the mutants transformed with the binary vector pPZP211 containing a ProAAO3-AAO3 insert. Seeds were germinated on MS medium (MS) or medium supplemented with either 200 mm NaCl or 400 mm mannitol. The picture was taken at 10 d after sowing. B, Percentage of seeds that germinated and developed green cotyledons under osmotic stress for wild-type, aao3-2, aao3-3, and representative transformed lines (T3 progeny) of aao3-2 and aao3-3. Seeds were sowed on MS agar plates supplemented either with 200 mm NaCl (white bars) or 400 mm mannitol (black bars). Approximately 200 seeds were sowed and scored 5 (NaCl) or 8 (mannitol) d later. Germination in MS medium was between 95% and 100%. C, The enhanced transpiration rate of both aao3-2 and aao3-3 plants is complemented by a wild-type AAO3 transgene. Loss of fresh weight was measured in detached rosette leaves of wild type, aao3-2, aao3-3, and T1 transformed lines of aao3-2 and aao3-3. Four leaves per individual were excised, and fresh weight was determined at ambient conditions (25°C and approximately 40% relative humidity) after 60 min. The percentage of fresh weight lost is indicated.