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. 2008 Oct;20(10):2729–2745. doi: 10.1105/tpc.108.061515

Figure 3.

Figure 3.

Coexpression of ABI5 and PKABA1 Is Sufficient to Block Seed Germination, and PKABA1-Dependent Activation of ABI5 Is Associated with ABI5 Phosphorylation.

(A) Protein gel blot analysis of HA-ABI5 and HA-PKABA1 protein levels in wild-type (Wassilewskija) and WT/35S:HA-ABI5 plants transformed with the ind:HA-PKABA1 DNA construct. HA-PKABA1 protein levels could be detected only in the presence of the inducer (50 μM 17β-estradiol). Protein extracts were isolated from plant material at 48 h after seed imbibition under normal conditions with or without inducer. Ponceau S staining is shown as a loading control.

(B) Mobility differences of HA-ABI5 protein isolated from WT/35S:HA-ABI5 lines transformed with a ind:HA-PKABA1 DNA construct. Same protein gel blot analysis as in Figure 2B under the indicated germination conditions. λ phosphatase was inactivated by heat (65°C for 20 min) or left active (30°C for 30 min). The gel image is duplicated at bottom with a dashed line to facilitate the comparison of protein mobility in each lane.

(C) Plant material used in (A) at 120 h after seed imbibition. Excess GA (50 μM) in the medium did not promote germination when PKABA1 protein was present.