Figure 4.

Autophagy-dependent mobilization of stromal proteins into the vacuole by RCBs in rice. A, Schematic representation illustrating the construct for CT-sGFP expression. Fusion proteins consisting of the transit peptide (TP) of OsRBCS2 and sGFP were expressed under the control of the CaMV 35S promoter. B, Visualization of RCBs in rice leaves. The first leaf of CT-sGFP-expressing rice was excised and observed immediately (0 h) or after incubation for 20 h in the presence of 1 μm concanamycin A without (+concA) or with (+concA+wort) 5 μm wortmannin. C, RCB accumulation is not observed in OsATG7 knockout plants. First leaves of Osatg7-1 or the corresponding wild-type plant (OsATG7+/+) expressing CT-sGFP were excised and incubated for 20 h in the presence of 1 μm concanamycin A (+concA). GFP fluorescence appears green, and chlorophyll autofluorescence appears magenta. In merged images, overlapping signals appear white. Bars = 10 μm.