Figure 3.

Map-Based Cloning of BUI1.

(A) The BUI1 locus was mapped on chromosome 7 between two SSR markers, RM1132 and RM505.

(B) Fine-mapping of BUI1. BUI1 was narrowed to a 60-kb region between two makers, 7WB8 and 7WB16, on a single BAC (AP004275), which contains three predicted genes.

(C) Sequence comparison revealed a substitution of A to G in one intron of the gene FH5/Os07g0596300 in bui1. WT, wild type.

(D) Detection of altered splicing in bui1 by RT-PCR analysis. Rice UBI1 was used as an internal control.

(E) RNA gel blot analysis to confirm the length of the FH5/BUI1 transcript. Total RNA (10 μg) extracted from the wild type and bui1 was used for the analysis (shown below the blot). M, RNA ladder.

(F) Confirmation of the full-length FH5/BUI1 transcript by RT-PCR. The primer pairs (P1 and P2) used for RT are indicated in (C).

(G) Complementation test of the FH5/BUI1 gene. One representative line (1300-BUI1) of complementation is shown. Bar = 2 cm.

(H) Expression pattern of FH5 revealed by RNA gel blot. YS, Young seedling; FL, flower; RA, rachis; LF, leaf; LS, leaf sheath; ND, node; IN, internode; RT, root.