Figure 1.

T-DNA Insertions in the fim5-1 and fim5-2 Lines Generated Null fim5 Alleles.

(A) Structure of the FIM5 gene (At5g35700). A total of 14 exons and 14 introns are present in the gene. Introns, exons, and untranslated regions are defined by gray boxes, black boxes, and black lines, respectively. The two T-DNA insertion lines, CS856909 (designated as fim5-1) and CS853476 (designated as fim5-2), contained insertions in the 10th intron and 13th exon, respectively.

(B) RT-PCR results demonstrated that the fim5-1 and fim5-2 lines contain fim5 null alleles. Transcript expression levels in the insertion lines were determined by RT-PCR using three independent pairs of primers. eIF4A was used as an internal control. The first pair of primers (F5f1 and F5r1) was designed to amplify the FIM5 full-length cDNA, which was not present in fim5-1 and fim5-2 mutant plants. The second and third primer pairs, F5f2/F5r2 and F5f3/F5r3, were used to amplify partial transcript 1 and partial transcript 2, respectively. RT-PCR results showed that no partial FIM5 cDNAs were detected in fim5-1 and fim5-2 mutant plants. Lane 1, water (negative control; CK); lane 2, wild-type (WT) Col-0; lane 3, fim5-1; lane 4, fim5-2.