Molecular Characterization of the brd2 Mutation

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Molecular Characterization of the brd2 Mutation Molecular Characterization of the brd2 Mutation.(A) The brd2 mutation was mapped at ∼25 centimorgan on chromosome 10, near the map position of Dim/dwf1.(B) Schematic diagram of the structure of Dim/dwf1 and the site of the mutation in the brd2 allele. Molecular Characterization of the brd2 Mutation.(A) The brd2 mutation was mapped at ∼25 centimorgan on chromosome 10, near the map position of Dim/dwf1.(B) Schematic diagram of the structure of Dim/dwf1 and the site of the mutation in the brd2 allele. Closed boxes represent exons, and open boxes indicate introns. The first Met (ATG) and the stop codon (TAA) are located in exons 2 and 3, respectively. The single base deletion in the brd2 allele is indicated by an arrow, and the premature stop codon is marked with an asterisk.(C) The deduced amino acid sequence of Dim/dwf1. The mutation site is marked with a closed triangle, and the FAD binding domain is underlined.(D) Levels of Dim/dwf1 transcripts in wild-type (left) and brd2 (right) plants. Ten micrograms of total RNA was loaded per lane. Ethidium bromide–stained rRNA bands were monitored as a loading control.(E) Molecular complementation. Transgenic plants derived from mutant callus transformed with the Dim/dwf1 gene had the wild-type phenotype, in contrast with the dwarf phenotype of the mutant (right), and transformants containing the empty vector had the dwarf phenotype (left).(F) Organ-specific expression of the Dim/dwf1 gene in the wild type. RT, root; SA, shoot apex; LB, leaf blade; LS, leaf sheath; ST, stem; PA, panicle; FL, flower. Five micrograms of total RNA was loaded per lane, and ethidium bromide–stained rRNA bands were monitored as a loading control.(G) The effect of BL on Dim/dwf1 expression. Total RNA was isolated from wild-type, brd1-1, and d61-2 seedlings either treated for 10 d with a final concentration of 1 μM BL (+) or untreated (−); 7.5 μg was loaded per lane. Zhi Hong et al. Plant Cell 2005;17:2243-2254 ©2005 by American Society of Plant Biologists