Creation of <i>mALS1</i> T-DNA transgenic lines.

(A) A schematic representation of the T-DNA construct inserted into the plant genome. The T-DNA, in which mALS1 is fused with NPTII, is driven by a native ALS1 promoter. Grey boxes flanking the LB and RB indicate plant DNA. HPT, hygromycin phosphotransferase; NPTII, neomycin phosphotransferase II, Nos, terminator of nopaline synthase; pA, poly-A site; T2A, 2A peptide from Thosea asigna virus capsid protein. Four nonsynonymous mutations in mALS1 are shown. (B) Location of T-DNA insertion sites in the DMRH S5 28–5 genome identified by inverse PCR approach. Size of each chromosome is proportional to its physical size. Black lines indicate locations of T-DNA insertion in transgenic lines. Note that the true location of the T-DNA insertion site in als1-17 is unknown due to multiple mapping of reads derived from the inverse-PCR fragment to the genome. Full information of T-DNA insertion sites is provided in S1 Table.