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. 2017 Dec 22;8:2182. doi: 10.3389/fpls.2017.02182

FIGURE 2.

FIGURE 2

PRE1 positional cloning and pre1-1 complementation. (A) The PRE1 locus was mapped to BAC clone X7J on chromosome 1. PRE1/At1g58360/AAP1 is composed of 6 exons (black boxes) and 5 introns (lines). BAC names and their regions spanned are indicated. DNA sequencing revealed a 19-bp deletion in the first exon of PRE1/AAP1 in pre1-1, resulting in truncation of PRE1/AAP1. The T-DNA insertion site in pre1-2 is indicated with a triangle. (B) Quantification of proline sensitivity and (C) average primary root length (C) in 9-day-old seedlings of wild type, mutant, and complemented lines treated with 0.5× MS medium supplemented with 45 mM L-proline (Pro). (D) The proline content of 12-day-old seedlings of wild type, mutant, and complemented lines treated with 5 mM Pro for 24 h. (E) The relative expression of PRE1/AAP1 in 12-day-old seedlings of wild type, mutant, and complemented lines. Col, wild-type Columbia; pre1-1 PRE1-GFP, pre1-1 transformed PRE1 cDNA fused with GFP under the control of the PRE1/AAP1 promoter; pre1-1 gPRE1, pre1-1 transformed with a genomic fragment of PRE1/AAP1. Experiments were repeated a minimum of three times with similar results. The values are means ± standard error (n = 6). Different letters above columns indicate significant difference (P < 0.05, Duncan’s multiple range test).