Figure 2
Map-Based Cloning of gns2. (A) Schematic representation of the mapping strategy for identifying the gns2 mutation. New molecular markers used in this study are presented in Additional file 1: Table S1. (B) A dCAPS marker distinguishing gns2 and WT. A deletion of a guanine (39 bp from ATG) was detected in the first exon of AtELP3, which allowed the development of a dCAPS marker to genotypically distinguish the mutant from the wild type (Additional file 1: Table S1). (C) Structure of the GNS2/ELO3/AtELP3 gene and the positions of the gns2 and elo3-1 mutations [39]. Boxes denote the translated regions and lines between boxes denote introns. (D) Ten-day-old seedlings of WT, npr1, Atelp3-10 npr1, and two complementation lines Com(n)#1 and Com(n)#2 grown on half-strength MS agar medium containing 0.26 mM SA. Com(n), 35S::AtELP3 Atelp3-10 npr1 transgenic plants. (E) Four-week-old soil-grown WT, npr1, Atelp3-10 npr1, and Com(n) plants. (F) Free SA levels in Psm ES4326-infected WT, npr1, Atelp3-10 npr1, and Com(n) plants. Plants were inoculated with Psm ES4326 (OD600 = 0.001). Twenty-four hours later, the inoculated leaves were collected for SA measurement. Data represent the mean of four independent samples with SD. Different letters above the bars indicate significant differences (p < 0.05, Student’s t-test). (G) Total SA levels in Psm ES4326-infected plants. The experiment was carried out as in (F). (H) Growth of Psm ES4326 in WT, npr1, Atelp3-10 npr1, and Com(n) plants. Cfu, colony-forming units. Leaves of four-week-old plants were inoculated with Psm ES4326 (OD600 = 0.0001). The in planta bacterial titers were determined immediately and three days postinoculation. Cfu, colony-forming units. Data represent the mean of eight independent samples with SD. Different letters above the bars indicate significant differences (p < 0.05, Student’s t-test).