Fig. 6

Analysis of anther wax and cutin in the wild type and fad2–3. (a) Wax constituents in the wild-type and fad2–3. (b) Cutin monomers in the wild-type and fad2–3. C23 ALK, tricosane; C25 ALK, pentacosane; C27 ALK, heptacosane; C28 ALK, octacosane; C29 ALK, nonacosane; C31 ALK, hentriacontane; C35 ALK, pentatriacontane. C16:0 FA, hexadecanoic acid; C18:0 FA, octadecanoic acid; C18:1 FA, 9-octadecenoic acid; C18:2 FA, 9,12-octadecadienoic acid; C18:3 FA, 9,12,15-octadecatrienoic acid; C20 FA, eicosanoic acid; C22 FA, docosanoic acid; C24 FA, tetracosanoic. C16:0 DCA, hexadecane-1,16-dioic acid; C18:1 DCA, α, ω-octadecenoic acid; C18:2 DCA, α, ω-octadecadiendioic acid; triOH C18:1 FA, 9,10,18-trihydroxy octadecenoic acid; 9,10 Epoxy 18-OH acid, 9,10-epoxy-18-OH-C18:1; DW, dry weight. The wax of anther at mature pollen stage was analyzed according to Jung et al. [26]. The wax monomer was derivatized with 1 ml BFTSA in 1 ml pyridine (1:1) for 40 min at 70 °C before GC-MS analysis. The constituent analyses were performed using GCMS-QP2020 with a DB-1 column. Each compound was quantified against the internal standard by automatic integrating the peak areas. The protocol for lipid polyester analysis was performed according to Li-Beisson et al. [50]. The cutin monomer fraction was derivatized with BFTSA/pyridine (1:1) for 60 min at 70 °C. The constituent were analyzed using GCMS-QP2020 with a DB-1 column. The GC-MS was conducted according to Li-Beisson et al. [50] with helium carrier gas at 2 ml/min. Each compound was quantified on the basis of their total ion current as described by Li-Beisson et al. [50]. Error bars are standard errors. Values represent the means ± SE, n = 3. Asterisks denote significant differences to wild-type (WT) as determined by Student’s t test: ***p < 0.001; **p < 0.01; *p < 0.05