FIGURE 10.

Identification and characterization of Atsuc7 T-DNA insertion lines. (A) Genomic organization of AtSUC7. Introns and untranslated regions are shown as black lines; exon regions containing coding sequences are represented by numbered gray bars. Arrows indicate the primers used for PCRs shown in (B,C). The positions of the insertion in the T-DNA lines are marked. LB, left border; RB, right border. (B) PCR products obtained from genomic DNA preparations of homozygous Atsuc7.3 and WT plants. Primer combinations for the detection of wild type (WT) and mutant allele (m) are listed in Supplementary Table 2. (C) RT-PCR analyses of pollen tube RNA obtained from a homozygous Atsuc7.3 mutant and a WT plant with primers (Supplementary Table 3) amplifying either the AtSUC7 sequence traversing, upstream of or downstream of the insertion. WT genomic DNA was used a control for genomic contaminations. (D) Length of main roots of 12-day-old Atsuc7.3 and WT seedlings on MS-0 or MS medium supplemented with 2% (w/v) sucrose. Means of five biological replicates ± SE are shown. n > 50 for each genotype. (E) Average number of seeds/silique ± SD of Atsuc7.3 and WT plants after self-pollination. n > 55 siliques/genotype. (F) Pollen tube lengths of WT and Atsuc7.3 in vitro. Pollen were grown on medium with different sucrose concentrations for 8 h. Bars represent mean values of three biological replicates ± SE (n > 500 in total per sucrose concentration for each genotype). (G) Genotypes of F1 descendants of a cross-pollination experiment with heterozygous Atsuc7.3/AtSUC7 pollen and pistils from a WT plant. Bars represent mean values (±SE) of WT and heterozygous plants in the F1 generation resulting from seven independent crossings (n = 89 in total).