Characterization and complementation of the HAP1/hap1-2 mutant.

(A) HAP1 gene structure. Exons were shown as colored boxes; introns were shown as straight lines; UTRs were shown as gray boxes. The T-DNA insertion in hap1-1 and hap1-2 was indicated by red arrow. (B) rtPCR results showed the expression of a full-length transcript (AtMago), a truncated transcript (AtMagoΔC) and GAPDH in qrt, qrt;HAP1/hap1-2, qrt;sr45-1 and qrt;upf3-1 mutant plants. Total RNAs were extracted from the inflorescence tissue. The exon composition of the transcript and the position of used primers were shown next to the corresponding rtPCR results. (C) GUS stained pollen grains from qrt, qrt;HAP1/hap1-1 and qrt;HAP1/hap1-2 showing the identity of mutant pollen grains. Scale bar = 20 μm. (D) Seed number per silique in qrt and qrt;HAP1/hap1-2 plants. Scale bar = 1 mm. The quantification was done with sixteen siliques. Student t-test was used for statistical analysis. Error bars represent standard deviations. (E) HAP1 gene complemented hap1-2 mutant phenotype. The mature plants, GUS-stained pollen grains and seed yield per plant were shown in qrt, qrt;HAP1/hap1-2, HAP1;qrt; hap1-2. (F) qPCR results showing that the AtMago gene expression level was fully recovered in the transgenic plant. GAPDH was used as control. Three biological replicates were used in the analysis. Error bar showed standard deviation. Statistical significance compared to qrt was measured by student t-test (p