Figure 4.

Isolation of the kin17-1 mutant line. A, Scheme representing the KIN17 gene and the translated protein with its conserved domains defined. The T-DNA insertion in the kin17-1 line is indicated by an inverted triangle and corresponding flanking sequences. The positions of the oligonucleotides used to genotype the lines are shown by arrows (F, I, and R). The Zn finger domain (ZFD), Kin17, and KOW domains are indicated with squares. aa, Amino acids. B, Discrimination of homozygous kin17-1 plants. kin17-1 plants were self-crossed, and the offspring was genotyped with the oligonucleotide combinations indicated in A: for the wild-type (WT) allele, F + R; for the insertion allele, F + I. C, The kin17-1 line is a knockdown mutant. Total RNA from 6-d-old seedlings of the wild type, kin17-1, and pKIN17::KIN17 in the kin17-1 mutant background grown on 1/2 MS medium supplemented with Suc and BCS was prepared and reverse transcribed to cDNA. KIN17 mRNA relative levels were determined by qPCR and normalized to ACTIN1 and ELONGATION FACTOR1α levels. Error bars correspond to sd values of at least three independent biological replicates. The asterisk indicates a statistically significant difference from the wild type (P < 0.05).