Identification of Pha13 involvement in <i>PhaPR1</i> expression by CymMV-based VIGS system and sequence analysis.

<p>(A) Expression level of <i>Pha13</i>, <i>PhaPR1</i>, and CymMV were analyzed by RT-PCR from leaves of <i>P</i>. <i>aphrodite</i> inoculated with buffer (Mock), or infiltrated with agrobacterium carrying pCymMV (Cy) or pCymMV-Pha13 (Pha13-VIGS). <i>PhaUbiquitin 10</i> (PhaUBQ10) was used as a loading control, and the relative expression level of corresponding genes are indicated. (B) Domains of Pha13. Open rectangle indicates the entire protein. A20 (black rectangle) and AN1 (diagonally-striped rectangle), nuclear localization signal (open square with vertical lines), and 21-nucleotide position (short black line) used for designing hairpin RNA of <i>Pha13</i> (hpPha13) are indicated. (C) Sequence alignment of A20/AN1 zinc finger domains of Pha13 with stress-associated proteins from <i>Arabidopsis thaliana</i> (AtSAP5, accession number: AT3G12630), and <i>Oryza sativa</i> (OsSAP3, accession number: LOC_Os01g56040.1; OsSAP5, accession number: LOC_Os02g32840.1) are shown. Conserved amino acid sequences are indicated with a black box. Conserved cysteine (C) and histidine (H) are indicated with a black triangle. (D) Primary sequence organization of A20 and AN1 domains. Xn: the number of amino acid residues between zinc ligands. (B to D) The mutation positions for domain functional analysis are indicated with red triangle.</p>