Truncation analysis of the AmidP in cotyledons of transgenic Arabidopsis seedlings.

<p>A. Promoter truncation, the primers binding sites and endonucleases sites are illustrated. The 1504 bp cloned sequence for the AmidP includes 4 nucleotides which are added to form an <i>Eco</i>RI acting site. All the fused structures are obtained by ligation of the pFGC-DR and the PCR products precut by <i>Eco</i>RI and <i>Nco</i>I respectively, except that of the P3-DR which is self-ligated with the P1-DR digested by <i>Bam</i>HI and <i>Bgl</i>II. B-G. GUS activities in the transgenic Arabidopsis cotyledons of P1-DR (B), P2-DR (C), P3-DR (D), P4-DR (E), P5-DR (F), and P6-DR (G).</p

Tandem copies of <i>VVE</i> (or the reverse complement orientation) motif on the strength of GUS activities.

<p>A. The number and orientation of <i>VVE</i> tandems is illustrated (not to scale). <i>VVE</i>, represented with black arrows, is placed upstream of the minimal 35S promoter (−65; black box) to drive <i>uidA</i> (blank box) expression (see “Materials and methods”). B. Quantitative GUS activity analysis of the tandem construction in leaves of 10 d transgenic Arabidopsis seedlings. pFGC-DR and pFGC-MiniGUS were used as positive and negative controls, respectively. GUS activity in pFGC-DR transgenic seedlings was assigned as 100%. GUS activity is replicated three times of each collection of seedlings from independent transgenic lines (indicated in parentheses) per construction. Error bars are standard deviations.</p

The AmidP drives the GUS expression in the vascular vein of leaves resembling a pattern of the sink-to-source transition.

<p>A. GUS expression is detected in cotyledons and the distal tip of young leaves of 10-d seedlings. B. The AmidP drives expression in the germinating seed joint of above- and under-ground part. C–D. GUS activity is detected in sepals of flowers (C), as shown in an amplified flower indicated with a red arrow (D). E–J. X-Gluc staining is detected throughout the vascular veins of a cotyledon (E), expanded source leaves (F, G, and H) and progresses basipetally down transition leaves (I and J).</p

The construction and results of the 5′- and 3′- deletions of <i>VVE</i> motif between −1500 and −1324 of the AmidP.

<p>A. Sequence and element site analysis of the <i>VVE</i> motif in the AmidP. 4 nucleotides (grey box) are arbitrarily added to form an <i>Eco</i>RI acting site. Element sites for known transcription factors indicated as arrows are detected by AthaMap web tools (see “Materials and methods”). Vertical dotted line indicated deletion sites. B. Schematic diagram of the chimeric constructs. The numbers above the bars indicate the residual region of the <i>VVE</i> motif after 5′- or 3′- deletions. All the fused constructs are obtained by ligation of the pFGC-MiniGUS and the PCR products precut by <i>Eco</i>RI and <i>Bam</i>HI respectively. C–K. Representative histochemical stained cotyledon demonstrates the strength and specificity of GUS activities in the transgenic Arabidopsis of 5M1 (C), 5M2 (D), 5M3 (E), 5M4 (F), 3M1 (G), 3M2 (H), 3M3 (I), 3M4 (J), and 3M5 (K).</p

Various primers used in this experiment.

<p>Various primers used in this experiment.</p