Microarray data from field grown plants.

<p>Microarrays performed on tissue sampled throughout the day from two <i>AtBBX32</i>-expressing lines (lines 1 and 2 from <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0030717#pone-0030717-t001" target="_blank">Table 1</a>) in the field demonstrate 219 genes show 2–8 fold changes (8-fold is maximum change observed) in abundance in both transgenic events relative to the control and that the majority of these changes occur around ZT 0 (6 am). Dark bar represents genes increased in abundance and light bar represents genes decreased in abundance. All changes significant at a false discovery rate of 5 percent.</p

<i>AtBBX32</i> extends the reproductive period between R3 and R7 developmental stages in soybean resulting in a delay in final maturity compared to control.

<p>The timing of reproductive development was measured according to standard methods <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0030717#pone.0030717-Fehr1" target="_blank">[16]</a> in ten field plot replicates for each line. R1 is the initiation of flowering. R3 is the onset of pod development. R7 is the beginning of maturation. R8 is the stage where 95 percent of the pods are physiologically mature. The number of days to reach each developmental stage was calculated on a whole plot basis and the mean is indicated below, where units are days after planting.</p><p>*p<0.05.</p

<i>AtBBX32</i> transgenic soybean plants increase yield components.

<p>Both field and growth chamber grown plants show increases in plant height and pod number in the transgenic lines relative to controls. Node number, flower number, seed number, and 100 seed weight were also increased in growth chamber grown plants. Growth chamber experiments were performed in a 10:14 hour photoperiod (Light∶Dark) with 900 mE of light. Data was collected from ten plant replicates that were randomized among entries in the chamber. Field grown plants were grown under standard agronomic conditions and ambient light. All differences between the transgenic lines and control are significant to p<0.05 unless otherwise indicated as (ns) not significant.</p

<i>AtBBX32</i> transgenic soybean plants demonstrate improved grain yield over non-transgenic controls.

<p>Mean yield values (kilograms per hectare) and percent improvement over controls for transgenic plots are shown for three growing seasons. The difference in the day of flowering (DOF) between the transgenic lines and control was calculated to determine delta DOF. The difference in day of final maturity (MAT) was examined in transgenic lines and compared to control to determine delta MAT (units = days). The low yielding event 4 produced no detectable transcript. N represents the number of environments tested. p-values were based on the difference between the transgenic lines and wildtype control.</p><p>*p≤0.05,</p><p>**p≤0.01.</p

Expression of <i>AtBBX32</i> in soybean affects the transcript abundance of central clock components near ZT 0.

<p>Levels of both central clock components <i>GmLCL2</i> (A) and <i>GmTOC1</i> (B) were assayed by quantigene RNA extraction and expression analysis from V2 leaf tissue harvested from soybean plants grown in a controlled environment. Growth chamber experiment was performed in a 14:10 hour photoperiod (Light∶Dark) with 650 mE of light. p-values based on the difference between both transgenic lines and wildtype control. * p≤0.05. Where error bars are not visible they are smaller than the data points.</p

<i>AtBBX32</i> expression in soybean delays leaf senescence and brown pod maturity.

<p>A) Field grown soybeans were visually assessed and scored every few days late in the season on a whole plot basis according to green leaf color. Leaf senescence was rated on a 1–9 scale based on whole plot appearance. 9 = dark green, no yellow leaves on the top canopy; 5 = 40 percent change yellow leaves, 10 percent change fallen leaves; 1 = more than 95 percent change fallen leaves. B) The same soybean plots were visually inspected for the appearance of brown pods and the percentage of the plot containing brown pods was determined. * Event 1 significantly different from control at p<0.05, † Event 2 significantly different from control at p≤0.05.</p