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. 2020 Mar 25;9:e52786. doi: 10.7554/eLife.52786

Figure 4. The FBP can be used to build reporters of hormone signaling dynamics in planta.

(A) Schematic of the Luz expression cassette driven by the ABA-responsive AtRAB18 promoter. (B) Bar plots summarizing mean luminescence signal observed from N. benthamaina leaves co-infiltrated with the pRAB18:Luz FBP variant along with increasing concentrations of the hormone ABA and then imaged after three days. Black bars represent standard deviation (n = 3). (C) Luminescence signal observed from N. benthamaina leaves infiltrated with an FBP and then either kept moist (blue background) or allowed to desiccate to trigger an ABA signal (orange background). Gray and red dots represent data from independent leaves infiltrated with FBP with a 35S or pRAB18 driven Luz respectively. Representative images of watered and un-watered leaves infiltrated with the pRAB18:Luz FBP with luminescence signal overlaid on a bright field picture. Warmer colors represent higher signals. (D) Still images from the start and end of Video Figure 1, where one leaf each on two N. benthamiana plants were agro-infiltrated with FBPs that had either a 35S or pRAB18 driven Luz. One plant was allowed to desiccate (bottom two leaves labeled unwatered) while the other was kept watered (top two leaves labeled watered). The infiltrated leaves are highlighted with a dashed yellow line. A paired bright field image is inset in corner of each image.

Figure 4—source data 1. Raw data for Figure 4.

Figure 4.

Figure 4—figure supplement 1. An FBP with pRAB18 driving Luz expression shows increased luminescence in unwatered conditions.

Figure 4—figure supplement 1.

Time course luminescence data from N. benthamiana leaves infiltrated with a pRAB18:Luz FBP variant either kept moist (n = 3, blue) or allowed to dry out (n = 6, orange). The solid lines are mean luminescence and light outlines represent standard deviation.
Figure 4—video 1. Time lapse movie of Nicotiana benthamiana transiently expressing an FBP with pRAB18 driving Luz expression in watered and unwatered conditions with parallel positive controls.
Download video file (1.5MB, mp4)
Time lapse imaging was performed with our RaspberryPi controlled DSLR imaging platform to visualize ABA induced FBP expression in plants exposed to drought stress. We infiltrated one leaf each on two N. benthamiana plants with FBPs that had either a 35S (+control) or pRAB18 driven Luz. One plant was allowed to desiccate while the other was kept watered. Bioluminescence in the leaves as then imaged over three days. The treatments of the leaves are labeled with white legends.