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We report a systematic comparison of 19 plant promoters and 20 promoter-terminator combinations in two expression systems: agroinfiltration in Nicotiana benthamiana leaves, and Nicotiana tabacum BY-2 plant cell packs. The set of promoters tested comprised those not present in previously published work, including several computationally predicted synthetic promoters validated here for the first time. The expression of EGFP driven by different promoters varied by more than two orders of magnitude and was largely consistent between two tested Nicotiana systems. We confirmed previous reports of significant modulation of expression by terminators, as well as synergistic effects of promoters and terminators. Additionally, we observed non-linear effects of gene dosage on expression level. The dataset presented here can inform the design of genetic constructs for plant engineering and transient expression assays.

The discovery of the bioluminescence pathway in the fungus Neonothopanus nambi enabled engineering of eukaryotes with self-sustained luminescence. However, the brightness of luminescence in heterologous hosts was limited by performance of the native fungal enzymes. Here we report optimized versions of the pathway that enhance bioluminescence by one to two orders of magnitude in plant, fungal and mammalian hosts, and enable longitudinal video-rate imaging. Improvements to the fully genetically encoded Neonothopanus nambi bioluminescence pathway enhance autobioluminescence by up to two orders of magnitude in plants and other species, enabling novel applications of bioluminescence imaging in biology.

Jasmonic and salicylic acids are the major hormones involved in plant response to pests and pathogens. Here, we engineered autoluminescent plants that report activity of these hormones with up to 53-fold contrast. Using consumer-grade cameras, we imaged reporter Arabidopsis thaliana and Nicotiana benthamiana plants throughout normal development, and in response to attacks of pests and pathogens.Competing Interest StatementThis study was partially funded by Planta (planta.bio) and Light Bio (light.bio).