Affiliation: | 1. College of Agriculture and Biotechnology, Zhejiang University, Hangzhou, China ZJU-Hangzhou Global Scientific and Technological Innovation Center, Zhejiang University, Hangzhou, China These authors have equal contributions to this study.;2. College of Agriculture and Biotechnology, Zhejiang University, Hangzhou, China These authors have equal contributions to this study.;3. College of Agriculture and Biotechnology, Zhejiang University, Hangzhou, China;4. College of Agriculture and Biotechnology, Zhejiang University, Hangzhou, China Hainan Institute of Zhejiang University, Sanya, China;5. College of Agriculture and Biotechnology, Zhejiang University, Hangzhou, China ZJU-Hangzhou Global Scientific and Technological Innovation Center, Zhejiang University, Hangzhou, China;6. Marine Agriculture Research Center, Tobacco Research Institute of Chinese Academy of Agricultural Sciences, Qingdao, China;7. Key Laboratory of Structural Biology of Zhejiang Province, School of Life Sciences, Westlake University, Westlake Laboratory of Life Sciences and Biomedicine, Institute of Biology, Westlake Institute for Advanced Study, Hangzhou, China;8. State Key Laboratory of Subtropical Silviculture, Zhejiang A&F University, Hangzhou, China |
Abstract: | The fungal bioluminescence pathway (FBP) was identified from glowing fungi, which releases self-sustained visible green luminescence. However, weak bioluminescence limits the potential application of the bioluminescence system. Here, we screened and characterized a C3′H1 (4-coumaroyl shikimate/quinate 3′-hydroxylase) gene from Brassica napus, which efficiently converts p-coumaroyl shikimate to caffeic acid and hispidin. Simultaneous expression of BnC3′H1 and NPGA (null-pigment mutant in A. nidulans) produces more caffeic acid and hispidin as the natural precursor of luciferin and significantly intensifies the original fungal bioluminescence pathway (oFBP). Thus, we successfully created enhanced FBP (eFBP) plants emitting 3 × 1011 photons/min/cm2, sufficient to illuminate its surroundings and visualize words clearly in the dark. The glowing plants provide sustainable and bio-renewable illumination for the naked eyes, and manifest distinct responses to diverse environmental conditions via caffeic acid biosynthesis pathway. Importantly, we revealed that the biosynthesis of caffeic acid and hispidin in eFBP plants derived from the sugar pathway, and the inhibitors of the energy production system significantly reduced the luminescence signal rapidly from eFBP plants, suggesting that the FBP system coupled with the luciferin metabolic flux functions in an energy-driven way. These findings lay the groundwork for genetically creating stronger eFBP plants and developing more powerful biological tools with the FBP system. |