Abstract
Arabidopsis Cytochrome P450 85A2 (AtCYP85A2) was introduced to Arabidopsis thaliana seeds using a seed-specific promoter, pAt5g54000. GUS (β-Glucuronidase) activity and RT-PCR analysis demonstrated that AtCYP85A2 overexpression only occurred in seeds of a transgenic plant, pAt5g54000-AtCYP85A2::Col-0. A crude enzyme solution prepared pAt5g54000-AtCYP85A2::Col-0 seeds successfully catalyzed the conversion of castasterone (CS) to brassinolide (BL), which was not detected in wild-type seeds. Furthermore, a higher level of CS and BL was detected in pAt5g54000-AtCYP85A2::Col-0 seeds compared to untransformed seeds, thus demonstrating that seed-specific overexpression of AtCYP85A2 efficiently encoded a bi-functional enzyme for brassinosteroids 6-oxidase/brassinolide synthase to generate CS and BL in seeds of pAt5g54000-AtCYP85A2::Col-0. Compared to the wild type, pAt5g54000-AtCYP85A2::Col-0 produced substantially larger seeds with a high concentration of nutrients due to an enhancement in brassinosteroids signaling. Additionally, pAt5g54000-AtCYP85A2::Col-0 exhibited superior seed germination, seedling and rosette plant growth, and flower and silique formation, indicating that seed-specific AtCYP85A2 expression activates overall vegetative and reproductive growth and development in A. thaliana.
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Acknowledgements
We would like to thank Prof. Jeong Sheop Shin (Korea University, Seoul, Republic of Korea) for providing the seed-specific promoter construct pAt5g54000/pBI101. This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korean government’s Ministry of Science, ICT, and Future Planning (2021R1A2C1007516).
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S-KK, JR, MHY, and C-HP planned the experiments; MHY, C-HP, and YL carried out the experiments; S-KK, JR, MHY, and C-HP analyzed the data; S-KK, JR, MHY, and C-HP wrote and revised the manuscript for publication. All authors agreed on the contents of this manuscript and declare no conflicts of interest.
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Yeon, M.H., Park, CH., Lee, Y.E. et al. Seed-Specifically Overexpressed Arabidopsis Cytochrome P450 85A2 Promotes Vegetative and Reproductive Growth and Development of Arabidopsis thaliana. J. Plant Biol. 65, 75–86 (2022). https://doi.org/10.1007/s12374-021-09340-3
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DOI: https://doi.org/10.1007/s12374-021-09340-3