Rational design of novel fluorescent enzyme biosensors for direct detection of strigolactones
- Author
- Rebecca J. Chesterfield, Jason H. Whitfield, Benjamin Pouvreau, Da Cao (UGent) , Kirill Alexandrov, Christine A. Beveridge and Claudia E. Vickers
- Organization
- Abstract
- Strigolactones are plant hormones and rhizosphere signaling molecules with key roles in plant development, mycorrhizal fungal symbioses, and plant parasitism. Currently, sensitive, specific, and high-throughput methods of detecting strigolactones are limited. Here, we developed genetically encoded fluorescent strigolactone biosensors based on the strigolactone receptors DAD2 from Petunia hybrida, and HTL7 from Striga hermonthica. The biosensors were constructed via domain insertion of circularly permuted GFP. The biosensors exhibited loss of cpGFP fluorescence in vitro upon treatment with the strigolactones 5-deoxystrigol and orobanchol, or the strigolactone analogue rac-GR24, and the ShHTL7 biosensor also responded to a specific antagonist. To overcome biosensor sensitivity to changes in expression level and protein degradation, an additional strigolactone-insensitive fluorophore, LSSmOrange, was induded as an internal normalization control. Other plant hormones and karrikins resulted in no fluorescence change, demonstrating that the biosensors report on compounds that specifically bind the SL receptors. The DAD2 biosensor likewise responded to strigolactones in an in vivo protoplast system, and retained strigolactone hydrolysis activity. These biosensors have applications in high-throughput screening for agrochemical compounds, and may also have utility in understanding strigolactone mediated signaling in plants.
- Keywords
- Biochemistry, Genetics and Molecular Biology (miscellaneous), Biomedical Engineering, General Medicine, SUICIDAL GERMINATION, PROTEIN SWITCHES, HORMONE-RECEPTOR, DWARF14, PERCEPTION, ARABIDOPSIS, DEGRADATION, INHIBITION, REPRESSOR, OROBANCHE
Citation
Please use this url to cite or link to this publication: http://hdl.handle.net/1854/LU-8758345
- MLA
- Chesterfield, Rebecca J., et al. “Rational Design of Novel Fluorescent Enzyme Biosensors for Direct Detection of Strigolactones.” ACS SYNTHETIC BIOLOGY, vol. 9, no. 8, 2020, pp. 2107–18, doi:10.1021/acssynbio.0c00192.
- APA
- Chesterfield, R. J., Whitfield, J. H., Pouvreau, B., Cao, D., Alexandrov, K., Beveridge, C. A., & Vickers, C. E. (2020). Rational design of novel fluorescent enzyme biosensors for direct detection of strigolactones. ACS SYNTHETIC BIOLOGY, 9(8), 2107–2118. https://doi.org/10.1021/acssynbio.0c00192
- Chicago author-date
- Chesterfield, Rebecca J., Jason H. Whitfield, Benjamin Pouvreau, Da Cao, Kirill Alexandrov, Christine A. Beveridge, and Claudia E. Vickers. 2020. “Rational Design of Novel Fluorescent Enzyme Biosensors for Direct Detection of Strigolactones.” ACS SYNTHETIC BIOLOGY 9 (8): 2107–18. https://doi.org/10.1021/acssynbio.0c00192.
- Chicago author-date (all authors)
- Chesterfield, Rebecca J., Jason H. Whitfield, Benjamin Pouvreau, Da Cao, Kirill Alexandrov, Christine A. Beveridge, and Claudia E. Vickers. 2020. “Rational Design of Novel Fluorescent Enzyme Biosensors for Direct Detection of Strigolactones.” ACS SYNTHETIC BIOLOGY 9 (8): 2107–2118. doi:10.1021/acssynbio.0c00192.
- Vancouver
- 1.Chesterfield RJ, Whitfield JH, Pouvreau B, Cao D, Alexandrov K, Beveridge CA, et al. Rational design of novel fluorescent enzyme biosensors for direct detection of strigolactones. ACS SYNTHETIC BIOLOGY. 2020;9(8):2107–18.
- IEEE
- [1]R. J. Chesterfield et al., “Rational design of novel fluorescent enzyme biosensors for direct detection of strigolactones,” ACS SYNTHETIC BIOLOGY, vol. 9, no. 8, pp. 2107–2118, 2020.
@article{8758345,
abstract = {{Strigolactones are plant hormones and rhizosphere signaling molecules with key roles in plant development, mycorrhizal fungal symbioses, and plant parasitism. Currently, sensitive, specific, and high-throughput methods of detecting strigolactones are limited. Here, we developed genetically encoded fluorescent strigolactone biosensors based on the strigolactone receptors DAD2 from Petunia hybrida, and HTL7 from Striga hermonthica. The biosensors were constructed via domain insertion of circularly permuted GFP. The biosensors exhibited loss of cpGFP fluorescence in vitro upon treatment with the strigolactones 5-deoxystrigol and orobanchol, or the strigolactone analogue rac-GR24, and the ShHTL7 biosensor also responded to a specific antagonist. To overcome biosensor sensitivity to changes in expression level and protein degradation, an additional strigolactone-insensitive fluorophore, LSSmOrange, was induded as an internal normalization control. Other plant hormones and karrikins resulted in no fluorescence change, demonstrating that the biosensors report on compounds that specifically bind the SL receptors. The DAD2 biosensor likewise responded to strigolactones in an in vivo protoplast system, and retained strigolactone hydrolysis activity. These biosensors have applications in high-throughput screening for agrochemical compounds, and may also have utility in understanding strigolactone mediated signaling in plants.}},
author = {{Chesterfield, Rebecca J. and Whitfield, Jason H. and Pouvreau, Benjamin and Cao, Da and Alexandrov, Kirill and Beveridge, Christine A. and Vickers, Claudia E.}},
issn = {{2161-5063}},
journal = {{ACS SYNTHETIC BIOLOGY}},
keywords = {{Biochemistry,Genetics and Molecular Biology (miscellaneous),Biomedical Engineering,General Medicine,SUICIDAL GERMINATION,PROTEIN SWITCHES,HORMONE-RECEPTOR,DWARF14,PERCEPTION,ARABIDOPSIS,DEGRADATION,INHIBITION,REPRESSOR,OROBANCHE}},
language = {{eng}},
number = {{8}},
pages = {{2107--2118}},
title = {{Rational design of novel fluorescent enzyme biosensors for direct detection of strigolactones}},
url = {{http://doi.org/10.1021/acssynbio.0c00192}},
volume = {{9}},
year = {{2020}},
}
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