Abstract
Molecular biomarkers such as microRNAs (miRNAs) play important roles in regulating various developmental processes in plants. Understanding these pathways will help bioengineer designing organisms for efficient biomass accumulation. Current methods for RNA analysis require sample extraction and multi-step sample analysis, hindering work in field studies. Recent work in the incorporation of nanomaterials for plant bioengineering research is leading the way of an agri-tech revolution. As an example, surface-enhanced Raman scattering (SERS)–based sensors can be used to monitor RNA in vivo. However, the use of SERS in the field has been limited due to issues with observing Raman signal over complex background. To this end, shifted-excitation Raman difference spectroscopy (SERDS) offers an effective solution to extract the SERS signal from high background based on a physical approach. In this manuscript, we report the first application of SERDS on SERS sensors. We investigated this technique on SERS sensor developed for the detection of a microRNA biomarker, miR858. We tested the technique on in vitro samples and validated the technique by detecting the presence of exogenous miR858 in plants directly under ambient light in a growth chamber. The possibility of moving the detection of nucleic acid targets outside the constraints of laboratory setting enables numerous important bioengineering applications. Such applications can revolutionize biofuel production and agri-tech through the use of nanotechnology-based monitoring of plant growth, plant health, and exposure to pollution and pathogens.
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Acknowledgments
We would like to thank Tai-Ping Sun and Rodolfo Zentella from the Duke Biology Department for letting us use the growth chamber for these studies.
Funding
This material is based upon work supported by the US Department of Energy Office of Science, under Award Number DE-SC0019393. The authors received financial support by the Federal Ministry of Education and Research (BMBF) under contract 031A564C. Parts of the dual-wavelength diode laser turn-key system were developed within the project RaMBo (Raman Messsystem zur ortsspezifischen Bodenanalytik) within the consortium I4S (Intelligence for Soil) within the funding measure BonaRes (Soil as a Sustainable Resource for the Bioeconomy).
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Published in the topical collection Advances in Direct Optical Detection with guest editors Antje J. Baeumner, Günter Gauglitz, and Jiri Homola.
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Strobbia, P., Odion, R.A., Maiwald, M. et al. Direct SERDS sensing of molecular biomarkers in plants under field conditions. Anal Bioanal Chem 412, 3457–3466 (2020). https://doi.org/10.1007/s00216-020-02544-5
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DOI: https://doi.org/10.1007/s00216-020-02544-5