智慧农业 (Mar 2022)
Quantitative Determination of Plant Hormone Abscisic Acid Using Surface Enhanced Raman Spectroscopy
Abstract
Plant hormone Abscisic Acid (ABA) plays an important role in regulating plant growth. However, the content of ABA in plant tissues is very low, and rapid and sensitive detection methods are urgently needed. In this study, a rapid and quantitative ABA detection method was established based on aptamer recognition and surface-enhanced Raman spectroscop (SERS). The gold nanoparticles modified by ABA aptamer had the characteristics of SERS signal enhancement and selective recognition, realizing the rapid and sensitive detection of trace ABA in complex plant sample matrix. When ABA molecules appeared in detect system, the aptamer would specifically bind with ABA molecules, and the aptamer folded into G-tetrad structure at same time, which wrapped ABA molecules in the tetrad structure, shortened the distance between ABA molecules and gold nanoparticles, and the enhanced and stable ABA molecules SERS signal were obtained. Under the condition of optimized aptamer concentration at 0.12 µmol/L, different concentrations of ABA solutions in the detection system were detected. Within the concentration range of 0.1－100 µmol/L, the SERS intensity of ABA presented a good linear relationship with the concentration. The detection limit of this method was 0.1 µmol/L and the linear correlation coefficient R2 was 0.9855. The repeatability test of 20 points randomly on SERS substrate showed that the relative standard deviation (RSD) was 6.71%, indicating the stability of SERS substrate was well. Furthermore, the substrate of gold nanoparticles modified by the ABA aptamer terminal with sulfhydryl group (SH-Apt) could be stored in the refrigerator for more than half a year, indicating that the substrate has good stability. Once the preparation of the synthesized SH-Apt modified gold nanoparticles was completed. It could be used on demand without the need to prepare SERS substrate for every detection. In this sense, the constructed aptamer SERS biosensor could realize the rapid and quantitative detection of ABA. The method was used for the determination of ABA in wheat leaves, and the result was in good agreement with the Enzyme Linked Immunosorbent Assay (ELISA) (The max relative error was 9.13%). This biosensor is an exploratory study on the detection of plant hormones by SERS, and the results of the study will have important reference value for the subsequent quantitative and on-site detection of ABA, as well as the detection of other plant hormones.
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