Rapid Detection and Quantification of Plant Innate Immunity Response Using Raman Spectroscopy

Abstract

We have developed a rapid Raman spectroscopy-based method for the detection and quantification of early innate immunity responses in Arabidopsis and Choy Sum plants. Arabidopsis plants challenged with flg22 and elf18 elicitors could be differentiated from mock-treated plants by their Raman spectral fingerprints. From the difference Raman spectrum and the value of p at each Raman shift, we derived the Elicitor Response Index (ERI) as a quantitative measure of the response whereby a higher ERI value indicates a more significant elicitor-induced immune response. Among various Raman spectral bands contributing toward the ERI value, the most significant changes were observed in those associated with carotenoids and proteins. To validate these results, we investigated several characterized Arabidopsis pattern-triggered immunity (PTI) mutants. Compared to wild type (WT), positive regulatory mutants had ERI values close to zero, whereas negative regulatory mutants at early time points had higher ERI values. Similar to elicitor treatments, we derived an analogous Infection Response Index (IRI) as a quantitative measure to detect the early PTI response in Arabidopsis and Choy Sum plants infected with bacterial pathogens. The Raman spectral bands contributing toward a high IRI value were largely identical to the ERI Raman spectral bands. Raman spectroscopy is a convenient tool for rapid screening for Arabidopsis PTI mutants and may be suitable for the noninvasive and early diagnosis of pathogen-infected crop plants.

Plant phenotyping relevance

植物の免疫応答をRamanスペクトルから定量化する指標を開発し、変異体・病原体感染で検証した中心的なフェノタイピング手法研究。

Code and data availability

The supplied blocks describe Raman spectral data collection and ERI/IRI analysis, but contain no public deposit of the spectral datasets, analysis code, or trained models. The only URLs are generic tools (Primer-BLAST, ImageJ) and the article's supplementary material link, which is not stated to contain datasets or the

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