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. 2024 Sep 13;25(18):9903.
doi: 10.3390/ijms25189903.

NMR-Based Metabolomic Analysis of Biotic Stress Responses in the Traditional Korean Landrace Red Pepper (Capsicum annuum var. annuum, cv. Subicho)

Affiliations

NMR-Based Metabolomic Analysis of Biotic Stress Responses in the Traditional Korean Landrace Red Pepper (Capsicum annuum var. annuum, cv. Subicho)

Gi-Un Seong et al. Int J Mol Sci. .

Abstract

Korean landrace red peppers (Capsicum annuum var. Subicho), such as the traditional representative Subicho variety, are integral to Korean foods and are often consumed raw or used as a dried powder for cuisine. However, the known vulnerability of local varieties of landrace to biotic stresses can compromise their quality and yield. We employed nuclear magnetic resonance (NMR) spectroscopy coupled with a multivariate analysis to uncover and compare the metabolomic profiles of healthy and biotic-stressed Subicho peppers. We identified 42 metabolites, with significant differences between the groups. The biotic-stressed Subicho red peppers exhibited lower sucrose levels but heightened concentrations of amino acids, particularly branched-chain amino acids (valine, leucine, and isoleucine), suggesting a robust stress resistance mechanism. The biotic-stressed red peppers had increased levels of TCA cycle intermediates (acetic, citric, and succinic acids), nitrogen metabolism-related compounds (alanine, asparagine, and aspartic acid), aromatic amino acids (tyrosine, phenylalanine, and tryptophan), and γ-aminobutyric acid. These findings reveal the unique metabolic adaptations of the Subicho variety, underscoring its potential resilience to biotic stresses. This novel insight into the stress response of the traditional Subicho pepper can inform strategies for developing targeted breeding programs and enhancing the quality and economic returns in the pepper and food industries.

Keywords: Capsicum annuum; NMR; Subicho; biotic stress; metabolomics; red pepper; stress resistance.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
Characteristic 1H NMR spectrum (700 MHz) of the quality control sample prepared by pooling equal volumes of all the red pepper extracts: (a) the full spectrum (0–10 ppm), (b) the aliphatic or allylic region (0.5–3 ppm), (c) the alkene or heteroatom-attached region (3–5.6 ppm), and (d) the alkene or aromatic region (5.6–9.5 ppm). α-Glc—α-Glucose; β-Glc—β-Glucose; Suc—Sucrose; Fru—Fructose; Tre—Trehalose; Thr—Threonine; Ser—Serine; Asn—Asparagine; Gln—Glutamine; Ala—Alanine; Gly—Glycine; Pro—Proline; Val—Valine; Leu—Leucine; Ile—Isoleucine; Asp—Aspartic acid; GABA—γ-Aminobutyric acid; Arg—Arginine; Phe—Phenylalanine; Trp—Tryptophan; Tyr—Tyrosine; AcOH—Acetic acid; FoA—Formic acid; MA—Malic acid; SA—Succinic acid; GA—Glutaric acid; CA—Citric acid; QA—Quinic acid; HBA—Hydrobenzoic acid; Cin—Cinnamate; Chol—Choline; TG—Trigonelline; MEA—Ethanolamine; Ino—Inosine; 3-IS—3-Indoxylsulfate; Urd—Uridine; Ac-Tyr—Acetyl-tyrosine; NADP+—Nicotinamide adenine dinucleotide phosphate; ST—Sterols; GL—Glycerol; FAs—Fatty acids; and BA—Butyric acid.
Figure 2
Figure 2
OPLS-DA score plots between healthy Subicho and biotic-stressed Subicho red peppers derived from the 1H NMR spectra (700 MHz).
Figure 3
Figure 3
OPLS-DA score (a) and coefficients or loading plots (b) between the healthy Subicho and biotic-stressed Subicho red peppers to identify metabolites responsible for the metabolic differentiation. The color code in the loading plot corresponds to the correlation between the variables. All the OPLS-DA models were generated with one predictive and one orthogonal component. Their reliability and predictability are indicated by R2X, R2Y, and Q2. Abbreviations of the names of the assigned metabolites are listed in Table S1.
Figure 4
Figure 4
Relative quantification of the individual metabolites in red peppers, and a comparison of relative metabolite contents between the healthy Subicho and biotic-stressed Subicho red peppers. Asterisks represent statistical significance using the Student’s t-test (* p < 0.05, ** p < 0.01, *** p < 0.001). OPLS-DA score plots between healthy Subicho and biotic-stressed Subicho red peppers derived from the 1H NMR spectra (700 MHz).
Figure 5
Figure 5
Schematic metabolic flux between the healthy Subicho and biotic-stressed Subicho red pepper. Asterisks represent statistical significance using the Student’s t-test (* p < 0.05, ** p < 0.01).

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