Wheat (Triticum aestivum L.) is known to be negatively affected by heat stress, and its production is threatened by global warming, particularly in arid regions. Thus, efforts to better understand the molecular responses of wheat to heat stress are required. In the present study, Fourier transform infrared (FTIR) spectroscopy, coupled with chemometrics, was applied to develop a protocol that monitors chemical changes in common wheat under heat stress. Wheat plants at the three-leaf stage were subjected to heat stress at a 42 °C daily maximum temperature for 3 days, and this led to delayed growth in comparison to that of the control. Measurement of FTIR spectra and their principal component analysis showed partially overlapping features between heat-stressed and control leaves. In contrast, supervised machine learning through linear discriminant analysis (LDA) of the spectra demonstrated clear discrimination of heat-stressed leaves from the controls. Analysis of LDA loading suggested that several wavenumbers in the fingerprinting region (400–1800 cm−1) contributed significantly to their discrimination. Novel spectrum-based biomarkers were developed using these discriminative wavenumbers that enabled the successful diagnosis of heat-stressed leaves. Overall, these observations demonstrate the versatility of FTIR-based chemical fingerprints for use in heat-stress profiling in wheat.

Heat stress is one of the major environmental constraints for wheat production; thus, a comprehensive understanding of the metabolomic responses of wheat is required for breeding heat-tolerant varieties. In this study, the metabolome responses of heat-tolerant genotypes Imam and Norin 61, and susceptible genotype Chinese Spring were comparatively analyzed using Fourier transform infrared (FTIR) spectroscopy in combination with chemometric data mining techniques. Principal component analysis of the FTIR data suggested a spectral feature partially overlapping between the three genotypes. FTIR spectral biomarker assay showed similar heat responses between the genotypes for markers Fm482 and Fm1502, whereas genotype-dependent variations were observed for other markers. The markers Fm1251 and Fm1729 showed contrasting behaviors between heat-tolerant and susceptible genotypes, suggesting that these markers may potentially serve as a tool for distinguishing heat-tolerant genotypes. Linear discriminant analysis (LDA) of the spectra demonstrated a clear separation between the three genotypes in terms of the heat stress effect. Analysis of LDA coefficients identified several spectral regions that were potentially responsible for the discrimination of FTIR spectra between different genotypes and environments. These results suggest that a combination of FTIR and chemometrics can be a useful technique for characterizing the metabolic behavior of diverse wheat genotypes under heat stress.