The chemical method for the determination of the resistant starch (RS) content in grains is time-consuming and labor intensive. Near-infrared (NIR) and attenuated total reflectance mid-infrared (ATR-MIR) spectroscopy are rapid and nondestructive analytical techniques for determining grain quality. This study was the first report to establish and compare these two spectroscopic techniques for determining the RS content in wheat grains. Calibration models with four preprocessing techniques based on the partial least squares (PLS) algorithm were built. In the NIR technique, the mean normalization + Savitzky–Golay smoothing (MN + SGS) preprocessing technique had a higher coefficient of determination ( R c 2  = 0.672; R p 2  = 0.552) and a relative lower root mean square error value (RMSEC = 0.385; RMSEP = 0.459). In the ATR-MIR technique, the baseline preprocessing method exhibited a better performance regarding to the values of coefficient of determination ( R c 2  = 0.927; R p 2  = 0.828) and mean square error value (RMSEC = 0.153; RMSEP = 0.284). The validation of the developed best NIR and ATR-MIR calibration models showed that the ATR-MIR best calibration model has a better RS prediction ability than the NIR best calibration model. Two high grain RS content wheat mutants were screened out by the ATR-MIR best calibration model from the wheat mutant library. There was no significant difference between the predicted values and chemical measured values in the two high RS content mutants. It proved that the ATR-MIR model can be a perfect substitute in RS measuring. All the results indicated that the ATR-MIR spectroscopy with improved screening efficiency can be used as a fast, rapid, and nondestructive method in high grain RS content wheat breeding.

Short-term heat stress can affect the growth of rice (Oryza sativa L.) seedlings, subsequently decreasing yields. Determining the dynamic response of rice seedlings to short-term heat stress is highly important for accelerating research on rice heat tolerance. Here, we observed the seedling characteristics of two contrasting cultivars (T11: heat-tolerant and T15: heat-sensitive) after different durations of 42 °C heat stress. The dynamic transcriptomic changes of the two cultivars were monitored after 0 min, 10 min, 30 min, 1 h, 4 h, and 10 h of stress. The results indicate that several pathways were rapidly responding to heat stress, such as protein processing in the endoplasmic reticulum, glycerophospholipid metabolism, and plant hormone signal transduction. Functional annotation and cluster analysis of differentially expressed genes at different stress times indicate that the tolerant cultivar responded more rapidly and intensively to heat stress compared to the sensitive cultivar. The MAPK signaling pathway was found to be the specific early-response pathway of the tolerant cultivar. Moreover, by combining data from a GWAS and RNA-seq analysis, we identified 27 candidate genes. The reliability of the transcriptome data was verified using RT-qPCR on 10 candidate genes and 20 genes with different expression patterns. This study provides valuable information for short-term thermotolerance response mechanisms active at the rice seedling stage and lays a foundation for breeding thermotolerant varieties via molecular breeding.

Resistant starch (RS) shows several health benefits. Enhancing the RS content of wheat is of major commercial importance. However, knowledge regarding the RS synthesis mechanism in wheat remains limited. In this study, the dynamic accumulation of RS during the filling process in two wheat (Triticum aestivum L.) genotypes with contrasting RS contents (H242, high RS content; H189, low RS content) were investigated. The results demonstrate that beyond 25 days after pollination (DAP), the RS content of H189 was relatively stable, but that of H242 continued to increase. Secondary accumulation was observed in the high-RS-content wheat genotype. A comparative transcriptome analysis between H242 and H189 at 20 DAP and 35 DAP showed that the differentially expressed genes were mainly involved in glycerolipid metabolism, glycerophospholipid metabolism and glucuronate interconversions. Furthermore, weighted gene coexpression network analysis suggested that lipid metabolic pathways such as the glycerophospholipid metabolism pathway might be involved in RS synthesis, and lipid-related genes upregulated beyond 25 DAP resulted in RS secondary accumulation. This work provides insight into the characteristics and mechanisms of RS synthesis.

DNA methylation and histone modification enable plants to rapidly adapt to adverse temperature stresses, including low temperature (LT) and high temperature (HT) stress. In this study, we conducted physiological, epigenetic, and transcriptomic analyses of barley seedlings grown under control (22°C), mild low temperature (MLT, 14°C) and HT (38°C) conditions to elucidate the underlying molecular mechanisms. Compared to MLT, HT implies greater deleterious effects on barley seedlings' growth. The methylation‐sensitive amplification polymorphism analysis showed that MLT induced more DNA methylation and HT more DNA demethylation compared to control. Besides, the higher levels of H3K9ac and H3K4me3 under HT compared to MLT stresses might lead to the loosening of chromatin and, subsequently, the activation of gene expression. Consistently, the transcriptome analysis revealed that there were more differentially expressed genes (DEGs) in plants subjected to HT stress than MLT stress compared to control. The common and unique pathways of these DEGs between MLT and HT were also analyzed. Transcription factors, such as ERF, bHLH, NAC, HSF, and MYB, were most involved in MLT and HT stress. The underlying gene regulation networks of epigenetic modulation‐related genes were further explored by weight gene co‐expression network analysis. Our study provides new insights into the understanding of epigenetic regulation responses to temperature stress in barley, which will lead to improved strategies for the development of cold‐ and heat‐tolerant barley varieties for sustainable barley production in a climate‐changing world.