Metabolic profiling of Oryza sativa seedlings under chilling stress using nanoliter electrospray ionization mass spectrometry.

Low temperatures significantly impact on rice (Oryza sativa) yield and quality. Traditional metabolomic techniques, often involving time-consuming chromatography-mass spectrometry procedures, are currently in use. This study investigated metabolomic responses of rice seedlings under low-temperature stress using nanoliter electrospray ionization mass spectrometry (nanoESI-MS) in combination with multivariate analysis. Results revealed distinct metabolic profiles in 'Qiutianxiaoting' (japonica) and '93-11' (indica) rice seedlings. Among the 36 identified compounds in rice, seven key metabolites, comprising l-glutamic acid, asparagine, tryptophan, citric acid, α-linolenic acid, malic acid, and inositol, were identified as responsive to cold stress. Notably, malic acid content reached 1332.40 µg/g dry weight in Qiutianxiaoting and 1444.13 µg/g in 93-11. Both the qualitative and quantitative results of nanoESI-MS were further confirmed through gas chromatography-mass spectrometry validation. The findings highlight the potential of nanoESI-MS for rapidly characterizing crucial metabolites across diverse plant species under exposure to stress.

Metabolic Profiling of Oryza sativa L. Triggered by Chilling Stress Using Ultraperformance Liquid Chromatography Coupled with Quadrupole/Time-of-Flight Mass Spectrometry (UPLC-QTOF-MS) with Transcriptome Analysis.

Low temperature, a major abiotic stress, often causes molecular changes in crops, which leads to metabolic disturbances and probably affects crop yield. In this study, chilling stress induced distinct metabolic profiles associated with transcriptome regulation, exhibiting great metabolic differences between Qiutianxiaoting (japonica) and 93-11 (indica). In total, 41 and 58 differential metabolites were screened and identified in Qiutianxiaoting and 93-11, respectively. Five key metabolites were screened in response to chilling stress, which were involved or related to different metabolic pathways. Moreover, starch and sucrose metabolism, aminoacyl-tRNA biosynthesis, and phenylpropanoid biosynthesis were significantly enriched in Qiutianxiaoting to maintain cellular homeostasis. Aminoacyl-tRNA biosynthesis and antioxidation metabolism were significantly enriched in 93-11, but disorders of the metabolome and transcriptome occurred at recovery stage. The results could provide some useful information for in-depth understanding of cold-resistant mechanisms, as well as reference for the selection and breeding of rice varieties.

Differential Antioxidant Compounds and Activities in Seedlings of Two Rice Cultivars Under Chilling Treatment.

Variations in antioxidant compounds were examined in seedlings of two rice cultivars (Qiutianxiaoting and 93-11) exposed to low temperature (4°C) for 0, 12, 36, and 48 h. Antioxidant activity was identified by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP) assays. The concentrations of total phenols, flavonoids, chlorophyll, and anthocyanins (ACNs) were determined by spectrophotometry. In addition, high-performance liquid chromatography (HPLC) was used to reveal the changes in phenolic compound concentrations in rice seedlings under chilling treatment. Results showed that antioxidant concentrations and antioxidant activity after chilling treatment were higher in 93-11 compared to Qiutianxiaoting, reaching the highest level at 36 h chilling treatment in 93-11. Phenolic compounds in Qiutianxiaoting decreased between 12 and 36 h but then increased at 48 h, whereas the corresponding levels in 93-11 increased as chilling time increased. Moreover, 10 phenolic compounds were detected and quantified by HPLC, of which gallic acid and caffeic acid tended to only exist in 93-11, whereas rutin was observed only in Qiutianxiaoting. The results of this study could be leveraged to optimize the antioxidant potential of rice in the context of healthy food choices.

[In silico cloning and bioinformatics analysis of HSP21 in Manihot esculenta].

HSP21 gene is a key gene to respond high temperature stress in plant and plays an important role in preventing protein denaturation, protecting cell structure and maintaining normal growth and development. Therefore, cloning HSP21 gene is the basis for revealing the molecular mechanism of resistance to high temperature stress in cassava. To obtain cassava HSP21 homologous gene and analyze the properties of predicted protein, electronic cloning technology was used to assemble and derivate new gene in this study, and bioinformatics analysis method was used to analyze the primary to highest structure, hydrophilicity/hydrophobicity, signal peptide, protein homology and phylogenetic evolution of expressed protein. HSP21 gene was 969 bp, its open reading frame was 705 bp, and the predicted protein contains 234 amino acids. The predicted protein is a non-transmembrane protein that is alkaline and hydrophilic, and is mainly localized in the chloroplast. Through multiple sequence alignment and phylogenetic analysis, it was found that the cassava HSP21 protein has high homology with other plants such as Hevea brasiliensis, Ricinus communis, and Jatropha curcas. The results could provide reference for the study of cloning and transformation of this gene.