Overexpressing GhADC2-WT-Silencing GhADC2 - Non-target Metabolite Mass Spectrometry Detection and Analysis

The following 3 QC tests perform segmented scanning, along with the secondary spectra obtained from the experimental samples, to determine the metabolites. The QC tests inserted during sample detection are used to evaluate the stability of the system throughout the entire experiment and to conduct data quality control analysis. Firstly, the original files (in .raw format) obtained from the mass spectrometry detection are imported into the Compound Discoverer 3.1 (hereinafter referred to as CD3.1) software for spectral processing and database search, to obtain the qualitative and quantitative results of the metabolites.

Then, quality control of the data is carried out to ensure the accuracy and reliability of the data results. Next, multivariate statistical analysis of the metabolites is conducted, including principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA), etc., to reveal the differences in metabolic patterns among different groups. Hierarchical clustering (HCA) and metabolite correlation analysis are used to reveal the relationships between samples and between metabolites and metabolites.

Finally, the biological significance related to the metabolites is explained through functional analysis of metabolic pathways and other functions.