A structure-guided screening strategy for the discovery and identification of potential gibberellins from plant samples using liquid chromatography-mass spectrometry assisted by chemical isotope labeling.

Gibberellins (GAs) play crucial roles in plant growth and development, and their regulatory functions rely on complex metabolic networks and signaling pathways. Therefore, the exploration of GAs metabolic network is of great importance. However, limited GAs have been found in given plant species, which makes it difficult to comprehensively study the GAs metabolic network. Herein, a structure-guided strategy for GAs screening based on liquid chromatography-mass spectrometry analysis assisted by chemical isotope labeling (CIL-LC-MS) was developed. In the proposed strategy, N,N-dimethyl ethylenediamine (DMED) and its isotopologue d4-DMED were used to label GAs. In light of the characteristic fragmentation patterns exhibited by the labeled GAs, four principles were summarized to screen the potential GAs from plant tissues. Subsequently, the MS/MS fragmentation behavior and quantitative structure-retention relationship (QSRR) model were employed to assist in deciphering structures of GA candidates. With this strategy, thirty potential GAs were screened out and identified from five plant species. Seven of them were confirmed by the authentic standards. Twenty-two of them have not been reported before in the five plant species used in this study, including thirteen that have been reported in other plant species and nine that have never been reported in any plant species. Noteworthily, a total of nine potential GAs were speculated to be novel 16, 17-double hydrated GAs, which indicated that the 16, 17-double hydration may be a ubiquitous metabolic pathway of GAs in plants. This study was the first attempt to establish a structure-guided screening strategy for GAs. Our findings have enriched the GA species in plants and expanded the GAs family, which may be helpful for study of the metabolic pathway and physiological function of GAs.