Coronatine (COR) is a new type of plant growth regulator that is produced by
Pseudomonas syringae
pathovars and plays an important role in modulating plant growth, development, and tolerance to multiple stresses. However, the factors affecting COR production are not very clear. In this study, the effects of FeCl
3
on COR production were researched. The data-independent acquisition (DIA) approach, which is a proteomic quantitative analysis method, was applied to quantitatively trace COR production and proteomic changes in
P. syringae
pv.
tomato
DC3000 under different FeCl
3
culture conditions. The results showed that COR production increased with the addition of FeCl
3
and that there was significant upregulation in the expression of proteins related to COR synthesis and regulation. In addition, FeCl
3
also affected the expression of related proteins involved in various metabolic pathways such as glycolysis and the tricarboxylic acid cycle. Moreover, various precursors such as isoleucine and succinate semialdehyde, as well as other related proteins involved in the COR synthesis pathway, were significantly differentially expressed. Our findings revealed the dynamic regulation of COR production in response to FeCl
3
at the protein level and showed the potential of using the DIA method to track the dynamic changes of the
P. syringae
pv.
tomato
DC3000 proteome during COR production, providing an important reference for future research on the regulatory mechanism of COR biosynthesis and theoretical support for COR fermentation production.