Fig. 1: c-di-GMP promoted intestinal colonization of L. plantarum.

a, b High-resolution mass spectra (a) and triple-quadrupole mass spectra in a multiple reaction monitoring model (b) of c-di-GMP extracted from L. plantarum WCFS1. Left panel: c-di-GMP standard; right panel: extracted from L. plantarum WCFS1. The experiments were performed three times with similar results. c–g Overexpression of active c-di-GMP synthetase WspR or the active c-di-GMP phosphodiesterase RocR influenced the acid- (c) and bile salt (d) tolerance, intracellular c-di-GMP level (e), adhesive ability to mucus-secreting HT-29 cells (f), and colonization ability within mice colon (g) of L. plantarum. The genes rocR and wspR were cloned into the pLH01 plasmid and then electroporated into the wild-type strain of L. plantarum WCFS1. WT(pLH01) is the wild-type strain containing pLH01 plasmid as a control vector. Data are presented as mean values ± standard deviations (SD). Error bars indicate the SD. Dots represent individual data. Statistical significance in (c–g) was determined using one-way ANOVA (e–g) with Tukey’s multiple-comparison test. In (c, d, f, g): n = 5 biological replicates. In (e): n = 3 biological replicates. Source data are provided as a Source Data file.