Quorum sensing (QS) regulates various physiological processes in a cell density–dependent mode via cell–cell communication. Stenotrophomonas rhizophila DSM14405^T having the diffusible signal factor (DSF)-QS system, is a plant growth-promoting rhizobacteria (PGPR) that enables host plants to tolerate saline-alkaline stress. However, the regulatory mechanism of DSF-QS in S. rhizophila is not fully understood. In this study, we used S. rhizophila DSM14405^T wild-type (WT) and an incompetent DSF production rpfF-knockout mutant to explore the regulatory role of QS in S. rhizophila growth, stress responses, biofilm formation, and colonization under saline-alkaline stress. We found that a lack of DSF-QS reduces the tolerance of S. rhizosphere ΔrpfF to saline-alkaline stress, with a nearly 25-fold reduction in the ΔrpfF population compared with WT at 24 h under stress. Transcriptome analysis revealed that QS helps S. rhizophila WT respond to saline-alkaline stress by enhancing metabolism associated with the cell wall and membrane, oxidative stress response, cell adhesion, secretion systems, efflux pumps, and TonB systems. These metabolic systems enhance penetration defense, Na⁺ efflux, iron uptake, and reactive oxygen species scavenging. Additionally, the absence of DSF-QS causes overexpression of biofilm-associated genes under the regulation of sigma 54 and other transcriptional regulators. However, greater biofilm formation capacity confers no advantage on S. rhizosphere ΔrpfF in rhizosphere colonization. Altogether, our results show the importance of QS in PGPR growth and colonization; QS gives PGPR a collective adaptive advantage in harsh natural environments.