Soil salinity is a growing threat to agricultural sustainability, particularly in arid and semi-arid regions. Understanding how salinity affects soil organic matter (OM) is critical for improving land management and maintaining soil health. This study addresses these challenges by exploring the molecular-level impact of salinity on OM dynamics. Salinity exerts a depth-dependent influence on lignin and microbial lipid biomarkers, which are used to trace plant inputs and microbial activity, respectively. For lignin biomarkers, in the surface layer (0–20 cm), higher salinity levels are associated with increased Syringyl/Vanillyl (S/V) and Cinnamyl/Vanillyl (C/V) ratios, suggesting enhanced preservation of syringyl (S) and cinnamyl (C) units. In the middle layer (−20 to −60 cm), higher salinity correlates with elevated SVC (total lignin phenols), Acid/aldehyde (Ad/Al) ratios, and other markers of selective lignin degradation. For lipid biomarkers, salinity modulates microbial adaptation and turnover, as seen in variations in i17 (iso-C17), a17 (anteiso-C17), and unsaturation indices such as C16:1/C16, reflecting Gram-positive and Gram-negative bacterial activity. These trends indicate that salinity stress alters microbial lipid profiles, leading to reduced turnover and enhanced preservation in deeper, more anoxic environments. Principal Component Analysis (PCA) revealed depth- and salinity-driven patterns that distinguish between surface microbial transformations and deep-layer molecular preservation. Correlation analysis of Principal Components (PCs) with salinity revealed that higher salinity favored molecular stability in deeper layers, while lower salinity was associated with microbial transformations in surface layers. These findings underscore salinity’s critical role in OM stabilization and turnover, and provide a molecular framework to guide sustainable management of saline soils.