Rubberwood (Hevea brasiliensis (Willd. ex A. Juss.) Müll. Arg), a key economic species in tropical regions, presents significant processing challenges due to the wide variation in tension wood (TW) content. This study systematically compared the chemical composition and structural features of fibres from tension wood and opposite wood zones in rubberwood, aiming to elucidate the formation mechanisms of tension wood and provide theoretical support for its targeted modification and high-value utilization. Analyses included histochemical staining, chemical analysis, FTIR, thermogravimetric analysis (TGA), and thioacidolysis-GC-MS. The results showed that TW fibers featured a prominently thickened gelatinous layer (G-layer) composed of highly crystalline cellulose, with significantly reduced lignin content compared to NW. Thioacidolysis revealed that TW lignin not only exhibited a lower total content but also an increased proportion of syringyl (S) units, suggesting the adaptive regulation mechanisms within the lignin biosynthesis pathway. FTIR spectroscopy showed stronger O-H and aliphatic C-H stretching bands in TW, consistent with its higher cellulose content. The TGA results showed higher mass loss at 220–390 °C and lower char yield for TW, indicating cellulose-dominated decomposition behavior. This study elucidated the synergistic effect of S-unit enrichment and linkage rearrangement in TW lignin, underscoring that the G-layer’s high cellulose and low lignin architecture dictates thermal response. These findings improve the understanding of tension wood formation and provide practical strategies for enhancing the processing and utilization of plantation rubberwood.