Single species-based altitudinal transects may provide a new understanding of the variabilities in sedimentary wax-derived n-alkane hydrogen isotope (δH) values caused by altitude and complex climatic change linked with the growth of mountains. We investigated Kobresia pygmaea (Kobresia), Quercus aquifolioides (Quercus) and Berberis thunbergii DC (Berberis) along three altitudinal transects on the Tibetan Plateau (TP), i.e., the southern TP, the Longmen Mountains (LM; eastern TP) and the Qilian Mountains (QL; northeastern TP). Here we present 47 plant δH values: these include 14 Kobresia, 27 Berberis and 6 Quercus samples, which are accompanied by comparisons with nine new soil δH values from the QL, and 105 previously-published δH values for surface soils along the first two transects. Our data show that altitude is the dominant factor in determining three plant δH values. However, we observed substantial differences in the δH values and their ε ratios for Kobresia, Quercus and Berberis for different climatic regimes and along these three transects. Significantly, for Kobresia along the LM and QL transects, ∆δH = -84.3‰/km (r = 0.94; p < 0.05; n = 4) and - 65.5‰/km (r = 0.74; p < 0.01; n = 10), and ∆ε = -80.4‰/km (r = 0.93; p < 0.05) and -56.7‰/km (r = 0.66; p < 0.01), respectively, were three or four times as large as for the soil δH values observed along these altitudinal gradients. Overall, the altitudinal lapse rate (ALR) of δH values and their ε ratios varies between species, with Kobresia being the most negative and Berberis the least negative, potentially resulting from the strong response of monocotyledoneae Kobresia δH values to cooling with increasing altitude, and the relative influence of cryosphere meltwater at higher altitudes. Thus, impact of climate change on the sedimentary δH values should therefore be fully taken into account during reconstructions of paleoaltitudes.