A range of annual legume genotypes comprising one line of Trifolium subterraneum, four lines of T. michelianum, 11 of T. resupinatum var. resupinatum, and one line of T. resupinatum var. majus were grown in glasshouses under temperature regimes of 10-15°C and 16-21°C. Dry matter (DM) weights of stem, leaf, and flower tissues were measured when plants had six nodes, at first flower appearance, and at senescence. All samples were scanned by near-infrared reflectance spectroscopy (NIRS). One-third of the samples, covering the range of spectral characteristics, were analysed for in vitro digestible organic matter (DOMD), organic matter, crude protein (CP), neutral detergent fibre (NDF), lignin, cellulose, and the hemicellulosic polysaccharide monomers arabinose, xylose, mannose, galactose, and rhamnose. These data were used to develop calibration equations from which the composition of the remaining samples was predicted by NIRS. The higher temperature resulted in plants reaching respective phenological stages earlier, but did not affect either DM yields of total plant, stem, leaf, and petiole tissues or the proportions of each fraction. In vitro DOMD and arabinose and galactose levels decreased, while lignin, cellulose, NDF, xylose, mannose, and rhamnose levels increased with advancing maturity. In vitro DOMD was positively associated with contents of CP, arabinose, galactose, and the arabinose/xylose ratio and was negatively associated with contents of lignin, cellulose, NDF, xylose, mannose, and rhamnose. Lignin contents were highly correlated with levels of both xylose and mannose. Stems were more digestible than leaves in subterranean clover and T. resupinatum var. majus. The study also demonstrated that NIRS can be used routinely as a quick, inexpensive, and reliable laboratory technique to predict feed components of annual Trifolium legumes.