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Insufficient resources of plant fibre for developing bio-based products and processing fibrous plant stalks to high quality and quantity fibres are two challenges of the natural fibre industry. Considering the first challenge, two alternative plant fibres including canola and sweet clover fibres were characterized and compared to hemp and flax fibres which are two traditional fibres. Their characteristics, including microstructural, surface, thermal, and mechanical properties, were measured. Mechanical behaviour of fibres was also simulated using the Discrete Element Method (DEM). Considering the second challenge, fibre extraction experiments were conducted on both traditional and alternative fibrous plants. In the experiments, hemp fibre was mechanically extracted using a decorticator. Mechanical extraction of canola fibres was not successful; therefore, canola fibre was extracted through retting. Results demonstrated that the microstructural, surface, and thermal properties of the alternative fibres (canola and sweet clover) were in the same ranges with the traditional fibres (hemp and flax). However, the alternative fibres had lower Young’s modulus, while the traditional fibres had higher stiffness and tensile strength. Simulation results indicated that the DEM model generated comparable results to the tensile test experiments in terms of Young’s modulus and tensile strength. For mechanical extraction of hemp fibre using the decorticator, the most effective parameters, in terms of quantity and quality of the fibres, were the retting condition of the feed stalks, the number of pass of stalks through the decorticator rollers, and the feed amount. For retting extraction of canola fibre using chemical solution, when the time, temperature, and NaHCO3 concentration were set to 37.8 h, 57.7 ºC, and 5.62%, respectively, the highest quantity and quality canola fibres were achieved. The results from this study have important implications to applications of plant fibres.