Due to the high availability of lignocellulosic biomass, which can be obtained from terrestrial plants, agricultural waste biomass, and the agro-food, paper or wood industries, its use for energy production by methane fermentation is economically and environmentally justified. However, due to their complex structures, lignocellulosic substrates have a low conversion factor to biogas. Therefore, scientists are still working on the development of new methods of the pre-treatment of lignocellulosic materials that will increase the biogas productivity from lignocellulosic biomass. The presented research focuses on the use of a pulsed electric field (PEF) to disintegrate rapeseed straw prior to the methane fermentation process. Scanning electron microscopy observation showed that, in the disintegrated sample, the extent of damage to the plant tissue was more severe than in the control sample. In the sample disintegrated for 7 min, the chemical oxygen demand increased from 4146 ± 75 mg/L to 4920 ± 60 mg/L. The best result was achieved with a 5-min PEF pre-treatment. The methane production reached 290.8 ± 12.1 NmL CH4/g VS, and the biogas production was 478.0 ± 27.5 NmL/g VS; it was 14% and 15% higher, respectively, compared to the control sample.