This study evaluated the influence of four thermal pretreatment techniques—autoclaving, hot air oven treatment, hot water immersion, and microwave irradiation—on Parthenium hysterophorus biomass to improve its biodegradability and biogas generation potential under batch anaerobic digestion. Among the investigated methods, hot air oven pretreatment at 110 °C for 90 minutes exhibited the most significant enhancement in biomass solubilization, as indicated by a 51.5% rise in soluble chemical oxygen demand (sCOD) and an increase in volatile fatty acids (VFAs) compared with the untreated control. These compositional improvements facilitated faster hydrolysis and led to a 25.73% higher cumulative methane yield in biochemical methane potential (BMP) assays. Structural analysis revealed pronounced alterations in the lignocellulosic matrix, with reductions in hemicellulose and partial delignification improving substrate accessibility. Complementary characterization using field emission scanning electron microscopy (FESEM), X-ray diffraction (XRD), and Fourier-transform infrared spectroscopy (FTIR) confirmed the disruption of crystalline cellulose regions and modification of functional groups, supporting the observed biochemical improvements. Collectively, the results demonstrate that hot air oven pretreatment is a practical and energy-efficient approach for enhancing the digestibility of P. hysterophorus biomass, promoting its utilization as a sustainable feedstock for renewable biogas production and environmental management of this invasive weed.