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Agro-waste having lignocellulosic biomass is considered most effective (heating value 16 MJ/kg) for energy production through anaerobic digestion (AD). However, recalcitrant lignocellulosic fraction in agro-waste obstructs its biotransformation and is a rate-limiting step of the process. This study investigated the effects of hydrothermal and thermal-alkaline pretreatment on anaerobic co-digestion of wheat straw (WS). The hydrothermal pretreatment of WS revealed that 60 min was the best pretreatment time to achieve the highest substrate solubilization. It was employed for thermal-alkali pretreatment at variable temperatures and NaOH doses. Thermal-alkali pretreatment at 125°C-7% NaOH shows the highest (34%) biogas yield of 662 mL/gVS, followed by 646 mL/gVS biogas yield at 150°C-1% NaOH assay (31% higher) over control. Although the 125°C-7% NaOH assay achieved the highest biogas yield, the 150°C …