Abstract: To support the implementation of the national rural revitalization and dual-carbon target strategies, and improve the output and clean energy utilization of biomass in rural areas, this paper proposes a multi-hydraulic retention time(HRT) optimal operation method for rural biomass power generation system under uncertain temperature. First, accurate models are established for processes such as raw material pretreatment, anaerobic digestion for biogas production, biogas purification and storage, biogas power generation and waste heat recovery, and temperature enhancement for aiding production. The model quantifies the energy consumption for aiding production and the increment in electricity generation in rural biomass production and power generation. Then, considering the influence of environmental uncertainties on the efficiency of anaerobic digestion for biogas production, a rolling optimization method across multiple HRTs is proposed to address the difficulty in decision-making regarding the aiding methods and operating variables. The method uses the most recent HRT as the control domain, updates variable data such as environmental temperature and solar radiation through rolling time window shifting, and aims to maximize the annual total electricity generation to generate operation plans for rural biomass power generation systems. Finally, simulation is conducted using a large-scale rural biomass power generation system in a northern region of China as an example. The results demonstrate that the proposed scheme effectively lowers system energy consumption, achieving a 14.66% increase in total electricity generation over the complete operational for cycle compared to waste heat recovery alone.