Introduction: The number of traffic accidents involving truck drivers remains high, and strategies to eliminate the probability of such accidents have been proposed, among which enhancing the safety climate has attracted much interest. The main purpose of the current study was to validate the Chinese version of the safety climate scale for truck drivers and apply it to investigate the mediating effect of safety climate between truck driver personality and driving behavior. Method: A total of 389 male truck drivers completed the Big Five Inventory, the Chinese version of the trucking safety climate scale, the driver behavior questionnaire and the positive driver behavior scale. Results: The reliability and validity of the Chinese version of the organization-level safety climate scale and the group-level safety climate scale were confirmed through factor analysis. More importantly, a path analysis revealed that the organization-level safety climate mediated the effects of agreeableness and neuroticism on aggressive violations, ordinary violations and lapses, while the group-level safety climate mediated the influences of agreeableness, neuroticism and openness on positive behavior and all four kinds of aberrant driving behaviors. Conclusions: This study introduced the trucking safety climate scale into China and stressed the significance of improving both the organizational and the group levels of safety climate to reduce accidents involving professional truck drivers. Practical Applications: First, the adapted safety climate scale for Chinese truck drivers contributes to further investigating the role that safety climate plays in the safety problem of truck drivers in China. Moreover, the critical impacts of both levels of the trucking safety climate serve as reminders for relevant companies to not only pay attention to establishing an organization-level safety climate but also invest more effort into promoting the group-level safety climate.