Based on the panel data of 30 provincial administrative regions in China from 2010 to 2023, this paper empirically examines the impact of artificial intelligence application on the competitiveness of regional innovation ecosystems. The research results show that artificial intelligence applications is conducive to enhancing the competitiveness of regional innovation ecosystems. The heterogeneity test results indicate that the empowerment effect of artificial intelligence application on the competitiveness of regional innovation ecosystems is more significant in southern regions and regions with higher levels of human capital. The mediation path test confirms that the flow of R&D capital and R&D personnel plays a mediating role in the relationship between artificial intelligence application and the competitiveness of regional innovation ecosystems. The spatial spillover effect test finds that artificial intelligence application can effectively enhance the innovation ecosystem competitiveness of the province itself and its neighboring provinces. This paper deeply analyzes the path through which artificial intelligence application affects the competitiveness of regional innovation ecosystems, with the aim of providing theoretical references and practical inspirations for China’s scientific and technological innovation to achieve.