[Objective] This study aimed to investigate the regulatory mechanisms of carotenoid biosynthesis in Neurospora crassa. [Method] Gene knockout mutants pro- ducing less carotenoid were screened from 6 087 mutants; the yield of carotenoid and asexual spore was measured; finally fluorescent quantitative real-time PCR was adopted to analyze the transcription of genes related to carotenoid synthesis and asexual sporulation, [Result] Six knockout mutants produced less carotenoid. In one of them, the yield of both carotenoid and asexual spore reduced, because the gene which encodes an ATP-dependent chromatin remodelling complex ATPase chain ISW1 was knocked out. This gene was named /ca-1 in this study. And the /ca-1 deletion result- ed in a reduction of 88% in conidial production and a decrease of 81% in carotenoid production. [Conclusion] The Ica-1 positively regulates the carotenoid syn- thesis and asexual sporulation in N. crassa.