The paralytic shellfish toxin producing dinoflagellate Gymnodinium catenatum is a globally distributed species and often forms massive blooms. However, the physiological and molecular responses of G. catenatum to nitrate starvation have not been thoroughly investigated. Our results showed that multiple forms of N could be utilized by G. catenatum under nitrate-deficient conditions. Nitrate deficiency adversely affected the growth, cellular Chlorophyll a (Chl a) content, and toxin production of G. catenatum. Transcriptomic analysis revealed significant down-regulation of gene expressions involved in the light reaction of photosynthesis, while genes related to fatty acids synthesis and antioxidation were significantly upregulated in the N-depleted cultures. Our results suggested that excess carbon was channeled into lipid synthesis for energy storage, and antioxidant reactions were upregulated to eliminate toxic peroxides caused by nitrate limitation. These findings highlight the adaptative strategy of G. catenatum in low-nitrate environments, which are crucial factors driving its bloom formation.