Nowadays, the extract of seaweeds has drawn attention as a rich source of bioactive metabolites. Seaweeds are known for their biologically active compounds whose antibacterial and antifungal activities have been documented. This research aimed to study the profile of phenolic compounds using the HPLC method and determine biologically active compounds using the GC-MS method and the antifungal activity of Gracilariopsis persica against plant pathogenic fungi. G. persica was collected from its natural habitat in Suru of Bandar Abbas, Iran, dried, and extracted by methanol. The quantitative results on phenolic compounds using the HPLC method showed that the most abundant compounds in G. persica were rosmarinic acid (20.9 ± 0.41 mg/kg DW) and quercetin (11.21 ± 0.20 mg/kg DW), and the least abundant was cinnamic acid (1.4 ± 0.10 mg/kg DW). The GC-MS chromatography revealed 50 peaks in the methanolic extract of G. persica, implying 50 compounds. The most abundant components included cholest-5-en-3-ol (3 beta) (27.64%), palmitic acid (17.11%), heptadecane (7.71%), and palmitic acid methyl ester (6.66%). The antifungal activity of different concentrations of the extract was determined in vitro. The results as to the effect of the alga extract at the rates of 200, 400, 600, 800, and 1000 μL on the mycelial growth of four important plant pathogenic fungi, including Botrytis cinerea, Aspergillus niger, Penicillium expansum, and Pyricularia oryzae, revealed that the mycelial growth of all four fungi was lower at higher concentrations of the alga extract. However, the extract concentration of 1000 μL completely inhibited their mycelial growth. The antifungal activity of this alga may be related to the phenolic compounds, e.g., rosmarinic acid and quercetin, as well as compounds such as palmitic acid, oleic acid, and other components identified using the GC-MS method whose antifungal effects have already been confirmed.