This study investigated the phytochemical constituents of methanolic rhizome extracts of Alpinia calcarata through the preliminary phytochemical screening, HPLC analysis and its antifungal properties. A.calcarata, Zingiberaceae, is an important rhizomatous perennial medicinal herb and native of India. The phytochemical screening on qualitative analysis showed that the rhizomes of A. calcarata are rich in sterols, anthraquinones etc. which are popular phytochemical constituents and HPLC analysis recorded that it contains five major compounds. Still, there is no proper information on the evaluation of rhizomes of A.calcarata for their antifungal activity. So, methanolic extracts of A. calcarata rhizomes were tested against five important species of pathogenic fungi such as Aspergillus flavus (15.0±0.08), A. niger (13.0±0.07), A. fumigatus (13.0±0.10), Rhizopus stolonifer (11.0±0.13) and Candida albicans (14.0±0.05). The test fungi were mainly associated with seed biodeterioration during storage. Extracts of the rhizome of A.calcarata have recorded significant antifungal activity against species tested. Among these five fungal strains, A. flavus recorded high susceptibility (15.0±0.08).