Objectives: The objectives of the study were the gas chromatography-mass spectrometry (GC-MS) identification and comparison of the chemical constituents, evaluation of the antifungal and anticancer activities of two species of genus Chenopodium, for example, Chenopodium ambrosioides (Ca) and Chenopodium botrys (Cb) growing in Kashmir, Himalayan region.Methods: The hydrodistilled essential oil of Ca and Cb was subjected to GC-MS analysis and antifungal activity for minimum inhibitory concentration (MIC) determination against different human pathogenic fungal strains using broth microdilution assay in 96-well microtiter plates as per the protocol of the clinical and laboratory standards institute (2008 M27-A3). The anti-proliferative ability of the essential oils was also evaluated against the two cell lines MCF-7 (human mammary carcinoma cells) and A549 (Human lung adenocarcinoma epithelial cells).Results: A total of 34 compounds identified in Ca with α-terpinene (37.17%), isoascaridole (20.48%), and ascaridole (14.83%) as the key compounds. The key compounds of Cb were shyobunol (18.91%), and hedycaryol (9.51%), germacrene-D-4-ol (8.57%), with 65 identified compounds. Both the species were found to have comparable antifungal activities against human pathogenic fungi with MIC80 values in the range of 0.031 mg/ml– 0.256 mg/ml for Ca and 0.031 mg/ml–0.126 mg/ml in case of Cb. Maximum anti-proliferative activity was observed at 125 μg/ml concentration in A549 cell line, while as the oils inhibited the growth of MCF-7 cell line at a lower concentration of 31.25 μg/ml.Conclusion: The essential oils of Ca and Cb were found to have potent anticancer and antifungal activities and can have potential use as a natural fungicide
