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Powdery mildew in cucumber is caused by the Podosphaera xanthii. No strategy for improving disease resistance can be successful in the absence of thorough insights into the physiological and biochemical responses of cucumber plants to powdery mildew. Therefore, a field experiment was executed to evaluate five commercial cucumber varieties (V 1 : Dynasty, V 2 : Long green, V 3 :Desi Kheera, V 4 :Thamin II, V 5 :Cucumber 363) for their inherent immunity to powdery mildew. Upon inoculating cucumber plants with Podosphaera xanthii , we noted differential responses among the varieties. Compared to other varieties, V 1 and V 2 showed higher values (P ≤ 0.05) for chlorophyll-a under control and pathogen-attacked plants respectively. The minimum value of anthocyanin content (−53.73%) was recorded in V 3 as compared to other varieties post pathogen infection. All pathogen-infected cucumber varieties showed a considerable (P ≤ 0.05) loss in flavonoid content except V 2. The maximum destruction for Phenolics under powdery mildew (179%) were recorded in V 4 , whereas V 1 exhibited maximum phenolic content under control conditions. In pathogen-infected plants, the minimum AsA was recorded in V 5 as compared to all other varieties. Pathogen invasion impacted significantly (P ≤ 0.05) the activity of superoxide dismutase (SOD). Besides, cucumber plants after pathogen inoculation resulted in a considerable (P ≤ 0.05) increase of peroxidase (POD) activity in V 1 (5.02%), V 2 (7.5%), and V 3 (11%) in contrast to V 4. Our results confirmed that cucumber varieties perform differently, which was brought on by distinct metabolic and physiological modifications that have an impact on growth and development. The changes in different attributes were correlated with cucumber resistance against powdery mildew. The results would help us fully harness the potential of these varieties to trigger disease management initiatives and defense responses. • Powdery mildew disease in cucumber is pervasive and devastating. • Control of powdery mildew needs improved resistance in cucumber. • We estimated physio-biochemical responses of cucumber for immunity to powdery mildew. • Cucumber varieties performed differently by distinct metabolic modifications. • Different attributes were correlated with cucumber resistance to powdery mildew. [ABSTRACT FROM AUTHOR]