Thirty one wheat germplasm lines were screened under natural epiphytotic conditions against stripe rust at University Research Farm, Chatha, during Rabi, 2014-15. On the basis of final rust severity (FRS), area under rust progress curve (AURPC) and coefficient of infection (CI), eight lines (Raj 4037, Sonara 64, NP 823, HPW 42, K9351, NIAW 301, PBW 12, and HUW 213) exhibited partial resistance to the disease while as on the basis of infection rate (r) six lines (NP 825, HP 1633, K8434, PBW 12, Ajanta and K9533) showed partial resistance to the disease. Field response of these lines against stripe rust showed that 14 genotypes (Sonara 64, Utkalia, NI 5439, NIAW 301, PBW 12, HUW 213, Ajanta, NP 823, K8434, K9533, Sharbati Sonara, Raj 4037, HP 1633, HPW 42 and K9351) were moderately resistant, 16 (Tawa, KRL, RW 346, HD 2643, HS 1097, NP 825, WH 291, HUW 12, PBW 226, NI 179, NI 5439, K9644, HD1925, PBW 65, PV 18 and GW 503) were moderately susceptible and one genotype (Agra Local) was susceptible. Assessment of losses was also calculated at different growth stages and it was observed that losses at flowering stage and dough stage were highest in one line (Agra Local). Nine wheat germplasm lines (Sonara 64, K9351, HP 1633, Raj 4037, Sharbati Sonara, K9533, K8434, NP 823 and Ajanta) amplifying a band of 523 bp fragment indicating presence of Yr18 gene.