The aphid Sitobion avenae F. is one of the most harmful pests of wheat growth in the world. A primary field screening test was carried out to evaluate the S. avenae resistance of 527 wheat landraces from Shaanxi. The results indicated that 25 accessions (4.74%) were resistant to S. avenae in the three consecutive seasons, of which accession S849 was highly resistant, and seven accessions were moderately resistant. The majority of S. avenae resistant accessions come from Qinling Mountains. Then, the genetic variability of a set of 33 accessions (25 S. avenae resistant and 8 S. avenae susceptible) originating from Qinling Mountains have been assessed by 20 morphological traits and 99 simple sequence repeat markers (SSRs). Morphological traits and SSRs displayed a high level of genetic diversity within 33 accessions. The clustering of the accessions based on morphological traits and SSR markers showed significant discrepancy according to the geographical distribution, resistance to S. avenae and species of accessions. The highly and moderately resistant landrace accessions were collected from the middle and the east part of Qinling Mountains with similar morphology characters, for example slender leaves with wax, lower leaf area, and high ear density. These S. avenae resistant landraces can be used in wheat aphid resistance breeding as valuable resources.