Cold stress is one of the main limiting factors for autumn planting of chickpea. The probability of presence of cold tolerant genes is considered to be higher in accessions originating from cold and highlands areas due to natural selection process. With this purpose, 812 Desi chickpea accessions of the National Plant Gene Bank of Iran (NPGBI) originated from cold and high altitude regions (altitude more than 1200 m) were screened for cold tolerance. The experiment was set up as augmented design and carried out in dryland agricultural research station in Maragheh, Iran, in 2018-19 cropping season. The reaction of these accessions to the natural cold temperatures in the region was investigated in comparison to tolerant cultivars (Saeed and Saral), and susceptible line (ILC533). Accessions were scored based on the percentage of frost resistance ratio (FRR). Considerable variation was observed among accessions, while the average FRR was estimated 30% and 50% in Saeed and Saral cultivars, respectively, and ILC533 was completely killed by frost. Frost resistance ratio ranged between 0 and 70 percent in NPGBI Desi chickpea accessions. In total, 114 accessions could survive natural cold temperatures and completed their life cycle. The accessions number KC215724, KC215443, KC215712, KC215905, KC215911, KC216223, KC215458, KC215468 and KC215856 showed 60% FRR, and accession number KC215724 obtained 70% FRR, and were more cold tolerant than the other accessions. Therefore, these accessions can be used for being selected and released as new cold tolerant cultivars as well as in chickpea breeding programs for target environments.