Turmeric (Curcuma longa) is a high value export oriented important commercial crop among the spices. The production was declined due to several biotic and abiotic stresses. Among biotic stresses, root-knot nematode, Meloidogyne incognita is a major threat to turmeric cultivation. Seventy cultivars were screened to identify the resistance to root-knot nematode, Meloidogyne incognita. The result revealed that cultivars ‘Dugirala’, ‘PTS-31’, ‘Ansitapani’, ‘PTS-42’, ‘PTS-47’noted as fully resistant;‘361 Gorakhpur’, ‘328 Sugandham’, ‘PTS-21’ rated as moderately resistant and rest other cultivars were susceptible. The cultivar ‘328 Sugandham’ was moderately resistant to root-knot nematode. This was further confirmed through DNA amplification studies with ISSR markers. The similarity matrix was obtained after multivariate analysis using Nei and Li’s coefficient and the matrix value was ranged from 0.35 to 0.89, with a mean value of 0.62. The two cultivars ‘Dugirala’ and ‘361 Gorakhpur’ with 48% similarity with other 21 cultivars. Both the cultivars were resistance to root knot nematode (RKN) having indexed ranged from 2.0 to 3.0. The five cultivars i.e. ‘Tu No.4’, ‘Tu No.1’, ‘Erode local’, ‘TC-4’ and ‘ ‘Phulbani Wild’ were 78% similarity and susceptible to RKN having index from 4.0 to 5.0. Cultivars ‘Dugirala’, ‘328 Sugandham’ and ‘PTS-47’ exhibited resistance to both root knot nematodes. This investigation as an understanding of the level and partitioning of genetic variation within the cultivars with resistant/susceptible to root knot nematode disease would provide an important input into determining efficient management strategies for breeding program.