Alternaria blight (Alternaria brassicae) causes severe foliar damage to Indian mustard in Uttarakhand. Ten (10) isolates of A. brassicae were collected from different hosts and characterized for cultural, morphological, pathogenic and molecular variations. A. brassicae colonies varied in their cultural behaviour ranging from cottony, flurry to feathery, with smooth to rough margins. Colour of colonies ranged between white, off white to light brown. Colony growth varied from slow, medium to fast with fast growth in isolate KM and least in JD on the potato dextrose agar (PDA) medium. Significant morphological variations in conidia length, conidia width, and number of horizontal septa were observed in the isolates. Average conidial size ranged from 105 to 135 × 10 to 20 μm. Isolates exhibited variations in disease index, number and size of the lesions. The dendrogram analysis, based on molecular (random amplification of polymorphic DNA, RAPD) basis revealed two groups at 15% similarity coefficient. Group I was composed of seven isolates namely, VR, DV, P7, LM. P10, KR and ND with 18% similarity (82% dissimilarity) while group II was composed of only three isolates namely, JD, KA and AS with only 24% similarity (76% dissimilarity).Keywords: Pathogenic variability, Brassica, Alternaria brassicae, RAPD-PCRAfrican Journal of Biotechnology, Vol. 13(3), pp. 441-448, 15 January, 201