An improvement in the concentration of vitamin A in adapted yellow maize varieties grown in Africa can have a positive impact on the dietary intakes in regions where maize is a staple food. The present study was designed to identify heterotic groups and divergent parents for developing new pro-vitamin A enriched maize lines. Ten Simple Sequence Repeats (SSR) markers were used to generate DNA profiles among thirteen commonly grown yellow maize lines across south western Nigeria and three high pro-vitamin A lines from International Institute of Tropical Agriculture (IITA), Ibadan. The result obtained estimated 100% polymorphism among the ten SSR markers with polymorphic information content that ranged from 0.28 to 0.71 on an average of 0.50. Genetic similarity coefficients among the 16 maize lines varied from 0.28 to 0.92 GS with an average of 0.63 GS. Four well defined groups were identified at 0.65 GS with an IITA line, PVA8, solely, formed a group. The study identified PVA8 and its most three distant relatives as potential divergent parents that could serve as important genetic resources for broadening the genetic base of the presently assessed IAR&T maize collections and also to develop new maize lines with higher level of pro-vitamin A content.