Multiple mating by females with different males (polyandry) is difficult to explain in many taxa because it carries significant costs to females, yet benefits are often hard to identify. Polyandry is a derived trait in social insects, the evolutionary origins of which remain unclear. One of several leading hypotheses for its evolution is that it improves division of labour by increasing intra-colonial genetic diversity. Division of labour is a key player in the ecological success of social insects, and in many successful species of ants is based on morphologically distinct castes of workers, each with their own task specialisations. Atta leaf-cutting ants exhibit one of the most extreme and complicated forms of morphologically specialised worker castes and have been reported to be polyandrous but with relatively low mating frequencies (~2.5 on average). Here, we show for the first time that there is a significant genetic influence on worker size in Atta colombica leaf-cutting ants. We also provide the first estimate of the mating frequency of Atta cephalotes (four matings) and, by analysing much higher within-colony sample sizes, find that Atta are more polyandrous than previously thought (approximately six to seven matings). The results show that high polyandry and a genetic influence on worker caste are present in both genera of leaf-cutting ants and add weight to the hypothesis that division of labour is a potential driver of the evolution of polyandry in this clade of ants.