Most legumes can engage in symbiosis with N-fixing bacteria called rhizobia. This symbiosis, called nodulation, evolved from the more widespread symbiosis that most land plants form with arbuscular mycorrhiza, which is reflected in a common requirement of certain genes for both these symbioses. One key nodulation gene, Nodule Inception (NIN), has been intensively studied. Mutants in NIN are unable to form nodules, which has made it difficult to identify downstream genes under the control of NIN. The analysis of data from our recent transcriptomics study revealed that some genes with an altered expression of nin during nodulation are upregulated in mycorrhizal roots. In addition, another study reported the decreased colonization of nin roots by arbuscular mycorrhiza. We therefore investigated a role for NIN in mycorrhiza formation. Our time course study, using two nin alleles with differing genetic backgrounds, suggests that that loss of NIN does not affect colonization of Medicago truncatula roots, either in the presence or absence of rhizobia. This, and recent phylogenetic analyses showing that the loss of NIN is correlated with loss of nodulation in the FaFaCuRo clade, but not with the ability to form mycorrhiza, argue against NIN being required for arbuscular mycorrhization in legumes.