Heme Oxygenase (HO) is an enzyme universally found in animals, plants and microbes. In plants, the role of heme oxygenase in the synthesis of the phytochrome chromophore is well recognized and has been extensively studied; however its role in regulating reactive oxygen species (ROS) in plants is just beginning to be explored, particularly in legumes. Legumes interact with Rhizobium bacteria to form symbiotic nitrogen fixing nodules. ROS plays an important role in the development of roots as well as symbiotic nodules. In the model legume Medicago truncatula, ROS in the root is regulated in part by the LATD/NIP gene. The M. truncatula giraffe mutant has a deletion that removes the entire HO coding sequence. We have found that the M. truncatula GIRAFFE HO regulates expression of some of the LATD/NIP-regulated ROS genes such as RESPRATORY BURST OXIDASE HOMOLOG C (RBOHC) and a cell wall peroxidase (cwPRX2) in seedlings. This means that the wild-type function of GIRAFFE is to up-regulate expression of RBOHC and cwPRX2 in roots, in contrast to LATD/NIP, which down-regulates them. We also found that LATD/NIP and GIRAFFE do not regulate expression of each other in seedlings. Given that the highest expression of GIRAFFE HO is in a senescing nodule, we tested the expression of ROS-regulatory enzymes in senescing nodules. We found that GIRAFFE up-regulates expression of RBOHC during nitrate-induced nodule senescence. At present, with changing climatic conditions and exposure to various environmental stresses that can alter ROS homeostasis, characterizing the role of GIRAFFE in the antioxidant machinery of legumes can be useful in improving crop productivity and for enhancing soil fertility.