Evolution of hydrogen gas (H2) during N2 reduction in root nodules results in inefficient use of energy needed for N2 fixation. Cultivars of chickpea (Cicer arietinum L.) were inoculated with Rhizobium strains with and without genes for uptake hydrogenase (Hup) activity. H2 evolution, acetylene reduction activity, and uptake hydrogenase (Hup) activity were assayed on the resulting nodules. The Hup– strains produced higher plant yields than the Hup+ strains. The +N controls produced significantly higher yields than the –N controls and plants inoculated with Rhizobium strains. Hydrogen uptake activity by Rhizobium strains was influenced by the cultivar characteristics. Expression of the plasmid-borne hup genes (pHU52) of Bradyrhizobium japonicum was modified by the host cultivar. The average nodule fresh weight and shoot and root dry weights of the cultivars significantly increased following inoculation with the transconjugant Hup+Rhizobium strain. Thus, biological N2 fixation may be enhanced by selecting Rhizobium strains that are appropriately matched to the particular cultivar. Incorporation of transconjugant Hup+ genes can increase rhizobial activity.