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N accumulation, nodulation, and acetylene reduction activity were measured at frequent intervals during the growth of two chickpea genotypes, and N2 fixation was estimated by an isotope-dilution method, using safflower as a non-N2-fixing reference. Safflower was more efficient at N uptake than both the chickpea genotypes for at least the first 50 days and thus could not be used as an accurate reference control. We recommend that further work should employ non-nodulatiog genotypes of chickpea as reference plants and use slow-release forms of 15N fertilizer. Direct genotype comparison by isotope dilution estimated that genotype K 850 fixed 16–18 kg ha–1 more N than G 130, and this difference was supported by the greater nodule mass and acetylene reduction activity in the K 850 cultivar. Inoculation with an ineffective chickpea Rhizobium sp. led to 69% nodulation on cultivar K 850 but only 33% on G 130. While nodule weight, N uptake, and acetylene reduction activity decreased with inoculation in K 850, the isotope dilutions were similar for both inoculation treatments. The lack of a significant effect on N2 fixation was ascribed to the partial success of inoculant establishment

Evaluation of 15N-isotope dilution for measurement of nitrogen fixation in chickpea (Cicer arietinum L.)