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Nitrogen (N) fixation was estimated for three Acacia senegal (L.) (A. senegal) Willd. varieties (A. senegal var. senegal, kerensis, and leiorhachis) growing naturally in different sites in the dryland areas of Kenya. The quantities of N2 fixed were estimated by the 15N natural abundance method, using leaves as the sampling material. Balanites aegyptiaca (B. aegyptiaca) was selected as the reference species growing in the same area. Soil samples were also collected under A. senegal trees for nodule assessment. Leaf 15N natural abundance values (δ15N) were significantly different between A. senegal and B. aegyptiaca. These values averaged 6.35, 4.67, and 3.03% for A. senegal var. kerensis, leiorhachis, and senegal, respectively, and were lower than those of the adjacent reference species. There were also significant differences in the amount of N2 fixed (%Ndfa) among the varieties. A. senegal var. senegal showed the highest levels of N2 fixation with a mean of 36% while A. senegal var. kerensis and leiorhachis had equal estimates of 25%. However, no nodules were observed in the collected soil samples. Leaf N values were significantly different among the varieties with a mean of 2.73, 2.46, and 4.03% for A. senegal var. kerensis, leiorhachis, and senegal, respectively. This study shows that the three varieties of A. senegal are able to fix N2 in their natural ecosystems and the differences could probably be due to soil properties and nutrient availability under the different environments. The species can hence be utilized as plantations in agriculture and land rehabilitation programs.