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Impact of free-living diazotrophs, Azospirillum lipoferum and Gluconacetobacter azotocaptans, on growth and nitrogen utilization by wheat (Triticum aestivum cv. Lillian)



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Nitrogen (N) is an essential plant nutrient, widely applied as N-fertilizer to improve yields of agriculturally important crops. An alternative to fertilizer use could be the exploitation of plant growth-promoting bacteria, capable of enhancing growth and yield of many plant species. Azospirillum and Gluconacetobacter are root colonizing, free-living, N2-fixing bacteria (diazotrophs) with the potential to transfer fixed N to associated plants. The purpose of this study was to evaluate the agronomic efficiency of two diazotrophs, Azospirillum lipoferum and Gluconacetobacter azotocaptans, inoculated onto wheat. Physiological parameters and yield components were evaluated. The objectives of this study were to: 1) determine the survival of each diazotroph species on wheat seeds over time; 2) determine the survival of A. lipoferum and G. azotocaptans inoculated on wheat seed treated with a fungicide seed treatment, Dividend® XL RTA®; 3) determine if inoculation of wheat with the diazotrophs under controlled conditions causes an increase in dry matter, N2-fixation and N uptake; 4) determine if fertilizer N applied at three levels influences atmospheric N2-fixation by A. lipoferum or G. azotocaptans; 5) determine if inoculation of wheat with A. lipoferum or G. azotocaptans under field conditions causes any increase in dry matter, N2-fixation and N uptake; 6) determine if N-fertilization levels under field conditions influenced N2-fixation by A. lipoferum or G. azotocaptans. In order to meet these objectives lab, growth chamber, and field studies were completed. Laboratory investigations revealed that the decline in recovery of colony forming units (CFU) of G. azotocaptans was not significantly different (P<0.05) for any seed treatment. There was a general decrease in CFU over time regardless of seed treatment. Analysis of the recovered CFU of A. lipoferum over time showed that there was a significant difference (P<0.05) between both the non-sterilized seed and the Dividend® XL RTA® treated seed when compared sterilized seed. Recovery of CFU on sterilized seed declined at a more rapid rate compared to the other two seed treatments. Gluconacetobacter azotocaptans and A. lipoferum were not negatively influenced by the Dividend® XL RTA® seed treatment. Also, both diazotrophs were able to compete with other microorganisms that may have been on the seed coat of unsterilized seeds. Azospirillum lipoferum and G. azotocaptans were able to fix atmospheric N, but, there were no significant (P<0.05) differences between the diazotroph species. Additions of fertilizer N enhanced N2-fixation, in both the growth chamber and field studies. As the amount of fertilizer N increased, so did the %Ndfa and N uptake. In the growth chamber study, inoculated wheat, and fertilized with 12.2 and 24.5 µg N g-1 had the highest %Ndfa of 25.5%, and wheat fertilized with 24 µg N g-1 had the highest N uptake (1.3 g pot-1) at maturity. In the field study, inoculated wheat fertilized with of 80 kg N ha-1 had significantly higher (P<0.05) %Ndfa (10.5%) compared to wheat grown with the other fertilizer levels, which also corresponded to the highest N uptake in wheat plants (47 kg ha-1). The diazotrophs also affected the partitioning of N in the wheat plants differently. Wheat inoculated with A. lipoferum had significantly higher (P<0.05) amounts of N accumulated in heads of plants, and wheat inoculated with G. azotocaptans had significantly higher (P<0.05) amounts of N accumulated in stems of plants. However, this trend was not evident in the field study.



diazotrophs, Azospirillum lipoferum, Gluconacetobacter azotocaptans, nitrogen fixation, dry matter, %Ndfa, growth chamber, field experiment, fertilizer application



Master of Science (M.Sc.)


Agricultural and Bioresource Engineering


Soil Science


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