Plant growth is frequently limited by nitrogen. Plants generally obtain nitrogen from soil reserves of nitrate or ammonium (so-called mineral nitrogen) but these reserves are often scarce.
Natural ecosystems can ‘run down’ with respect to nitrogen through soil leaching and fire. Relative abundance of nitrogen-fixing species will then increase. For example, a walk from east to west across Fraser Island, Queensland, will take you across progressively older and more nitrogen deficient sand dunes, and from rainforest to heathland.
Agriculture and horticulture, with its harvest of nitrogen-rich grains or leaves, removes site nitrogen that must be replaced by further mineralisation of soil nitrogen, import of mineral nitrogen (fertiliser) or fixation of atmospheric nitrogen (N2).
The earth’s atmosphere is rich in N2 (about 78% N2) which is very unreactive, due to its stable triple bond. No known eukaryotes have the ability to fix nitrogen, i.e. to reduce atmospheric nitrogen into a usable form like ammonia (NH3). Hydrogen (H2) will react with N2 at very high temperatures and pressures on a catalyst by the Haber-Bosch process, where the pressures needed are 10–100 MPa, the temperatures are 400–550 ºC, and the catalyst is Fe. Then:
\[ N_{2} + 3H_{2} \rightarrow 2NH_{3} \tag{1} \]
Large quantities of ammonia are produced by this method for industrial and agricultural use. Some nitrogen is also fixed in the atmosphere during lightning strikes (Fowler et al. 2013).
Amazingly, some bacteria have the ability to catalyse this reaction with an enzyme complex called nitrogenase donating at least four pairs of electrons to every N2 molecule to effect reduction to two NH4+ and at least one H2. The reaction takes place at ambient conditions, catalysed by the Fe–Mo-containing enzyme, nitrogenase.
\[N_{2} + 16ATP + 8e^– + 10H^+ \rightarrow 2NH_{4}^+ + H_{2} + 16ADP + 6P_{i} \tag{2}\]
Biological N2 fixation is energetically expensive even though it occurs at ambient conditions — estimates fall between 3 and 7 g carbon respired gram of nitrogen fixed (Layzell 1992). Photoassimilate consumed to support N2 fixation is unavailable for other processes such as growth. Consider a crop fertilised with 140 kg N ha–1. An N2 fixer could replace this fertiliser, but only at a cost of at least 420 kg C ha–1. As most plant dry matter contains 40% carbon, this is equivalent to a loss of one tonne of dry matter per hectare! However, in natural ecosystems where no nitrogen fertiliser is applied, and with rising costs of synthetic nitrogen fertiliser, which consumes about 2% of global fossil fuels annually, biological nitrogen fixation confers a distinct advantage to plants that associate with N2-fixing bacteria (Figure 4.39). In fact, our use of nitrogen fertilisers in agriculture has expanded so enormously that pollution of aquatic habitats by nitrogen run-off has moved beyond safe limits for our planet (Steffen et al. 2015). During the last 30 years, research into biological nitrogen fixation, especially in the symbiosis of legumes with rhizobia, has advanced to a point where transfer of this symbiosis to non-legume crops is now becoming a serious goal (Rogers and Oldroyd 2014).
4.4-Ch-Fig-4.39.png
Figure 4.39 Benefit of nitrogen fixation in legumes depending on N fertilisation. Total nitrogen content (A) and plant biomass (B) of subclover (Trifolium subterraneum) plants grown in a glasshouse for four weeks in the presence and absence of nitrogen fertiliser (10 mM potassium nitrate, ‘N’) and/or the rhizobium symbiont (Rhizobium leguminosarum bv. trifolii, ‘R’). Plants benefit from rhizobial inoculation only in the absence of N fertilizer, as shown by the increases in N content and biomass. Rhizobia confer a similar benefit to the unfertilised subclover plants as a 10 mM nitrate addition does, although this varies with different rhizobial strains and with legume species. (Data modified from C.-H. Goh et al., Plant Cell Environ, 2015)