Biological N
2 fixation, a major pathway for new nitrogen (N) input to terrestrial ecosystems, largely determines the dynamics of ecosystem structure and functions under global change. Nevertheless, the responses of N
2 fixation to multiple global change factors remain poorly understood. Here,
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Biological N
2 fixation, a major pathway for new nitrogen (N) input to terrestrial ecosystems, largely determines the dynamics of ecosystem structure and functions under global change. Nevertheless, the responses of N
2 fixation to multiple global change factors remain poorly understood. Here, saplings of two N
2-fixing plant species,
Alnus cremastogyne and
Cajanus cajan, were grown at rural and urban sites, respectively, with the latter representing an environment with changes in multiple factors occurring simultaneously. Symbiotic N
2 fixation per unit of nodule was significantly higher at the urban site than the rural site for
A. cremastogyne, but the rates were comparable between the two sites for
C. cajan. The nodule investments were significantly lower at the urban site relative to the rural site for both species. Symbiotic N
2 fixation per plant increased by 31.2 times for
A. cremastogyne, while that decreased by 88.2% for
C. cajan at the urban site compared to the rural site. Asymbiotic N
2 fixation rate in soil decreased by 46.2% at the urban site relative to the rural site. The decrease in symbiotic N
2 fixation per plant for
C. cajan and asymbiotic N
2 fixation in soil was probably attributed to higher N deposition under the urban conditions, while the increase in symbiotic N
2 fixation per plant for
A. cremastogyne was probably related to the higher levels of temperature, atmospheric CO
2, and phosphorus deposition at the urban site. The responses of N
2 fixation to multiple global change factors and the underlying mechanisms may be divergent either between symbiotic and asymbiotic forms or among N
2-fixing plant species. While causative evidence is urgently needed, we argue that these differences should be considered in Earth system models to improve the prediction of N
2 fixation under global change.
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