Summary: | In the karst habitats with nitrate-abundant and ammonium-rare soil, the bicarbonate supply plays a crucial role in both inorganic carbon and nitrogen assimilation in various plant species. Consequently, two carbon sources, carbon dioxide (CO<sub>2</sub>) and bicarbonate (HCO<sub>3</sub><sup>−</sup>), and two nitrogen sources, namely nitrate (NO<sub>3</sub><sup>−</sup>) and ammonium (NH<sub>4</sub><sup>+</sup>) are available for plants. However, variations in the absorption and utilization of nitrate, ammonium, and inorganic carbon during bicarbonate supply in different plants are not well-depicted. In this study, we evaluated the nitrate/ammonium use efficiency and their contributions to the total nitrogen assimilation/utilization capacity at different bicarbonate levels using a bidirectional stable nitrogen isotope tracer approach. The inorganic carbon assimilation, such as the photosynthesis, carbon/nitrogen enzymatic activities, carbon/nitrogen content, nitrogen assimilation/utilization capacity, and nitrate/ammonium contributions to plant growth, were also evaluated to decipher the responses of both carbon and nitrogen metabolism to bicarbonate supply in karst habitats. With the increasing bicarbonate level, <i>Orychophragmus violaceus</i> (<i>Ov</i>) was found to be more available for nitrate to total nitrogen assimilation and utilization than <i>Bn</i> (<i>Brassica napus</i>). Further, it enhanced the contributions of nitrate and nitrogen accumulation/utilization capacity and inorganic carbon assimilation, increasing photosynthesis, carbon/nitrogen enzymatic activities, and carbon accumulation, and promoted the growth in <i>Ov</i>. Though the highest bicarbonate level was conducive to ammonium utilization and water use efficiency in both <i>Ov</i> and <i>Bn</i>, it inhibited total inorganic carbon and nitrogen assimilation, leading to growth suppression in <i>Bn</i> compared to <i>Ov</i>. Moreover, considering the optimistic responses of both carbon and nitrogen assimilation to the high bicarbonate supply in nitrate-abundant, as well as ammonium-rare, environments, we conclude that <i>Ov</i> was more adaptable to the karst habitats. This study provides a novel approach to elucidate the responses of nitrate/ammonium utilization and inorganic carbon assimilation to bicarbonate. Furthermore, the current study reveals the complex interactions among different carbon–nitrogen metabolism pathways in various plants and their adaptations to karst habitats.
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