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Aims <p>Both nitrogen (N) and phosphorus (P) are essential nutrients for plant growth but are often limiting in high-yielding agricultural production systems. This study investigated the synergistic effect of nitrogen fixation and organic phosphate mineralization by rhizospheric microorganisms, thereby promoting the growth of host plants.</p> Methods <p>After assessing phosphate-solubilizing activities of five <i>Pseudomonas</i> strains and identifying putative phosphatase genes <i>in silico</i>, selected genes were expressed in nitrogen-fixing <i>P. stutzeri</i> A1501, and the resulting recombinants were evaluated for growth, nitrogenase activity, organic P solubilization (pure and rice co-culture), root colonization, and rice growth-promotion under different N regimes.</p> Results <p>Nine heterologous phosphatase genes were introduced into A1501, and the resulting recombinant strains displayed significantly elevated extracellular phosphatase activity. Most engineered strains maintained nitrogenase activity comparable to A1501, except A15NapD (37% increase) and A1510ACP (35% decrease). In lecithin solubilization assays, recombinant strains A1510ACP, A15PAALP1, A15NapA and A15PhoC significantly increased available phosphorus by 9.2–110.98% relative to A1501. In rice co-culture, these four strains enhanced organic phosphorus hydrolysis, leading to higher phosphorus availability compared with A1501, with A1510ACP also displaying superior root colonization. Pot experiments further demonstrated that inoculation with these four recombinant strains generally promoted rice growth under both nitrogen-depleted and nitrogen-supplemented conditions. In particular, A1510ACP exerted a significant growth‑promoting effect under nitrogen‑supplemented conditions.</p> Conclusion <p>This study presents a successful example of engineering a nitrogen-fixing bacterium with enhanced organic phosphate-solubilizing capacity for enhanced growth of host rice, highlighting the potential of multifunctional engineered strains as the new generation of biological fertilizers.</p>

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Engineering a root-associated bacterium Pseudomonas stutzeri A1501 with nitrogen fixation and phosphate solubilization activities for enhanced growth of host rice

  • 期刊论文

  • 发表日期：

    2026年7月16日

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  • Changyan Yin,
  • Dongqi Wang,
  • Bodan Su,
  • Yaoyao Liu,
  • Yuhua Zhan,
  • Wei Lu,
  • Haichao Feng,
  • Yongliang Yan,
  • Xiubin Ke,
  • Min Lin

摘要

Aims

Both nitrogen (N) and phosphorus (P) are essential nutrients for plant growth but are often limiting in high-yielding agricultural production systems. This study investigated the synergistic effect of nitrogen fixation and organic phosphate mineralization by rhizospheric microorganisms, thereby promoting the growth of host plants.

Methods

After assessing phosphate-solubilizing activities of five Pseudomonas strains and identifying putative phosphatase genes in silico, selected genes were expressed in nitrogen-fixing P. stutzeri A1501, and the resulting recombinants were evaluated for growth, nitrogenase activity, organic P solubilization (pure and rice co-culture), root colonization, and rice growth-promotion under different N regimes.

Results

Nine heterologous phosphatase genes were introduced into A1501, and the resulting recombinant strains displayed significantly elevated extracellular phosphatase activity. Most engineered strains maintained nitrogenase activity comparable to A1501, except A15NapD (37% increase) and A1510ACP (35% decrease). In lecithin solubilization assays, recombinant strains A1510ACP, A15PAALP1, A15NapA and A15PhoC significantly increased available phosphorus by 9.2–110.98% relative to A1501. In rice co-culture, these four strains enhanced organic phosphorus hydrolysis, leading to higher phosphorus availability compared with A1501, with A1510ACP also displaying superior root colonization. Pot experiments further demonstrated that inoculation with these four recombinant strains generally promoted rice growth under both nitrogen-depleted and nitrogen-supplemented conditions. In particular, A1510ACP exerted a significant growth‑promoting effect under nitrogen‑supplemented conditions.

Conclusion

This study presents a successful example of engineering a nitrogen-fixing bacterium with enhanced organic phosphate-solubilizing capacity for enhanced growth of host rice, highlighting the potential of multifunctional engineered strains as the new generation of biological fertilizers.