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Adaptation of Plasmid-ID Technology for Evaluation of N 2 -Fixing Effectiveness and Competitiveness for Root Nodulation in the Sinorhizobium-Medicago System.

Environmental microbiology · 1 Jun 2025 · 10.1111/1462-2920.70118

Abstract

Maximising the nitrogen fixation occurring in rhizobia-legume associations represents an opportunity to sustainably reduce nitrogen fertiliser inputs in agriculture. High-throughput measurement of symbiotic traits has the potential to accelerate the identification of elite rhizobium/legume associations and enable novel research approaches. Plasmid-ID technology, recently deployed in Rhizobium leguminosarum, facilitates the concurrent assessment of rhizobium nitrogen-fixing effectiveness and competitiveness for root nodulation. This study adapts Plasmid-ID technology to function in Sinorhizobium species that are central models for studying rhizobium-legume associations and form economically important symbioses with alfalfa. New Sino-Plasmid-IDs were developed and tested for stability and their ability to measure competitiveness for root nodulation and nitrogen-fixing effectiveness. Rhizobial competitiveness is measured by identifying strain-specific nucleotide barcodes using next-generation sequencing, whereas effectiveness is measured by GFP fluorescence driven by the synthetic nifH promoter. Sino-Plasmid-IDs allow researchers to efficiently study competitiveness and effectiveness in a multitude of Sinorhizobium strains simultaneously.

Plant phenotyping relevance

Sinorhizobium向けにPlasmid-ID技術を適応・開発し、根粒形成競争性と窒素固定有効性を同時に測定する方法を安定性・性能とともに検証しており、植物共生形質の取得法が中心である。

abstractHigh-throughput measurement of symbiotic traits has the potential to accelerate the identification of elite rhizobium/legume associations
abstractThis study adapts Plasmid-ID technology to function in Sinorhizobium species
abstractNew Sino-Plasmid-IDs were developed and tested for stability and their ability to measure competitiveness for root nodulation and nitrogen-fixing effectiveness.
abstractSino-Plasmid-IDs allow researchers to efficiently study competitiveness and effectiveness in a multitude of Sinorhizobium strains simultaneously.

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