Journal
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
Volume 117, Issue 18, Pages 9822-9831Publisher
NATL ACAD SCIENCES
DOI: 10.1073/pnas.1921225117
Keywords
nodulation; legume; rhizobium; competition; nitrogen-fixing
Categories
Funding
- Biotechnology and Biological Sciences Research Council [BB/L011484/1, BB/N003608/1, BB/N013387/1]
- Mexican Government Grant Consejo Nacional de Ciencia y Tecnologia [266954/399852]
- University of Oxford Disability Advisory Service
- BBSRC [BB/N013387/1, BB/L011484/1, BB/K006134/1, BB/N003608/1, BB/T001801/1] Funding Source: UKRI
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Legumes tend to be nodulated by competitive rhizobia that do not maximize nitrogen (N-2) fixation, resulting in suboptimal yields. Rhizobial nodulation competitiveness and effectiveness at N-2 fixation are independent traits, making their measurement extremely time-consuming with low experimental throughput. To transform the experimental assessment of rhizobial competitiveness and effectiveness, we have used synthetic biology to develop reporter plasmids that allow simultaneous high-throughput measurement of N-2 fixation in individual nodules using green fluorescent protein (GFP) and barcode strain identification (Plasmid ID) through next generation sequencing (NGS). In a proof-of-concept experiment using this technology in an agricultural soil, we simultaneously monitored 84 different Rhizobium leguminosarum strains, identifying a supercompetitive and highly effective rhizobial symbiont for peas. We also observed a remarkable frequency of nodule coinfection by rhizobia, with mixed occupancy identified in similar to 20% of nodules, containing up to six different strains. Critically, this process can be adapted to multiple Rhizobium-legume symbioses, soil types, and environmental conditions to permit easy identification of optimal rhizobial inoculants for field testing to maximize agricultural yield.
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