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Optimizing Rhizobium-legume symbioses by simultaneous measurement of rhizobial competitiveness and N2 fixation in nodules.


ABSTRACT: Legumes tend to be nodulated by competitive rhizobia that do not maximize nitrogen (N2) fixation, resulting in suboptimal yields. Rhizobial nodulation competitiveness and effectiveness at N2 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 N2 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 ?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.

SUBMITTER: Mendoza-Suarez MA 

PROVIDER: S-EPMC7211974 | biostudies-literature | 2020 May

REPOSITORIES: biostudies-literature

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Optimizing <i>Rhizobium-</i>legume symbioses by simultaneous measurement of rhizobial competitiveness and N<sub>2</sub> fixation in nodules.

Mendoza-Suárez Marcela A MA   Geddes Barney A BA   Sánchez-Cañizares Carmen C   Ramírez-González Ricardo H RH   Kirchhelle Charlotte C   Jorrin Beatriz B   Poole Philip S PS  

Proceedings of the National Academy of Sciences of the United States of America 20200421 18


Legumes tend to be nodulated by competitive rhizobia that do not maximize nitrogen (N<sub>2</sub>) fixation, resulting in suboptimal yields. Rhizobial nodulation competitiveness and effectiveness at N<sub>2</sub> 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 simultaneo  ...[more]

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