Genomic resources for identification of the minimal N2 -fixing symbiotic genome

Environmental Microbiology
George diCenzoTurlough M Finan

Abstract

The lack of an appropriate genomic platform has precluded the use of gain-of-function approaches to study the rhizobium-legume symbiosis, preventing the establishment of the genes necessary and sufficient for symbiotic nitrogen fixation (SNF) and potentially hindering synthetic biology approaches aimed at engineering this process. Here, we describe the development of an appropriate system by reverse engineering Sinorhizobium meliloti. Using a novel in vivo cloning procedure, the engA-tRNA-rmlC (ETR) region, essential for cell viability and symbiosis, was transferred from Sinorhizobium fredii to the ancestral location on the S. meliloti chromosome, rendering the ETR region on pSymB redundant. A derivative of this strain lacking both the large symbiotic replicons (pSymA and pSymB) was constructed. Transfer of pSymA and pSymB back into this strain restored symbiotic capabilities with alfalfa. To delineate the location of the single-copy genes essential for SNF on these replicons, we screened a S. meliloti deletion library, representing > 95% of the 2900 genes of the symbiotic replicons, for their phenotypes with alfalfa. Only four loci, accounting for < 12% of pSymA and pSymB, were essential for SNF. These regions will serve as ou...Continue Reading

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Citations

May 18, 2016·Environmental Microbiology·Juan Sanjuán
Apr 12, 2017·Molecular Plant-microbe Interactions : MPMI·George C diCenzoTurlough M Finan
Aug 11, 2017·Microbiology and Molecular Biology Reviews : MMBR·George C diCenzo, Turlough M Finan
Sep 12, 2018·Canadian Journal of Microbiology·George C diCenzoAlessio Mengoni
Mar 23, 2018·G3 : Genes - Genomes - Genetics·George C diCenzoTurlough M Finan
Nov 28, 2017·Molecular Plant-microbe Interactions : MPMI·Yuan Hui LiuWen Feng Chen
Aug 28, 2020·Frontiers in Microbiology·Luis Alfredo Bañuelos-VazquezSusana Brom
Jan 2, 2021·Proceedings of the National Academy of Sciences of the United States of America·Barney A GeddesTurlough M Finan

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