Document Type
Article
Publication Date
3-2019
Keywords
JGM
JAX Source
G3 (Bethesda) 2019 Mar; 9(3):651-661
Volume
9
Issue
3
First Page
651
Last Page
661
ISSN
2160-1836
PMID
30705119
DOI
https://doi.org/10.1534/g3.118.200928
Abstract
The reciprocal interaction between rhizosphere bacteria and their plant hosts results in a complex battery of genetic and physiological responses. In this study, we used insertion sequencing (INSeq) to reveal the genetic determinants responsible for the fitness of Pseudomonas aeruginosa PGPR2 during root colonization. We generated a random transposon mutant library of Pseudomonas aeruginosa PGPR2 comprising 39,500 unique insertions and identified genes required for growth in culture and on corn roots. A total of 108 genes were identified as contributing to the fitness of strain PGPR2 on roots. The importance in root colonization of four genes identified in the INSeq screen was verified by constructing deletion mutants in the genes and testing them for the ability to colonize corn roots singly or in competition with the wild type. All four mutants were affected in corn root colonization, displaying 5- to 100-fold reductions in populations in single inoculations, and all were outcompeted by the wild type by almost 100-fold after seven days on corn roots in mixed inoculations of the wild type and mutant. The genes identified in the screen had homology to genes involved in amino acid catabolism, stress adaptation, detoxification, signal transduction, and transport. INSeq technology proved a successful tool to identify fitness factors in P aeruginosa PGPR2 for root colonization.
Recommended Citation
Sivakumar R,
Ranjani J,
Vishnu U,
Jayashree S,
Lozano G,
Miles J,
Broderick N,
Guan C,
Gunasekaran P,
Handelsman J,
Rajendhran J.
Evaluation of INSeq To Identify Genes Essential for G3 (Bethesda) 2019 Mar; 9(3):651-661
Comments
This open access article is licensed under a Creative Commons Attribution 4.0 International License