Gluconacetobacter diazotrophicus Gene Fitness during Diazotrophic Growth

Erin M. Schwister, Benjamin R. Dietz, Carolann M Knutson, Neil E. Olszewski, Brett M. Barney

Research output: Contribution to journalArticlepeer-review

2 Scopus citations

Abstract

Plant growth-promoting (PGP) bacteria are important to the development of sustainable agricultural systems. PGP microbes that fix atmospheric nitrogen (diazotrophs) could minimize the application of industrially derived fertilizers and function as a biofertilizer. The bacterium Gluconacetobacter diazotrophicus is a nitrogen-fixing PGP microbe originally discovered in association with sugarcane plants, where it functions as an endophyte. It also forms endophyte associations with a range of other agriculturally relevant crop plants. G. diazotrophicus requires microaerobic conditions for diazotrophic growth. We generated a transposon library for G. diazotrophicus and cultured the library under various growth conditions and culture medium compositions to measure fitness defects associated with individual transposon inserts (transposon insertion sequencing [Tn-seq]). Using this library, we probed more than 3,200 genes and ascertained the importance of various genes for diazotrophic growth of this microaerobic endophyte. We also identified a set of essential genes. IMPORTANCE Our results demonstrate a succinct set of genes involved in diazotrophic growth for G. diazotrophicus, with a lower degree of redundancy than what is found in other model diazotrophs. The results will serve as a valuable resource for those interested in biological nitrogen fixation and will establish a baseline data set for plant free growth, which could complement future studies related to the endophyte relationship.

Original languageEnglish (US)
JournalApplied and environmental microbiology
Volume88
Issue number23
DOIs
StatePublished - Dec 2022

Bibliographical note

Funding Information:
This work was supported by grants from the National Institute of Food and Agriculture (project no. MIN-12-070 and MIN-12-081) and award no. 2020-67019-31148 through the United States Department of Agriculture. C.M.K. was further supported through funding from the National Science Foundation (CBET-1437758) and a BBE Graduate Fellowship from the Department of Bioproducts and Biosystems Engineering.

Publisher Copyright:
Copyright © 2022 American Society for Microbiology. All Rights Reserved.

Keywords

  • Gluconacetobacter diazotrophicus
  • Tn-seq
  • gene essentiality
  • nitrogen fixation

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