Article
 

Genetics of germination-arrest factor (GAF) production by Pseudomonas fluorescens WH6: identification of a gene cluster essential for GAF biosynthesis

Public Deposited
default representative image
https://ir.library.oregonstate.edu/concern/articles/08612t467

Descriptions

Attribute NameValues
Creator
Abstract
  • The genetic basis of the biosynthesis of the germination-arrest factor (GAF) produced by Pseudomonas fluorescens WH6, and previously identified as 4-formylaminooxyvinylglycine, has been investigated here. In addition to inhibiting the germination of a wide range of grassy weeds, GAF exhibits a selective antimicrobial activity against the bacterial plant pathogen Erwinia amylovora. We utilized the in vitro response of E. amylovora to GAF as a rapid screen for loss-of-function GAF phenotypes generated by transposon mutagenesis. A Tn5 mutant library consisting of 6364 WH6 transformants was screened in this Erwinia assay, resulting in the identification of 18 non-redundant transposon insertion sites that led to loss of GAF production in WH6, as confirmed by TLC analysis. These insertions mapped to five different genes and four intergenic regions. Three of these genes, including two putative regulatory genes (gntR and iopB homologues), were clustered in a 13 kb chromosomal region containing 13 putative ORFs. A GAF mutation identified previously as affecting an aminotransferase also maps to this region. We suggest that three of the genes in this region (a carbamoyltransferase, an aminotransferase and a formyltransferase) encode the enzymes necessary to synthesize dihydroGAF, the putative immediate precursor of GAF in a proposed GAF biosynthetic pathway. RT-qPCR analyses demonstrated that mutations in the gntR and iopB regulatory genes, as well as in a prtR homologue identified earlier as controlling GAF formation, suppressed transcription of at least two of the putative GAF biosynthetic genes (encoding the aminotransferase and formyltransferase) located in this 13 kb region.
  • Keywords: Rhizosphere bacteria, Sequence, Antimetabolite, Aeruginosa, Rhizobitoxine, Pathway, Construction, Family, Amino acids
Resource Type
DOI
Date Available
Date Issued
Citation
  • Halgren, A., Maselko, M., Azevedo, M., Mills, D., Armstrong, D., & Banowetz, G. (2013). Genetics of germination-arrest factor (GAF) production by pseudomonas fluorescens WH6: Identification of a gene cluster essential for GAF biosynthesis. Microbiology (Reading, England), 159(Pt 1), 36-45. doi: 10.1099/mic.0.062166-0
Journal Title
Journal Volume
  • 159
Journal Issue/Number
  • 1
Academic Affiliation
Rights Statement
Funding Statement (additional comments about funding)
  • Support from the United States Department of Agriculture Cooperative State Research, Education and Extension Service (USDA CSREES) Grass Seed Cropping Systems for Sustainable Agriculture Special Grant Program and from the Oregon State University Agricultural Research Foundation is gratefully acknowledged by D.M and D.A.
Publisher
Peer Reviewed
Language
Replaces

Relationships

Parents:

This work has no parents.

Items

Thumbnail Title Date Uploaded Visibility Actions
file details MillsDalliceBotanyPlantPathologyGeneticsGerminationArrest.pdf 10/27/2017 Public
file details MillsDalliceBotanyPlantPathologyGeneticsGerminationArrestSupplementary.pdf 10/27/2017 Public