• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of aemPermissionsJournals.ASM.orgJournalAEM ArticleJournal InfoAuthorsReviewers
Appl Environ Microbiol. Feb 1991; 57(2): 535–538.
PMCID: PMC182744

Rapid In Situ Assay for Indoleacetic Acid Production by Bacteria Immobilized on a Nitrocellulose Membrane

Abstract

We have developed a new assay that differentiates between indoleacetic acid (IAA)-producing and -nonproducing bacteria on a colony plate lift. Medium supplemented with 5 mM L-tryptophan is inoculated with isolates of interest, overlaid with a nitrocellulose membrane, and then incubated until bacterial colonies reach 1 to 2 mm in diameter. The membrane is removed to a filter paper saturated with Salkowski reagent and incubated until distinct red haloes form around the colonies. The colorimetric reaction to IAA is limited to a region immediately surrounding each colony, is specific to isolates producing IAA, occurs within 1 h after the membrane is placed in the reagent, and is sensitive to as little as 50 pmol of IAA in a 2-mm2 spot. We have used this assay for quantifying epiphytic and endophytic populations of IAA-producing isolates of Pseudomonas syringae subsp. savastanoi and for detecting IAA-producing colonies of other pseudomonads and Erwinia herbicola. The assay provides a rapid and convenient method to screen large numbers of bacteria.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (908K), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Comai L, Kosuge T. Involvement of plasmid deoxyribonucleic acid in indoleacetic acid synthesis in Pseudomonas savastanoi. J Bacteriol. 1980 Aug;143(2):950–957. [PMC free article] [PubMed]
  • Comai L, Kosuge T. Cloning characterization of iaaM, a virulence determinant of Pseudomonas savastanoi. J Bacteriol. 1982 Jan;149(1):40–46. [PMC free article] [PubMed]
  • Fett WF, Osman SF, Dunn MF. Auxin production by plant-pathogenic pseudomonads and xanthomonads. Appl Environ Microbiol. 1987 Aug;53(8):1839–1845. [PMC free article] [PubMed]
  • Gordon SA, Weber RP. COLORIMETRIC ESTIMATION OF INDOLEACETIC ACID. Plant Physiol. 1951 Jan;26(1):192–195. [PMC free article] [PubMed]
  • Kado CI, Heskett MG. Selective media for isolation of Agrobacterium, Corynebacterium, Erwinia, Pseudomonas, and Xanthomonas. Phytopathology. 1970 Jun;60(6):969–976. [PubMed]
  • Kosuge T, Heskett MG, Wilson EE. Microbial synthesis and degradation of indole-3-acetic acid. I. The conversion of L-tryptophan to indole-3-acetamide by an enzyme system from Pseudomonas savastanoi. J Biol Chem. 1966 Aug 25;241(16):3738–3744. [PubMed]
  • Lindow SE, Hirano SS, Barchet WR, Arny DC, Upper CD. Relationship between Ice Nucleation Frequency of Bacteria and Frost Injury. Plant Physiol. 1982 Oct;70(4):1090–1093. [PMC free article] [PubMed]
  • Yamada T, Palm CJ, Brooks B, Kosuge T. Nucleotide sequences of the Pseudomonas savastanoi indoleacetic acid genes show homology with Agrobacterium tumefaciens T-DNA. Proc Natl Acad Sci U S A. 1985 Oct;82(19):6522–6526. [PMC free article] [PubMed]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...