Plant and Soil

, Volume 161, Issue 1, pp 31–41 | Cite as

Rapid identification ofRhizobium species based on cellular fatty acid analysis

  • B. D. W. Jarvis
  • S. W. Tighe
Article

Abstract

As understanding of the evolutionary relationships between strains and species of root nodule bacteria increases the need for a rapid identification method that correlates well with phylogenetic relationships is clear. We have examined 123 strains ofRhizobium: R. fredii (19),R. galegae (20),R. leguminosarum (22),R. loti (17),R. meliloti (21), andR. tropici (18) and six unknowns. All strains were grown on modified tryptone yeast-extract (TY) agar, as log phase cultures, scraped from the agar, lysed, and the released fatty acids derivatized to their corresponding methyl esters. The methyl esters were analysed by gas-chromatography using the MIDI/Hewlett-Packard Microbial Identification System. All species studied contained 16:0, 17:0, 18:0 and 19cyclow9C fatty acids but onlyR loti andR tropici produced 12:0 3 OH,13:0 iso 3 OH,18:1w9C and 15:0 iso 3 OH,17:0 iso 3 OH and 20:2w6,9C fatty acids respectively. Principal component analysis was used to show that strains could be divided into clusters corresponding to the six species. Fatty acid profiles for each species were developed and these correctly identified at least 95% of the strains belonging to each species. A dendrogram is presented showing the relationships betweenRhizobium species based on fatty acid composition. The data base was used to identify unknown soil isolates as strains ofRhizobium lacking a symbiotic plasmid and a bacterium capable of expressing a symbiotic plasmid fromR. leguminosarum asSphingobacterium spiritovorum.

Key words

FAME fatty acid analysis rapid identification Rhizobium 

Abbreviations

16SrRNA

16S ribosomal ribonucleic acid

MIS

Microbial identification system

FAME

Fatty acid methyl ester

FID

Flame ionization detector

TY

Tryptone yeast extract

ECL

Equivalent carbon length

ED

Euclidean distance and

UPGMA

Unweighted pair group method with arithmetric averages

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Copyright information

© Kluwer Academic Publishers 1994

Authors and Affiliations

  • B. D. W. Jarvis
    • 1
  • S. W. Tighe
    • 1
    • 2
  1. 1.Department of Microbiology and GeneticsMassey UniversityPalmerston NorthNew Zealand
  2. 2.Analytical Services Inc.USA

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