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Applied and Environmental Microbiology, November 2004, p. 6619-6627, Vol. 70, No. 11
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.11.6619-6627.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Differentiation of Micromonospora Isolates from a Coastal Sediment in Wales on the Basis of Fourier Transform Infrared Spectroscopy, 16S rRNA Sequence Analysis, and the Amplified Fragment Length Polymorphism Technique

Hongjuan Zhao,1,2 Yankuba Kassama,1 Michael Young,1 Douglas B. Kell,1,2 and Royston Goodacre1,2*

Institute of Biological Sciences, University of Wales, Aberystwyth, Ceredigion, Wales,1 Department of Chemistry, University of Manchester Institute of Sciences and Technology, Manchester, United Kingdom2

Received 25 February 2004/ Accepted 29 June 2004

A number of actinomycetes isolates were recovered from coastal sediments in Aberystwyth (Wales, United Kingdom) with standard isolation techniques. Most of them were putatively assigned to the genera Streptomyces and Micromonospora on the basis of their morphological characteristics, and there appeared to be no difference whether the isolation media contained distilled water or seawater. A group of 20 Micromonospora isolates was selected to undergo further polyphasic taxonomic investigation. Three approaches were used to analyze the diversity of these isolates, 16S rDNA sequencing, fluorescent amplified fragment length polymorphism (AFLP), and Fourier transform infrared spectroscopy (FT-IR). The 16S rDNA sequence analysis confirmed that all of these isolates should be classified to the genus Micromonospora, and they were analyzed with a group of other Micromonospora 16S rDNA sequences available from the Ribosomal Database Project. The relationships of the 20 isolates were observed after hierarchical clustering, and almost identical clusters were obtained with these three techniques. This has obvious implications for high-throughput screening for novel actinomycetes because FT-IR spectroscopy, which is a rapid and reliable whole-organism fingerprinting method, can be applied as a very useful dereplication tool to indicate which environmental isolates have been cultured previously.

* Corresponding author. Mailing address: School of Chemistry, The University of Manchester, Faraday Bldg., P.O. Box 88, Sackville St., Manchester M60 1QD, United Kingdom. Phone: 44 (0) 161 200 4480. Fax: 44 (0) 161 200 4519. E-mail: R.Goodacre{at}manchester.ac.uk.

Applied and Environmental Microbiology, November 2004, p. 6619-6627, Vol. 70, No. 11
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.11.6619-6627.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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