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Applied and Environmental Microbiology, October 2002, p. 5042-5050, Vol. 68, No. 10
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.10.5042-5050.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Molecular Analysis of a Bacterial Chitinolytic Community in an Upland Pasture{dagger}

A. C. Metcalfe,1 M. Krsek,1 G. W. Gooday,2 J. I. Prosser,2 and E. M. H. Wellington1*

Department of Biological Sciences, University of Warwick, Coventry CV4 7AL,1 Department of Molecular and Cell Biology, University of Aberdeen, Institute of Medical Sciences, Foresterhill, Aberdeen AB25 2ZD, United Kingdom2

Received 27 February 2002/ Accepted 9 July 2002

The effects of agricultural-improvement treatments on the chitinolytic activity and diversity of a microbial community were investigated within an upland pasture. The treatments of interest were lime and treated sewage sludge, both commonly applied to pasture land to improve fertility. Burial of chitin-containing litter bags at the field site resulted in enrichment of bacteria according to 16S rRNA fingerprinting. Chitinolytic-activity measurements showed that the highest activity occurred in those bags recovered from sludge-amended plots, which correlated well with increased counts of actinobacteria in samples from these chitin bags. Our findings suggest that sewage sludge increases the fertility of the soil in terms of chitinase activity. Ten clone libraries were constructed from family 18 subgroup A chitinases, PCR amplified from litter bags buried in soil in July 2000 or in September 2000, in a separate study. Analysis of these libraries by restriction fragment length polymorphism and sequencing showed that they were dominated by actinobacterium-like chitinase sequences. This suggests that actinobacteria have an important chitinolytic function in this soil ecosystem. Our findings showed that sludge application increased chitinolytic activity but decreased the diversity of chitinases present.

* Corresponding author. Mailing address: Department of Biological Sciences, University of Warwick, Gibbet Hill Rd., Coventry CV4 7AL, United Kingdom. Phone: 44 2476522431. Fax: 44 2476523701. E-mail: ewellington{at}bio.warwick.ac.uk.

{dagger} This paper is dedicated to the memory of Graham Gooday, who died earlier this year.

Applied and Environmental Microbiology, October 2002, p. 5042-5050, Vol. 68, No. 10
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.10.5042-5050.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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