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Applied and Environmental Microbiology, March 2000, p. 1195-1201, Vol. 66, No. 3
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Selected Chitinase Genes in Cultured and Uncultured Marine Bacteria in the alpha - and gamma -Subclasses of the Proteobacteria

Matthew T. Cottrell, Daniel N. Wood, Liying Yu, and David L. Kirchman*

College of Marine Studies, University of Delaware, Lewes, Delaware 19958

Received 14 October 1999/Accepted 29 December 1999

PCR primers were patterned after chitinase genes in four gamma -proteobacteria in the families Alteromonadaceae and Enterobacteriaceae (group I chitinases) and used to explore the occurrence and diversity of these chitinase genes in cultured and uncultured marine bacteria. The PCR results from 104 bacterial strains indicated that this type of chitinase gene occurs in two major groups of marine bacteria, alpha - and gamma -proteobacteria, but not the Cytophaga-Flavobacter group. Group I chitinase genes also occur in some viruses infecting arthropods. Phylogenetic analysis indicated that similar group I chitinase genes occur in taxonomically related bacteria. However, the overall phylogeny of chitinase genes did not correspond to the phylogeny of 16S rRNA genes, possibly due to lateral transfer of chitinase genes between groups of bacteria, but other mechanisms, such as gene duplication, cannot be ruled out. Clone libraries of chitinase gene fragments amplified from coastal Pacific Ocean and estuarine Delaware Bay bacterioplankton revealed similarities and differences between cultured and uncultured bacteria. We had hypothesized that cultured and uncultured chitin-degrading bacteria would be very different, but in fact, clones having nucleotide sequences identical to those of chitinase genes of cultured alpha -proteobacteria dominated both libraries. The other clones were similar but not identical to genes in cultured gamma -proteobacteria, including vibrios and alteromonads. Our results suggest that a closer examination of chitin degradation by alpha -proteobacteria will lead to a better understanding of chitin degradation in the ocean.

* Corresponding author. Mailing address: College of Marine Studies, University of Delaware, 700 Pilottown Rd., Lewes, DE 19958. Phone: (302) 645-4375. Fax: (302) 645-4028. E-mail: kirchman{at}udel.edu.

Applied and Environmental Microbiology, March 2000, p. 1195-1201, Vol. 66, No. 3
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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